Labetalol Hydrochloride / Official Monographs JP XVI Nitrile (1 in 2)

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1014 Labetalol Hydrochloride / Official Monographs JP XVI nitrile (1 in 2). When the procedure is run with 5 mLofthis solution under the above operating conditions, leucomycin Labetalol Hydrochloride A5 and josamycin are eluted in this order with the resolution between these peaks being not less than 5. ラベタロール塩酸塩 System repeatability: When the test is repeated 6 times with 5 mL of the sample solution under the above operating conditions, the relative standard deviation of the peak area of leucomycin A5 is not more than 1.0z. Purity (1) Clarity and color of solution—Dissolve 1.0 g of Kitasamycin Tartrate in 10 mL of water: the solution is clear and colorless or light yellow. (2) Heavy metals <1.07>—Proceed with 1.0 g of Kitasamycin Tartrate according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Stand- ard Lead Solution (not more than 30 ppm). Water <2.48> Not more than 3.0z (0.1 g, volumetric titration, direct titration). Assay Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics

<4.02> according to the following conditions. C19H24N2O3.HCl: 364.87 (i) Test organism—Bacillus subtilis ATCC 6633 2-Hydroxy-5-{(1RS)-1-hydroxy-2-[(1RS)-1-methyl- (ii) Culture medium—Use the medium i in 1) Medium 3-phenylpropylamino]ethyl}benzamide monohydrochloride for test organism [5] under (1) Agar media for seed and base 2-Hydroxy-5-{(1RS)-1-hydroxy-2-[(1SR)-1-methyl- layer. 3-phenylpropylamino]ethyl}benzamide monohydrochloride (iii) Standard solutions—Weigh accurately an amount of [32780-64-6]

Leucomycin A5 RS, equivalent to about 30 mg (potency), dissolve in 10 mL of methanol, add water to make exactly Labetalol Hydrochloride, when dried, contains 100 mL, and use this solution as the standard stock solution. not less than 98.5z and not more than 101.0z of Keep the standard stock solution at not exceeding 59C, and C19H24N2O3.HCl. use within 3 days. Take exactly a suitable amount of the Description Labetalol Hydrochloride occurs as a white standard stock solution before use, add phosphate buffer so- crystalline powder. lution, pH 8.0 to make solutions so that each mL contains 30 It is freely soluble in methanol, and sparingly soluble in mg (potency) and 7.5 mg (potency), and use these solutions as water and in ethanol (99.5). the high concentration standard solution and the low con- It dissolves in 0.05 mol/L sulfuric acid TS. centration standard solution, respectively. Melting point: about 1819C (with decomposition). (iv) Sample solutions—Weigh accurately an amount of Kitasamycin Tartrate, equivalent to about 30 mg (potency), Identification (1) Determine the absorption spectrum of a and dissolve in water to make exactly 100 mL. Take exactly a solution of Labetalol Hydrochloride in 0.05 mol/L sulfuric suitable amount of this solution, add phosphate buffer solu- acid TS (1 in 20,000) as directed under Ultraviolet-visible tion, pH 8.0 to make solutions so that each mL contains 30 Spectrophotometry <2.24>, and compare the spectrum with mg (potency) and 7.5 mg (potency), and use these solutions as the Reference Spectrum: both spectra exhibit similar intensi- the high concentration sample solution and the low concen- ties of absorption at the same wavelengths. tration sample solution, respectively. (2) Determine the infrared absorption spectrum of Labetalol Hydrochloride as directed in the potassium chlo- Containers and storage Containers—Tight containers. ride disc method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. (3) A solution of Labetalol Hydrochloride (1 in 50) responds to the Qualitative Tests <1.09> for chloride. pH <2.54> The pH of a solution prepared by dissolving 0.5 g of Labetalol Hydrochloride in 50 mL of water is between 4.0 and 5.0. Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Labetalol Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.8 g of Labetalol Hy- drochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add JP XVI Official Monographs / Labetalol Hydrochloride Tablets 1015 methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions Labetalol Hydrochloride Tablets as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a ラベタロール塩酸塩錠 plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 2-propanol, water, Labetalol Hydrochloride Tablets contain not less and ammonia solution (28) (25:15:8:2) to a distance of about than 93.0z and not more than 107.0z of the labeled 10 cm, and air-dry the plate. Allow the plate to stand in amount of labetalol hydrochloride (C H N O .HCl: iodine vapor for 30 minutes: the spots other than the princi- 19 24 2 3 364.87). pal spot from the sample solution do not exceed 2 in number and are not more intense than the spot obtained from the Method of preparation Prepare as directed under Tablets, standard solution. with Labetalol Hydrochloride. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, Identification (1) To a quantity of powdered Labetalol 3 hours). Hydrochloride Tablets equivalent to 5 mg of Labetalol Hy- drochloride according to the labeled amount, add 100 mL of Residue on ignition <2.44> Not more than 0.1z (1 g). 0.05 mol/L sulfuric acid TS, shake, and filter. Determine the Isomer ratio Dissolve 5 mg of Labetalol Hydrochloride in absorption spectrum of the filtrate as directed under Ultravi- 0.7 mL of a solution of n-butylboronic acid in anhydrous olet-visible Spectrophotometry <2.24>: it exhibits a maxi- pyridine (3 in 250), allow to stand for 20 minutes, and use mum between 300 nm and 304 nm. this solution as the sample solution. Perform the test with 2 (2) To a quantity of powdered Labetalol Hydrochloride mL of the sample solution as directed under Gas Chromatog- Tablets equivalent to 0.25 g of Labetalol Hydrochloride ac- raphy <2.02> according to the following conditions. Deter- cording to the labeled amount, add 25 mL of methanol, mine the areas of two adjacent peaks, Aa and Ab,whereAa is shake vigorously for 30 minutes, filter, and use the filtrate as the peak area of the shorter retention time and Ab is the peak the sample solution. Separately, dissolve 10 mg of labetalol area of the longer retention time, using the automatic in- hydrochloride in 1 mL of methanol, and use this solution as tegration method: the ratio Ab/(Aa ＋ Ab) is between 0.45 the standard solution. Perform the test using these solutions and 0.55. as directed under Thin-layer Chromatography <2.03>. Spot 5 Operating conditions— mL each of the sample solution and standard solution on a Detector: A hydrogen flame-ionization detector. plate of silica gel with fluorescent indicator for thin-layer Column:Afusedsilicacolumn0.53mmininsidediameter chromatography. Develop the plate with a mixture of ethyl and 25 m in length, coated inside with methyl silicone poly- acetate, 2-propanol, water, and ammonia solution (28) mer for gas chromatography in 5 mm thickness. (25:15:8:2) to a distance of about 10 cm, and air-dry the Column temperature: A constant temperature of about plate. Examine under ultraviolet light (main wavelength: 254 2909C. nm): the principal spot obtained from the sample solution Injection port temperature: A constant temperature of and the spot obtained from the standard solution show the about 3509C. same Rfvalue. Detector temperature: A constant temperature of about Uniformity of dosage units <6.02> Perform the test accord- 3509C. ing to the following method: it meets the requirement of the Carrier gas: Helium. Content uniformity test. Flow rate: Adjust the flow rate so that the retention time To 1 tablet of Labetalol Hydrochloride Tablets add 5 mL of the peak showing earlier elution of the two peaks of of 0.5 mol/L sulfuric acid TS and 30 mL of water, shake vig- labetalol is about 9 minutes. orously for 30 minutes, add water to make exactly 50 mL, System suitability— and filter. Discard the first 5 mL of the filtrate, pipet 4 mL System performance: Proceed with 2 mLofthesampleso- of the subsequent filtrate, add 0.05 mol/L sulfuric acid TS lution under the above conditions: the resolution between to make exactly V mL so that each mL contains about 40 mg the two labetalol peaks is not less than 1.5. of labetalol hydrochloride (C H N O .HCl), and use this System repeatability: Repeat the test 6 times under the 19 24 2 3 solution as the sample solution. Separately, weigh accurately above conditions with 2 mL of the sample solution: the rela- about 20 mg of labetalol hydrochloride for assay, previously tive standard deviation of the ratio of the peak area of dried at 1059C for 3 hours, and dissolve in 0.05 mol/L sulfu- labetalol with the shorter retention time to that of the longer ricacidTStomakeexactly50mL.Pipet5mLofthissolu- retention time is not more than 2.0z. tion, add 0.05 mol/L sulfuric acid TS to make exactly 50 Assay Weigh accurately about 0.3 g of Labetalol Hydro- mL, and use this solution as the standard solution. Deter- chloride, previously dried, dissolve in 100 mL of a mixture mine the absorbances, AT and AS, of the sample solution of acetic anhydride and acetic acid (100) (7:3), and titrate and standard solution at 302 nm as directed under Ultravio- <2.50> with 0.1 mol/L perchloric acid VS (potentiometric let-visible Spectrophotometry <2.24>. titration). Perform a blank determination in the same man- Amount (mg) of labetalol hydrochloride (C H N O .HCl) ner and make any necessary correction. 19 24 2 3 ＝ MS × AT/AS × V/40 Each mL of 0.1 mol/L perchloric acid VS MS: Amount (mg) of labetalol hydrochloride for assay ＝ 36.49 mg of C19H24N2O3.HCl Dissolution <6.10> When the test is performed at 50 revolu- Containers and storage Containers—Tight containers. tions per minute according to the Paddle method, using 900 1016 Lactic Acid / Official Monographs JP XVI mL of water as the dissolution medium, the dissolution rate in 30 minutes of Labetalol Hydrochloride Tablets is not less Lactic Acid than 75z. Start the test with 1 tablet of Labetalol Hydrochloride 乳酸 Tablets, withdraw not less than 20 mL of the medium at specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Dis- card the first 10 mL of the filtrate, pipet V mL of the subse- C3H6O3:90.08 quent filtrate, and add water to make exactly V?mL so that (2RS)-2-Hydroxypropanoic acid each mL contains about 50 mg of labetalol hydrochloride [50-21-5]

(C19H24N2O3.HCl) according to the labeled amount, and use this solution as the sample solution. Separately, weigh accu- Lactic Acid is a mixture of lactic acid and lactic an- rately about 50 mg of labetalol hydrochloride for assay, pre- hydride. viously dried at 1059C for 3 hours, and dissolve in water to It contains not less than 85.0z and not more than make exactly 100 mL. Pipet 10 mL of this solution, add 92.0z of C3H6O3. water to make exactly 100 mL, and use this solution as the Description Lactic Acid occurs as a clear, colorless or light standard solution. Perform the test with the sample solution yellow, viscous liquid. It is odorless or has a faint, unpleas- and standard solution as directed under Ultraviolet-visible ant odor. Spectrophotometry <2.24>, and determine the absorbances, It is miscible with water, with ethanol (95) and with diethyl A and A ,at302nm. T S ether. Dissolution rate (z) with respect to the labeled amount It is hygroscopic. 20 of labetalol hydrochloride (C19H24N2O3.HCl) Specific gravity d 20: about 1.20 ＝ M × A /A × V?/V × 1/C × 90 S T S Identification A solution of Lactic Acid (1 in 50) changes

MS: Amount (mg) of labetalol hydrochloride for assay blue litmus paper to red and responds to the Qualitative C: Labeled amount (mg) of labetalol hydrochloride Tests <1.09> for lactate. (C H N O .HCl) in 1 tablet 19 24 2 3 Purity (1) Chloride <1.03>—Perform the test with 1.0 g of Assay Weigh accurately not less than 20 Labetalol Hydro- Lactic Acid. Prepare the control solution with 1.0 mL of chloride Tablets, and powder. Weigh accurately a portion of 0.01 mol/L hydrochloric acid VS (not more than 0.036z). the powder, equivalent to about 1 g of labetalol hydrochlo- (2) Sulfate <1.14>—Perform the test with 2.0 g of Lactic ride (C19H24N2O3.HCl), add 100 mL of 0.5 mol/L sulfuric Acid. Prepare the control solution with 0.40 mL of 0.005 acid TS and 600 mL of water, shake vigorously for 30 mol/L sulfuric acid VS (not more than 0.010z). minutes, add water to make exactly 1000 mL, and filter. Dis- (3) Heavy metals <1.07>—To 2.0 g of Lactic Acid add 10 card the first 5 mL of the filtrate, pipet 5 mL of the subse- mL of water and 1 drop of phenolphthalein TS, and add am- quent filtrate, and add 0.05 mol/L sulfuric acid TS to make monia TS dropwise until a pale red color appears. Add 2 mL exactly 25 mL. Pipet 5 mL of this solution, add 0.05 mol/L of dilute acetic acid and water to make 50 mL, and perform sulfuric acid TS to make exactly 25 mL, and use this solution the test using this solution as the test solution. Prepare the as the sample solution. Separately, weigh accurately about control solution from 2.0 mL of Standard Lead Solution and 40 mg of labetalol hydrochloride for assay, previously dried 2 mL of dilute acetic acid, and dilute with water to 50 mL at 1059C for 3 hours, and dissolve in 0.05 mol/L sulfuric (not more than 10 ppm). acid TS to make exactly 100 mL. Pipet 5 mL of this solution, (4) Iron <1.10>—Prepare the test solution with 4.0 g of add 0.05 mol/L sulfuric acid TS to make exactly 50 mL, and Lactic Acid according to Method 1, and perform the test use this solution as the standard solution. Perform the test according to Method A. Prepare the control solution with with the sample solution and standard solution as directed 2.0 mL of Standard Iron Solution (not more than 5 ppm). under Ultraviolet-visible Spectrophotometry <2.24>, and de- (5) Sugars—To 1.0 g of Lactic Acid add 10 mL of water, termine the absorbances, AT and AS,at302nm. and neutralize with sodium hydroxide TS. Boil the mixture with 10 mL of Fehling's TS for 5 minutes: no red precipitate Amount (mg) of labetalol hydrochloride (C H N O .HCl) 19 24 2 3 is produced. ＝ MS × AT/AS × 25 (6) Citric, oxalic, phosphoric and L-tartaric acid—To

MS: Amount (mg) of labetalol hydrochloride for assay 1.0gofLacticAcidadd1.0mLofwater,followedby40mL of calcium hydroxide TS. Boil the mixture for 2 minutes: no Containers and storage Containers—Tight containers. change occurs. (7) Glycerin or mannitol—Shake 10 mL of Lactic Acid with 12 mL of diethyl ether: no turbidity is produced. (8) Volatile fatty acids—Warm Lactic Acid: it does not produce any acetic acid-like or butyric acid-like odor. (9) Cyanide—Transfer 1.0 g of Lactic Acid to a Nessler tube, add 10 mL of water and 1 drop of phenolphthalein TS, add dropwise a solution of sodium hydroxide (1 in 10) by shaking until a pale red color develops, add 1.5 mL of a solution of sodium hydroxide (1 in 10) and water to make 20 mL, and heat in a water bath for 10 minutes. Cool, add JP XVI Official Monographs / L-Lactic Acid 1017 dropwise dilute acetic acid until a red color of the solution water bath for 15 minutes. Cool, and adjust to pH 7.0 with 1 disappears, add 1 drop of dilute acetic acid, add 10 mL of mol/L hydrochloric acid VS. Dissolve 5.0 g of hexaammoni- phosphate buffer solution, pH 6.8, and 0.25 mL of sodium um heptamolybdate tetrahydrate in this solution, add water toluensulfonchloramide TS, stopper immediately, mix to make exactly 50 mL, and determine the optical rotation gently, and allow to stand for 5 minutes. To the solution add usinga100-mmcell. 15 mL of pyridine-pyrazolone TS and water to make 50 mL, Purity (1) Chloride <1.03>—Perform the test with 1.0 g of andallowtostandat259C for 30 minutes: the solution has L-Lactic Acid. Prepare the control solution with 1.0 mL of no more color than the following control solution. 0.01 mol/L hydrochloric acid VS (not more than 0.036z). Control solution: Pipet 1.0 mL of Standard Cyanide Solu- (2) Sulfate <1.14>—Perform the test with 2.0 g of L-Lac- tion, and add water to make exactly 20 mL. Transfer 1.0 mL tic Acid. Prepare the control solution with 0.40 mL of 0.005 of this solution to a Nessler tube, add 10 mL of water and 1 mol/L sulfuric acid VS (not more than 0.010z). drop of phenolphthalein TS, and then proceed as described (3) Heavy metal <1.07>—To 2.0 g of L-Lactic Acid add above. 10 mL of water and 1 drop of phenolphthalein TS, and add (10) Readily carbonizable substances—Superimpose ammonia TS dropwise until a pale red color appears. Add 2 slowly 5 mL of Lactic Acid, previously kept at 159C, upon 5 mL of dilute acetic acid and water to make 50 mL, and per- mL of sulfuric acid for readily carbonizable substances, pre- form the test using this solution as the test solution. Prepare viously kept at 159C, and allow to stand at 159Cfor15 the control solution from 2.0 mL of Standard Lead Solution minutes: no dark color develops at the zone of contact. and 2 mL of dilute acetic acid, and dilute with water to 50 Residue on ignition <2.44> Not more than 0.1z (1 g). mL (not more than 10 ppm). (4) Iron <1.10>—Prepare the test solution with 4.0 g of Assay Weigh accurately about 3 g of Lactic Acid, transfer L-Lactic Acid according to Method 1, and perform the test in a conical flask, add accurately measured 40 mL of 1 according to Method A. Prepare the control solution with mol/L sodium hydroxide VS, invert a watch glass over the 2.0 mL of Standard Iron Solution (not more than 5 ppm). flask, and heat on a water bath for 10 minutes. Titrate <2.50> (5) Sugars—To 1.0 g of L-Lactic Acid add 10 mL of the excess sodium hydroxide with 0.5 mol/L sulfuric acid VS water, and neutralize with sodium hydroxide TS. Boil the immediately (indicator: 2 drops of phenolphthalein TS). Per- mixture with 10 mL of Fehling's TS for 5 minutes: no red form a blank determination. precipitate is produced. Each mL of 1 mol/L sodium hydroxide VS (6) Citric, oxalic, phosphoric and L-tartaric acid—To

＝ 90.08 mg of C3H6O3 1.0 g of L-Lactic Acid add 1.0 mL of water, followed by 40 mL of calcium hydroxide TS. Boil the mixture for 2 minutes: Containers and storage Containers—Tight containers. no change occurs. (7) Glycerin or mannitol—Shake 10 mL of L-Lactic Acid with 12 mL of diethyl ether: no turbidity is produced. L-Lactic Acid (8) Volatile fatty acids—Warm L-Lactic Acid: it does not produce any acetic acid-like or butyric acid-like odor. L-乳酸 (9) Cyanide—Transfer 1.0 g of L-Lactic Acid to a Ness- ler tube, add 10 mL of water and 1 drop of phenolphthalein TS, add dropwise a solution of sodium hydroxide (1 in 10) while shaking until a pale red color develops, then add 1.5

C3H6O3: 90.08 mL of a solution of sodium hydroxide (1 in 10) and water to (2S)-2-Hydroxypropanoic acid make 20 mL, and heat in a water bath for 10 minutes. After [79-33-4] cooling, add dropwise dilute acetic acid until a red color of the solution disappears, add 1 drop of dilute acetic acid, and L-Lactic Acid is a mixture of L-lactic acid and 10 mL of phosphate buffer solution, pH 6.8, and 0.25 mL of L-lactic anhydride. sodium toluenesulfonchloramide TS, stopper immediately, It contains not less than 85.0z and not more than mix gently, and allow to stand for 5 minutes. To the solution 92.0z of C3H6O3. add 15 mL of pyridine-pyrazolone TS and water to make 50 mL, and allow to stand at 259C for 30 minutes: the solution Description L-Lactic Acid occurs as a clear, colorless or has no more color than the following control solution. light yellow, viscous liquid. It is odorless or has a faint, no Control solution: Pipet 1.0 mL of Standard Cyanide Solu- unpleasant odor. tion, and add water to make 20 mL. Transfer 1.0 mL of this It is miscible with water, with ethanol (99.5) and with solution to a Nessler tube, add 10 mL of water and 1 drop of diethyl ether. phenolphthalein TS, and then proceed as described above. It is hygroscopic. 20 (10) Readily carbonizable substances—Superimpose Specific gravity d 20: about 1.20 slowly 5 mL of L-Lactic Acid, previously kept at 159C, upon Identification A solution of L-Lactic Acid (1 in 50) changes 5 mL of sulfuric acid for readily carbonizable substances, the color of blue litmus paper to red, and responds to the previously kept at 159C,andallowtostandat159Cfor15 Qualitative Tests <1.09> for lactate. minutes: no dark color develops at the zone of contact.

20 Optical rotation <2.49> [a]D : －46 – －529 Weigh accu- Residue on ignition <2.44> Not more than 0.1z (1 g). rately an amount of L-Lactic Acid, equivalent to about 2 g of Assay Weigh accurately about 3 g of L-Lactic Acid, trans- L-lactic acid (C H O ), add exactly 25 mL of 1 mol/L so- 3 6 3 fer in a conical flask, add accurately measured 40 mL of 1 dium hydroxide VS, cover with a watch glass, and heat on a 1018 Anhydrous Lactose / Official Monographs JP XVI mol/L sodium hydroxide VS, invert a watch glass over the of Anhydrous Lactose in 10 mL of hot water: the solution is flask, and heat on a water bath for 10 minutes. Titrate <2.50> clear, and colorless or nearly colorless. Determine the absor- the excess sodium hydroxide with 0.5 mol/L sulfuric acid VS bance at 400 nm of this solution as directed under Ultravio- immediately (indicator: 2 drops of phenolphthalein TS). Per- let-visible Spectrophotometry <2.24>, using water as the con- form a blank determination. trol solution: not more than 0.04. (2) Acidity or alkalinity—Dissolve 6 g of Anhydrous Each mL of 1 mol/L sodium hydroxide VS Lactose by heating in 25 mL of freshly boiled and cooled ＝ 90.08 mg of C H O 3 6 3 water, and after cooling, add 0.3 mL of phenolphthalein TS: Containers and storage Containers—Tight containers. the solution is colorless, and not more than 0.4 mL of 0.1 mol/L sodium hydroxide VS is required to produce a pale red color or red color. Anhydrous Lactose (3) Heavy metals <1.07>—Proceed with 4.0 g of Anhy- drous Lactose according to Method 2, and perform the test. 無水乳糖 Prepare the control solution with 2 mL of Standard Lead So-

lution (not more than 5 ppm). (4) Proteins and light absorbing substances—Dissolve 1.0 g of Anhydrous Lactose in water to make 100 mL, and use this solution as the sample solution. Determine the absorbances as directed under Ultraviolet-visible Spectropho- tometry <2.24>, using water as the control solution: not more than 0.25 at between 210 nm and 220 nm, and not more than 0.07 at between 270 nm and 300 nm. Loss on drying <2.41> Not more than 0.5z (1 g, 809C, 2 hours). Water <2.48> Not more than 1.0z (1 g, volumetric titration, direct titration. Use a mixture of methanol for Karl

C12H22O11: 342.30 Fischer method and formamide for Karl Fischer method b-D-Galactopyranosyl-(1→4)-b-D-glucopyranose (2:1) instead of methanol for Karl Fischer method). (b-lactose) Residue on ignition <2.44> Not more than 0.1z (1 g). b-D-Galactopyranosyl-(1→4)-a-D-glucopyranose (a-lactose) Microbial limit <4.05> The acceptance criteria of TAMC [63-42-3, Anhydrous Lactose] and TYMC are 102 CFU/g and 5 × 101 CFU/g, respectively. Salmonella and Escherichia coli are not observed. This monograph is harmonized with the European Phar-  macopoeia and the U.S. Pharmacopeia. The parts of the text Isomer ratio Place 1 mg of Anhydrous Lactose in a 5-mL  that are not harmonized are marked with symbols ( ). screw capped reaction vial for gas chromatography, add 0.45 mL of dimethylsulfoxide, stopper, and shake well. Add 1.8 Anhydrous Lactose is b-lactose or a mixture of b- mL of a mixture of pyridine and trimethylsilylimidazole lactose and a-lactose. (18:7), seal the vial tightly with a screw cap, and mix gently. The relative quantities of a-lactose and b-lactose in Allow to stand for 20 minutes, and use this solution as the Anhydrous Lactose is labeled as the isomer ratio. sample solution. Perform the test with 2 mLofthesample solution as directed under Gas Chromatography <2.02> Description Anhydrous Lactose occurs as white crystals according to the following conditions. Determine the peak or powder. areas of a-lactose and b-lactose, A and A , and calculate the It is freely soluble in water, and practically insoluble in a b contents (z)ofa-lactose and b-lactose in Anhydrous Lac- ethanol (99.5).  tose by the following equations. Identification Determine the infrared absorption spec- Content (z)ofa-lactose ＝ A /(A ＋ A ) × 100 trum of Anhydrous Lactose, previously dried, as directed in a a b Content (z)ofb-lactose ＝ A /(A ＋ A ) × 100 the potassium bromide disk method under Infrared Spectro- b a b photometry <2.25>, and compare the spectrum with the Ref- Operating conditions— erence Spectrum or the spectrum of Anhydrous Lactose RS: Detector: A hydrogen flame-ionization detector. both spectra exhibit similar intensities of absorption at the Injection port temperature: A constant temperature of same wave numbers. about 2759C. Detector temperature: A constant temperature of about Optical rotation <2.49> [a]20: ＋54.4 – ＋55.99. Weigh ac- D 2759C. curately about 10 g of Anhydrous Lactose, calculated on the Column: A glass column 4 mm in inside diameter and 90 anhydrous basis, dissolve in 80 mL of water warmed to cm in length, packed with siliceous earth for gas chromatog- 509C, and add 0.2 mL of ammonia TS after cooling. After raphy coated at the ratio of 3z with 25z phenyl-25z standing for 30 minutes, add water to make exactly 100 mL, cyanopropyl-methylsilicone polymer for gas chromatogra- and determine the optical rotation of this solution in a phy. 100-mm cell. Column temperature: A constant temperature of about Purity (1) Clarity and color of solution—Dissolve 1.0 g 2159C. JP XVI Official Monographs / Lactose Hydrate 1019

Carrier gas: Helium. standing for 30 minutes, add water to make exactly 100 mL, Flow rate: A constant flow rate of about 40 mL per and determine the optical rotation of this solution in a minute. 100-mm cell. System suitability— Purity (1) Clarity and color of solution—Dissolve 1.0 g System performance: Prepare a solution with 1 mg of a of Lactose Hydrate in 10 mL of hot water: the solution is mixture of a-lactose and b-lactose (1:1) in the same manner clear, and colorless or nearly colorless. Determine the absor- as for preparing the sample solution, and proceed with 2 mL bance at 400 nm of this solution as directed under Ultravio- of this solution under the above operating conditions, and let-visible Spectrophotometry <2.24>, using water as the con- determine the retention times of the peaks of a-lactose and trol solution: not more than 0.04. b-lactose: the relative retention time of a-lactose with respect (2) Acidity or alkalinity—Dissolve 6 g of Lactose Hy- to that of b-lactose is about 0.7 with the resolution between drate by heating in 25 mL of freshly boiled and cooled these peaks being not less than 3.0. water, and after cooling, add 0.3 mL of phenolphthalein TS: Containers and storage Containers—Well-closed contain- the solution is colorless, and not more than 0.4 mL of 0.1 ers. mol/L sodium hydroxide VS is required to produce a pale red color or red color. (3) Heavy metals <1.07>—Dissolve 4.0 g of Lactose Hy- Lactose Hydrate drate in 20 mL of warm water, add 1 mL of 0.1 mol/L hydrochloric acid TS and water to make 50 mL. Proceed with Lactose this solution according to Method 1, and perform the test. Prepare the control solution with 1 mL of 0.1 mol/L hydro- 乳糖水和物 chloric acid TS and 2.0 mL of Standard Lead Solution (not

more than 5 ppm). (4) Proteins and light absorbing substances—Dissolve 1.0 g of Lactose Hydrate in water to make 100 mL, and use this solution as the sample solution. Determine the absorbances as directed under Ultraviolet-visible Spectrophotome- try <2.24>, using water as the control solution: not more than 0.25 at between 210 nm and 220 nm, and not more than 0.07 at between 270 nm and 300 nm. Loss on drying <2.41> Not more than 0.5z.Forthe granulated powder, not more than 1.0z (1 g, 809C,

C12H22O11.H2O: 360.31 2 hours). b-D-Galactopyranosyl-(1→4)-a-D-glucopyranose Water <2.48> 4.5–5.5z. For the granulated powder, monohydrate 4.0–5.5z (1 g, volumetric titration, direct titration. Use a [64044-51-5,Mixtureofa-andb-lactose monohydrate]  mixture of methanol for Karl Fischer method and for- This monograph is harmonized with the European Phar- mamide for Karl Fischer method (2:1) instead of methanol macopoeia and the U.S. Pharmacopeia. The parts of the text for Karl Fischer method). that are not harmonized are marked with symbols ( ).  Residue on ignition <2.44> Not more than 0.1z (1 g).

Lactose Hydrate is the monohydrate of b-D-galac- Microbial limit <4.05> The acceptance criteria of TAMC topyranosyl-(1→4)-a-D-glucopyranose. and TYMC are 102 CFU/g and 5 × 101 CFU/g, respectively.  It is a disaccharide obtained from milk, consist of Salmonella and Escherichia coli are not observed. one unit of glucose and one unit of galactose.  Containers and storage Containers—Well-closed contain- The label states the effect where it is the granulated ers. powder. Description Lactose Hydrate occurs as white, crystals, powder or granulated powder. It is freely soluble in water, and practically insoluble in ethanol (99.5). Identification Determine the infrared absorption spectrum of Lactose Hydrate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho- tometry <2.25>, and compare the spectrum with the Refer- ence Spectrum or the spectrum of Lactose Hydrate RS: both spectra exhibit similar intensities of absorption at the same wave numbers.

20 Optical rotation <2.49> [a]D : ＋54.4 – ＋55.99. Weigh accurately about 10 g of Lactose Hydrate, calculated on the anhydrous basis, dissolve in 80 mL of water warmed to 509C, and add 0.2 mL of ammonia TS after cooling. After 1020 Lactulose / Official Monographs JP XVI

Amount (mg) of galactose (C6H12O6) Lactulose ＝ MS × QTa/QSa M : Amount (mg) of galactose ラクツロース S

Amount (mg) of lactose (C12H22O11.H2O) ＝ MS × QTb/QSb

MS: Amount (mg) of lactose Hydrate Loss on drying <2.41> Not more than 35z (0.5 g, in vacuum, 809C, 5 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 1 g of Lactulose, add exactly 10 mL of the internal standard solution and water to make 50 mL, and use this solution as the sample solution. C H O : 342.30 12 22 11 Separately, weigh accurately about 0.5 g of Lactulose RS, b-D-Galactopyranosyl-(1→4)-D-fructose accurately about 80 mg of D-galactose and accurately about [4618-18-2] 40 mg of lactose monohydrate, add exactly 10 mL of the internal standard solution and water to make 50 mL, and use Lactulose is a solution of lactulose prepared by this solution as the standard solution. Perform the test with isomerizing lactose under the existing of alkaline and 20 mL each of the sample solution and standard solution as purified by ion-exchange resin. directed under Liquid Chromatography <2.01> according to It contains not less than 50.0z and not more than the following conditions, and calculate the ratios, QT and 56.0z of C12H22O11. QS, of the peak height of lactulose to that of the internal Description Lactulose occurs as a clear, colorless or light standard, respectively. yellow, viscous liquid. It is odorless, and has a sweet taste. Amount (mg) of C H O It is miscible with water and with formamide. 12 22 11 ＝ MS × QT/QS Identification (1) To 0.7 g of Lactulose add 10 mL of M : Amount (mg) of Lactulose RS water, 10 mL of a solution of hexaammonium heptamolyb- S date tetrahydrate (1 in 25) and 0.2 mL of acetic acid (100), Internal standard solution—A solution of D-mannitol (1 in and heat in a water bath for 5 to 10 minutes: a blue color de- 20). velops. Operating conditions— (2) Mix 0.3 g of Lactulose and 30 mL of water, add 16 Detector: A differential refractometer. mL of 0.5 mol/L iodine TS, then immediately add 2.5 mL of Column: A stainless steel column 8 mm in inside diameter 8 mol/L sodium hydroxide TS, allow to stand for 7 minutes, and 50 cm in length, packed with gel type strong acid ion- and add 2.5 mL of diluted sulfuric acid (3 in 20). To this so- exchange resin for liquid chromatography (degree of cross- lution add a saturated solution of sodium sulfite heptahy- linkage: 6z)(11mminparticlediameter). drate until the solution turns light yellow, then add 3 drops Column temperature: A constant temperature of about of methyl orange TS, neutralize with a solution of sodium 759C. hydroxide (4 in 25), and add water to make 100 mL. To 10 Mobile phase: Water. mL of this solution add 5 mL of Fehling's TS, and boil for 5 Flow rate: Adjust the flow rate so that the retention time minutes: a red precipitate is produced. of lactulose is about 18 minutes. System suitability— pH <2.54> To 2.0 g of Lactulose add water to make 15 mL: System performance: When the procedure is run with 10 the pH of the solution is between 3.5 and 5.5. mL of the standard solution under the above operating con- 20 Specific gravity <2.56> d 20: 1.320 – 1.360 ditions, lactulose and the internal standard are eluted in this order with the resolution between these peaks being not less Purity (1) Heavy metals <1.07>—Proceed with 5.0 g of than 8. Lactulose according to Method 4, and perform the test. Pre- System repeatability: When the test is repeated 6 times pare the control solution with 2.5 mL of Standard Lead So- with 20 mL of the standard solution under the above operat- lution (not more than 5 ppm). ing conditions, the relative standard deviation of the ratios (2) Arsenic <1.11>—Prepare the test solution with 1.0 g of the peak heights of lactulose, galactose and lactose to the of Lactulose according to Method 1, and perform the test height of the internal standard are not more than 2.0z,re- (not more than 2 ppm). spectively. (3) Galactose and lactose—Determine the heights of the peaks corresponding to galactose and lactose respectively, on Containers and storage Containers—Tight containers. the chromatogram obtained in Assay from the sample solution and the standard solution, and calculate the ratios of the peak heights of galactose and lactose to that of the internal standard from the sample solution, QTa and QTb,andthen from the standard solution, QSa and QSb: it contains galactose of not more than 11z, and lactose of not more than 6z. JP XVI Official Monographs / Lanatoside C Tablets 1021

20 Optical rotation <2.49> [a]D : ＋32 – ＋359(after drying, Lanatoside C 0.5 g, methanol, 25 mL, 100 mm). Loss on drying <2.41> Not more than 7.5z (0.5 g, in vacu- ラナトシド C um, phosphorus (V) oxide, 609C, 4 hours). Residue on ignition <2.44> Not more than 0.5z (0.1 g). Assay Weigh accurately about 50 mg each of Lanatoside C and Lanatoside C RS, previously dried, and dissolve in methanol to make exactly 25 mL. Pipet 5 mL each of these solutions, add methanol to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Pipet 5 mL each of the sample solution and standard solution into 25-mL light-resistant, volumetric flasks, and add 5 mL of 2,4,6-trinitrophenol TS and 0.5 mL of a solution of sodium hydroxide (1 in 10), shake well, and add methanol to make 25 mL. Allow these solutions to stand between 189Cand229C for 25 minutes, and determine the

absorbances, AT and AS, of the solutions at 485 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution prepared with 5 mL of methanol in the same manner as the blank solution.

Amount (mg) of C49H76O20 ＝ MS × AT/AS

C49H76O20: 985.12 MS: Amount (mg) of Lanatoside C RS 3b-[b-D-Glucopyranosyl-(1→4)-3-O-acetyl-2,6-dideoxy- Containers and storage Containers—Tight containers. b-D-ribo-hexopyranosyl-(1→4)-2,6-dideoxy-b-D-ribo- Storage—Light-resistant. hexopyranosyl-(1→4)-2,6-dideoxy-b-D-ribo- hexopyranosyloxy]-12b,14-dihydroxy-5b,14b-card- 20(22)-enolide [17575-22-3] Lanatoside C Tablets ラナトシド C 錠 Lanatoside C, when dried, contains not less than 90.0z and not more than 102.0z of C49H76O20. Lanatoside C Tablets contain not less than 90.0z Description Lanatoside C occurs as colorless or white crys- and not more than 110.0z of the labeled amount of tals or a white, crystalline powder. It is odorless. lanatoside C (C H O : 985.12). It is soluble in methanol, slightly soluble in ethanol (95), 49 76 20 and practically insoluble in water and in diethyl ether. Method of preparation Prepare as directed under Tablets, It is hygroscopic. with Lanatoside C. Identification Place 1 mg of Lanatoside C to a small test Identification (1) Shake a quantity of powdered Lanato- tube having an internal diameter of about 10 mm, dissolve in side C Tablets, equivalent to 1 mg of Lanatoside C according 1 mL of a solution of iron (III) chloride hexahydrate in ace- to the labeled amount, with 3 mL of diethyl ether, and filter. tic acid (100) (1 in 10,000), and underlay gently with 1 mL of Wash the residue with two 3-mL portions of diethyl ether, sulfuric acid: at the zone of contact of the two liquids, a andair-dry.Totheremainingresidueadd10mLofamix- brown ring is produced, and the color of the upper layer ture of chloroform and methanol (9:1), shake, and filter. near the contact zone gradually changes to blue through pur- Wash the residue with two 5-mL portions of a mixture of ple. Finally the color of the entire acetic acid layer changes chloroform and methanol (9:1), combine the filtrate and to blue-green through deep blue. washings, and evaporate on a water bath to a smaller volume. Transfer the solution to a small test tube having an Purity Related substances—Dissolve 10 mg of Lanatoside internal diameter of about 10 mm, further evaporate on a C in exactly 5 mL of methanol, and use this solution as the water bath to dryness, and proceed as directed in the Iden- sample solution. Separately, dissolve 1.0 mg of Lanatoside C tification under Lanatoside C. RS in exactly 5 mL of methanol, and use this solution as the (2) Perform the test with the sample solution and the standard solution. Perform the test as directed under Thin- standard solution obtained in the Assay as directed under layer Chromatography <2.03> with these solutions. Spot 20 Thin-layer Chromatography <2.03>. Spot 25 mLeachofthese mL each of the sample solution and standard solution on a solutions on a plate of silica gel for thin-layer chromatogra- plate of silica gel for thin-layer chromatography. Develop phy. Develop the plate with a mixture of dichloromethane, the plate with a mixture of dichloromethane, methanol and methanol and water (84:15:1) to a distance of about 13 cm, water (84:15:1) to a distance of about 13 cm, and air-dry the and air-dry the plate. Spray evenly dilute sulfuric acid on the plate. Spray evenly dilute sulfuric acid on the plate, and heat plate, and heat the plate at 1109C for 10 minutes: the spots the plate at 1109C for 10 minutes: any spots other than the obtained from the sample solution and standard solution principal spot from the sample solution are neither larger show a black color, and have the same Rf values. nor darker than the spot from the standard solution. 1022 Hydrous Lanolin / Official Monographs JP XVI

Uniformity of dosage unit <6.02> Perform the test accord- ties, FT, FS and FB, of the sample solution and the standard ing to the following method: it meets the requirement of the solution at 355 nm of the excitation wavelength and at 490 Content uniformity test. nm of the fluorescence wavelength as directed under Fluoro- Warm 1 tablet of Lanatoside C Tablets with 5 mL of metry <2.22>. water until the tablet is disintegrated, add 30 mL of ethanol Dissolution rate (z) with respect to the labeled amount (95), disperse finely the particles with the aid of ultrasonic of lanatoside C (C H O ) waves, add ethanol (95) to make exactly V mL of a solution 49 76 20 ＝ MS × (FT － FB)/(FS － FB) × V?/V × 1/C containing about 5 mgoflanatosideC(C49H76O20)ineach mL, and filter. Discard the first 10 mL of the filtrate, and MS: Amount (mg) of Lanatoside C RS use the subsequent filtrate as the sample solution. Sepa- C: Labeled amount (mg) of lanatoside C (C49H76O20)in1 rately, weigh accurately about 25 mg of Lanatoside C RS, tablet previously dried in vacuum over phosphorus (V) oxide at Assay Weigh accurately and powder not less than 20 609C for 4 hours, and dissolve in ethanol (95) to make ex- Lanatoside C Tablets. Weigh accurately a portion of the actly 100 mL. Pipet 2 mL of this solution, add 10 mL of powder, equivalent to about 5 mg of lanatoside C water, add ethanol (95) to make exactly 100 mL, and use this (C H O ), into a 100-mL light-resistant volumetric flask, solution as the standard solution. Pipet 2 mL each of the 45 76 20 add 50 mL of ethanol (95), and shake for 15 minutes. Then sample solution, the standard solution and diluted ethanol dilute with ethanol (95) to make exactly 100 mL. Filter this (95) (17 in 20) into three brown glass-stoppered test tubes T, solution, discard the first 20 mL of the filtrate, and use the S and B, previously containing exactly 10 mL of 0.012 w/vz subsequent filtrate as the sample solution. Separately, weigh L-ascorbic acid-hydrochloric acid TS, add exactly 1 mL each accurately about 5 mg of Lanatoside C RS, previously dried of dilute hydrogen peroxide TS immediately, shake vigor- in vacuum over phosphorus (V) oxide at 609C for 4 hours, ously, and allow to stand at a constant temperature between dissolve in ethanol (95) to make exactly 100 mL, and use this 259Cand309C for 40 minutes. Determine the fluorescence solution as the standard solution. Pipet 5 mL each of the intensities, F , F and F , of the subsequent solutions from T S B sample solution and standard solution into light-resistant, the sample solution and the standard solution and the glass-stoppered test tubes, add exactly 3 mL each of alkaline diluted ethanol (95) (17 in 20) at 355 nm of the excitation 2,4,6-trinitrophenol TS, shake well and allow these solutions wavelength and at 490 nm of the fluorescence wavelength as to stand between 229Cand289C for 25 minutes. Determine directed under Fluorometry <2.22>, respectively. the absorbances, AT and AS, of the subsequent sample solu- Amount (mg) of lanatoside C (C49H76O20) tion and the subsequent standard solution at 490 nm as di- ＝ MS × (FT － FB)/(FS － FB) × V/5000 rected under Ultraviolet-visible Spectrophotometry <2.24>, using a solution, prepared by the same manner with 5 mL of M : Amount (mg) of Lanatoside C RS S ethanol (95), as the blank. Dissolution <6.10> When the test is performed at 100 revo- Amount (mg) of lanatoside C (C H O ) lutions per minute according to the Paddle method, using 49 76 20 ＝ M × A /A 500 mL of diluted hydrochloric acid (3 in 500) as the dissolu- S T S tion medium, the dissolution rate in 60 minutes of Lanato- MS: Amount (mg) of Lanatoside C RS side C Tablets is not less than 65z. No retest requirement is Containers and storage Containers—Tight containers. applied to Lanatoside C Tablets. Storage—Light-resistant. Start the test with 1 tablet of Lanatoside C Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard the first Hydrous Lanolin 10 mL of the filtrate, pipet V mL of the subsequent filtrate, 加水ラノリン add the dissolution medium to make exactly V?mL so that each mL contains about 0.5 mg of lanatoside (C49H76O20)according to the labeled amount, and use this solution as the Hydrous Lanolin is Purified Lanolin to which water sample solution. Separately, dry Lanatoside C RS in vacuum is added. It contains not less than 70z and not more over phosphorus (V) oxide at 609C for 4 hours, weigh accu- than 75z of Purified Lanolin (as determined by the rately a portion of it, equivalent to 100 times an amount of test for Residue on evaporation). the labeled amount of lanatoside C (C H O ), dissolve in 49 76 20 Description Hydrous Lanolin is a yellowish white, oint- ethanol (95) to make exactly 100 mL. Pipet 1 mL of this so- ment-like substance, and has a slight, characteristic odor, lution, add the dissolution medium to make exactly 500 mL, which is not rancid. warm at 37 ± 0.59C for 60 minutes, and use this solution as It is soluble in diethyl ether and in cyclohexane, with the the standard solution. Pipet 3 mL each of the sample solu- separation of water. tion, the standard solution and the dissolution medium, and When melted by heating on a water bath, it separates into transfer to glass-stoppered brown test tubes T, S and B, a clear oily layer and a clear water layer. respectively. To these solutions add exactly 10 mL each of Melting point: about 399C. 0.012 w/vz L-ascorbic acid-hydrochloric acid TS, and shake. Immediately add exactly 0.2 mL each of diluted Identification Dissolve 1 g of Hydrous Lanolin in 50 mL of hydrogen peroxide TS (1 in 100), shake well, and allow to cyclohexane, and remove the separated water. Superimpose stand at a constant temperature between 309Cand379Cfor carefully 1 mL of the cyclohexane solution on 2 mL of sulfu- 45 minutes. Determine immediately the fluorescence intensi- ric acid: a red-brown color develops at the zone of contact, JP XVI Official Monographs / Purified Lanolin 1023 and sulfuric acid layer shows a green fluorescence. another separator, add 10 mL of diethyl ether, shake, and combine the diethyl ether layer and diethyl ether in the first Acid value <1.13> Not more than 1.0. separator. Shake the diethyl ether layer with 3 g of anhy- Iodine value 18 – 36 Heat a suitable amount of Hydrous drous sodium sulfate, and filter through dry filter paper. Lanolin on a water bath to remove its almost moisture, then Wash the separator and the filter paper with two 20-mL por- weigh accurately about 0.8 g of the treated Hydrous Lanolin tions of diethyl ether, combine the washings with the filtrate, in a glass-stoppered 500-mL flask, and add 10 mL of cyclo- evaporate on a water bath until the odor of diethyl ether is hexane to dissolve, and add exactly 25 mL of Hanus's TS, no longer perceptible, and dry in a desiccator (in vacuum, and mix well. If a clear solution is not obtained, add more silica gel) for 24 hours: the content is not less than 70z and cyclohexane to make clear, and allow the mixture to stand not more than 75z. for 1 hour between 209Cand309C in a light-resistant, well- Containers and storage Containers—Well-closed contain- closed container while occasional shaking. Add 20 mL of a ers. solution of potassium iodide (1 in 10) and 100 mL of water, Storage—Not exceeding 309C. shake, and titrate <2.50> the liberated iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Per- form a blank determination in the same manner. Purified Lanolin Iodine value ＝ (a － b) × 1.269/M Adeps Lanae Purificatus M: amount (g) of sample a: Volume (mL) of 0.1 mol/L sodium thiosulfate VS con- 精製ラノリン sumed in the blank determination b: Volume (mL) of 0.1 mol/L sodium thiosulfate VS con- sumedinthetitration Purified Lanolin is the purified product of the fat- like substance obtained from the wool of Ovis aries Purity (1) Acidity or alkalinity—To 5 g of Hydrous Linne(á Bovidae). Lanolin add 25 mL of water, boil for 10 minutes, and cool. Add water to restore the previous mass, and separate the Description Purified Lanolin is a light yellow to yellowish aqueous layer: the aqueous layer is neutral. brown, viscous, ointment-like substance, and has a faint, (2) Chloride <1.03>—To 2.0 g of Hydrous Lanolin add characteristic but not rancid odor. 40 mL of water, boil for 10 minutes, and cool. Add water to It is very soluble in diethyl ether and in cyclohexane, freely restore the previous mass, and filter. To 20 mL of the filtrate soluble in tetrahydrofuran and in toluene, and very slightly add 6 mL of dilute nitric acid and water to make 50 mL. Use soluble in ethanol (95). It is practically insoluble in water, this solution as the test solution, and perform the test. Pre- but miscible without separation with about twice its mass of pare the control solution with 1.0 mL of 0.01 mol/L hydro- water, retaining ointment-like viscosity. chloricacidVS(notmorethan0.036z). Melting point: 37 – 439C (3) Ammonia—To 10 mL of the aqueous layer obtained Identification Superimpose carefully 1 mL of a solution of in (1) add 1 mL of sodium hydroxide TS, and boil: the gas Purified Lanolin in cyclohexane (1 in 50) on 2 mL of sulfuric evolved does not turn moistened red litmus paper to blue. acid: a red-brown color develops at the zone of contact, and (4) Water-soluble organic substances—To 5 mL of the the sulfuric acid layer shows a green fluorescence. aqueous layer obtained in (1) add 0.25 mL of 0.002 mol/L potassium permanganate VS, and allow to stand for 5 Acid value <1.13> Not more than 1.0. minutes: the red color of the solution does not disappear. Iodine value 18 – 36 Weigh accurately about 0.8 g of (5) Petrolatum—Dissolve 1.0 g of the dried residue ob- Purified Lanolin in a glass-stoppered 500-mL flask, add 20 tained in the Residue on evaporation in 10 mL of a mixture mL of cyclohexane to dissolve, and add exactly 25 mL of of tetrahydrofuran and isooctane (1:1), and use this solution Hanus' TS, and mix well. If a clear solution is not obtained, as the sample solution. Add dissolve 20 mg of vaseline in 10 add more cyclohexane to make clear, and allow the mixture mL of a mixture of tetrahydrofuran and isooctane (1:1), and to stand for 1 hour between 209Cand309C in light-resistant, use this solution as the standard solution. Perform the test well-closed containers, with occasional shaking. Add 20 mL with these solutions as directed under Thin-layer Chroma- of a solution of potassium iodide (1 in 10) and 100 mL of tography <2.03>. Spot 25 mL each of the sample solution and water, shake, and titrate the liberated iodine with 0.1 mol/L standard solution on a plate of silica gel for thin-layer chro- sodium thiosulfate VS (indicator: 1 mL of starch TS). Per- matography. Develop the plate with isooctane to a distance form a blank determination. of about 10 cm, and air-dry the plate. Spray evenly diluted sulfuric acid (1 in 2) on the plate, heat the plate at 809Cfor5 Iodine value ＝ (a － b) × 1.269/M minutes, cool, and examine under ultraviolet light (main M: amount (g) of sample. wavelength: 365 nm): no fluorescent spot is observed in the a: Volume (mL) of 0.1 mol/L sodium thiosulfate VS used same level with the spot of standard solution. For this test in the blank determination. use a thin-layer plate previously developed with isooctane to b: Volume (mL) of 0.1 mol/L sodium thiosulfate VS used the upper end, dried in air, and heated at 1109Cfor60 in the titration of the sample. minutes. Purity (1) Acid or alkali—To 5 g of Purified Lanolin add Residue on evaporation Weigh accurately about 12.5 g of 25 mL of water, boil for 10 minutes, and cool. Add water to Hydrous Lanolin, dissolve in 50 mL of diethyl ether, place it restore the previous mass, and separate the aqueous layer: in a separator, transfer the separated aqueous layer to 1024 Lard / Official Monographs JP XVI the aqueous layer is neutral. Purity (1) Moisture and coloration—Melt 5 g of Lard by (2) Chloride <1.03>—To2.0gofPurifiedLanolinadd40 heating on a water bath: it forms a clear liquid, from which mL of water, boil for 10 minutes, and cool. Add water to re- no water separates. Observe the liquid in a layer 10 mm store the previous mass, and filter. To 20 mL of the filtrate thick: the liquid is colorless to slightly yellow. add 6 mL of dilute nitric acid and water to make 50 mL. Use (2) Alkalinity—To 2.0 g of Lard add 10 mL of water, this solution as the test solution, and perform the test. Pre- melt by warming on a water bath, and shake vigorously. pare the control solution with 1.0 mL of 0.01 mol/L hydro- After cooling, add 1 drop of phenolphthalein TS to the sepa- chloricacidVS(notmorethan0.036z). rated water layer: the layer is colorless. (3) Ammonia—To 10 mL of the aqueous layer obtained (3) Chloride <1.03>—To1.5gofLardadd30mLof in (1) add 1 mL of sodium hydroxide TS, and boil: the gas ethanol (95), boil for 10 minutes under a reflux condenser, evolved does not turn moistened red litmus paper to blue. and filter after cooling. To 20 mL of the filtrate add 5 drops (4) Water-soluble organic substances—To 5 mL of the of a solution of silver nitrate in ethanol (95) (1 in 50): the aqueous layer obtained in (1) add 0.25 mL of 0.002 mol/L opalescence of the mixture does not exceed that of the fol- potassium permanganate VS, and allow to stand for 5 lowing control solution. minutes: the red color of the solution does not disappear. Control solution: To 1.0 mL of 0.01 mol/L hydrochloric (5) Petrolatum—Dissolve 1.0 g of Purified Lanolin in 10 acid VS add ethanol (95) to make 20 mL, and add 5 drops of mL of a mixture of tetrahydrofuran and isooctane (1:1), and a solution of silver nitrate in ethanol (95) (1 in 50). use this solution as the sample solution. And dissolve 20 mg (4) Beef tallow—Dissolve 5 g of Lard in 20 mL of diethyl of vaseline in 10 mL of a mixture of tetrahydrofuran and ether, stopper lightly with absorbent cotton, and allow to isooctane (1:1), and use this solution as the standard solu- stand at 209C for 18 hours. Collect the separated crystals, tion. Perform the test with the sample solution as directed moisten them with ethanol (95), and examine under a micro- under Thin-layer Chromatography <2.03>. Spot 25 mLeach scope of 200 magnifications: the crystals are in the form of of the sample solution and standard solution on a plate of rhomboidal plates grouped irregularly, and do not contain silica gel for thin-layer chromatography. Develop the plate prisms or needles grouped in fan-shaped clusters. with isooctane to a distance of about 10 cm, and air-dry the Containers and storage Containers—Well-closed contain- plate. Spray evenly diluted sulfuric acid (1 in 2) on the plate, ers. heat the plate at 809C for 5 minutes, cool, and examine Storage—Not exceeding 309C. under ultraviolet light (main wavelength: 365 nm): no fluorescent spot is observable same level of the spot of standard solution. Use a thin-layer plate previously developed with isooctane to the upper end, dried in air, and heated at 1109C Latamoxef Sodium for 60 minutes. ラタモキセフナトリウム Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Total ash <5.01> Not more than 0.1z. Containers and storage Containers—Well-closed containers. Storage—Not exceeding 309C.

C20H18N6Na2O9S: 564.44 Disodium (6R,7R)-7-[2-carboxylato- Lard 2-(4-hydroxyphenyl)acetylamino]-7-methoxy-3-(1-methyl- 1H-tetrazol-5-ylsulfanylmethyl)-8-oxo-5-oxa- Adeps Suillus 1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate [64953-12-4] 豚脂 Latamoxef Sodium contains not less than 830 mg (potency) and not more than 940 mg (potency) per mg, Lard is the fat obtained from Sus scrofa Linneá var. calculated on the anhydrous basis. The potency of domesticus Gray (Suidae). Latamoxef Sodium is expressed as mass (potency) of Description Lard occurs as a white, soft, unctuous mass, latamoxef (C20H20N6O9S: 520.47). and has a faint, characteristic odor and a bland taste. Description Latamoxef Sodium occurs as white to light yel- It is freely soluble in diethyl ether and in petroleum ether, lowish white, powder or masses. very slightly soluble in ethanol (95), and practically insoluble It is very soluble in water, freely soluble in methanol, and in water. slightly soluble in ethanol (95). Melting point: 36 – 429C Congealing point of the fatty acids: 36 – 429C Identification (1) Determine the absorption spectrum of a solution of Latamoxef Sodium (3 in 100,000) as directed Acid value <1.13> Not more than 2.0. under Ultraviolet-visible Spectrophotometry <2.24>,and Saponification value <1.13> 195–203 compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same Iodine value <1.13> 46–70 wavelengths. JP XVI Official Monographs / Latamoxef Sodium 1025

(2) Determine the infrared absorption spectrum of use the peak area for 1-methyl-1H-tetrazole-5-thiol after Latamoxef Sodium as directed in the potassium bromide multiplying by its ralative response factor, 0.52. disk method under Infrared Spectrophotometry <2.25>,and Operating conditions— compare the spectrum with the Reference Spectrum: both Proceed as directed in the operating conditions in the spectra exhibit similar intensities of absorption at the same Assay. wave numbers. System suitability— (3) Determine the 1H spectrum of a solution of Latamox- System performance: Proceed as directed in the system ef Sodium in heavy water for nuclear magnetic resonance suitability in the Assay. spectroscopy (1 in 10) as directed under Nuclear Magnetic System repeatability: When the test is repeated 6 times Resonance Spectroscopy <2.21>, using sodium 3-trimethyl- with 5 mL of the standard solution under the above operating silylpropanesulfonate for nuclear magnetic resonance spec- conditions, the relative standard deviation of the peak area troscopy as an internal reference compound: it exhibits of latamoxef is not more than 2.0z. single signals, A and B, at around d 3.5 ppm and at around Water <2.48> Not more than 5.0z (0.5 g, volumetric titra- d 4.0 ppm. The ratio of the integrated intensity of these sig- tion, back titration). nals, A:B, is about 1:1. (4) Latamoxef Sodium responds to the Qualitative Tests Isomer ratio Dissolve 25 mg of Latamoxef Sodium in water <1.09> (1) for sodium salt. to make 50 mL, and use this solution as the sample solution. Perform the test with 5 mL of the sample solution as directed Optical rotation <2.49> [a]20: －32 – －409(0.5 g calculated D under Liquid Chromatography <2.01> according to the fol- on the anhydrous basis, phosphate buffer solution, pH 7.0, lowing conditions, and determine the areas, A and A ,of 50 mL, 100 mm). a b the two peaks in order of elution, which appear close to each pH <2.54> The pH of a solution obtained by dissolving other at the retention time of about 10 minutes: Aa/Ab is be- 1.0 g of Latamoxef Sodium in 10 mL of water is between 5.0 tween 0.8 and 1.4. and 7.0. Operating conditions— Detector: An ultraviolet absorption photometer (wave- Purity (1) Clarity and color of solution—Dissolve 1.0 g length: 254 nm). of Latamoxef Sodium in 10 mL of water: the solution is Column: A stainless steel column 4 mm in inside diameter clear and has no more color than the following control solu- and 15 cm in length, packed with octadecylsilanized silica gel tion. for liquid chromatography (10 mminparticlediameter). Control solution: To a mixture of 3.0 mL of Cobalt (II) Column temperature: A constant temperature of about Chloride CS and 36 mL of Iron (III) Chloride CS add 11 mL 259C. of diluted dilute hydrochloric acid (1 in 10). To 2.5 mL of Mobile phase: Dissolve 7.7 g of ammonium acetate in this solution add 7.5 mL of diluted dilute hydrochloric acid water to make 1000 mL. To 950 mL of this solution add 50 (1:10). mL of methanol. (2) Heavy metals <1.07>—Carbonize 1.0 g of Latamoxef Flow rate: Adjust the flow rate so that the retention time Sodium by heating gently, previously powdered if it is of the first eluted peak of latamoxef is about 8 minutes. masses. After cooling, add 10 mL of a solution of magne- System suitability— sium nitrate hexahydrate in ethanol (1 in 10), and burn the System performance: When the procedure is run with 5 mL ethanol. After cooling, add 1 mL of sulfuric acid. Proceed of the sample solution under the above operating conditions, according to Method 4, and perform the test. Prepare the the resolution between the two peaks of latamoxef is not less control solution with 2.0 mL of Standard Lead Solution (not than 3. more than 20 ppm). System repeatability: When the test is repeated 3 times (3) Arsenic <1.11>—Prepare the test solution by dissolv- with 5 mL of the sample solution under the above operating ing 1.0 g of Latamoxef Sodium in 20 mL of water, and per- conditions, the relative standard deviation of the area of the form the test (not more than 2 ppm). first eluted peak of latamoxef is not more than 2.0z. (4) Related substances—Dissolve an amount of Latamoxef Sodium, equivalent to about 25 mg (potency), in Assay Weigh accurately an amount of Latamoxef Sodium water to make exactly 50 mL, and use this solution as the and Latamoxef Ammonium RS, equivalent to about 25 mg sample solution. Pipet 2 mL of the sample solution, add (potency) each, dissolve in exactly 5 mL of the internal water to make exactly 100 mL, and use this solution as the standard solution, add water to make 50 mL, and use these standard solution. Perform the test with exactly 5 mLeachof solutions as the sample solution and standard solution. Per- the sample solution and standard solution as directed under form the test with 5 mL each of the sample solution and Liquid Chromatography <2.01> according to the following standard solution as directed under Liquid Chromatography conditions, and determine each peak area by the automatic <2.01> according to the following conditions, and calculate integration method: the peak area of 1-methyl-1H-tetrazole- the ratios, QT and QS, of the peak area of latamoxef to that 5-thiol, having the relative retention time of about 0.5 with of the internal standard. respect to the first eluted peak of the two peaks of latamo- Amount [mg (potency)] of latamoxef (C H N O S) xef, obtained from the sample solution is not larger than the 20 20 6 9 ＝ M × Q /Q × 1000 peak area of latamoxef from the standard solution, and the S T S peak area of decarboxylatamoxef, having the relative reten- MS: Amount [mg (potency)] of Latamoxef Ammonium tion time of about 1.7 with respect to the first peak of the RS two peaks of latamoxef, is not larger than 2 times that of Internal standard solution—A solution of m-cresol (3 in latamoxef from the standard solution. For this calculation, 200). 1026 Lauromacrogol / Official Monographs JP XVI

Operating conditions— (2) Unsaturated compound—Shake 0.5 g of Lauro- Detector: An ultraviolet absorption photometer (wave- macrogol with 10 mL of water, and add 5 drops of bromine length: 254 nm). TS: the color of the solution does not disappear. Column: A stainless steel column 4 mm in inside diameter Residue on ignition <2.44> Not more than 0.2z (1 g). and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Containers and storage Containers—Tight containers. Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 6.94 g of potassium dihydrogen Lenampicillin Hydrochloride phosphate, 3.22 g of disodium hydrogen phosphate dodecahydrate and 1.60 g of tetra n-butylammonium bromide in レナンピシリン塩酸塩 watertomakeexactly1000mL.To750mLofthissolution add 250 mL of methanol. Flow rate: Adjust the flow rate so that the retention time of latamoxef is about 7 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating conditions, latamoxef and the internal standard are eluted in this C H N O S.HCl: 497.95 order with the resolution between these peaks being not less 21 23 3 7 5-Methyl-2-oxo[1,3]dioxol-4-ylmethyl (2S,5R,6R)-6- than 5. [(2R)-2-amino-2-phenylacetylamino]-3,3-dimethyl-7- System repeatability: When the test is repeated 6 times oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylate with 5 mL of the standard solution under the above operating monohydrochloride conditions, the relative standard deviation of the ratios of [80734-02-7] the peak area of latamoxef to that of the internal standard is not more than 1.0z. Lenampicillin Hydrochloride is the hydrochloride of Containers and storage Containers—Tight containers. ampicillin methyloxodioxolenylmethyl ester. Storage—Not exceeding 59C. It contains not less than 653 mg (potency) and not more than 709 mg (potency) per mg, calculated on the anhydrous basis and corrected by the amount of the Lauromacrogol residual solvents. The potency of Lenampicillin Hy- drochloride is expressed as mass (potency) of ampicil- Polyoxyethylene Lauryl Alcohol Ether lin (C16H19N3O4S: 349.40). Description Lenampicillin Hydrochloride occurs as a white ラウロマクロゴール to light yellowish white powder. It is very soluble in water, in methanol and in ethanol (95), Lauromacrogol is a polyoxyethylene ether prepared and freely soluble in N,N-dimethylformamide. by the polymerization of ethylene oxide with laury al- Identification (1) Determine the infrared absorption spec- cohol. trum of Lenampicillin Hydrochloride as directed in the Description Lauromacrogol is a colorless or light yellow, potassium chloride disk method under Infrared Spectropho- clear liquid or a white, petrolatum-like or waxy solid. It has tometry <2.25>, and compare the spectrum with the Refer- a characteristic odor, and a somewhat bitter and slightly ence Spectrum or the spectrum of Lenampicillin Hydrochlo- irritative taste. ride RS: both spectra exhibit similar intensities of absorption It is very soluble in ethanol (95), in diethyl ether and in at the same wave numbers. carbon tetrachloride. (2) To 1 mL of a solution of Lenampicillin Hydrochlo- It is freely soluble or dispersed as fine oily drops in water. ride (1 in 100) add 0.5 mL of dilute nitric acid and 1 drop of silver nitrate TS: a white precipitate is formed. Identification (1) Shake well 0.5 g of Lauromacrogol 20 with 10 mL of water and 5 mL of ammonium thiocyanate- Optical rotation <2.49> [a]D : ＋174 – ＋1949(0.2 g calcu- cobalt nitrate TS, then shake with 5 mL of chloroform, and lated on the anhydrous basis and corrected on the amount of allow to stand: the chloroform layer becomes blue in color. residual solvent, ethanol (95), 20 mL, 100 mm). (2) Dissolve 0.35 g of Lauromacrogol in 10 mL of car- Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of bon tetrachloride, and perform the test as directed in the Lenampicillin Hydrochloride according to Method 2, and Solution method under Infrared Spectrophotometry <2.25> perform the test. Prepare the control solution with 2.0 mL of using a 0.1-mm fixed cell: it exhibits absorption at the wave Standard Lead Solution (not more than 10 ppm). numbers of about 1347 cm－1, 1246 cm－1 and 1110 cm－1. (2) Arsenic <1.11>—Prepare the test solution with 1.0 g Purity (1) Acidity—Transfer 10.0 g of Lauromacrogol of Lenampicillin Hydrochloride according to Method 3, and into a flask, and add 50 mL of neutralized ethanol. Heat on perform the test (not more than 2 ppm). a water bath nearly to boil, shaking once or twice while heat- (3) Free ampicillin—Weigh accurately about 0.1 g of ing. Cool, and add 5.3 mL of 0.1 mol/L sodium hydroxide Lenampicillin Hydrochloridein, dissolve in exactly 10 mL of VS and 5 drops of phenolphthalein TS: a red color develops. the internal standard solution, and use this solution as the JP XVI Official Monographs / Lenampicillin Hydrochloride 1027 sample solution. Separately, weigh accurately an amount of propanol and about 0.12 g of ethyl acetate, and add N,N- Ampicillin RS, equivalent to about 25 mg (potency), and dis- dimethylformamide to make exactly 100 mL. Pipet 1 mL solve in water to make exactly 100 mL. Pipet 2 mL of this and 3 mL of this solution, add exactly 1 mL each of the in- solution, add exactly 10 mL of the internal standard solu- ternal standard solution, add N,N-dimethylformamide to tion, and use this solution as the standard solution. The sam- make 5 mL, and use these solutions as the standard solution ple solution should be used to the following test immediately (1) and the standard solution (2), respectively. Perform the after the solution is prepared. Perform the test with 10 mL test with 4 mL each of the sample solution, standard solution each of the sample solution and standard solution as directed (1) and (2) as directed under Gas Chromatography <2.02> ac- under Liquid Chromatography <2.01> according to the fol- cording to the following conditions, and calculate the ratios, lowing conditions, and calculate the ratios, QT and QS,of QTa and QTb, of the peak height of 2-propanol and ethyl ace- the peak height of ampicillin to that of the internal standard: tate to that of the internal standard of the sample solution, the amount of ampicillin is not more than 1.0z. the ratios, QSa1 and QSb1, of the peak height of 2-propanol and ethyl acetate to that of the internal standard of the Amount (z) of ampicillin (C H N O S) 16 19 3 4 standard solution (1) and the ratios, Q and Q ,ofthe ＝ M /M × Q /Q × 2 Sa2 Sb2 S T T S peak height of 2-propanol and ethyl acetate to that of the

MS: Amount [mg (potency)] of Ampicillin RS internal standard of the standard solution (2). Calculate the MT: Amount (mg) of the sample amounts of 2-propanol and ethyl acetate by the following equations: not more than 0.7z and not more than 1.7z, Internal standard solution—A solution of anhydrous respectively. caffeine in the mobile phase (1 in 50,000). Operating conditions— Amount (z) of 2-propanol

Detector: An ultraviolet absorption photometer (wave- ＝ MSa/MT × (2QTa － 3QSa1 ＋ QSa2)/(QSa2 － QSa1) length: 230 nm). Amount (z) of ethyl acetate Column: A stainless steel column 4 mm in inside diameter ＝ M /M × (2Q － 3Q ＋ Q )/(Q － Q ) and 30 cm in length, packed with octadecylsilanized silica gel Sb T Tb Sb1 Sb2 Sb2 Sb1 for liquid chromatography (10 mm in particle diameter). MSa: Amount (g) of 2-propanol Column temperature: A constant temperature of about MSb: Amount (g) of ethyl acetate 259C. MT: Amount (g) of the sample Mobile phase: Dissolve 1.22 g of potassium dihydrogen Internal standard solution—A solution of cyclohexane in phosphate in water to make 900 mL, and add 100 mL of N,N-dimethylformamide (1 in 1000). acetonitrile. Operating conditions— Flow rate: Adjust the flow rate so that the retention time Detector: A hydrogen flame-ionization detector. of ampicillin is about 7 minutes. Column: A glass column 3 mm in inside diameter and 3 m System suitability— in length, packed with siliceous earth for gas chromatogra- System performance: When the procedure is run with 10 phy (180 – 250 mm in particle diameter) coated with tetra- mL of the standard solution under the above operating con- kishydroxypropylethylenediamine for gas chromatography ditions, ampicillin and the internal standard are eluted in this at the ratio of 10 to 15z. order with the resolution between these peaks being not less Column temperature: A constant temperature of about than 5. 809C. System repeatability: When the test is repeated 6 times Injection port temperature: A constant temperature of with 10 mL of the standard solution under the above operat- about 1609C. ing conditions, the relative standard deviation of the ratios Carrier gas: Nitrogen. of the peak height of ampicillin to that of the internal stand- Flow rate: Adjust the flow rate so that the retention time ard is not more than 5z. of the internal standard is about 1 minute. (4) Penicilloic acid—Weigh accurately about 0.1 g of System suitability— Lenampicillin Hydrochloride, dissolve in water to make ex- System performance: When the procedure is run with 4 mL actly 100 mL, and use this solution as the sample solution. of the standard solution (2) under the above operating condi- Pipet 10 mL of the sample solution, add 10 mL of potassium tions, the internal standard, ethyl acetate and 2-propanol are hydrogen phthalate buffer solution, pH 4.6 and exactly 10 eluted in this order, and the resolution between the peaks of mL of 0.005 mol/L iodine VS, allow to stand for exactly 15 the internal standard and ethyl acetate is not less than 2.0. minutes while protecting from exposure to light, and titrate System repeatability: When the test is repeated 3 times <2.50> with 0.01 mol/L sodium thiosulfate VS (indicator: 1 with 4 mL of the standard solution (2) under the above oper- mL of starch TS). Perform a blank determination, and make ating conditions, the relative standard deviation of the ratios any necessary correction: the amount of penicilloic acid of the peak height of ethyl acetate to that of the internal (C H N O S: 367.42) is not more than 3.0z. 16 21 3 5 standard is not more than 5.0z. Each mL of 0.01 mol/L sodium thiosulfate VS Water <2.48> Not more than 1.5z (1 g, volumetric titra- ＝ 0.45 mg of C H N O S 16 21 3 5 tion, direct titration). (5) Residual solvent <2.46>—Weigh accurately about Residue on ignition <2.44> Not more than 0.2z (1 g). 0.25 g of Lenampicillin Hydrochloride, dissolve in exactly 1 mL of the internal standard solution, add N,N-dimethylfor- Assay Weigh accurately an amount of Lenampicillin Hy- mamide to make 5 mL, and use this solution as the sample drochloride and Lenampicillin Hydrochloride RS, equivalent solution. Separately, weigh accurately about 80 mg of 2- to about 0.1 g (potency), dissolve each in the internal stand- 1028 L-Leucine / Official Monographs JP XVI ard solution to make exactly 10 mL, and use these solutions of L-Leucine, previously dried, as directed in the potassium as the sample solution and the standard solution. Perform bromide disk method under Infrared Spectrophotometry the test with 5 mL each of the sample solution and standard <2.25>, and compare the spectrum with the Reference Spec- solution as directed under Liquid Chromatography <2.01> trum: both spectra exhibit similar intensities of absorption at according to the following conditions, and calculate the the same wave numbers. ratios, Q and Q , of the peak area of lenampicillin to that T S Optical rotation <2.49> [a]20: ＋14.5 – ＋16.09(after dry- of the internal standard. D ing, 1 g, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm).

Amount [mg (potency)] of ampicillin (C16H19N3O4S) pH <2.54> Dissolve 1.0 g of L-Leucine in 100 mL of water: ＝ M × Q /Q × 1000 S T S the pH of this solution is between 5.5 and 6.5. M : Amount [mg (potency)] of Lenampicillin Hydrochlo- S Purity (1) Clarity and color of solution—Dissolve 0.5 g ride RS of L-Leucine in 10 mL of 1 mol/L hydrochloric acid TS: the Internal standard solution—A solution of ethyl aminobenzo- solution is clear and colorless. ate in the mobile phase (1 in 4000). (2) Chloride <1.03>—Dissolve 0.5 g of L-Leucine in 40 Operating conditions— mL of water and 6 mL of dilute nitric acid, and add water to Detector: An ultraviolet absorption photometer (wave- make 50 mL. Perform the test using this solution as the test length: 254 nm). solution. Prepare the control solution with 0.30 mL of 0.01 Column: A stainless steel column 6 mm in inside diameter mol/L hydrochloric acid VS (not more than 0.021z). and 15 cm in length, packed with octadecylsilanized silica gel (3) Sulfate <1.14>—Dissolve 0.6 g of L-Leucine in 40 mL for liquid chromatography (5 mminparticlediameter). of water and 1 mL of dilute hydrochloric acid, and add Column temperature: A constant temperature of about water to make 50 mL. Perform the test using this solution as 259C. the test solution. Prepare the control solution with 0.35 mL Mobile phase: Dissolve 9.53 g of potassium dihydrogen of 0.005 mol/L sulfuric acid VS (not more than 0.028z). phosphate in water to make exactly 700 mL, and add aceto- (4) Ammonium <1.02>—Perform the test with 0.25 g of nitrile to make exactly 1000 mL. L-Leucine. Prepare the control solution with 5.0 mL of Flow rate: Adjust the flow rate so that the retention time Standard Ammonium Solution (not more than 0.02z). of lenampicillin is about 6 minutes. (5) Heavy metals <1.07>—Proceed with 1.0 g of L-Leu- System suitability— cine according to Method 4, and perform the test. Prepare System performance: When the procedure is run with 5 mL the control solution with 2.0 mL of Standard Lead Solution of the standard solution under the above operating condi- (not more than 20 ppm). tions, lenampicillin and the internal standard are eluted in (6) Arsenic <1.11>—Prepare the test solution with 1.0 g this order with the resolution between these peaks being not of L-Leucine according to Method 2, and perform the test less than 10. (not more than 2 ppm). System repeatability: When the test is repeated 6 times (7) Related substances—Dissolve 0.10 g of L-Leucine in with 5 mL of the standard solution under the above operating water by warming, after cooling, add water to make 25 mL, conditions, the relative standard deviation of the ratios of and use this solution as the sample solution. Pipet 1 mL of the peak area of lenampicillin to that of the internal standard the sample solution, and add water to make exactly 50 mL. is not more than 1.0z. Pipet 5 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform Containers and storage Containers—Tight containers. the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin- L-Leucine layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance of L-ロイシン about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat at 809C for 5 minutes: the spots other than the principal spot from the sample solution are not more in-

C6H13NO2:131.17 tense than the spot from the standard solution. (2S)-2-Amino-4-methylpentanoic acid Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, [61-90-5] 3 hours).

L-Leucine, when dried, contains not less than 98.5z Residue on ignition <2.44> Not more than 0.1z (1 g). of C6H13NO2. Assay Weigh accurately about 0.13 g of L-Leucine, previ- Description L-Leucine occurs as white crystals or crystal- ously dried, and dissolve in 3 mL of formic acid, add 50 mL line powder. It is odorless or has a faint characteristic odor, of acetic acid (100), and titrate <2.50> with 0.1 mol/L per- and has a slightly bitter taste. chloric acid VS (potentiometric titration). Perform a blank It is freely soluble in formic acid, sparingly soluble in determination, and make any necessary correction. water, and practically insoluble in ethanol (95). Each mL of 0.1 mol/L perchloric acid VS It dissolves in dilute hydrochloric acid. ＝ 13.12 mg of C6H13NO2 Identification Determine the infrared absorption spectrum Containers and storage Containers—Well-closed contain- JP XVI Official Monographs / Levallorphan Tartrate Injection 1029 ers. with a mixture of methanol and ammonia TS (200:3) to a distance of about 10 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate: the spots other Levallorphan Tartrate than the principal spot from the sample solution are not more intense than the spot from the standard solution. レバロルファン酒石酸塩 Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum, phosphorus (V) oxide, 809C, 4 hours). Residue on ignition <2.44> Not more than 0.10z (1 g). Assay Weigh accurately about 0.5 g of Levallorphan Tar- trate, previously dried, dissolve in 30 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 2 drops of crystal violet TS). Perform a blank determination, and make any necessary correction.

C19H25NO.C4H6O6: 433.49 Each mL of 0.1 mol/L perchloric acid VS 17-Allylmorphinan-3-ol monotartrate ＝ 43.35 mg of C19H25NO.C4H6O6 [71-82-9] Containers and storage Containers—Well-closed containers. Levallorphan Tartrate, when dried, contains not less than 98.5z of C19H25NO.C4H6O6. Description Levallorphan Tartrate occurs as a white to Levallorphan Tartrate Injection pale yellow, crystalline powder. It is odorless. It is soluble in water and in acetic acid (100), sparingly レバロルファン酒石酸塩注射液 soluble in ethanol (95), and practically insoluble in diethyl ether. Levallorphan Tartrate Injection is an aqueous solu- Identification (1) Determine the absorption spectrum of a tion for injection. solution of Levallorphan Tartrate in 0.01 mol/L hydrochlo- It contains not less than 93.0z and not more than ric acid TS (1 in 10,000) as directed under Ultraviolet-visible 107.0z of the labeled amount of levallorphan tartrate Spectrophotometry <2.24>, and compare the spectrum with (C19H25NO.C4H6O6: 433.49). the Reference Spectrum: both spectra exhibit similar intensi- Method of preparation Prepare as directed under Injec- ties of absorption at the same wavelengths. tion, with Levallorphan Tartrate. (2) Determine the infrared absorption spectrum of Levallorphan Tartrate, previously dried, as directed in the Description Levallorphan Tartrate Injection is a clear, col- potassium bromide disk method under Infrared Spectropho- orless liquid. tometry <2.25>, and compare the spectrum with the Refer- pH: 3.0 – 4.5 ence Spectrum: both spectra exhibit similar intensities of ab- Identification Take an exact volume of Levallorphan Tar- sorption at the same wave numbers. trate Injection, equivalent to 3 mg of Levallorphan Tartrate (3) A solution of Levallorphan Tartrate (1 in 30) re- according to the labeled amount, add 5 mL of water and 2 sponds to the Qualitative Tests <1.09> (1)and(2)fortartrate. drops of dilute hydrochloric acid, and wash with five 15-mL 20 Optical rotation <2.49> [a]D : －37.0 – －39.29(after dry- portions of diethyl ether by a vigorous shaking. Take the ing, 0.2 g, water, 10 mL, 100 mm). water layer, evaporate the diethyl ether remained by warming on a water bath, and after cooling, add 0.01 mol/L pH <2.54> Dissolve 0.2 g of Levallorphan Tartrate in 20 hydrochloric acid TS to make 50 mL. Determine the absorp- mL of water: the pH of this solution is between 3.3 and 3.8. tion spectrum of this solution as directed under Ultraviolet- Melting point <2.60> 174–1789C visible Spectrophotometry <2.24>: it exhibits a maximum between 277 nm and 281 nm. Purity (1) Clarity and color of solution—Dissolve 0.2 g of Levallorphan Tartrate in 10 mL of water: the solution is Bacterial endotoxins <4.01> Less than 150 EU/mg. clear and colorless. Extractable volume <6.05> It meets the requirement. (2) Heavy metals <1.07>—Proceed with 1.0 g of Levallor- phan Tartrate according to Method 4, and perform the test. Foreign insoluble matter <6.06> Perform the test according Prepare the control solution with 2.0 mL of Standard Lead to Method 1: it meets the requirement. Solution (not more than 20 ppm). Insoluble particulate matter <6.07> It meets the require- (3) Related substances—Dissolve 0.20 g of Levallorphan ment. Tartrate in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to Sterility <4.06> Perform the test according to the Mem- make exactly 100 mL, and use this solution as the standard brane filtration method: it meets the requirement. solution. Perform the test with these solutions as directed Assay Take exactly a volume of Levallorphan Tartrate In- under Thin-layer Chromatography <2.03>. Spot 20 mLeach jection, equivalent to about 2 mg of levallorphan tartrate of the sample solution and standard solution on a plate of (C H NO.C H O ), add exactly 10 mL of the internal silica gel for thin-layer chromatography. Develop the plate 19 25 4 6 6 1030 Levodopa / Official Monographs JP XVI standard solution, and use this solution as the sample solu- Description Levodopa occurs as white or slightly grayish tion. Separately, weigh accurately about 0.1 g of levallor- white crystals or crystalline powder. It is odorless. phan tartrate for assay, previously dried at 809C for 4 hours It is freely soluble in formic acid, slightly soluble in water, on phosphorus (V) oxide under reduced pressure, and dis- and practically insoluble in ethanol (95). solve in water to make exactly 100 mL. Pipet 2 mL of this It dissolves in dilute hydrochloric acid. solution, add exactly 10 mL of the internal standard solu- The pH of a saturated solution of Levodopa is between tion, and use this solution as the standard solution. Perform 5.0 and 6.5. the test with 10 mL each of the sample solution and standard Melting point: about 2759C (with decomposition). solution as directed under Liquid Chromatography <2.01> Identification (1) To 5 mL of a solution of Levodopa (1 according to the following operating conditions, and calcu- in 1000) add 1 mL of ninhydrin TS, and heat for 3 minutes late the ratios, Q and Q , of the peak area of levallorphan T S in a water bath: a purple color develops. to that of the internal standard: (2) To 2 mL of a solution of Levodopa (1 in 5000) add 10

Amount (mg) of C19H25NO.C4H6O6 mL of 4-aminoantipyrine TS, and shake: a red color de- ＝ MS × QT/QS × 1/50 velops. (3) Dissolve 3 mg of Levodopa in 0.001 mol/L hydro- M : Amount (mg) of levallorphan tartrate for assay S chloric acid TS to make 100 mL. Determine the absorption Internal standard solution—Dissolve 0.04 g of isobutyl par- spectrum of the solution as directed under Ultraviolet-visible ahydroxybenzoate in 10 mL of ethanol (95), add water to Spectrophotometry <2.24>, and compare the spectrum with make 100 mL, and to 10 mL of this solution add water to the Reference Spectrum: both spectra exhibit similar intensi- make 100 mL. ties of absorption at the same wavelengths. Operating conditions— Absorbance <2.24> E 1z (280 nm): 136 – 146 (after drying, Detector: An ultraviolet absorption photometer (wave- 1cm 30 mg, 0.001 mol/L hydrochloric acid TS, 1000 mL). length: 280 nm). 20 Column: A stainless steel column 4.6 mm in inside diame- Optical rotation <2.49> [a]D : －11.5 – －13.09(after dry- ter and 15 cm in length, packed with octadecylsilanized silica ing, 2.5 g, 1 mol/L hydrochloric acid TS, 50 mL, 100 nm). gel for liquid chromatography (5 mm in particle diameter). Purity (1) Clarity and color of solution—Dissolve 1.0 g Column temperature: A constant temperature of about of Levodopa in 20 mL of 1 mol/L hydrochloric acid TS: the 409C. solution is clear and colorless. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in (2) Chloride <1.03>—Dissolve 0.5 g of Levodopa in 6 mL 500 mL of diluted phosphoric acid (1 in 1000), and adjust of dilute nitric acid, and add water to make 50 mL. Perform the pH to 3.0 with sodium hydroxide TS. To 300 mL of this the test using this solution as the test solution. Prepare the solution add 200 mL of acetonitrile. control solution with 0.3 mL of 0.01 mol/L hydrochloric Flow rate: Adjust the flow rate so that the retention time acid VS (not more than 0.021z). of levallorphan is about 12 minutes. (3) Sulfate <1.14>—Dissolve 0.40 g of Levodopa in 1 mL System suitability— of dilute hydrochloric acid and 30 mL of water, and add System performance: When the procedure is run with 10 water to make 50 mL. Perform the test using this solution as mL of the standard solution under the above operating con- the test solution. Prepare the control solution with 0.25 mL ditions, the internal standard and levallorphan are eluted in of 0.005 mol/L sulfuric acid VS (not more than 0.030z). this order with the resolution between these peaks being not (4) Heavy metals <1.07>—Proceed with 1.0 g of Levo- less than 5. dopa according to Method 2, and perform the test. Prepare System repeatability: When the test is repeated 6 times the control solution with 2.0 mL of Standard Lead Solution with 10 mL of the standard solution under the above operat- (not more than 20 ppm). ing conditions, the relative standard deviation of the ratios (5) Arsenic <1.11>—Dissolve 1.0 g of Levodopa in 5 mL of the peak area of levallorphan to that of the internal stand- of dilute hydrochloric acid, and perform the test with this so- ard is not more than 1.0z. lution as the test solution (not more than 2 ppm). Containers and storage Containers—Hermetic containers. (6) Related sulstances—Dissolve 0.10 g of Levodopa in 10 mL of sodium disulfite TS, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add so- Levodopa dium disulfite TS to make exactly 25 mL. Pipet 1 mL of this solution, add sodium disulfite TS to make exactly 20 mL, レボドパ and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro- matography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of 1- butanol, water, acetic acid (100) and methanol (10:5:5:1) to

C9H11NO4:197.19 a distance of about 10 cm, and air-dry the plate. Spray 3-Hydroxy-L-tyrosine evenly a solution of ninhydrin in acetone (1 in 50) on the [59-92-7] plate and heat at 909C for 10 minutes: the spots other than the principal spot from the sample solution are not more in- Levodopa, when dried, contains not less than 98.5z tense than the spot from the standard solution. of C9H11NO4. JP XVI Official Monographs / Levofloxacin Hydrate 1031

Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, Levofloxacin Hydrate according to Method 4, and perform 3 hours). the test. Prepare the control solution with 2.0 mL of Stand- ard Lead Solution (not more than 10 ppm). Residue on ignition <2.44> Not more than 0.1z (1 g). (2) Related substances—Conduct this procedure using Assay Weigh accurately about 0.3 g of Levodopa, previ- light-resistant vessels. Dissolve 50 mg of Levofloxacin Hy- ously dried, dissolve in 3 mL of formic acid, add 80 mL of drate in 10 mL of a mixture of water and methanol (1:1), acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric and use this solution as the sample solution. Pipet 1 mL of acid VS until the color of the solution changes from purple the sample solution, and add a mixture of water and metha- through blue-green to green (indicator: 3 drops of crystal nol (1:1) to make exactly 10 mL. Pipet 1 mL of this solution, violet TS). Perform a blank determination, and make any add a mixture of water and methanol (1:1) to make exactly necessary correction. 10 mL, and use this solution as the standard solution. Per- form the test with exactly 10 mL each of the sample solution Each mL of 0.1 mol/L perchloric acid VS and standard solution as directed under Liquid Chromatog- ＝ 19.72 mg of C H NO 9 11 4 raphy <2.01> according to the following conditions. Deter- Containers and storage Containers—Tight containers. mine each peak area of both solutions by the automatic in- Storage—Light-resistant. tegration method: the area of the peak having the relative retention time of about 1.2 with respect to levofloxacin obtained from the sample solution is not larger than 2/5 times Levofloxacin Hydrate the peak area of levofloxacin from the standard solution, and the area of each peak other than the peak of levofloxa- レボフロキサシン水和物 cin and other than the peak having the relative retention time of about 1.2 with respect to levofloxacin from the sample solution is not larger than 1/5 times the peak area of levofloxacin from the standard solution. Furthermore, the total area of the peaks other than the peak of levofloxacin and other than the peak having the relative retention time of about 1.2 with respect to levofloxacin from the sample solution is not larger than 3/10 times the peak area of levofloxacin from the 1 C18H20FN3O4. 2 H2O: 370.38 standard solution. (3S)-9-Fluoro-3-methyl-10-(4-methylpiperazin-1-yl)- Operating conditions— 7-oxo-2,3-dihydro-7H-pyrido[1,2,3-de][1,4]benzoxazine- Detector: An ultraviolet absorption photometer (wave- 6-carboxylic acid hemihydrate length: 340 nm). [138199-71-0] Column: A stainless steel column 4.6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica Levofloxacin Hydrate contains not less than gel for liquid chromatography (5 mminparticlediameter). 99.0z and not more than 101.0z of levofloxacin Column temperature: A constant temperature of about (C18H20FN3O4: 361.37), calculated on the anhydrous 459C. basis. Mobile phase: Dissolve 1.76 g of L-valine, 7.71 g of ammonium acetate and 1.25 g of Copper (II) sulfate pentahy- Description Levofloxacin Hydrate occurs as light yellowish drate in water to make 1000 mL. To this solution add 250 white to yellowish white crystals or crystalline powder. mL of methanol. It is freely soluble in acetic acid (100), sparingly soluble in Flow rate: Adjust the flow rate so that the retention time water and in methanol, and slightly soluble in ethanol (99.5). of levofloxacin is about 22 minutes. It dissolves in 0.1 mol/L hydrochloric acid TS. Time span of measurement: About 2 times as long as the It gradually turns dark light yellowish white on exposure retention time of levofloxacin, beginning after the solvent to light. peak. Melting point: about 2269C (with decomposition). System suitability— Identification (1) Determine the absorption spectrum of a Test for required detectability: Pipet 1 mL of the standard solution of Levofloxacin Hydrate in 0.1 mol/L hydrochloric solution, and add a mixture of water and methanol (1:1) to acid solution (1 in 150,000) as directed under Ultraviolet- make exactly 20 mL. Confirm that the peak area of levoflox- visible Spectrophotometry <2.24>, and compare the spectrum acin obtained from 10 mL of this solution is equivalent to 4 with the Reference Spectrum: both spectra exhibit similar in- to 6z of that of levofloxacin from 10 mL of the standard so- tensities of absorption at the same wavelengths. lution. (2) Determine the infrared absorption spectrum of System performance: Dissolve 10 mg of ofloxacin in 20 Levofloxacin Hydrate as directed in the potassium bromide mL of a mixture of water and methanol (1:1). To 1 mL of disk method under Infrared Spectrophotometry <2.25>,and this solution add a mixture of water and methanol (1:1) to compare the spectrum with the Reference Spectrum: both make 10 mL. When the procedure is run with 10 mLofthis spectra exhibit similar intensities of absorption at the same solution under the above operating conditions, the resolu- wave numbers. tion between the peak of levofloxacin and the peak having the relative retention time of about 1.2 with respect to Optical rotation <2.49> [a]20: －92 – －999(0.1 g calculated D levofloxacin is not less than 3. on the anhydrous basis, methanol, 10 mL, 100 mm). System repeatability: When the test is repeated 6 times Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of with 10 mL of the standard solution under the above operat- 1032 Levomepromazine Maleate / Official Monographs JP XVI ing conditions, the relative standard deviation of the peak 25-mL portions of diethyl ether. Combine all the diethyl area of levofloxacin is not more than 3.0z. ether extracts, evaporate the diethyl ether in a water bath at (3) Residual solvent Being specified separately. a temperature of about 359Cwiththeaidofacurrentofair: the residue melts <2.60> between 1289Cand1369C. Water <2.48> 2.1 – 2.7z (0.5 g, volumetric titration, direct 20 titration). Optical rotation <2.49> [a]D : －13.5 – －16.59(after drying, 0.5 g, chloroform, 20 mL, 200 mm). Residue on ignition <2.44> Not more than 0.1z (1 g). Purity (1) Clarity and color of solution—To 0.5 g of Assay Weigh accurately about 0.3 g of Levofloxacin Hy- Levomepromazine Maleate add 10 mL of methanol, and dis- drate, dissolve in 100 mL of acetic acid (100), and titrate solve by warming: the solution is clear, and colorless or pale <2.50> with 0.1 mol/L perchloric acid VS (potentiometric yellow. titration). Perform a blank determination in the same man- (2) Chloride <1.03>—Dissolve 0.5 g of Levomepromazine ner, and make any necessary correction. Maleate in 40 mL of methanol, and add 6 mL of dilute nitric Each mL of 0.1 mol/L perchloric acid VS acid and water to make 50 mL. Perform the test using this

＝ 36.14 mg of C18H20FN3O4 solution as the test solution. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS, 40 mL of Containers and storage Containers—Tight containers. methanol, 6 mL of dilute nitric acid and water to make 50 Storage—Light-resistant. mL (not more than 0.028z). (3) Heavy metals <1.07>—Proceedwith2.0gof Levomepromazine Maleate according to Method 2, and per- Levomepromazine Maleate form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). レボメプロマジンマレイン酸塩 Loss on drying <2.41> Not more than 0.5z (2 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 1 g of Levomepromazine Maleate, previously dried, and dissolve in a mixture of 40 mL of acetic acid (100) and 20 mL of acetone for nona-

C19H24N2OS.C4H4O4:444.54 queous titration. Titrate <2.50> with 0.1 mol/L perchloric (2R)-3-(2-Methoxy-10H-phenothiazin-10-yl)- acid VS until the color of the solution changes from red- N,N,2-trimethylpropylamine monomaleate purple through blue-purple to blue (indicator: 5 drops of [7104-38-3] bromocresol green-methylrosaniline chloride TS). Perform a blank determination, and make any necessary correction. Levomepromazine Maleate, when dried, contains Each mL of 0.1 mol/L perchloric acid VS not less than 98.0z of C19H24N2OS.C4H4O4. ＝ 44.45 mg of C19H24N2OS.C4H4O4 Description Levomepromazine Maleate occurs as white Containers and storage Containers—Tight containers. crystals or crystalline powder. It is odorless, and has a Storage—Light-resistant. slightly bitter taste. It is freely soluble in acetic acid (100), soluble in chloroform, sparingly soluble in methanol, slightly soluble in ethanol (95) and in acetone, very slightly soluble in water, Levothyroxine Sodium Hydrate and practically insoluble in diethyl ether. レボチロキシンナトリウム水和物 Melting point: 184 – 1909C (with decomposition). Identification (1) Dissolve 5 mg of Levomepromazine Maleate in 5 mL of sulfuric acid: a red-purple color develops, which slowly becomes deep red-purple. To this solution add 1 drop of potassium dichromate TS: a brownish yellow-red color is produced.

(2) To 0.2 g of Levomepromazine Maleate add 5 mL of C15H10I4NNaO4.xH2O sodium hydroxide TS and 20 mL of diethyl ether, and shake Monosodium O-(4-hydroxy-3,5-diiodophenyl)-3,5-diiodo- well. Separate the diethyl ether layer, wash twice with 10-mL L-tyrosinate hydrate portions of water, add 0.5 g of anhydrous sodium sulfate, [25416-65-3] filter, evaporate the diethyl ether on a water bath, and dry the residue at 1059C for 2 hours: the residue melts <2.60> be- Levothyroxine Sodium Hydrate contains not less tween 1249Cand1289C. than 97.0z of levothyroxine sodium (C15H10I4NNaO4: (3) To 0.5 g of Levomepromazine Maleate add 5 mL of 798.85), calculated on the dried basis. water and 2 mL of ammonia solution (28), extract with three Description Levothyroxine Sodium Hydrate occurs as a 5-mL portions of chloroform, separate and evaporate the pale yellowish white to light yellow-brown powder. It is water layer to dryness. To the residue add 2 to 3 drops of odorless. dilute sulfuric acid and 5 mL of water, and extract with four It is slightly soluble in ethanol (95), and practically insolu- JP XVI Official Monographs / Levothyroxine Sodium Tablets 1033 bleinwaterandindiethylether. Combustion Method <1.06>, using a mixture of 10 mL of so- It dissolves in sodium hydroxide TS. dium hydroxide solution (1 in 100) and 1 mL of a freshly It is gradually colored by light. prepared sodium bisulfate solution (1 in 100) as the absorbing liquid, and prepare the test solution. Apply a small Identification (1) Heat 0.1 g of Levothyroxine Sodium amount of water to the upper part of apparatus A, pull out Hydrate over a flame: a purple gas evolves. C carefully, and wash C, B and the inner wall of A with 40 (2) To 0.5 mg of Levothyroxine Sodium Hydrate add 8 mL of water. To the test solution add 1 mL of bromine- mL of a mixture of water, ethanol (95), hydrochloric acid acetic acid TS, insert the stopper C, and shake vigorously for and sodium hydroxide TS (6:5:2:2), warm in a water bath 1 minute. Remove the stopper, rinse the stopper, the sample for 2 minutes, cool, and add 0.1 mL of sodium nitrite TS. holder and the inner wall of the flask with 40 mL of water, Allow to stand in a dark place for 20 minutes, and add 1.5 and add 0.5 mL of formic acid. Stopper the flask with C, mL of ammonia solution (28): a yellowish red color is pro- and shake vigorously for 1 minute again. Remove the stop- duced. per, and rinse the stopper, the sample holder and the inner (3) Determine the absorption spectrum of a solution of wall of the flask with 40 mL of water. Bubble the solution Levothyroxine Sodium Hydrate in dilute sodium hydroxide with enough nitrogen gas in the flask to remove the oxygen TS (1 in 10,000) as directed under Ultraviolet-visible Spectro- and excess bromine, add 0.5 g of potassium iodide to the so- photometry <2.24>, and compare the spectrum with the Ref- lution, and dissolve. Add immediately 3 mL of dilute sulfu- erence Spectrum: both spectra exhibit similar intensities of ric acid, mix, and allow to stand for 2 minutes. Titrate <2.50> absorption at the same wavelengths. the solution with 0.02 mol/L sodium thiosulfate VS (indica- (4) Moisten Levothyroxine Sodium Hydrate with sulfu- tor: 3 mL of starch TS). Perform a blank determination, and ric acid, and ignite: the residue responds to the Qualitative make any necessary correction. Tests <1.09> (1) and (2) for sodium salt. Each mL of 0.02 mol/L sodium thiosulfate VS Optical rotation <2.49> [a]20: －5–－69(0.3 g, calculated D ＝ 0.6657 mg of C H I NNaO on the dried basis, a mixture of ethanol (95) and sodium hy- 15 10 4 4 droxide TS (2:1), 10 mL, 100 mm). Containers and storage Containers—Tight containers. Storage—Light-resistant. Purity (1) Clarity and color of solution—Dissolve 0.3 g of Levothyroxine Sodium Hydrate in 10 mL of a mixture of ethanol (95) and sodium hydroxide TS (2:1) by warming: the solution is clear and pale yellow to pale yellow-brown in Levothyroxine Sodium Tablets color. レボチロキシンナトリウム錠 (2) Soluble halides—Dissolve 0.01 g of Levothyroxine Sodium Hydrate in 10 mL of water and 1 drop of dilute nitric acid, shake for 5 minutes, and filter. To the filtrate Levothyroxine Sodium Tablets contain not less than add water to make 10 mL, then add 3 drops of silver nitrate 90.0z and not more than 110.0z of the labeled TS, and mix: the solution has no more opalescence than the amount of levothyroxine sodium (C15H10I4NNaO4: following control solution. 798.85). Control solution: To 0.20 mL of 0.01 mol/L hydrochloric Method of preparation Prepare as directed under Tablets, acid VS add 10 mL of water and 1 drop of dilute nitric acid, with Levothyroxine Sodium Hydrate. and proceed as directed above. (3) Related substances—Dissolve 20 mg of Levothyro- Identification (1) Weigh a quantity of powdered Levo- xine Sodium Hydrate in 2 mL of a mixture of ethanol (95) thyroxine Sodium Tablets, equivalent to 0.5 mg of Levo- and ammonia solution (28) (14:1), and use this solution as thyroxine Sodium Hydrate according to the labeled amount, the sample solution. Pipet 1 mL of the sample solution, add add 8 mL of a mixture of water, ethanol (95), hydrochloric a mixture of ethanol (95) and ammonia solution (28) (14:1) acid and sodium hydroxide TS (6:5:2:2), warm in a water to make exactly 50 mL, and use this solution as the standard bath for 2 minutes, cool, and filter. To the filtrate add 0.1 solution. Perform the test with these solutions as directed mL of sodium nitrite TS, and allow to stand in a dark place under Thin-layer Chromatography <2.03>. Spot 5 mLeachof for 20 minutes. Add 1.5 mL of ammonia solution (28): a yel- the sample solution and standard solution on a plate of silica lowish red color develops. gel for thin-layer chromatography. Develop the plate with a (2) To a quantity of powdered Levothyroxine Sodium mixture of t-butanol, t-amyl alcohol, water, ammonia solu- Tablets, equivalent to 1 mg of Levothyroxine Sodium Hy- tion (28) and 2-butanone (59:32:17:15:7) to a distance of drate according to the labeled amount, add 10 mL of ethanol about 12 cm, and air-dry the plate. Spray evenly a solution (95), shake, filter, and use the filtrate as the sample solution. of 0.3 g of ninhydrin in 100 mL of a mixture of 1-butanol Dissolve 0.01 g of levothyroxine sodium for thin-layer chro- and acetic acid (100) (97:3) on the plate, and heat at 1009C matography in 100 mL of ethanol (95), and use this solution for 3 minutes: the red-purple spots other than the principal as the standard solution. Perform the test with these solu- spot from the sample solution are not more intense than the tions as directed under Thin-layer Chromatography <2.03>. spot from the standard solution. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Loss on drying <2.41> 7–11z (0.5 g, in vacuum, phospho- Develop the plate with a mixture of t-butanol, t-amyl alco- rus (V) oxide, 609C, 4 hours). hol, water, ammonia solution (28) and 2-butanone Assay Weigh accurately about 25 mg of Levothyroxine So- (59:32:17:15:7) to a distance of about 12 cm, and air-dry the dium Hydrate, and proceed as directed under Oxygen Flask plate. Spray a solution of 0.3 g of ninhydrin in 100 mL of a 1034 Lidocaine / Official Monographs JP XVI mixture of 1-butanol and acetic acid (100) (97:3) on the Dissolution Being specified separately. plate, and heat at 1009C for 3 minutes: the spots obtained Assay Weigh accurately and powder not less than 20 Levo- from the sample solution and the standard solution show a thyroxine Sodium Tablets. Weigh accurately a portion of the red-purple color, and has the same Rfvalue. powder, equivalent to about 3 mg of levothyroxine sodium

Purity Soluble halides—Weigh a quantity of powdered (C15H10I4NNaO4), into a crucible, and add potassium car- Levothyroxine Sodium Tablets, equivalent to 2.5 mg of bonate amounting to twice the mass of the powder. In the Levothyroxine Sodium Hydrate according to the labeled case that the weighed powder is less than 4 g, add 8 g of po- amount, add 25 mL of water, warm to 409C, shake for 5 tassium carbonate to the crucible. Mix well, and gently tap minutes, add 3 drops of dilute nitric acid, and filter. To the the crucible on the bench to compact the mixture. Overlay filtrate add 3 drops of silver nitrate TS, and mix: the solu- with 10 g of potassium carbonate, and compact again by tap- tion has no more opalescence than the following control so- ping. Heat the crucible strongly at a temperature between lution. 6759Cand7009C for 25 minutes. Cool, add 30 mL of water, Control solution: To 0.25 mL of 0.01 mol/L hydrochloric heat gently to boiling, and filter into a flask. To the residue acid VS add 25 mL of water and 3 drops of dilute nitric acid, add 30 mL of water, boil, and filter into the same flask. and proceed as directed above. Rinse the crucible and the char on the funnel with hot water until the filtrate measures 300 mL. Add slowly 7 mL of Uniformity of dosage units <6.02> Perform the test accord- freshly prepared bromine TS and diluted phosphoric acid ing to the following method: it meets the requirement of the (1 in 2) in the ratio of 3.5 mL to 1 g of the added potassium Content uniformity test. carbonate, and boil until starch-potassium iodide paper is no Place 1 tablet of Levothyroxine Sodium Tablets in a glass- longer colored blue by the evolved gas. Wash the inside of stoppered centrifuge tube, add exactly 10 mL of 0.01 mol/L the flask with water, and continue boiling for 5 minutes. sodium hydroxide TS, warm at 509C for 15 minutes, and During the boiling add water from time to time to maintain a shake vigorously for 20 minutes. Centrifuge this solution, volume of not less than 250 mL. Cool, add 5 mL of a solu- pipet 5 mL of the supernatant liquid, add exactly 1 mL of tion of phenol (1 in 20), again rinse the inside of the flask the internal standard solution, and use this solution as the with water, and allow to stand for 5 minutes. Add 2 mL of sample solution. Perform the test with 20 mLofthesample diluted phosphoric acid (1 in 2) and 5 mL of potassium solution as directed under Liquid Chromatography <2.01> iodide TS, and titrate <2.50> immediately the liberated iodine according to the following conditions, and calculate the ratio with 0.01 mol/L sodium thiosulfate VS (indicator: 3 mL of of the peak area of levothyroxine to that of the internal starch TS). Perform a blank determination, and make any standard. Calculate the mean value from the ratios of each necessary correction. peak area of 10 samples: the deviation (z)ofthemeanvalue and the ratio of each peak area should be not more than Each mL of 0.01 mol/L sodium thiosulfate VS

15z. When the deviation (z) is more than 15z,and1sam- ＝ 0.3329 mg of C15H10I4NNaO4 ple shows not more than 25z, perform another test with 20 Containers and storage Containers—Tight containers. samples. Calculate the deviation (z) of the mean value of Storage—Light-resistant. the 30 samples used in the 2 tests and the ratio of each peak area: there should be not more than 1 sample with the deviation more than 15z but not more than 25z, and no sample should deviate by more than 25z. Lidocaine Internal standard solution—A solution of ethinylestradiol in リドカイン a mixture of acetonitrile and diluted phosphoric acid (1 in 10) (9:1) (3 in 40,000). Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: a constant wavelength between 220 nm and 230 nm). Column: A stainless steel column 4 to 6 mm in inside diameter and 10 to 25 cm in length, packed with octadecyl- C14H22N2O: 234.34 silanized silica gel. 2-Diethylamino-N-(2,6-dimethylphenyl)acetamide Column temperature: A constant temperature at about [137-58-6] 259C. Mobile phase: A mixture of methanol, water and phos- Lidocaine, when dried, contains not less than 99.0z phoric acid (1340:660:1). of C14H22N2O. Flow rate: Adjust the flow rate so that the retention time Description Lidocaine occurs as white to pale yellow crys- of levothyroxine is about 9 minutes. tals or crystalline powder. Selection of column: To 5 mL of a solution of It is very soluble in methanol and in ethanol (95), soluble levothyroxine sodium in 0.01 mol/L sodium hydroxide TS in acetic acid (100) and in diethyl ether, and practically in- (1 in 200,000) add 1 mL of the internal standard solution. soluble in water. Proceed with 20 mL of this solution under the above operat- It dissolves in dilute hydrochloric acid. ing conditions, and calculate the resolution. Use a column giving elution of levothyroxine and the internal standard in Identification (1) Dissolve 40 mg of Lidocaine in 10 mL this order with the resolution between these peaks being not of 1 mol/L hydrochloric acid TS, and add water to make 100 less than 2.0. mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, JP XVI Official Monographs / Lidocaine Injection 1035 and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the Lidocaine Injection same wavelengths. (2) Determine the infrared absorption spectrum of Lidocaine Hydrochloride Injection Lidocaine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the リドカイン注射液 spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Lidocaine Injection is an aqueous injection. Melting point <2.60> 66–699C It contains not less than 95.0z and not more than 105.0z of the labeled amount of lidocaine hydrochlo- Purity (1) Clarity and color of solution—Dissolve 1.0 g ride (C H N O.HCl: 270.80). of Lidocaine in 2 mL of dilute hydrochloric acid, and add 14 22 2 water to make 10 mL: the solution is clear and colorless to Method of preparation Prepare as directed under Injec- light yellow. tions, with Lidocaine and an equivalent amount of Hydro- (2) Chloride <1.03>—Dissolve 0.6 g of Lidocaine in 6 mL chloric Acid. of dilute nitric acid, add water to make 50 mL, and perform No preservative is added in the case of intravenous injec- the test using this solution as the test solution. Prepare the tions. control solution with 0.70 mL of 0.01 mol/L hydrochloric Description Lidocaine Injection is a colorless, clear liquid. acid VS (not more than 0.041z). pH: 5.0 – 7.0 (3) Sulfate <1.14>—Dissolve 0.5 g of Lidocaine in 5 mL of dilute hydrochloric acid, add water to make 50 mL, and Identification To a volume of Lidocaine Injection, equiva- perform the test using this solution as the test solution. Pre- lent to 20 mg of lidocaine hydrochloride (C14H22N2O.HCl) pare the control solution with 1.0 mL of 0.005 mol/L sulfu- according to the labeled amount, add 1 mL of sodium hy- ric acid VS, 5 mL of dilute hydrochloric acid and water to droxide TS, and extract with 20 mL of hexane. To 10 mL of make 50 mL (not more than 0.096z). the hexane extract add 20 mL of 1 mol/L hydrochloric acid (4) Heavy metals <1.07>—Carbonize 2.0 g of Lidocaine TS, and shake vigorously. Determine the absorption spec- by gentle ignition. After cooling, add 10 mL of a solution of trum of the water layer as directed under Ultraviolet-visible magnesium nitrate hexahydrate in ethanol (95) (1 in 10), and Spectrophotometry <2.24>: it exhibits a maximum between fire the ethanol to burn. After cooling, add 1 mL of sulfuric 261 nm and 265 nm. acid, proceed according to Method 4, and perform the test. Bacterial endotoxins <4.01> Less than 1.0 EU/mg. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Extractable volume <6.05> It meets the requirement. (5) Related substances—Dissolve 0.10 g of Lidocaine in Foreign insoluble matter <6.06> Perform the test according 2 mL of methanol, and use this solution as the sample solu- to Method 1: it meets the requirement. tion. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard Insoluble particulate matter <6.07> It meets the require- solution. Perform the test with these solutions as directed ment. under Thin-layer Chromatography <2.03>. Spot 10 mLeach Sterility <4.06> Perform the test according to the Mem- of the sample solution and standard solution on a plate of brane filtration method: it meets the requirement. silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 2- Assay To an exactly measured volume of Lidocaine Injec- butanone, water and formic acid (5:3:1:1) to a distance of tion, equivalent to about 0.1 g of lidocaine hydrochloride about 10 cm, air-dry the plate, and dry more at 809Cfor30 (C14H22N2O.HCl), add exactly 10 mL of the internal stand- minutes. After cooling, examine under ultraviolet light ard solution and 0.001 mol/L hydrochloric acid TS to make (main wavelength: 254 nm): the spots other than the princi- 50 mL, and use this solution as the sample solution. Sepa- pal spot from the sample solution are not more intense than rately, weigh accurately about 85 mg of lidocaine for assay, the spot from the standard solution. previously dried in a desiccator (in vacuum, silica gel) for 24 hours, dissolve in 0.5 mL of 1 mol/L hydrochloric acid TS Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- and a suitable volume of 0.001 mol/L hydrochloric acid TS, um, silica gel, 24 hours). and add exactly 10 mL of the internal standard solution, Residue on ignition <2.44> Not more than 0.1z (1 g). then add 0.001 mol/L hydrochloric acid TS to make 50 mL, and use this solution as the standard solution. Perform the Assay Dissolve about 0.5 g of Lidocaine, previously dried test with 5 mL each of the sample solution and standard solu- and accurately weighed, in 20 mL of acetic acid (100), and tion as directed under Liquid Chromatography <2.01> ac- titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 1 cording to the following conditions, and calculate the ratios, drop of crystal violet TS) until the color of the solution Q and Q , of the peak area of lidocaine to that of the inter- changes from purple to blue-green through blue. Perform a T S nal standard. blank determination, and make any necessary correction. Amount (mg) of lidocaine hydrochloride Each mL of 0.1 mol/L perchloric acid VS (C14H22N2O.HCl) ＝ 23.43 mg of C14H22N2O ＝ MS × QT/QS × 1.156 Containers and storage Containers—Tight containers. MS: Amount (mg) of lidocaine for assay 1036 Limaprost Alfadex / Official Monographs JP XVI

Internal standard solution—A solution of benzophenone in sulfuric acid to each of the residue, and shake them for 5 methanol (1 in 4000). minutes: the solution obtained from the sample solution (1) Operating conditions— develops an orange-yellow color while the solution from the Detector: An ultraviolet absorption photometer (wave- sample solution (2) does not develop any color. length: 254 nm). (2) Dissolve 20 mg of Limaprost Alfadex in 5 mL of Column: A stainless steel column 4 mm in inside diameter water, add 5 mL of ethyl acetate, shake, centrifuge, and and 15 cm in length, packed with octadecylsilanized silica gel evaporate the solvent of the upper layer under reduced pres- for liquid chromatography (10 mm in particle diameter). sure. Dissolve the residue in 2 mL of ethanol (95), 5 mL of Column temperature: A constant temperature of about 1,3-dinitrobenzene TS, add 5 mL of a solution of potassium 259C. hydroxide in ethanol (95) (17 in 100) while ice-cooling, and Mobile phase: Dissolve 2.88 g of sodium lauryl sulfate in allow to stand in a dark place while ice-cooling for 20 1000 mL of a mixture of 0.02 mol/L phosphate buffer solu- minutes: a purple color develops. tion, pH 3.0 and acetonitrile (11:9). (3) To 50 mg of Limaprost Alfadex add 1 mL of iodine Flow rate: Adjust the flow rate so that the retention time TS, dissolve by heating in a water bath, and allow to stand: a of lidocaine is about 6 minutes. dark blue precipitate is formed. System suitability— (4) Determine the absorption spectrum of a solution of System performance: When proceed with 5 mLofthe Limaprost Alfadex in dilute ethanol (3 in 10,000) as directed standard solution under the above operating conditions, under Ultraviolet-visible Spectrophotometry <2.24>: it does lidocaine and the internal standard are eluted in this order not exhibit a maximum between 200 nm and 400 nm. To 10 with the resolution between these peaks being not less than 6. mL of this solution add 1 mL of potassium hydroxide- System repeatability: When the test is repeated 6 times ethanol TS, and allow to stand for 15 minutes. Determine with 5 mL of the standard solution under the above operating the absorption spectrum of this solution as directed under conditions, the relative standard deviation of the ratios of Ultraviolet-visible Spectrophotometry <2.24>,andcompare the peak area of lidocaine to that of the internal standard is the spectrum with the Reference Spectrum: both spectra not more than 1.0z. exhibit similar intensities of absorption at the same wavelengths. Containers and storage Containers—Hermetic containers. 20 Optical rotation <2.49> [a]D : ＋125 – 1359(0.1 g, calculated on the anhydrous basis, dilute ethanol, 20 mL, 100 Limaprost Alfadex mm). Purity Related substances—Perform the test immediately リマプロストアルファデクス after preparation of the sample solution. Dissolve 0.10 g of Limaprost Alfadex in 2 mL of water, add 1 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add dilute ethanol to make exactly 100 mL, and use this solution as the standard solution (1). Pipet 3 mL of the standard solution (1), add dilute ethanol to make exactly 10 mL, and use this solution as the standard solution (2). Perform the test with exactly 3 mLeachofthe

C22H36O5･xC36H60O30 sample solution and standard solutions (1) and (2) as di- (2E)-7-{(1R,2R,3R)-3-Hydroxy-2-[(1E,3S,5S)-3- rected under Liquid Chromatography <2.01> according to hydroxy-5-methylnon-1-en-1-yl]- the following operating conditions, and determine each peak 5-oxocyclopentyl}hept-2-enoic acid-a-cyclodextrin area by the automatic integration method: the area of the [100459-01-6, limaprost:alfadex ＝ 1:1; clathrate compound] peak of 17-epi-isomer, having the relative retention time of about 1.1 with respect to limaprost, and the area of the peak Limaprost Alfadex is a a-cyclodextrin clathrate of 11-deoxy substance, having the relative retention time of compound of limaprost. about 2.1, are not larger than the peak area of limaprost It contains not less than 2.8z and not more than from the standard solution (2), and the area of the peak 3.2z of limaprost (C22H36O5: 380.52), calculated on other than the principal peak and the peaks mentioned above the anhydrous basis. is not larger than 1/3 times the peak area of limaprost from the standard solution (2). The total area of the peaks other Description Limaprost Alfadex occurs as a white powder. than limaprost from the samples solution is not larger than It is freely soluble in water, slightly soluble in methanol, the peak area of limaprost from the standard solution (1). very slightly soluble in ethanol (99.5), and practically insolu- Operating conditions— ble in ethyl acetate. Detector, column, column temperature, mobile phase and It is hygroscopic. flow rate: Proceed as directed in the operating conditions in Identification (1) Dissolve 20 mg of Limaprost Alfadex in the Assay. 5 mL of water, add 5 mL of ethyl acetate, shake, centrifuge, Time span of measurement: About 3 times as long as the and use the upper layer as the sample solution (1). Sepa- retention time of limaprost beginning after the solvent peak. rately, to 20 mg of Limaprost Alfadex add 5 mL of ethyl System suitability— acetate, shake, centrifuge, and use the supernatant liquid as Test for required detectability: To exactly 1 mL of the the sample solution (2). Evaporate the solvent of the sample standard solution (1) add dilute ethanol to make exactly 10 solutions (1) and (2) under reduced pressure, add 2 mL of mL. Confirm that the peak area of limaprost obtained from JP XVI Official Monographs / Lincomycin Hydrochloride Hydrate 1037

3 mL of this solution is equivalent to 8 to 12z of that from 3 mL of the standard solution (1). Lincomycin Hydrochloride Hydrate System performance: Proceed as directed in the system suitability in the Assay. リンコマイシン塩酸塩水和物 System repeatability: When the test is repeated 6 times with 3 mL of the standard solution (1) under the above conditions, the relative standard deviation of the peak area of limaprost is not more than 2.0z. Water <2.48> Not more than 6.0z (0.2 g, volumetric titration, direct titration). Assay Weigh accurately about 0.1 g of Limaprost Afladex, dissolve in 5 mL of water, add exactly 5 mL of the internal C18H34N2O6S.HCl.H2O: 461.01 standard solution, and use this solution as the sample solu- Methyl 6,8-dideoxy-6-[(2S,4R)-1-methyl-4- tion. Separately, weigh accurately about 3 mg of Limaprost propylpyrrolidine-2-carboxamido]-1-thio-D-erythro-a-D- RS, dissolve in 5 mL of water, add exactly 5 mL of the inter- galacto-octopyranoside monohydrochloride monohydrate nal standard solution, and use this solution as the standard [7179-49-9] solution. Perform the test with 3 mL each of the sample solution and the standard solution as directed under Liquid Lincomycin Hydrochloride Hydrate is the hydro- Chromatography <2.01> according to the following condi- chloride of a substance having antibacterial activity tions, and calculate the ratios, QT and QS, of the peak area produced by the growth of Streptomyces lincolnensis of limaprost to that of the internal standard. var. lincolnensis. It contains not less than 825 mg (potency) per mg, Amount (mg) of limaprost (C H O ) ＝ M × Q /Q 22 36 5 S T S calculated on the anhydrous basis. The potency of MS: Amount (mg) of Limaprost RS Lincomycin Hydrochloride Hydrate is expressed as mass (potency) of lincomycin (C H N O S: 406.54). Internal standard solution—A solution of propyl parahy- 18 34 2 6 droxybenzoate in ethanol (95) (1 in 4000). Description Lincomycin Hydrochloride Hydrate occurs as Operating conditions— white, crystals or crystalline powder. Detector: An ultraviolet absorption photometer (wave- It is freely soluble in water and in methanol, sparingly length: 215 nm). soluble in ethanol (95), and very slightly soluble in aceto- Column: A stainless steel column 4.6 mm in inside diame- nitrile. ter and 15 cm in length, packed with octadecylsilanized silica Identification (1) Determine the infrared absorption spec- gel for liquid chromatography (5 mm in particle diameter). trum of Lincomycin Hydrochloride Hydrate as directed in Column temperature: A constant temperature of about the paste method under Infrared Spectrophotometry <2.25>, 259C. and compare the spectrum with the Reference Spectrum or Mobile phase: A mixture of 0.02 mol/L potassium dihy- the spectrum of Lincomycin Hydrochloride RS: both spectra drogen phosphate TS, acetonitrile for liquid chromatogra- exhibit similar intensities of absorption at the same wave phy and 2-propanol for liquid chromatography (9:5:2). numbers. Flow rate: Adjust the flow rate so that the retention time (2) A solution of Lincomycin Hydrochloride Hydrate of limaprost is about 12 minutes. (1 in 100) responds to the Qualitative Tests <1.09> (2) for System suitability— chloride. System performance: When the procedure is run with 3 mL 20 of the standard solution under the above operating condi- Optical rotation <2.49> [a]D : ＋135 – ＋1509(0.5 g, water, tions, the internal standard and limaprost are eluted in this 25 mL, 100 mm). order with the resolution between these peaks being not less pH <2.54> Dissolve 0.10 g of Lincomycin Hydrochloride than 7. Hydrate in 1 mL of water: 3.0 – 5.5. System repeatability: When the test is repeated 6 times with 3 mL of the standard solution under the above operating Purity (1) Clarity and color of solution—Dissolve 1.0 g conditions, the relative standard deviation of the ratios of of Lincomycin Hydrochloride Hydrate in 10 mL of water: the peak area of limaprost to that of the internal standard is the solution is clear and colorless. not more than 1.0z. (2) Heavy metals <1.07>—Proceed with 2.0 g of Linco- mycin Hydrochloride Hydrate according to Method 4, and Containers and storage Containers—Tight containers. perform the test. Prepare the control solution with 1.0 mL of Storage—Light-resistant, at a temperature not exceeding Standard Lead Solution (not more than 5 ppm). －109C. (3) Lincomycin B—Perform the test with 20 mLofthe sample solution obtained in the Assay as directed under Liq- uid Chromatography <2.01> according to the following conditions, and determine the peak areas of lincomycin and lincomycin B, having the relative retention time of about 0.5 with respect to lincomycin, by the automatic integration method: the peak area of lincomycin B is not more than 5.0z of the sum of the peak areas of lincomycin and linco- 1038 Lincomycin Hydrochloride Injection / Official Monographs JP XVI mycin B. Operating conditions— Lincomycin Hydrochloride Injection Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in リンコマイシン塩酸塩注射液 the Assay. System suitability— Lincomycin Hydrochloride Injection is an aqueous Test for required detectability: Measure exactly 1 mL of injection. the sample solution, and add the mobile phase to make ex- It contains not less than 93.0z and not more actly 20 mL. Confirm that the peak area of lincomycin ob- than 107.0z of the labeled potency of lincomycin tained from 20 mL of this solution is equivalent to 3.5 to (C H N O S: 406.54). 6.5z of that from 20 mL of the sample solution. 18 34 2 6 System performance, and system repeatability: Proceed as Method of preparation Prepare as directed under Injec- directed in the system suitability in the Assay. tions, with Lincomycin Hydrochloride Hydrate. Water <2.48> 3.0 – 6.0z (0.5 g, volumetric titration, direct Description Lincomycin Hydrochloride Injection is a clear, titration). colorless liquid. Assay Weigh accurately an amount of Lincomycin Hydro- Identification To a volume of Lincomycin Hydrochloride chloride Hydrate and Lincomycin Hydrochloride RS, Injection, equivalent to 30 mg (potency) of Lincomycin Hy- equivalent to about 10 mg (potency), dissolve each in the drochloride Hydrate according to the labeled amount, add mobile phase to make exactly 10 mL, and use these solutions 30 mL of water, and use this solution as the sample solution. as the sample solution and standard solution. Perform the Separately, dissolve 10 mg (potency) of Lincomycin Hydro- test with exactly 20 mL each of the sample solution and chloride RS in 10 mL of water, and use this solution as the standard solution as directed under Liquid Chromatography standard solution. Perform the test with these solutions as <2.01> according to the following conditions, and determine directed under Thin-layer Chromatography <2.03>. Spot 5 the peak areas, AT and AS, of lincomycin. mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Dissolve Amount [mg (potency)] of lincomycin (C H N O S) 18 34 2 6 150 g of ammonium acetate in 800 mL of water, adjust the ＝ M × A /A × 1000 S T S pH to 9.6 with ammonia solution (28), and add water to

MS: Amount [mg (potency)] of Lincomycin Hydrochlo- make 1000 mL. To 80 mL of this solution add 40 mL of 2- ride RS propanol and 90 mL of ethyl acetate, shake, develop the plate with the upper layer of this solution to a distance of Operating conditions— about 15 cm, and air-dry the plate. Spray evenly a solution Detector: An ultraviolet absorption photometer (wave- of potassium permanganate (1 in 1000) on the plate: the length: 210 nm). principal spots from the sample solution and standard solu- Column: A stainless steel column 4 mm in inside diameter tion show the same Rf value. and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mminparticlediameter). pH <2.54> 3.5–5.5 Column temperature: A constant temperature of about Bacterial endotoxins <4.01> Less than 0.50 EU/mg (po- 469C. tency). Mobile phase: To 13.5 mL phosphoric acid add water to make 1000 mL, and adjust the pH to 6.0 with ammonia TS. Extractable volume <6.05> It meets the requirement. To 780 mL of this solution add 150 mL of acetonitrile and Foreign insoluble matter <6.06> Perform the test according 150 mL of methanol. to Method 1: it meets the requirement. Flow rate: Adjust the flow rate so that the retention time of lincomycin is about 9 minutes. Insoluble particulate matter <6.07> It meets the require- System suitability— ment. System performance: When the procedure is run with 20 Sterility <4.06> Perform the test according to the Mem- mL of the standard solution under the above operating con- brane filtration method: it meets the requirement. ditions, the number of theoretical plates and the symmetrical factor of the peak of lincomycin are not less than 4000 and Assay Pipet a volume of Lincomycin Hydrochloride Injec- not more than 1.3, respectively. tion, equivalent to about 0.3 g (potency) of Lincomycin Hy- System repeatability: When the test is repeated 6 times drochloride Hydrate, add the mobile phase to make exactly with 20 mL of the standard solution under the above operat- 30 mL. Pipet 2 mL of this solution, add the mobile phase to ing conditions, the relative standard deviation of the peak make exactly 20 mL, and use this solution as the sample so- area of lincomycin is not more than 2.0z. lution. Separately, weigh accurately an amount of Lincomy- cin Hydrochloride RS, equivalent to 20 mg (potency), dis- Containers and storage Containers—Tight containers. solve in the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Lincomycin Hydrochloride Hy- drate.

Amount [mg (potency)] of lincomycin (C18H34N2O6S) ＝ MS × AT/AS × 15 JP XVI Official Monographs / Liothyronine Sodium 1039

MS: Amount [mg (potency)] of Lincomycin Hydrochlo- no more color than the following control solution. ride RS Control solution: Weigh exactly 0.111 g of potassium iodide, and dissolve in water to make 1000 mL. Pipet 1 mL Containers and storage Containers—Hermetic containers. of this solution, add 10 mL of dilute hydroxide TS, 14 mL of water and 5 mL of dilute sulfuric acid, and mix. Filter the mixture into a Nessler tube, and perform the test with the fil- Liothyronine Sodium trate in the same manner as for the sample. (3) Related substances—Dissolve 0.15 g of Liothyronine リオチロニンナトリウム Sodium in 5 mL of diluted ammonia TS (1 in 3), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add diluted ammonia TS (1 in 3) to make exactly 50 mL, and use this solution as the standard solution. Per- form the test with these solutions as directed under Thin- layer Chromatography <2.03>. Spot 1 mLeachofthesample C H I NNaO : 672.96 15 11 3 4 solution and standard solution on a plate of silica gel for Monosodium O-(4-hydroxy-3-iodophenyl)-3,5-diiodo- thin-layer chromatography. Develop the plate with a mixture L-tyrosinate of t-butanol, t-amyl alcohol, water, ammonia solution (28) [55-06-1] and 2-butanone (59:32:17:15:7) to a distance of about 12 cm, and air-dry the plate. Spray evenly a solution of 0.3 g of nin- Liothyronine Sodium contains not less than 95.0z hydrin in 100 mL of a mixture of 1-butanol and acetic acid of C H I NNaO , calculated on the dried basis. 15 11 3 4 (100) (97:3) on the plate, and dry the plate at 1009Cfor3 Description Liothyronine Sodium occurs as a white to light minutes: the spots other than the principal spot from the brown powder. It is odorless. sample solution are not more intense than the spot from the It is slightly soluble in ethanol (95), and practically insolu- standard solution. bleinwaterandindiethylether. Loss on drying <2.41> Not more than 4.0z (0.2 g, 1059C, It dissolves in sodium hydroxide TS and in ammonia TS. 2 hours). Identification (1) To 5 mL of a solution of Liothyronine Assay Weigh accurately about 25 mg of Liothyronine So- Sodium in ethanol (95) (1 in 1000) add 1 mL of ninhydrin dium, and proceed as directed under Oxygen Flask Combus- TS, and warm in a water bath for 5 minutes: a purple color tion Method <1.06>, using a mixture of 10 mL of a solution develops. of sodium hydroxide (1 in 100) and 1 mL of a freshly pre- (2) Heat 0.02 g of Liothyronine Sodium with a few drops pared solution of sodium bisulfate (1 in 100) as the absorb- of sulfuric acid over a flame: a purple gas is evolved. ing liquid, and prepare the test solution. Apply a small (3) Determine the absorption spectrum of a solution of amount of water to the upper part of apparatus A, pull out Liothyronine Sodium in ethanol (95) (1 in 10,000) as directed C carefully, and wash C, B and the inner wall of A with 40 under Ultraviolet-visible Spectrophotometry <2.24>,and mL of water. To the test solution add 1 mL of bromine- compare the spectrum with the Reference Spectrum: both acetic acid TS, insert the stopper C, and shake vigorously for spectra exhibit similar intensities of absorption at the same 1 minute. Remove the stopper, rinse the stopper, the sample wavelengths. holder and the inner wall of the flask with 40 mL of water, (4) Ignite 0.02 g of Liothyronine Sodium until thor- and add 0.5 mL of formic acid. Stopper the flask with C, oughly charred. After cooling, add 5 mL of water to the and shake vigorously for 1 minute again. Remove the stop- residue, shake, and filter: the filtrate responds to the per, and rinse the stopper, the sample holder and the inner Qualitative Tests <1.09> (1) for sodium salt. wall of the flask with 40 mL of water again. Bubble the solu- 20 Optical rotation <2.49> [a]D : ＋18 – ＋229(0.2 g, calcu- tion with enough nitrogen gas in the flask to remove the lated on the dried basis, a mixture of ethanol (95) and 1 oxygen and excess bromine, add 0.5 g of potassium iodide to mol/L hydrochloric acid TS (4:1), 10 mL, 100 mm). the solution, and dissolve. Add immediately 3 mL of dilute sulfuric acid, mix, and allow to stand for 2 minutes. Titrate Purity (1) Soluble halide—To 10 mg of Liothyronine So- <2.50> the solution with 0.02 mol/L sodium thiosulfate VS dium add 10 mL of water and 1 drop of dilute nitric acid, (indicator: 3 mL of starch TS). Perform a blank determina- shake for 5 minutes, and filter. Add water to the filtrate to tion, and make any necessary correction. make 10 mL, and mix with 3 drops of silver nitrate TS: the solution shows no more turbidity than the following control Each mL of 0.02 mol/L sodium thiosulfate VS solution. ＝ 0.7477 mg of C15H11I3NNaO4 Control solution: To 0.35 mL of 0.01 mol/L hydrochloric Containers and storage Containers—Tight containers. acid VS add 1 drop of dilute nitric acid and water to make 10 Storage—Light-resistant. mL, and add 3 drops of silver nitrate TS. (2) Iodine and iodide—Dissolve 0.10 g of Liothyronine Sodium in 10 mL of dilute sodium hydroxide TS and 15 mL of water, add 5 mL of dilute sulfuric acid, and allow to stand for 10 minutes with occasional shaking. Filter the mixture into a Nessler tube, add 10 mL of chloroform and 3 drops of a solution of potassium iodate (1 in 100) to the filtrate, mix for 30 seconds, and allow to stand: the chloroform layer has 1040 Liothyronine Sodium Tablets / Official Monographs JP XVI

and 1 sample shows not more than 25z, perform another Liothyronine Sodium Tablets test with 20 samples. Calculate the deviation (z)ofeach ratio of the peak area from the mean value of the 30 samples リオチロニンナトリウム錠 used in the two tests: there should be not more than 1 sample with the deviation more than 15z but not more than 25z, and no sample should deviate by more than 25z. Liothyronine Sodium Tablets contain not less than Internal standard solution—A solution of propylpara- 90.0z and not more than 110.0z of the labeled hydroxybenzoate in a mixture of methanol and diluted phos- amount of liothyronine sodium (C H I NNaO : 15 11 3 4 phoric acid (1 in 10) (9:1) (1 in 250,000). 672.96). Operating conditions— Method of preparation Prepare as directed under Tablets, Detector: An ultraviolet absorption photometer (wave- with Liothyronine Sodium. length: 225 nm). Column: A stainless steel column 4.6 mm in inside diame- Identification (1) To a glass-stoppered centrifuge tube ter and 15 cm in length, packed with octadecylsylanized add a portion of finely powdered Liothyronine Sodium silica gel for liquid chromatography (5 mminparticlediame- Tablets, equivalent to 0.1 mg of Liothyronine Sodium ac- ter). cording to the labeled amount, add 30 mL of dilute sodium Column temperature: A constant temperature of about hydroxide TS, shake vigorously, and centrifuge. Transfer 259C. the supernatant liquid to a separator, add 10 mL of dilute Mobile phase: Diluted methanol (57 in 100). hydrochloric acid, and extract with two 20-mL portions of Flow rate: Adjust the flow rate so that the retention time ethyl acetate. Filter each extract successively through absor- of liothyronine is about 9 minutes. bent cotton previously overlaid with 8 g of anhydrous so- System suitability— dium sulfate. Evaporate the filtrate on a water bath to dry- System performance: To 5 mL of a solution of liothyro- ness with the aid of a current of nitrogen. Dissolve the nine sodium in 0.01 mol/L sodium hydroxide TS (1 in residue in 0.5 mL of methanol, and use this solution as the 2,000,000) add 1 mL of the internal standard solution, and sample solution. Separately, dissolve 10 mg of liothyronine use this solution as the solution for system suitability test. sodium for thin-layer chromatography in methanol to make When the procedure is run with 200 mL of this solution 50 mL, and use this solution as the standard solution. Per- under the above operating conditions, the internal standard form the test with these solutions as directed under Thin- and liothyronine are eluted in this order with the resolution layer Chromatography <2.03>. Spot 20 mLeachofthesample between these peaks being not less than 2.0. solution and standard solution on a plate of silica gel for System repeatability: When the test is repeated 6 times thin-layer chromatography. Develop the plate with a mixture with 200 mL of the solution for system suitability test under of t-butanol, t-amyl alcohol, water, ammonia solution (28) the above operating conditions, the relative standard devia- and 2-butanone (59:32:17:15:7) to a distance of about 12 cm, tion of the ratios of the peak area of liothyronine to that of and air-dry the plate. Spray evenly a solution of 0.3 g of nin- the internal standard is not more than 1.0z. hydrin in 100 mL of a mixture of 1-butanol and acetic acid (100) (97:3) on the plate, and dry the plate at 1009Cfor3 Assay Weigh accurately not less than 20 Liothyronine So- minutes: the spots obtained from the sample solution and dium Tablets, and finely powder. Place an accurately the standard solution show a red-purple color, and has the weighed portion of the powder, equivalent to about 50 mgof same Rf value. liothyronine sodium (C15H11I3NNaO4), in an agate mortar, (2) The colored solution obtained in the Assay is blue in add 1 g of powdered potassium carbonate, and mix well. color. Transfer the mixture cautiously to a porcelain crucible, and compact the contents by gently tapping the crucible on a Uniformity of dosage units <6.02> Perform the test accord- table. Add an additional 1.5 g of powdered potassium car- ing to the following method: it meets the requirement of the bonate to the same agate mortar, mix well with any content Content uniformity test. adhering to the mortar, cautiously overlay the mixture on Place 1 tablet of Liothyronine Sodium Tablets in a glass- the top of the same porcelain crucible, and compact the stoppered centrifuge tube, add exactly 10 mL of 0.01 mol/L charge again in the same manner. Ignite the combined mix- sodium hydroxide TS, warm at 509C for 15 minutes, and ture in the crucible between 6759C and 7009Cfor30 shake vigorously for 20 minutes. Centrifuge for 5 minutes, minutes. Cool, add a few mL of water to the crucible, heat and filter the supernatant liquid, if necessary. Pipet a gently to boiling, and filter the contents of the crucible definite volume of this solution, and add a volume of 0.01 through a glass filter (G4) into a 20-mL volumetric flask. mol/L sodium hydroxide TS to prepare a definite volume of Wash the residue with water, and combine the washings with a solution containing about 0.5 mg of liothyronine sodium the filtrate. Cool, add water to make 20 mL, and use this so- (C H I NNaO ) per mL. Pipet 5 mL of this solution, add 15 11 3 4 lution as the sample solution. Separately, weigh accurately exactly 1 mL of the internal standard solution, and use this about 75 mg of potassium iodide for assay, previously dried solution as the sample solution. Perform the test with 200 mL at 1059C for 4 hours, and dissolve in water to make exactly of the sample solution as directed under Liquid Chromatog- 200 mL. Measure exactly 5 mL of the solution, and add a so- raphy <2.01> according to the following conditions, and cal- lution of potassium carbonate (1 in 8) to make exactly 100 culate the ratio of the peak area of the liothyronine to that of mL. To 2 mL of this solution, exactly measured, add a solu- the internal standard. Calculate the mean value of the ratios tion of potassium carbonate (1 in 8) to make exactly 20 mL, of each peak area of 10 samples: the deviation (z)ofeach and use the solution as the standard solution. Pipet 5 mL ratio of the peak area from the mean value should be not each of the sample solution and the standard solution into more than 15z. When the deviation (z) is more than 15z, JP XVI Official Monographs / Lisinopril Hydrate 1041 glass-stoppered test tubes, add 3.0 mL of diluted sulfuric with the Reference Spectrum: both spectra exhibit similar in- acid (4 in 25) and 2.0 mL of potassium permanganate TS, tensities of absorption at the same wave numbers. and heat on a water bath for 15 minutes. Cool, add 1.0 mL Optical rotation <2.49> [a]25: －43.0 – －47.09(0.25 g cal- of diluted sodium nitrite TS (1 in 10), swirl to mix, and add D culated on the anhydrous basis, 0.25 ml/L zinc acetate 1.0 mL of a solution of ammonium amidosulfate (1 in 10). buffer solution, pH 6.4, 25 mL, 100 mm). Allow to stand at room temperature for 10 minutes with occasional shaking. Then add 1.0 mL of potato starch TS and Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of 1.0 mL of a freshly prepared, diluted potassium iodide TS (1 Lisinopril Hydrate according to Method 4, and perform the in 40), swirl to mix, and transfer each solution to a 20-mL test. Prepare the control solution with 2.0 mL of Standard volumetric flask. Rinse the test tube with water, collect the Lead Solution (not more than 10 ppm). washings in the volumetric flask, add water to make 20 mL, (2) Related substances—Dissolve about 0.10 g of and allow to stand for 10 minutes. Perform the test with Lisinopril Hydrate in 50 mL of water, and use this solution these solutions as directed under Ultraviolet-visible Spectro- as the sample solution. Pipet 3 mL of the sample solution, photometry <2.24>, using a solution prepared with 5 mL of add water to make exactly 200 mL, and use this solution as potassium carbonate (1 in 8) in the same manner as the sam- the standard solution. Perform the test with exactly 15 mL ple solution as the blank. Determine the absorbances, AT each of the sample solution and standard solution as directed and AS, of the subsequent solutions of the sample solution under Liquid Chromatography <2.01> according to the fol- and the standard solution at the wavelength of maximum ab- lowing conditions, and determine each peak area by the au- sorption at about 600 nm, respectively. tomatic integration method: the area of the peak, having the relative retention time of about 1.2 with respect to lisinopril, Amount (mg) of liothyronine sodium (C H I NNaO ) 15 11 3 4 is not larger than 1/5 times the peak area of lisinopril from ＝ M × A /A × 1/2000 × 1.351 S T S the standard solution, the area of the peak other than

MS: Amount (mg) of potassium iodide for assay lisinopril and the peak mentioned above is not larger than 2/15 times the peak area of lisinopril from the standard solu- Containers and storage Containers—Tight containers. tion, and the total area of the peaks other than lisinopril is Storage—Light-resistant. not larger than the peak area of lisinopril from the standard solution. Operating conditions— Lisinopril Hydrate Detector: An ultraviolet absorption photometer (wavelength: 215 nm). リシノプリル水和物 Column: A stainless steel column 4.0 mm in inside diameter and 20 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mminparticlediameter). Column temperature: A constant temperature of about 609C. Mobile phase A: Diluted 0.05 mol/L sodium dihydrogen phosphate TS (1 in 2). Mobile phase B: A mixture of diluted 0.05 mol/L sodium dihydrogen phosphate TS (1 in 2) and acetonitrile for liquid C H N O ･2H O: 441.52 21 31 3 5 2 chromatography (3:2). (2S)-1-{(2S)-6-Amino-2-[(1S)-1-carboxy- Flowing of the mobile phase: Control the gradient by mix- 3-phenylpropylamino]hexanoyl}pyrrolidine-2-carboxylic acid ing the mobile phases A and B as directed in the following dihydrate table. [83915-83-7]

Lisinopril Hydrate contains not less than 98.5z Time after injection Mobile phase A Mobile phase B of sample (min) (volz) (volz) and not more than 101.0z of lisinopril (C21H31N3O5: 405.49), calculated on the anhydrous basis. 0–10 90→ 50 10 → 50 Description Lisinopril Hydrate occurs as a white crystalline 10–25 50 50 powder, having a slight characteristic odor. It is soluble in water, sparingly soluble in methanol, and Flow rate: About 1.5 mL per minute. practically insoluble in ethanol (99.5). Time span of measurement: About 2.5 times as long as the Melting point: about 1609C (with decomposition). retention time of lisinopril beginning after the solvent peak. Identification (1) Determine the absorption spectrum of a System suitability— solution of Lisinopril Hydrate in methanol (1 in 1000) as di- Test for required detectability: Measure exactly 2.5 mL of rected under Ultraviolet-visible Spectrophotometry <2.24>, the standard solution, and add water to make exactly 50 mL. and compare the spectrum with the Reference Spectrum: Confirm that the peak area of lisinopril obtained with 15 mL both spectra exhibit similar intensities of absorption at the of this solution is equivalent to 3.5 to 6.5z of that with 15 same wavelengths. mL of the standard solution. (2) Determine the infrared absorption spectrum of System performance: To 10 mg of Lisinopril Hydrate and Lisinopril Hydrate as directed in the paste method under In- 2 mL of a solution of anhydrous caffeine (1 in 1000) add frared Spectrophotometry <2.25>, and compare the spectrum water to make 200 mL. When the procedure is run with 15 1042 Lisinopril Tablets / Official Monographs JP XVI mL of this solution under the above operating conditions, peak area of lisinopril from the standard solution. lisinopril and caffeine are eluted in this order with the resolu- Operating conditions— tion between these peaks being not less than 6. Proceed as directed in the operating conditions in the System repeatability: When the test is repeated 6 times Purity (2) under Lisinopril Hydrate. with 15 mL of the standard solution under the above operat- System suitability— ing conditions, the relative standard deviation of the peak Test for required detectability: To exactly 2.5 mL of the area of lisinopril is not more than 2.0z. standard solution add water to make exactly 50 mL. Con- firm that the peak area of lisinopril obtained with 15 mLof Water <2.48> Not less than 8.0z and not more than 9.5z this solution is equivalent to 3.5 to 6.5z of that with 15 mL (0.3 g, volumetric titration, back titration). of the standard solution. Residue on ignition <2.44> Not more than 0.1z (1 g). System performance: Proceed as directed in the system suitability in the Purity (2) under Lisinopril Hydrate. Assay Weigh accurately about 0.66 g of Lisinopril Hy- System repeatability: When the test is repeated 6 times drate, dissolve in 80 mL of water, and titrate <2.50> with 0.1 with 15 mL of the standard solution under the above operat- mol/L sodium hydroxide VS (potentiometric titration). Per- ing conditions, the relative standard deviation of the peak form a blank determination in the same manner, and make area of lisinopril is not more than 2.0z. any necessary correction. Uniformity of dosage units <6.02> Perform the test accord- Each mL of 0.1 mol/L sodium hydroxide VS ing to the following method: it meets the requirement of the ＝ 40.55 mg of C H N O 21 31 3 5 Content uniformity test. Containers and storage Containers—Well-closed contain- To 1 tablet of Lisinopril Tablets add exactly 5 mL each of ers. the internal standard solution per every 1 mg of lisinopril

(C21H31N3O5), shake for 20 minutes, centrifuge, and use the supernatant liquid as the sample solution. Hereafter, Lisinopril Tablets proceed as directed in the Assay. Amount (mg) of lisinopril (C H N O ) リシノプリル錠 21 31 3 5 ＝ MS × QT/QS × C/10 M : Amount (mg) of lisinopril for assay, calculated on the Lisinopril Tablets contain not less than 95.0z and S anhydrous basis not more than 105.0z of the labeled amount of C: Labeled amount (mg) of lisinopril (C H N O )in1 lisinopril (C H N O : 405.49). 21 31 3 5 21 31 3 5 tablet Method of preparation Prepare as directed under Tablets, Internal standard solution—A solution of anhydrous with Lisinopril Hydrate. caffeine (1 in 20,000). Identification To an amount of powdered Lisinopril Dissolution <6.10> When the test is performed at 50 revolu- Tablets, equivalent to 10 mg of lisinopril (C H N O ), add 21 31 3 5 tions per minute according to the Paddle method, using 900 10 mL of methanol, shake for 20 minutes, filter, and use the mL of water as the dissolution medium, the dissolution rate filtrate as the sample solution. Separately, dissolve 10 mg of of a 5 mg tablet in 60 minutes and that of a 10-mg tablet in lisinopril in 10 mL of methanol, and use this solution as the 90 minutes is not less than 80z, and that of a 20-mg tablet standard solution. Perform the test with these solutions as in 90 minutes is not less than 75z. directed under Thin-layer Chromatography <2.03>. Spot 30 Start the test with 1 tablet of Lisinopril Tablets, withdraw mL each of the sample solution and standard solution on a not less than 20 mL of the medium at the specified minute plate of silica gel for thin-layer chromatography. Develop after starting the test, and filter through a membrane filter the plate with a mixture of acetonitrile, acetic acid (100), with a pore size not exceeding 0.5 mm. Discard the first 10 water and ethyl acetate (2:2:1:1) to a distance of about 10 mL of the filtrate, pipet V mL of the subsequent filtrate, add cm, and air-dry the plate. Spray evenly ninhydrin TS on the water to make exactly V?mL so that each mL contains about plate, and heat at 1209C: the principal spot with the sample 5.6 mg of lisinopril (C H N O ) according to the labeled solution and the spot with the standard solution show a red- 21 31 3 5 amount, and use this solution as the sample solution. Sepa- purple color and their Rf values are the same. rately, weigh accurately about 15 mg of lisinopril for assay, Purity Related substances—Powder not less than 20 separately determined the water <2.48> inthesamemanner Lisinopril Tablets. Take a portion of the powder, equivalent as Lisinopril Hydrate, and dissolve in water to make exactly to about 25 mg of lisinopril (C21H31N3O5), add 25 mL of 100 mL. Pipet 2 mL of this solution, add water to make ex- water, shake for 20 minutes, filter, and use the filtrate as the actly 50 mL, and use this solution as the standard solution. sample solution. Pipet 3 mL of the sample solution, add Perform the test with exactly 50 mL each of the sample solu- water to make exactly 200 mL, and use this solution as the tion and standard solution as directed under Liquid Chroma- standard solution. Perform the test with exactly 15 mLeach tography <2.01> according to the following conditions, and of the sample solution and standard solution as directed determine the peak areas, AT and AS, of lisinopril. under Liquid Chromatography <2.01> according to the fol- Dissolution rate (z) with respect to the labeled amount lowing conditions, and determine each peak area by the of lisinopril (C H N O ) automatic integration method: the peak area of lisinopril 21 31 3 5 ＝ M × A /A × V?/V × 1/C × 36 diketopiperazine, having the relative retention time of about S T S

2.0 with respect to lisinopril, is not larger than 2/3 times the MS: Amount (mg) of lisinopril for assay, calculated on the JP XVI Official Monographs / Lithium Carbonate 1043

anhydrous basis than 7.

C: Labeled amount (mg) of lisinopril (C21H31N3O5)in1 System repeatability: When the test is repeated 6 times tablet with 10 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of Operating conditions— the peak area of lisinopril to that of the internal standard is Detector, column temperature, and mobile phase: Proceed not more than 1.0z. as directed in the operating conditions in the Assay. Column: A stainless steel column 4.6 mm in inside diame- Containers and storage Containers—Well-closed contain- ter and 15 cm in length, packed with octadecylsilanized silica ers. gel for liquid chromatography (5 mm in particle diameter). Flow rate: Adjust the flow rate so that the retention time of lisinopril is about 7 minutes. Lithium Carbonate System suitability— System performance: When the procedure is run with 50 炭酸リチウム mL of the standard solution under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of lisinopril are not less than 1000 and not Li2CO3:73.89 more than 1.5, respectively. System repeatability: When the test is repeated 6 times Lithium Carbonate, when dried, contains not less with 50 mL of the standard solution under the above operat- than 99.5z of Li2CO3. ing conditions, the relative standard deviation of the peak Description Lithium Carbonate occurs as a white, crystal- area of lisinopril is not more than 2.0z. line powder. It is odorless. Assay Weigh accurately the mass of not less than 20 It is sparingly soluble in water, slightly soluble in hot Lisinopril Tablets, and powder. Weigh accurately a portion water, and practically insoluble in ethanol (95) and in diethyl of the powder, equivalent to about 5 mg of lisinopril ether.

(C21H31N3O5), add exactly 25 mL of the internal standard It dissolves in dilute acetic acid. solution, shake for 20 minutes, centrifuge, and use the su- The pH of a solution dissolved 1.0 g of Lithium Car- pernatant liquid as the sample solution. Separately, weigh bonate in 100 mL or water is between 10.9 and 11.5. accurately about 10 mg of lisinopril for assay, separately de- Identification (1) Perform the test as directed under termined the water <2.48> in the same manner as Lisinopril Flame Coloration Test <1.04> (1) with Lithium Carbonate: a Hydrate, add exactly 50 mL of the internal standard solution persistent red color appears. to dissolve, and use this solution as the standard solution. (2) Dissolve 0.2 g of Lithium Carbonate in 3 mL of Perform the test with 10 mL each of the sample solution and dilute hydrochloric acid, and add 4 mL of sodium hydroxide standard solution as directed under Liquid Chromatography TS and 2 mL of disodium hydrogen phosphate TS: a white <2.01> according to the following conditions, and calculate precipitate is produced. To the precipitate add 2 mL of dilute the ratios, Q and Q , of the peak area of lisinopril to that of T S hydrochloric acid: it dissolves. the internal standard. (3) A solution of Lithium Carbonate (1 in 100) responds

Amount (mg) of lisinopril (C21H31N3O5) to the Qualitative Tests <1.09> for carbonate. ＝ M × Q /Q × 1/2 S T S Purity (1) Clarity and color of solution—Dissolve 0.10 g

MS: Amount (mg) of lisinopril for assay, calculated on the of Lithium Carbonate in 10 mL of water by warming: the so- anhydrous basis lution is clear and colorless. (2) Acetic acid-insoluble substances—Take 1.0 g of Internal standard solution—A solution of anhydrous Lithium Carbonate, dissolve in 40 mL of dilute acetic acid, caffeine (1 in 20,000). filter the insoluble substances using filter paper for quantita- Operating conditions— tive analysis, wash with five 10-mL portions of water, and Detector: An ultraviolet absorption photometer (wave- ignite the insoluble substances together with the filter paper length: 215 nm). to incinerate: the mass of the residue is not more than 1.5 Column: A stainless steel column 4.0 mm in inside diame- mg. ter and 20 cm in length, packed with octadecylsilanized silica (3) Chloride <1.03>—To 0.40 g of Lithium Carbonate gel for liquid chromatography (7 mm in particle diameter). add 10 mL of water and 7 mL of dilute nitric acid, and dis- Column temperature: A constant temperature of about solve by heating to boil. After cooling, add 6 mL of dilute 609C. nitric acid, and dilute with water to make 50 mL. Perform Mobile phase: A mixture of diluted 0.05 mol/L sodium di- the test using this solution as the test solution. Prepare the hydrogen phosphate TS (1 in 2) and acetonitrile for liquid control solution with 0.25 mL of 0.01 mol/L hydrochloric chromatography (19:1). acid VS (not more than 0.022z). Flow rate: Adjust the flow rate so that the retention time (4) Sulfate <1.14>—To 0.40 g of Lithium Carbonate add of lisinopril is about 6 minutes. 10 mL of water and 4 mL of dilute hydrochloric acid, and System suitability— dissolve by heating to boil. After cooling, add 1 mL of dilute System performance: When the procedure is run with 10 hydrochloric acid, and dilute with water to make 50 mL. mL of the standard solution under the above operating con- Perform the test using this solution as the test solution. Pre- ditions, lisinopril and the internal standard are eluted in this pare the control solution with 0.40 mL of 0.005 mol/L sulfu- order with the resolution between these peaks being not less ricacidVS(notmorethan0.048z). 1044 Lithium Carbonate / Official Monographs JP XVI

(5) Heavy metals <1.07>—To 4.0 g of Lithium Carbonate pale red color persists for 30 seconds: the amount of calcium add 5 mL of water, gradually add 10 mL of hydrochloric (Ca: 40.08) is not more than 0.05z. acid while mixing, and dissolve. Evaporate the solution on a Each mL of 0.02 mol/L potassium permanganate VS waterbathtodryness.Totheresidueadd10mLofwater, ＝ 2.004 mg of Ca and dissolve. Place the solution in a Nessler tube, add 1 drop of phenolphthalein TS, add ammonia TS until the solution (10) Magnesium—To 3.0 mL of solution A obtained in shows a slight red color, then add 2 mL of dilute acetic acid, (7) add 0.2 mL of a solution of titan yellow (1 in 1000) and and dilute with water to make 50 mL. Perform the test using water to make 20 mL, then add 5 mL of sodium hydroxide this solution as the test solution. Prepare the control solution (3 in 20), and allow to stand for 10 minutes: the solution has as follows: Evaporate 10 mL of hydrochloric acid on a water no more color than the following control solution. bath to dryness. To the residue add 10 mL of water, and dis- Control solution: Dissolve 49.5 mg of magnesium sulfate solve. Place the solution in a Nessler tube, add 1 drop of heptahydrate, previously dried at 1059C for 2 hours and phenolphthalein TS, add ammonia TS until the solution heated at 4509C for 3 hours, in water to make 1000 mL. To shows a pale red color, then add 2.0 mL of Standard Lead this solution add 3 mL of solution B obtained in (7), 0.2 mL Solution and 2 mL of dilute acetic acid, and dilute with of a solution of titanium yellow (1 in 1000) and water to water to make 50 mL (not more than 5 ppm). make 20 mL, and proceed in the same manner. (6) Iron <1.10>—Prepare the test solution with 1.0 g of (11) Potassium—Dissolve 1.0 g of Lithium Carbonate in Lithium Carbonate according to Method 2 using 11 mL of water to make 100 mL, and use this solution as the sample dilute hydrochloric acid, and perform the test according to solution. To 5 mL of the sample solution add 1.0 mL of Method B. Prepare the control solution with 1.0 mL of dilute acetic acid, shake, add 5 mL of a solution of sodium Standard Iron Solution (not more than 10 ppm). tetraphenylborate (1 in 30), shake immediately, and allow to (7) Aluminum—To 5.0 g of Lithium Carbonate add stand for 10 minutes: the solution has no more turbidity than 20 mL of water, add gradually 15 mL of hydrochloric acid the following control solution. while stirring, and evaporate to dryness on a water bath. Control solution: Dissolve 9.5 mg of potassium chloride in To the residue add 50 mL of water to dissolve, filter if neces- water to make 1000 mL. To 5 mL of this solution add 1.0 sary, and assign this solution as solution A. Separately, mL of dilute acetic acid, shake, and proceed in the same evaporate 15 mL of hydrochloric acid to dryness on a water manner. bath, then proceed in the same manner, and assign the solu- (12) Sodium—Weigh accurately about 0.8 g of Lithium tion so obtained as solution B. To 10 mL of solution A add Carbonate, dissolve in water to make exactly 100 mL, and 10 mL of water and 5 mL of acetic acid-sodium acetate use this solution as the sample stock solution. Measure ex- buffer solution, pH 4.5, and shake. Add 1 mL of a solution actly 25 mL of the sample stock solution, add water to make of L-ascorbic acid (1 in 100), 2 mL of aluminon TS and exactly 100 mL, and use this solution as the sample solution water to make 50 mL, shake well, and allow to stand for 10 (1). Separately, weigh accurately 25.4 mg of sodium chlo- minutes: the solution has no more color than the following ride, dissolve in water to make exactly 1000 mL, and use this control solution. solution as the standard solution. Measure exactly 25 mL of Control solution: Dissolve 0.1758 g of aluminum potas- the sample stock solution, add exactly 20 mL of the standard sium sulfate dodecahydrate in water to make 1000 mL. To solution, then add water to make exactly 100 mL, and use 1.0 mL of this solution add 10 mL of solution B obtained in this solution as the sample solution (2). Determine emission (7) and water to make 20 mL, add 5 mL of acetic acid-sodi- intensities of sodium using a flame photometer with the um acetate buffer solution, pH 4.5, and proceed in the same sample solution (1) and the sample solution (2) under the fol- manner. lowing conditions. Adjust the wavelength dial to 589 nm, (8) Barium—To 20 mL of solution A obtained in (7) add atomize the sample solution (2) into the flame, then adjust

6 mL of water, 0.5 mL of dilute hydrochloric acid, 3 mL of the sensitivity so that the emission intensity LS shows 100 ethanol (95) and 2 mL of potassium sulfate TS, and allow to adjustment, and determine emission intensity LT of the sam- stand for 1 hour: the solution has no more turbidity than the ple solution (1). Then, make the other conditions identical, following control solution. change the wavelength dial to 580 nm, determine emission

Control solution: Dissolve 17.8 mg of barium chloride di- intensity LB of the sample solution (1): the amount of so- hydrate in water to make 1000 mL. To 6 mL of this solution dium, calculated from the following equation, is not more add 20 mL of solution B obtained in (7), 0.5 mL of dilute than 0.05z. hydrochloric acid and 3 mL of ethanol (95), and proceed in Amount (z) of sodium (Na) thesamemanner. ＝ (L － L )/(L － L ) × M?/M × 100 (9) Calcium—Weigh accurately about 5 g of Lithium T B S T Carbonate, add 50 mL of water and 15 mL of hydrochloric M: Amount (mg) of the sample in 25 mL of the sample acid, and dissolve. Remove carbon dioxide from the solution stock solution by boiling, add 5 mL of ammonium oxalate TS, then make M?: Amount (mg) of sodium in 20 mL of the standard so- alkaline with ammonia TS, and allow to stand for 4 hours. lution Filter the produced precipitate through a glass filter (G4), (13) Arsenic <1.11>—Prepare the test solution with 1.0 g wash with warm water until the turbidity of the washing is of Lithium Carbonate, add 2 mL of water and 3 mL of hy- not produced with calcium chloride TS within 1 minute. drochloric acid, and perform the test (not more than 2 ppm). Transfer the precipitate and the glass filter into a beaker, add water until the glass filter is covered with water, then Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, add 3 mL of sulfuric acid, heat between 709Cand809C, and 3 hours). titrate with 0.02 mol/L potassium permanganate VS until a JP XVI Official Monographs / Losartan Potassium 1045

Assay Weigh accurately about 1 g of Lithium Carbonate, dilute nitric acid and water to make 50 mL. Perform the test previously dried, add exactly 100 mL of water and 50 mL of using this solution as the test solution. Prepare the control 0.5 mol/L sulfuric acid VS, remove carbon dioxide by boil- solution with 0.20 mL of 0.01 mol/L hydrochloric acid VS ing gently, cool, and titrate <2.50> the excess sulfuric acid (not more than 0.014z). with 1 mol/L sodium hydroxide VS until the color of the so- (2) Heavy metals <1.07>—Proceedwith1.0gofLora- lution changes from red to yellow (indicator: 3 drops of zepam according to Method 2, and perform the test. Prepare methyl red TS). Perform a blank determination. the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Each mL of 0.5 mol/L sulfuric acid VS (3) Arsenic <1.11>—Prepare the test solution with 1.0 g ＝ 36.95 mg of Li CO 2 3 of Lorazepam according to Method 3, and perform the test Containers and storage Containers—Well-closed contain- (not more than 2 ppm). ers. (4) Related substances—Dissolve 0.10 g of Lorazepam in 20 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ethanol (95) Lorazepam to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as di- ロラゼパム rected under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform, 1,4-dioxane and acetic acid (100) (91:5:4) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot from the sample solution are not more intense than the spot from the standard solution. C15H10Cl2N2O2: 321.16 (3RS)-7-Chloro-5-(2-chlorophenyl)-3-hydroxy- Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- 1,3-dihydro-2H-1,4-benzodiazepin-2-one um, [846-49-1] 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.3z (1 g). Lorazepam, when dried, contains not less than 98.5z of C15H10Cl2N2O2. Assay Weigh accurately about 0.4 g of Lorazepam, previously dried, dissolve in 50 mL of acetone, and titrate <2.50> Description Lorazepam occurs as a white, crystalline pow- with 0.1 mol/L tetrabutylammonium hydroxide VS (poten- der. It is odorless. tiometric titration). Perform a blank determination, and It is sparingly soluble in ethanol (95) and in acetone, make any necessary correction. slightly soluble in diethyl ether, and practically insoluble in water. Each mL of 0.1 mol/L tetrabutylammonium It is gradually colored by light. hydroxide VS ＝ 32.12 mg of C H Cl N O Identification (1) To 0.02 g of Lorazepam add 15 mL of 15 10 2 2 2 dilute hydrochloric acid, boil for 5 minutes, and cool: the so- Containers and storage Containers—Tight containers. lution responds to the Qualitative Tests <1.09> for primary Storage—Light-resistant. aromatic amines. (2) Determine the absorption spectrum of a solution of Lorazepam in ethanol (95) (1 in 200,000) as directed under Losartan Potassium Ultraviolet-visible Spectrophotometry <2.24>,andcompare the spectrum with the Reference Spectrum: both spectra ex- ロサルタンカリウム hibit similar intensities of absorption at the same wavelengths. (3) Determine the infrared absorption spectrum of Lorazepam, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. (4) Perform the test with Lorazepam as directed under Flame Coloration Test <1.04> (2): a green color appears. C22H22ClKN6O: 461.00 1z Absorbance <2.24> E 1cm (229 nm): 1080 – 1126 (after dry- Monopotassium 5-{[4?-(2-butyl-4-chloro-5-hydroxymethyl- ing, 1 mg, ethanol (95), 200 mL). 1H-imidazol-1-yl)methyl]biphenyl-2-yl}-1H-tetrazol-1-ide [124750-99-8] Purity (1) Chloride <1.03>—To 1.0 g of Lorazepam add 50 mL of water, allow to stand for 1 hour with occasional Losartan Potassium contains not less than 98.5z shaking, and filter. To 25 mL of the filtrate add 6 mL of and not more than 101.0z of C22H22ClKN6O, calcu- 1046 Losartan Potassium / Official Monographs JP XVI lated on the anhydrous basis. Time after injection Mobile phase A Mobile phase B Description Losartan Potassium occurs as a white crystal- of sample (min) (volz) (volz) line powder. It is very soluble in water, and freely soluble in methanol 0–25 75→ 10 25 → 90 and in ethanol (99.5). 25–35 10 90 Identification (1) Determine the absorption spectrum of a solution of Losartan Potassium in methanol (1 in 100,000) as Flow rate: 1.0 mL per minute. directed under Ultraviolet-visible Spectrophotometry <2.24>, Time span of measurement: 35 minutes after injection of and compare the spectrum with the Reference Spectrum or the sample. the spectrum of a solution of Losartan Potassium RS pre- System suitability— pared in the same manner as the sample solution: both spec- Test for required detectability: Pipet 1 mL of the standard tra exhibit similar intensities of absorption at the same wave- solution, and add methanol to make exactly 10 mL. Confirm lengths. that the peak area of losartan obtained from 10 mLofthis (2) Determine the infrared absorption spectrum of solution is equivalent to 7 to 13z of that of losartan from 10 Losartan Potassium as directed in the potassium bromide mL of the standard solution. disk method under Infrared Spectrophotometry <2.25>,and System performance: When the procedure is run with 10 compare the spectrum with the Reference Spectrum or the mL of the standard solution under the above operating con- spectrum of Losartan Potassium RS: both spectra exhibit ditions, the number of theoretical plates and the symmetry similar intensities of absorption at the same wave numbers. factor of the peak of losartan are not less than 10,000 and (3) Losartan Potassium responds to the Qualitative Tests not more than 1.3, respectively. <1.09> (1) for potassium salt. System repeatability: When the test is repeated 6 times (4) Perform the test with Losartan Potassium as directed with 10 mL of the standard solution under the above operat- under Flame Coloration Test <1.04> (2): a green color ap- ing conditions, the relative standard deviation of the peak pears. area of losartan is not more than 2.0z. (3) Residual solvent—Being specified separately. Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Losartan Potassium according to Method 2, and perform the Water <2.48> Not more than 0.5z (0.25 g, volumetric test. Prepare the control solution with 2.0 mL of Standard titration, direct titration). Lead Solution (not more than 10 ppm). Assay Weigh accurately about 25 mg each of Losartan (2) Related substances—Dissolve 30 mg of Losartan Potassium and Losartan Potassium RS (separately, deter- Potassium in 100 mL of methanol, and use this solution as mine the water <2.48> in the same manner as Losartan Potas- the sample solution. Pipet 1 mL of this solution, add metha- sium), dissolve separately in methanol to make exactly 100 nol to make exactly 100 mL, and use this solution as the mL, and use these solutions as the sample solution and the standard solution. Perform the test with exactly 10 mLeach standard solution, respectively. Perform the test with exactly of the sample solution and standard solution as directed 10 mL each of the sample solution and standard solution as under Liquid Chromatography <2.01> according to the fol- directed under Liquid Chromatography <2.01> according to lowing conditions. Determine each peak area of both solu- the following conditions, and determine the peak areas, AT tions by the automatic integration method: the area of each and AS, of losartan in each solution. peak other than the peaks of solvent and losartan obtained from the sample solution is not larger than 1/10 times the Amount (mg) of losartan potassium (C22H22ClKN6O) peak area of losartan from the standard solution, and the ＝ MS × AT/AS total area of the peaks other than the peak of losartan from MS: Amount (mg) of Losartan Potassium RS, calculated the sample solution is not larger than 3/10 times the peak on the anhydrous basis area of losartan from the standard solution. Operating conditions— Operating conditions— Detector: An ultraviolet absorption photometer (wave- Detector: An ultraviolet absorption photometer (wavelength: 220 nm). length: 254 nm). Column: A stainless steel column 4 mm in inside diameter Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticlediameter). for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about Column temperature: A constant temperature of about 259C. 359C. Mobile phase A: Diluted phosphoric acid (1 in 1000). Mobile phase: A mixture of diluted phosphoric acid (1 in Mobile phase B: Acetonitrile. 1000) and acetonitrile (3:2). Flowing of the mobile phase: Control the gradient by mix- Flow rate: Adjust the flow rate so that the retention time ing the mobile phases A and B as directed in the following of losartan is about 6 minutes. table. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of losartan are not less than 5500 and not more than 1.4, respectively. JP XVI Official Monographs / Loxoprofen Sodium Hydrate 1047

System repeatability: When the test is repeated 6 times tions as directed under Thin-layer Chromatography <2.03>. with 10 mL of the standard solution under the above operat- Spot 10 mL each of the sample solution and standard solu- ing conditions, the relative standard deviation of the peak tion on a plate of silica gel with fluorescent indicator for area of losartan is not more than 1.0z. thin-layer chromatography. Develop the plate with a mixture of 1,2-dichloroethane and acetic acid (100) (9:1) to a distance Containers and storage Containers—Tight containers. of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot from the sample solution are not more in- Loxoprofen Sodium Hydrate tense than the spot from the standard solution. ロキソプロフェンナトリウム水和物 Water <2.48> 11.0 – 13.0z (0.2 g, volumetric titration, direct titration). Assay Weigh accurately about 60 mg of Loxoprofen So- dium Hydrate, and dissolve in diluted methanol (3 in 5) to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 10 mL of the internal standard solution, add diluted methanol (3 in 5) to make 100 mL, and use this solution as C H NaO .2H O: 304.31 15 17 3 2 the sample solution. Separately, weigh accurately about 50 Monosodium 2-{4-[(2- mg of Loxoprofen RS, previously dried in a desiccator (in oxocyclopentyl)methyl]phenyl}propanoate dihydrate vacuum, 609C) for 3 hours, and dissolve in diluted methanol [80382-23-6] (3 in 5) to make exactly 100 mL. Pipet 5 mL of this solution, proceed in the same manner as directed for the preparation Loxoprofen Sodium Hydrate contains not less than of the sample solution, and use so obtained solution as the 98.5z of loxoprofen sodium (C H NaO : 268.28), 15 17 3 standard solution. Perform the test with 10 mLeachofthe calculated on the anhydrous basis. sample solution and standard solution as directed under Liq- Description Loxoprofen Sodium Hydrate occurs as white uid Chromatography <2.01> according to the following con- to yellowish white crystals or crystalline powder. ditions, and calculate the ratios, QT and QS, of the peak area It is very soluble in water and in methanol, freely soluble of loxoprofen to that of the internal standard. in ethanol (95), and practically insoluble in diethyl ether. Amount (mg) of loxoprofen sodium (C H NaO ) A solution of Loxoprofen Sodium Hydrate (1 in 20) does 15 17 3 ＝ M × Q /Q × 1.089 not show optical rotation. S T S

The pH of a solution of Loxoprofen Sodium Hydrate in MS: Amount (mg) of Loxoprofen RS freshly boiled and cooled water (1 in 20) is between 6.5 and Internal standard solution—A solution of ethyl benzoate in 8.5. diluted methanol (3 in 5) (7 in 50,000). Identification (1) Determine the absorption spectrum of a Operating conditions— solution of Loxoprofen Sodium Hydrate (1 in 55,000) as di- Detector: An ultraviolet absorption photometer (wave- rected under Ultraviolet-visible Spectrophotometry <2.24>, length: 222 nm). and compare the spectrum with the Reference Spectrum: Column: A stainless steel column 4.6 mm in inside diame- both spectra exhibit similar intensities of absorption at the ter and 15 cm in length, packed with octadecylsilanized silica same wavelengths. gel for liquid chromatography (5 mminparticlediameter). (2) Determine the infrared absorption spectrum of Lox- Column temperature: A constant temperature of about oprofen Sodium Hydrate as directed in the potassium bro- 409C. mide disk method under the Infrared Spectrophotometry Mobile phase: A mixture of methanol, water, acetic acid <2.25>, and compare the spectrum with the Reference Spec- (100) and triethylamine (600:400:1:1). trum: both spectra exhibit similar intensities of absorption at Flow rate: Adjust the flow rate so that the retention time the same wave numbers. of loxoprofen is about 7 minutes. (3) A solution of Loxoprofen Sodium Hydrate (1 in 10) System suitability— responds to the Qualitative Tests <1.09> for sodium salt. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con- Purity (1) Clarity and color of solution—Dissolve 1.0 g ditions, loxoprofen and the internal standard are eluted in of Loxoprofen Sodium Hydrate in 10 mL of water: the solu- this order with the resolution between these peaks being not tion is clear and colorless or pale yellow. The color is not less than 10. darker than that of diluted Matching Fluid for Color A (1 in System repeatability: When the test is repeated 5 times 2). with 10 mL of the standard solution under the above operat- (2) Heavy metals <1.07>—Proceed with 2.0 g of Lox- ing conditions, the relative standard deviation of the ratios oprofen Sodium Hydrate according to Method 2, and per- of the peak area of loxoprofen to that of the internal stand- form the test. Prepare the control solution with 2.0 mL of ard is not more than 1.0z. Standard Lead Solution (not more than 10 ppm). (3) Related substances—Dissolve 1.0 g of Loxoprofen Containers and storage Containers—Tight containers. Sodium Hydrate in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solu- 1048 L-Lysine Acetate / Official Monographs JP XVI

L-cystine, L-valine, L-methionine, L-isoleucine, L-leucine, L-Lysine Acetate L-tyrosine, L-phenylalanine, L-lysine hydrochloride, ammonium chloride, L-histidine and L-arginine, dissolve in 0.1 L-リシン酢酸塩 mol/L hydrochloric acid TS to make exactly 1000 mL, and use this solution as the standard stock solution. Pipet 5 mL of this solution, add 0.02 mol/L hydrochloric acid to make exactly 100 mL. Pipet 4 mL of this solution, add 0.02 mol/L hydrochloric acid to make exactly 50 mL, and use this solu-

C6H14N2O2.C2H4O2:206.24 tion as the standard solution. Perform the test with exactly (2S)-2,6-Diaminohexanoic acid monoacetate 20 mL each of the sample solution and standard solution as [57282-49-2] directed under Liquid Chromatography <2.01> according to the following conditions. Based on the peak heights of the L-Lysine Acetate, when dried, contains not amino acids obtained from the sample solution and standard less than 98.5z and not more than 101.0z of solution, determine the mass of the amino acids other than C6H14N2O2.C2H4O2. lysine contained in 1 mL of the sample solution, and calculate the mass percent: the amount of each amino acids other Description L-Lysine Acetate occurs as white crystals or than lysine is not more than 0.1z. crystalline powder. It has a characteristic odor and a slightly Operating conditions— acid taste. Detector: A visible spectrophotometer (wavelength: 570 It is very soluble in water, freely soluble in formic acid, nm). and practically insoluble in ethanol (99.5). Column: A stainless steel column 4.6 mm in inside diame- It is deliquescent. ter and 8 cm in length, packed with strongly acidic ion- Identification (1) Determine the infrared absorption spec- exchange resin for liquid chromatography (Na type) com- trum of L-Lysine Acetate as directed in the potassium bro- posed with a sulfonated polystyrene copolymer (3 mmin mide disk method under Infrared Spectrophotometry <2.25>, particle diameter). and compare the spectrum with the Reference Spectrum: Column temperature: A constant temperature of about both spectra exhibit similar intensities of absorption at the 579C. same wave numbers. Chemical reaction bath temperature: A constant tempera- (2) A solution of L-Lysine Acetate (1 in 20) responds to ture of about 1309C. the Qualitative Tests <1.09> (2) for acetate. Color developing time: About 1 minute. Mobile phase: Prepare mobile phases A, B, C, D and E Optical rotation <2.49> [a]20: ＋8.5 – ＋10.09(after drying, D according to the following table, and to each phase add 0.1 2.5 g, water, 25 mL, 100 mm). mL of capric acid. pH <2.54> Dissolve 1.0 g of L-Lysine Acetate in 10 mL of water: the pH of the solution is between 6.5 and 7.5. Mobile Mobile Mobile Mobile Mobile phase A phase B phase C phase D phase E Purity (1) Clarity and color of solution—Dissolve 1.0 g Citric acid of L-Lysine Acetate in 10 mL of water: the solution is color- 19.80 g 22.00 g 12.80 g 6.10 g — less and clear. monohydrate (2) Chloride <1.03>—Perform the test with 0.5 g of - Trisodium citrate L dihydrate 6.19 g 7.74 g 13.31 g 26.67 g — Lysine Acetate. Prepare the control solution with 0.30 mL of Sodium chlo- 0.01 mol/L hydrochloric acid VS (not more than 0.021z). ride 5.66 g 7.07 g 3.74 g 54.35 g — (3) Sulfate <1.14>—Perform the test with 0.6 g of L- Sodium hydroxide ————8.00g Lysine Acetate. Prepare the control solution with 0.35 mL of Ethanol (99.5) 130 mL 20 mL 4 mL — 100 mL 0.005 mol/L sulfuric acid VS (not more than 0.028z). Thiodiglycol 5 mL 5 mL 5 mL — — (4) Ammonium <1.02>—Perform the test with 0.25 g of Benzyl alcohol — — — 5 mL — L-Lysine Acetate. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). Lauromacrogol solution (1 in 4) 4 mL 4 mL 4 mL 4 mL 4 mL (5) Heavy metals <1.07>—Proceed with 1.0 g of L-Lysine Appropriate Appropriate Appropriate Appropriate Appropriate Acetate according to Method 4, and perform the test. Pre- Water amount amount amount amount amount pare the control solution with 1.0 mL of Standard Lead So- lution (not more than 10 ppm). Total volume 1000 mL 1000 mL 1000 mL 1000 mL 1000 mL (6) Iron <1.10>—Prepare the test solution with 1.0 g of L- Lysine Acetate according to Method 1, and perform the test Changing mobile phases: Proceed with 20 mL of the stand- according to Method A. Prepare the control solution with ard solution under the above operating conditions: aspartic 1.0 mL of Standard Iron Solution (not more than 10 ppm). acid, threonine, serine, glutamic acid, glycine, alanine, cys- (7) Related substances—Weigh accurately about 0.5 g of tine, valine, methionine, isoleucine, leucine, tyrosine, L-Lysine Acetate, dissolve in 0.5 mL of hydrochloric acid phenylalanine, lysine, ammonia, histidine and arginine are and water to make exactly 100 mL. Pipet 10 mL of this solu- eluted in this order. Switchover the mobile phases A, B, C, tion, add 0.02 mol/L hydrochloric acid to make exactly 50 D and E in sequence so that the resolution between the peaks mL, and use this solution as the sample solution. Separately, of isoleucine and leucine is not less than 1.2. weigh accurately 2.5 mol amounts of L-aspartic acid, Reaction reagents: Dissolve 204 g of lithium acetate dihy- L-threonine, L-serine, L-glutamic acid, glycine, L-alanine, drate in water, and add 123 mL of acetic acid (100), 401 mL JP XVI Official Monographs / L-Lysine Hydrochloride 1049 of 1-methoxy-2-propanol, and water to make 1000 mL, gas ties of absorption at the same wave numbers. If any differ- with nitrogen for 10 minutes, and use this solution as the so- ence appears between the spectra, dissolve L-Lysine Hydro- lution (I). Separately, to 979 mL of 1-methoxy-2-propanol chloride in water, evaporate the water to dryness at 609C, add 39 g of ninhydrin, gas with nitrogen for 5 minutes, add and repeat the test with the residue. 81 mg of sodium borohydride, gas the solution with nitrogen (2) A solution of L-Lysine Hydrochloride (1 in 10) re- for 30 minutes, and use this solution as solution (II). To 1 sponds to the Qualitative Tests <1.09> for chloride. volume of the solution (I) add 1 volume of the solution (II). Optical rotation <2.49> [a] : ＋19.0 – ＋21.59(after dry- Prepare before use. D ing, 2 g, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm). Mobile phase flow rate: 0.20 mL per minute. Reaction reagent flow rate: 0.24 mL per minute. pH <2.54> Dissolve 1.0 g of L-Lysine Hydrochloride in 10 System suitability— mL of water: the pH of this solution is between 5.0 and 6.0. System performance: When the procedure is run with 20 Purity (1) Clarity and color of solution—Dissolve 1.0 g mL of the standard solution under the above operating con- of L-Lysine Hydrochloride in 10 mL of water: the solution is ditions, the resolution between the peaks of glycine and ala- clear and colorless. nine is not less than 1.2. (2) Sulfate <1.14>—Perform the test with 0.6 g of System repeatability: When the test is repeated 6 times L-Lysine Hydrochloride. Prepare the control solution with with 20 mL of the standard solution under the above operat- 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than ing conditions, the relative standard deviation of the peak 0.028z). height of each amino acid in the standard solution is not (3) Ammonium <1.02>—Perform the test with 0.25 g of more than 5.0z, and the relative standard deviation of the L-Lysine Hydrochloride. Prepare the control solution with retention time is not more than 1.0z. 5.0 mL of Standard Ammonium Solution (not more than Loss on drying <2.41> Not more than 0.3z (1 g, 809C, 0.02z). 3 hours). (4) Heavy metals <1.07>—Proceed with 2.0 g of L-Lysine Hydrochloride according to Method 1, and perform the test. Residue on ignition <2.44> Not more than 0.1z (1 g). Prepare the control solution with 2.0 mL of Standard Lead Assay Weigh accurately about 0.1 g of L-Lysine Acetate, Solution (not more than 10 ppm). previously dried, dissolve in 3 mL of formic acid, add 50 mL (5) Arsenic <1.11>—Prepare the test solution with 1.0 g of acetic acid (100), and titrate <2.50> with 0.1 mol/L per- of L-Lysine Hydrochloride according to Method 1, and perchloric acid VS (potentiometric titration). Perform a blank form the test (not more than 2 ppm). determination in the same manner, and make any necessary (6) Related substances—Dissolve 0.10 g of L-Lysine Hy- correction. drochloride in 25 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add Each mL of 0.1 mol/L perchloric acid VS water to make exactly 50 mL, pipet 5 mL of this solution, ＝ 10.31 mg of C H N O .C H O 6 14 2 2 2 4 2 add water to make exactly 20 mL, and use this solution as Containers and storage Containers—Tight containers. the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a L-Lysine Hydrochloride plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-propanol and ammonia water Lysine Hydrochloride (28) (67:33) to a distance of about 10 cm, and dry the plate at 1009C for 30 minutes. Spray evenly the plate with a solution L-リシン塩酸塩 of ninhydrin in acetone (1 in 50) and heat at 809Cfor5 minutes: the spots other than the principal spot from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, C H N O .HCl: 182.65 6 14 2 2 3 hours). (2S)-2,6-Diaminohexanoic acid monohydrochloride [657-27-2] Residue on ignition <2.44> Not more than 0.1z (1 g).

Assay Weigh accurately about 0.1 g of L-Lysine Hydro- -Lysine Hydrochloride, when dried, contains not L chloride, previously dried, dissolve in 2 mL of formic acid, less than 98.5z of C H N O .HCl. 6 14 2 2 add exactly 15 mL of 0.1 mol/L perchloric acid VS, and heat Description L-Lysine Hydrochloride occurs as a white pow- on a water bath for 30 minutes. After cooling, add 45 mL of der. It is odorless, and has a slight, characteristic taste. acetic acid (100), and titrate <2.50> the excess perchloric acid It is freely soluble in water and in formic acid, and practi- with 0.1 mol/L sodium acetate VS (potentiometric titration). cally insoluble in ethanol (95). Perform a blank determination, and make any necessary correction. Identification (1) Determine the infrared absorption spectrum of L-Lysine Hydrochloride, previously dried, as di- Each mL of 0.1 mol/L perchloric acid VS rected in the potassium bromide disk method under Infrared ＝ 9.132 mg of C6H14N2O2.HCl Spectrophotometry <2.25>, and compare the spectrum with Containers and storage Containers—Tight containers. the Reference Spectrum: both spectra exhibit similar intensi- 1050 Lysozyme Hydrochloride / Official Monographs JP XVI

mL of this solution, add phosphate buffer solution, pH 6.2 Lysozyme Hydrochloride to them to make exactly 50 mL, and use these solutions as the standard solution (1) and the solution (2), respectively. リゾチーム塩酸塩 Keep the sample solution and the standard solutions in an ice-bath. Pipet 4 mL of substrate solution for lysozyme hydrochloride, previously warmed in a water bath of 359Cfor about 5 minutes, add exactly 100 mL of the sample solution, previously warmed in a water bath of 359C for about 3 minutes, and allow to stand at 359C for exactly 10 minutes, then add exactly 0.5 mL of 1 mol/L hydrochloric acid TS,

C616H963N193O182S10.xHCl and immediately shake. Determine the absorbance under Ul- [12650-88-3, egg white lysozyme] traviolet-visible Spectrophotometry <2.24>, AT, of this solution at 640 nm, using water as the blank. Determine the ab-

Lysozyme Hydrochloride is a hydrochloride of a sorbances, AS1 and AS2, of the solutions obtained with the basic polypeptide obtained from albumen of hen's standard solution (1) and the standard solution (2) in the egg, and has an activity to hydrolyze mucopolysaccha- same manner as the sample solution. rides. Amount [mg (potency)] of lysozyme per mg, It contains not less than 0.9 mg (potency) of lyso- calculated on the dried basis zyme per mg, calculated on the dried basis. ＝ MS/2MT × {(AS1 － AT)/(AS1 － AS2) ＋ 1} Description Lysozyme Hydrochloride occurs as white, M : Amount (mg) of Lysozyme RS, calculated on the crystals, or crystalline or amorphous powder. S dried basis. It is freely soluble in water, and practically insoluble in M : Amount (mg) of the sample, calculated on the dried ethanol (99.5). T basis. It is hygroscopic. The pH of a solution of Lysozyme Hydrochloride (3 in Containers and storage Containers—Tight containers. 200) is between 3.0 and 5.0. Identification (1) To 5 mL of a solution of Lysozyme Hy- drochloride in acetate buffer solution, pH 5.4 (1 in 500) add Macrogol 400 1 mL of ninhydrin TS, and heat for 10 minutes: a blue-purple color develops. Polyethylene Glycol 400 (2) Determine the absorption spectrum of a solution of マクロゴール 400 Lysozyme Hydrochloride in acetate buffer solution, pH 5.4 (1 in 10,000) as directed under Ultraviolet-visible Spectro- photometry <2.24>, and compare the spectrum with the Ref- Macrogol 400 is a polymer of ethylene oxide erence Spectrum: both spectra exhibit similar intensities of and water, represented by the formula HOCH2 absorption at the same wavelengths. (CH2OCH2)nCH2OH, in which the value of n ranges from 7 to 9. Purity (1) Clarity of solution—To 5 mL of a solution of Lysozyme Hydrochloride (3 in 200) add, if necessary, dilute Description Macrogol 400 occurs as a clear, colorless and hydrochloric acid to adjust the pH to 3: the solution is clear. viscous liquid. It has no odor or a slight, characteristic odor. (2) Heavy metals <1.07>—Proceed with 1.0 g of Lyso- It is miscible with water, with methanol, with ethanol (95) zyme Hydrochloride according to Method 2, and perform andwithpyridine. the test. Prepare the control solution with 2.0 mL of Stand- It is soluble in diethyl ether. ard Lead Solution (not more than 20 ppm). It is slightly hygroscopic. Congealing point: 4 – 89C Loss on drying <2.41> Not more than 8.0z (0.1 g, 1059C, Specific gravity d 20: 1.110 – 1.140 2 hours). 20 Identification Dissolve 50 mg of Macrogol 400 in 5 mL of Residue on ignition <2.44> Not more than 2.0z (0.5 g). dilute hydrochloric acid, add 1 mL of barium chloride TS, Nitrogen Perform the test as directed under Nitrogen De- shake, and filter, if necessary. To the filtrate add 1 mL of a termination <1.08>: the amount of nitrogen (N: 14.01) is be- solution of phosphomolybdic acid n-hydrate (1 in 10): a tween 16.8z and 18.6z,calculatedonthedriedbasis. yellow-green precipitate is formed. Assay Weigh accurately an amount of Lysozyme Hydro- pH <2.54> Dissolve 1.0 g of Macrogol 400 in 20 mL of chloride, equivalent to about 25 mg (potency), dissolve in water: the pH of this solution is between 4.0 and 7.0. phosphate buffer solution, pH 6.2 to make exactly 100 mL. Purity (1) Acidity—Dissolve 5.0 g of Macrogol 400 in Pipet 2 mL of this solution, add phosphate buffer solution, 20 mL of neutralized ethanol, and add 2 drops of phenol- pH 6.2 to make exactly 50 mL, and use this solution as the phthalein TS and 0.20 mL of 0.1 mol/L sodium hydroxide sample solution. Separately, weigh accurately an amount of VS: the solution is red in color. Lysozyme RS (separately determine its loss on drying <2.41> (2) Ethylene glycol and diethylene glycol—Dissolve 4.0 g in the same manner as Lysozyme Hydrochloride), equivalent of Macrogol 400 in water to make exactly 10 mL, and use to about 25 mg (potency), and dissolve in phosphate buffer this solution as the sample solution. Weigh accurately about solution, pH 6.2 to make exactly 100 mL. Pipet 1 mL and 2 50 mg each of ethylene glycol and diethylene glycol, dissolve JP XVI Official Monographs / Macrogol 1500 1051 in water to make exactly 100 mL, and use this solution as the in the test of the sample standard solution. Perform the test with exactly 2 mLeachof Average molecular mass is between 380 and 420. the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following con- Water <2.48> Not more than 1.0z (2 g, volumetric titra- ditions. Determine the peak heights, HTa and HSa,ofethyl- tion, direct titration). ene glycol of each solution, and the peak heights, H and Tb Residue on ignition <2.44> Not more than 0.1z (1 g). HSb, of diethylene glycol, and calculate the amount of ethylene glycol and diethylene glycol: the sum of the contents of Containers and storage Containers—Tight containers. ethylene glycol and diethylene glycol is not more than 0.25z. Amount (mg) of ethylene glycol Macrogol 1500 ＝ M × H /H × 1/10 Sa Ta Sa Polyethylene Glycol 1500 Amount (mg) of diethylene glycol

＝ MSb × HTb/HSb × 1/10 マクロゴール 1500

MSa: Amount (mg) of ethylene glycol for gas chromatography Macrogol 1500 is a mixture containing equal MSb: Amount (mg) of diethylene glycol for gas chroma- amounts of lower and higher polymers of ethylene tography oxide and water, represented by the formula HOCH2 (CH OCH ) CH OH, in which the value of n is 5 or 6 Operating conditions— 2 2 n 2 for the lower polymers and from 28 to 36 for the Detector: A hydrogen flame-ionization detector. higher. Column: A colum about 3 mm in inside diameter and about 1.5 m in length, packed with siliceous earth for gas Description Macrogol 1500 occurs as a white, smooth chromatography, 150 to 180 mm in particle diameter, coated petrolatum-like solid. It is odorless or has a faint, character- with D-sorbitol at the ratio of 12z. istic odor. Column temperature: A constant temperature of about It is very soluble in water, in pyridine and in diphenyl 1659C. ether, freely soluble in methanol, sparingly soluble in Carrier gas: Nitrogen or helium. ethanol (95), very slightly soluble in ethanol (99.5), and prac- Flow rate: Adjust the flow rate so that the retention time tically insoluble indiethylether. of diethylene glycol is about 8 minutes. Congealing point: 37 – 419C Selection of column: Proceed with 2 mL of the standard Identification Dissolve 50 mg of Macrogol 1500 in 5 mL of solution under the above operating conditions, and calculate dilute hydrochloric acid, add 1 mL of barium chloride TS, the resolution. Use a column clearly dividing peaks of ethyl- shake, and filter, if necessary. To the filtrate add 1 mL of a ene glycol and diethylene glycol in this order. solution of phosphomolybdic acid n-hydrate (1 in 10): a Detection sensitivity: Adjust the detection sensitivity so yellow-green precipitate is formed. that the peak height of diethylene glycol obtained from 2 mL of the standard solution composes about 80z of the full pH <2.54> Dissolve 1.0 g of Macrogol 1500 in 20 mL of scale. water: the pH of the solution is between 4.0 and 7.0. Average molecular mass Add 42 g of phthalic anhydride to Purity (1) Clarity and color of solution—Dissolve 5.0 g 300 mL of freshly distilled pyridine, exactly measured, in a of Macrogol 1500 in 50 mL of water: the solution is clear 1-L light-resistant glass-stoppered bottle. Shake the bottle and colorless. vigorously to dissolved the solid, and allow to stand for 16 (2) Acidity—Dissolve 5.0 g of Macrogol 1500 in 20 mL hours or more. Pipet 25 mL of this solution into an about of neutralized ethanol, and add 2 drops of phenolphthalein 200-mL glass-stoppered pressure bottle. Add about 1.5 g of TS and 0.20 mL of 0.1 mol/L sodium hydroxide VS: the so- Macrogol 400, accurately weighed, stopper the bottle, wrap lution is red in color. it securely with strong cloth, and immerse in a water bath, (3) Ethylene glycol and diethylene glycol—Place 50.0 g having a temperature of 98 ± 29C, to the level so that the of Macrogol 1500 in a distilling flask, add 75 mL of diphenyl mixture in the bottle soaks completely in water. Maintain the ether, warm to dissolve if necessary, distil slowly under a temperature of the bath at 98 ± 29C for 30 minutes. Re- reduced pressure of 0.13 to 0.27 kPa and take 25 mL of the move the bottle from the bath, and allow to cool in air to distillate in a 100-mL container with 1-mL graduation. To room temperature. Add exactly 50 mL of 0.5 mol/L sodium the distillate add exactly 20 mL of water, shake vigorously, hydroxide VS and 5 drops of a solution of phenolphthalein cool in ice water, congeal the diphenyl ether, and filtrate into in pyridine (1 in 100). Titrate <2.50> with 0.5 mol/L sodium a 25-mL volumetric flask. Wash the residue with 5.0 mL of hydroxide VS until a light red color remains for not less than ice-cold water, combine the washings with the filtrate, warm 15 seconds. Perform a blank determination. to room temperature, and add water to make 25 mL. Trans- fer this solution to a glass-stoppered flask, shake with 25.0 Average molecular mass ＝ (M × 4000)/(a － b) mL of freshly distilled acetonitrile, and use this solution as M: Amount (g) of sample the sample solution. Separately, to 62.5 mg of diethylene a: Volume (mL) of 0.5 mol/L sodium hydroxide VS used glycol add a mixture of water and freshly distilled aceto- in the blank determination nitrile (1:1) to make exactly 25 mL, and use this solution as b: Volume (mL) of 0.5 mol/L sodium hydroxide VS used the standard solution. Take exactly 10 mL each of the sam- 1052 Macrogol 4000 / Official Monographs JP XVI ple solution and the standard solution, and add to each from the bath, and allow to cool in air to room temperature. exactly 15 mL of cerium (IV) diammonium nitrate TS. Per- Add exactly 50 mL of 0.5 mol/L sodium hydroxide VS and 5 form the test with this solution as directed under Ultraviolet- drops of a solution of phenolphthalein in pyridine (1 in 100). visible Spectrophotometry <2.24> within 2 to 5 minutes: the Titrate <2.50> with 0.5 mol/L sodium hydroxide VS until a absorbance of the solution obtained from the sample solu- light red color remains for not less than 15 seconds. Perform tion at the wavelength of maximum absorption at about 450 a blank determination. nm is not larger than the absorbance of the solution obtained Average molecular mass ＝ (M × 4000)/(a － b) from the standard solution. M: Amount (g) of sample Water <2.48> Not more than 1.0z (2 g, volumetric titra- a: Volume (mL) of 0.5 mol/L sodium hydroxide VS con- tion, direct titration). sumed in the blank determination Residue on ignition <2.44> Not more than 0.1z (1 g). b: Volume (mL) of 0.5 mol/L sodium hydroxide VS consumed in the test of the sample Containers and storage Containers—Tight containers. Average molecular mass is between 2600 and 3800. Water <2.48> Not more than 1.0z (2 g, volumetric titra- Macrogol 4000 tion, direct titration). Polyethylene Glycol 4000 Residue on ignition <2.44> Not more than 0.2z (1 g). Containers and storage Containers—Well-closed contain- マクロゴール 4000 ers.

Macrogol 4000 is a polymer of ethylene oxide and water, represented by the formula HOCH2 Macrogol 6000 (CH2OCH2)nCH2OH, in which the value of n ranges from 59 to 84. Polyethylene Glycol 6000 Description Macrogol 4000 is a white, paraffin-like solid, マクロゴール 6000 occurring as flakes or powder. It is odorless or has a faint, characteristic odor. It is very soluble in water, freely soluble in methanol and Macrogol 6000 is a polymer of ethylene oxide in pyridine, and practically insoluble in ethanol (99.5) and in and water, represented by the formula HOCH2 diethyl ether. (CH2OCH2)nCH2OH, in which the value of n ranges Congealing point: 53 – 579C from 165 to 210. Identification Dissolve 50 mg of Macrogol 4000 in 5 mL of Description Macrogol 6000 is a white, paraffin-like solid, dilute hydrochloric acid, add 1 mL of barium chloride TS, occurring as flakes or powder. It is ordorless or has a faint, shake, and filter, if necessary. To the filtrate add 1 mL of a characteristic odor. solution of phosphomolybdic acid n-hydrate (1 in 10): a yel- It is very soluble in water, freely soluble in pyridine, and low-green precipitate is formed. practically insoluble in methanol, in ethanol (95), in ethanol (99.5) and in diethyl ether. pH <2.54> Dissolve 1.0 g of Macrogol 4000 in 20 mL of Congealing point: 56 – 619C water: the pH of this solution is between 4.0 and 7.5. Identification Dissolve 50 mg of Macrogol 6000 in 5 mL of Purity (1) Clarity and color of solution—A solution of dilute hydrochloric acid, add 1 mL of barium chloride TS, 5.0 g of Macrogol 4000 in 50 mL of water is clear and color- shake, and filter, if necessary. To the filtrate add 1 mL of a less. solution of phosphomolybdic acid n-hydrate (1 in 10): a (2) Acidity—Dissolve 5.0 g of Macrogol 4000 in 20 mL yellow-green precipitate is formed. of neutralized ethanol by warming, cool, and add 0.20 mL of 0.1 mol/L sodium hydroxide VS and 1 drop of phenol- pH <2.54> Dissolve 1.0 g of Macrogol 6000 in 20 mL of phthelein TS: the color of the solution is red. water: the pH of this solution is between 4.5 and 7.5. Average molecular mass Weigh accurately about 12.5 g of Purity (1) Clarity and color of solution—Dissolve 5.0 g Macrogol 4000, transfer to an about 200-mL glass-stoppered of Macrogol 6000 in 50 mL of water: the solution is clear pressure bottle, add about 25 mL of pyridine, dissolve by and colorless. warming, and allow to cool. Separately, pipet 300 mL of (2) Acidity—Dissolve 5.0 g of Macrogol 6000 in 20 mL freshly distilled pyridine into a 1000-mL light-resistant, of neutralized ethanol by warming, cool, and add 0.20 mL glass-stoppered bottle, add 42 g of phthalic anhydride, dis- of 0.1 mol/L sodium hydroxide VS and 1 drop of phenol- solve with vigorous shaking, and allow to stand for 16 hours phthalein TS: the color of the solution is red. or more. Pipet 25 mL of this solution, transfer to the former Average molecular mass Weigh accurately about 12.5 g of pressure bottle, stopper the bottle tightly, wrap it securely Macrogol 6000, transfer to an about 200-mL glass-stoppered with strong cloth, and immerse in a water bath, previously pressure bottle, add about 25 mL of pyridine, dissolve by heated at 98 ± 29C, to the level so that the mixture in the warming, and allow to cool. Separately, pipet 300 mL of bottle soaks completely in water. Maintain the temperature freshly distilled pyridine into a 1000-mL light-resistant, of the bath at 98 ± 29C for 30 minutes. Remove the bottle glass-stoppered bottle, add 42 g of phthalic anhydride, dis- JP XVI Official Monographs / Macrogol Ointment 1053 solve with vigorous shaking, and allow to stand for 16 hours phthalein TS: the color of the solution is red. or more. Pipet 25 mL of this solution, transfer to the former Average molecular mass Weigh accurately about 15 g of pressure bottle, stopper the bottle tightly, wrap it securely Macrogol 20000, transfer to an about 200-mL glass- with strong cloth, and immerse in a water bath, previously stoppered pressure bottle, add about 25 mL of pyridine, dis- heated at 98 ± 29C, to the level so that the mixture in the solve by warming, and allow to cool. Separately, pipet 300 bottle soaks completely in water. Maintain the temperature mL of freshly distilled pyridine into a 1000-mL light- of the bath at 98 ± 29C for 30 minutes. Remove the bottle resistant glass-stoppered bottle, add 42 g of phthalic anhy- from the bath, and allow to cool in air to room temperature. dride, dissolve with vigorous shaking, and allow to stand for Add exactly 50 mL of 0.5 mol/L sodium hydroxide VS and 5 16 hours or more. Pipet 25 mL of this solution, transfer to drops of a solution of phenolphthalein in pyridine (1 in 100). the former pressure bottle, stopper the bottle tightly, wrap it Titrate <2.50> with 0.5 mol/L sodium hydroxide VS until a securely with strong cloth, and immerse in a water bath, light red color remains for not less than 15 seconds. Perform having a temperature of 98 ± 29C, to the same depth as the a blank determination in the same manner. mixture in the bottle. Maintain the temperature of the bath Average molecular mass ＝ (M × 4000)/(a － b) at 98 ± 29C for 60 minutes. Remove the bottle from the bath, and allow to cool in air to room temperature. Add ex- M: Amount (g) of sample actly 50 mL of 0.5 mol/L sodium hydroxide VS and 5 drops a: Volume (mL) of 0.5 mol/L sodium hydroxide VS con- of a solution of phenolphthalein in pyridine (1 in 100). sumed in the blank determination Titrate <2.50> with 0.5 mol/L sodium hydroxide VS until a b: Volume (mL) of 0.5 mol/L sodium hydroxide VS con- light red color remains for not less than 15 seconds. Perform sumed in the test of the sample a blank determination. Average molecular mass is between 7300 and 9300. Average molecular mass ＝ (M × 4000)/(a － b) Water <2.48> Not more than 1.0z (2 g, volumetric titra- M: Amount (g) of sample tion, direct titration). a: Volume (mL) of 0.5 mol/L sodium hydroxide VS used Residue on ignition <2.44> Not more than 0.2z (1 g). in the blank determination b: Volume (mL) of 0.5 mol/L sodium hydroxide VS used Containers and storage Containers—Well-closed contain- in the test of the sample ers. Average molecular mass is between 15000 and 25000. Water <2.48> Not more than 1.0z (2 g, volumetric titra- Macrogol 20000 tion, direct titration). Polyethylene Glycol 20000 Residue on ignition <2.44> Not more than 0.2z (1 g). Containers and storage Containers—Well-closed contain- マクロゴール 20000 ers.

Macrogol 20000 is a polymer of ethylene oxide and water, represented by the formula HOCH2 Macrogol Ointment (CH2OCH2)nCH2OH, in which the value of n lies between 340 and 570. Polyethylene Glycol Ointment Description Macrogol 20000 occurs as white, paraffin-like マクロゴール軟膏 flakes or powder. It is ordorless or has a faint, characteristic odor. Itisfreelysolubleinwaterandinpyridine,andpractically Method of preparation insoluble in methanol, in ethanol (95), in dehydrated diethyl Macrogol 4000 500 g ether, in petroleum benzine and in macrogol 400. Macrogol 400 500 g Congealing point: 56 – 649C To make 1000 g Identification Dissolve 50 mg of Macrogol 20000 in 5 mL of dilute hydrochloric acid, add 1 mL of barium chloride TS, Melt Macrogol 4000 and Macrogol 400 by warming on a shake, and filter, if necessary. To the filtrate add 1 mL of a water bath at 659C, and mix well until it congeals. Less than solution of phosphomolybdic acid n-hydrate (1 in 10): a 100 g of Macrogol 4000 or Macrogol 400 may be replaced by yellow-green precipitate is formed. an equal amount of Macrogol 400 or Macrogol 4000 to prepare 1000 g of a proper soft ointment. pH <2.54> Dissolve 1.0 g of Macrogol 20000 in 20 mL of water: the pH of this solution is between 4.5 and 7.5. Description Macrogol Ointment is white in color. It has a faint, characteristic odor. Purity (1) Clarity and color of solution—Dissolve 5.0 g of Macrogol 20000 in 50 mL of water: the solution is clear Identification Dissolve 50 mg of Macrogol Ointment in 5 and colorless. mL of dilute hydrochloric acid, add 1 mL of barium chloride (2) Acidity—Dissolve 5.0 g of Macrogol 20000 in 20 mL TS, shake, filter if necessary, and add 1 mL of a solution of of neutralized ethanol by warming, cool, and add 0.20 mL phosphomolybdic acid n-hydrate (1 in 10) to the filtrate: a of 0.1 mol/L sodium hydroxide VS and 1 drop of phenol- yellow-green precipitate is formed. 1054 Magnesium Carbonate / Official Monographs JP XVI

Containers and storage Containers—Tight containers. mL of dilute hydrochloric acid. Add 250 mL of water and 5 mL of a solution of L-tartaric acid (1 in 5), then add 10 mL of a solution of 2,2?,2!-nitrilotrisethanol (3 in 10) and 10 mL Magnesium Carbonate of 8 mol/L potassium hydroxide TS, allow to stand for 5 minutes, and titrate <2.50> with 0.01 mol/L disodium dihy- 炭酸マグネシウム drogen ethylenediamine tetraacetate VS until the color of the solution changes form red-purple to blue (indicator: 0.1 g of NN indicator). Perform a blank determination, and make Magnesium Carbonate is a basic hydrated magne- any necessary correction. sium carbonate or a normal hydrated magnesium carbonate. Each mL of 0.01 mol/L disodium dihydrogen Magnesium Carbonate contains not less than 40.0z ethylenediamine tetraacetate VS and not more then 44.0z of magnesium oxide (MgO: ＝ 0.5608 mg of CaO 40.30). The content of calcium oxide (CaO: 56.08) is not more ``Heavy magnesium carbonate'' may be used as than 0.6z. commonly used name for Magnesium Carbonate (6) Acid-insoluble substances—Mix 5.0 g of Magnesium which shows the height of the precipitate below the Carbonate and 75 mL of water, add 10 mL of hydrochloric 12.0-mL graduation line in the Precipitation test. acid dropwise while stirring, boil for 5 minutes, and cool. Description Magnesium Carbonate occurs as white, friable Collect the insoluble residue using filter paper for quantita- masses or powder. It is odorless. tive analysis, wash well with water until the last washing It is practically insoluble in water, in ethanol (95), in shows no turbidity with silver nitrate TS, and ignite the diethyl ether and in 1-propanol. residue together with the filter paper: the mass of the residue It dissolves in dilute hydrochloric acid with effervescence. is not more than 2.5 mg. Its saturated solution is alkaline. Precipitation test Transfer 1.0 g of Magnesium Carbonate, Identification (1) Dissolve 1 g of Magnesium Carbonate previously sifted through a No. 100 (150 mm) sieve to a glass- in 10 mL of dilute hydrochloric acid, boil, then cool, neu- stoppered measuring cylinder with a 50-mL graduation line tralize with sodium hydroxide TS, and filter, if necessary: at 150 mm from the bottom, and add water to make 50 mL. the solution responds to the Qualitative Tests <1.09> for Shake vigorously for exactly 1 minute, allow to stand for 15 magnesium salt. minutes, and measure the height of the precipitate (in gradu- (2) Magnesium Carbonate responds to the Qualitative ationinml). Tests <1.09> (1) for carbonate. Assay Weigh accurately about 0.4 g of Magnesium Car- Purity (1) Soluble salts—To 2.0 g of Magnesium Car- bonate, dissolve in 10 mL of water and 3.5 mL of dilute hy- bonate add 40 mL of 1-propanol and 40 mL of water, heat drochloric acid, and add water to make exactly 100 mL. to boil with constant stirring, cool, and filter. Wash the Pipet 25 mL of the solution, add 50 mL of water and 5 mL residue with water, combine the washings with the filtrate, of ammonia-ammonium chloride buffer solution, pH 10.7, and add water to make exactly 100 mL. Evaporate 50 mL of and titrate <2.50> with 0.05 mol/L disodium dihydrogen thesolutiononawaterbathtodryness,anddryat1059Cfor ethylenediamine tetraacetate VS (indicator: 40 mg of erio- 1 hour: the mass of the residue does not exceed 10.0 mg. chrome black T-sodium chloride indicator). Perform a blank (2) Heavy metals <1.07>—Moisten 1.0 g of Magnesium determination, and make any necessary correction. From the Carbonate with 4 mL of water, dissolve by addition of 10 volume of 0.05 mol/L disodium dihydrogen ethylenediamine mL of dilute hydrochloric acid, and evaporate on a water tetraacetate VS consumed deduct the volume of 0.05 mol/L bath to dryness. Dissolve the residue in 35 mL of water, 2 disodium dihydrogen ethylenediamine tetraacetate VS cor- mL of dilute acetic acid, 1 drop of ammonia TS, filter, if responding to the content of calcium oxide (CaO) obtained necessary, wash the filter paper with water, combine the in the Purity (5). washings with the filtrate, and add water to make 50 mL, Each mL of 0.05 mol/L disodium dihydrogen and perform the test using this solution as the test solution. ethylenediamine tetraacetate VS Prepare the control solution as follows: evaporate 10 mL of ＝ 2.015 mg of MgO dilute hydrochloric acid on a water bath to dryness, add 2 mL of dilute acetic acid and 3.0 mL of Standard Lead Solu- Each mg of calcium oxide (CaO) tion, and dilute with water to make 50 mL (not more than 30 ＝ 0.36 mL of 0.05 mol/L disodium dihydrogen ppm). ethylenediamine tetraacetate VS (3) Iron <1.10>—Prepare the test solution with 0.10 g of Containers and storage Containers—Well-closed contain- Magnesium Carbonate according to Method 1, and perform ers. the test according to Method A. Prepare the control solution with 2.0 mL of Standard Iron Solution (not more than 200 ppm). (4) Arsenic <1.11>—Prepare the test solution with 0.40 g of Magnesium Carbonate, previously moistened with 1.5 mL of water, add 3.5 mL of dilute hydrochloric acid, and perform the test (not more than 5 ppm). (5) Calcium oxide—Weigh accurately about 0.6 g of Magnesium Carbonate, and dissolve in 35 mL of water and 6 JP XVI Official Monographs / Magnesium Oxide 1055

Magnesium Oxide

酸化マグネシウム

MgO: 40.30

Magnesium Oxide, when ignited, contains not less than 96.0z of MgO. When 5 g of Magnesium Oxide has a volume not more than 30 mL, it may be labeled heavy magnesium oxide. Description Magnesium Oxide occurs as a white powder or granules. It is odorless. It is practically insoluble in water, in ethanol (95) and in diethyl ether. It dissolves in dilute hydrochloric acid. It absorbs moisture and carbon dioxide in air. Identification A solution of Magnesium Oxide in dilute hydrochloric acid (1 in 50) responds to the Qualitative Tests <1.09> for magnesium salt. Purity (1) Alkali and soluble salts—Transfer 2.0 g of Magnesium Oxide to a beaker, add 100 mL of water, cover the beaker with a watch-glass, heat on a water bath for 5 minutes, and filter immediately. After cooling, to 50 mL of the filtrate add 2 drops of methyl red TS and 2.0 mL of 0.05 mol/L sulfuric acid VS: a red color develops. Evaporate 25 mL of the remaining filtrate to dryness, and dry the residue at 1059C for 1 hour: the mass of the residue is not more than 10 mg. tion, and make any necessary correction. (2) Carbonate—Boil 0.10 g of Magnesium Oxide with 5 Each mL of 0.01 mol/L disodium dihydrogen mL of water, cool, and add 5 mL of acetic acid (31): almost ethylenediamine tetraacetate VS no effervescence occurs. ＝ 0.5608 mg of CaO (3) Heavy metals <1.07>—Dissolve 1.0 g of Magnesium Oxide in 20 mL of dilute hydrochloric acid, and evaporate The mass of calcium oxide (CaO: 56.08) is not more than on a water bath to dryness. Dissolve the residue in 35 mL of 1.5z. water, add 1 drop of phenolphthalein TS, neutralize with (6) Arsenic <1.11>—Dissolve 0.20 g of Magnesium Oxide ammonia TS, add 2 mL of dilute acetic acid, and filter, if in 5 mL of dilute hydrochloric acid, and perform the test necessary. Wash the filter paper with water, add water to the with this solution as the test solution (not more than 10 combined washing and the filtrate to make 50 mL, and per- ppm). form the test using this solution as the test solution. Prepare (7) Acid-insoluble substances—Mix 2.0 g of Magnesium the control solution as follows: to 20 mL of dilute hydro- Oxide with 75 mL of water, add 12 mL of hydrochloric acid chloric acid add 1 drop of phenolphthalein TS, neutralize dropwise, while shaking, and boil for 5 minutes. Collect the with ammonia TS, and add 2 mL of dilute acetic acid, 4.0 insoluble residue using filter paper for quantitative analysis, mL of Standard Lead Solution and water to make 50 mL wash well with water until the last washing shows no tur- (not more than 40 ppm). bidity with silver nitrate TS, and ignite the residue together (4) Iron <1.10>—Prepare the test solution with 40 mg of with the filter paper: the mass of the ignited residue does not Magnesium Oxide according to Method 1, and perform the more than 2.0 mg. test according to Method A. Prepare the control solution (8) Fluoride—(i) Apparatus: Use a hard glass appa- with 2.0 mL of Standard Iron Solution (not more than 500 ratus as illustrated in the figure. Ground-glass joints may be ppm). used. (5) Calcium oxide—Weigh accurately about 0.25 g of (ii) Procedure: Transfer 5.0 g of Natural Aluminum Sili- Magnesium Oxide, previously ignited, dissolve in 6 mL of cate to the distilling flask A with the aid of 20 mL of water, dilute hydrochloric acid by heating. Cool, add 300 mL of add about 1 g of glass wool and 50 mL of diluted purified water and 3 mL of a solution of L-tartaric acid (1 in 5), then sulfuric acid (1 in 2), and connect A to the distillation appa- add 10 mL of a solution of 2,2?,2!-nitrilotrisethanol (3 in 10) ratus, previously washed with steam streamed through the and 10 mL of 8 mol/L potassium hydroxide TS, allow to steam introducing tube E. Connect the condenser C with the stand for 5 minutes, and titrate <2.50> with 0.01 mol/L diso- receiver D containing 10 mL of 0.01 mol/L sodium hydrox- dium dihydrogen ethylenediamine tetraacetate VS until the ide VS and 10 mL of water so that the lower end of C is color of the solution changes from red-purple to blue (indi- immersed in the solution. Heat A gradually until the temper- cator: 0.1 g of NN indicator). Perform a blank determina- ature of the solution in A reaches 1309C, then open the rub- 1056 Magnesium Silicate / Official Monographs JP XVI ber tube F, close the rubber tube G, boil water in the steam filtrate with ammonia TS: the solution responds to the generator B vigorously, and introduce the generated steam Qualitative Tests <1.09> for magnesium salt. into F. Simultaneously, heat A, and maintain the tempera- (2) Prepare a bead by fusing ammonium sodium hydro- ture of the solution in A between 1359Cand1459C. Adjust genphosphate tetrahydrate on a platinum loop. Place the the distilling rate to about 10 mL per minute. Collect about bead in contact with Magnesium Silicate, and fuse again: an 170 mL of the distillate, then stop the distillation, wash C infusible matter appears in the bead, which changes to an with a small quantity of water, combine the washings with opaque bead with a web-like structure upon cooling. the distillate, add water to make exactly 200 mL, and use this Purity (1) Soluble salts—Add 150 mL of water to 10.0 g solution as the test solution. Perform the test with the test of Magnesium Silicate, heat on a water bath for 60 minutes solution as directed in the procedure of determination for with occasional shaking, then cool, dilute with water to 150 fluoride under Oxygen Flask Combustion Method <1.06>. mL, and centrifuge. Dilute 75 mL of the resultant transpar- No corrective solution is used in this procedure. The content ent liquid with water to 100 mL, and use this solution as the of fluoride (F) is not more than 0.08z. sample solution. Evaporate 25 mL of the sample solution on Amount (mg) of fluoride (F: 19.00) in the test solution a water bath to dryness, and ignite the residue at 7009Cfor ＝ amount (mg) of fluoride in 5 mL of 2 hours: the mass of the ignited residue is not more than the standard solution 0.02 g.

× AT/AS × 200/V (2) Alkalinity—To 20 mL of the sample solution obtained in (1) add 2 drops of phenolphthalein TS and 1.0 mL Loss on ignition <2.43> Not more than 10z (0.25 g, 9009C, of 0.1 mol/L hydrochloric acid VS: no color develops. constant mass). (3) Chloride <1.03>—Take 10 mL of the sample solution Assay Ignite Magnesium Oxide to constant mass at 9009C, obtained in (1), add 6 mL of dilute nitric acid, dilute with weigh accurately about 0.2 g of the residue, dissolve in 10 water to 50 mL, and perform the test using this solution as mL of water and 4.0 mL of dilute hydrochloric acid, and the test solution. Prepare the control solution with 0.75 mL add water to make exactly 100 mL. Pipet 25 mL of this solu- of 0.01 mol/L hydrochloric acid VS (not more than tion, add 50 mL of water and 5 mL of ammonia-ammonium 0.053z). chloride buffer solution, pH 10.7, and titrate <2.50> with (4) Sulfate <1.14>—To the residue obtained in (1) add 0.05 mol/L disodium dihydrogen ethylenediamine tetra- about 3 mL of dilute hydrochloric acid, and heat on a water acetate VS (indicator: 40 mg of eriochrome black T-sodium bath for 10 minutes. Add 30 mL of water, filter, wash the chloride indicator). Perform a blank determination, and residue on the filter with water, combine the washings with make any necessary correction. the filtrate, and dilute to 50 mL with water. To 4 mL of the From the volume of 0.05 mol/L disodium dihydrogen solution add 1 mL of dilute hydrochloric acid and water to ethylenediamine tetraacetate VS consumed, deduct the make 50 mL. Perform the test using this solution as the test volume of 0.05 mol/L disodium dihydrogen ethylenediamine solution. Prepare the control solution with 1.0 mL of 0.005 tetraacetate VS corresponding to the content of calcium mol/L sulfuric acid VS (not more than 0.480z). oxide (CaO) obtained in the Purity (5). (5) Heavy metals <1.07>—To 1.0 g of Magnesium Silicate add 20 mL of water and 3 mL of hydrochloric acid, and boil Each mL of 0.05 mol/L disodium dihydrogen for 2 minutes. Filter, and wash the residue on the filter with ethylenediamine tetraacetate VS two 5-mL portions of water. Evaporate the combined filtrate ＝ 2.015 mg of MgO andwashingsonawaterbathtodryness,add2mLofdilute Each mg of calcium oxide (CaO) acetic acid to the residue, warm until solution is complete, ＝ 0.36 mL of 0.05 mol/L disodium dihydrogen filter, if necessary, add water to make 50 mL, and perform ethylenediamine tetraacetate VS the test using this solution as the test solution. Prepare the control solution with 3.0 mL of Standard Lead Solution, 2 Containers and storage Containers—Tight containers. mL of dilute acetic acid and water to make 50 mL (not more than 30 ppm). (6) Arsenic <1.11>—To 0.4 g of Magnesium Silicate add Magnesium Silicate 5 mL of dilute hydrochloric acid, heat gently to boiling while shaking well, cool rapidly, and centrifuge. Mix the residue ケイ酸マグネシウム with 5 mL of dilute hydrochloric acid with shaking, centrifuge, then add 10 mL of water to the residue, and repeat the Magnesium Silicate contains not less than 45.0z of extraction in the same manner. Concentrate the combined silicon dioxide (SiO2: 60.08) and not less than 20.0z extracts on a water bath to 5 mL. Use this solution as the test of magnesium oxide (MgO: 40.30), and the ratio of solution, and perform the test (not more than 5 ppm). percentage (z) of magnesium oxide to silicon dioxide Loss on ignition <2.43> Not more than 34z (0.5 g, 8509C, is not less than 2.2 and not more than 2.5. 3 hours). Description Magnesium Silicate occurs as a white, fine Acid-consuming capacity <6.04> Place about 0.2 g of Mag- powder. It is odorless and tasteless. nesium Silicate, accurately weighed, in a glass-stoppered It is practically insoluble in water, in ethanol (95) and in flask, add exactly 30 mL of 0.1 mol/L hydrochloric acid VS diethyl ether. and 20 mL of water, shake at 37 ± 29C for 1 hour, and Identification (1) Mix 0.5 g of Magnesium Silicate with cool. Pipet 25 mL of the supernatant liquid, and titrate 10 mL of dilute hydrochloric acid, filter, and neutralize the <2.50> the excess hydrochloric acid, while stirring well, with JP XVI Official Monographs / Magnesium Stearate 1057

0.1 mol/L sodium hydroxide VS until the pH becomes 3.5. Description Magnesium Stearate occurs as a white, light, 1 g of Magnesium Silicate, calculated on the anhydrous bulky powder. basis by making allowance for the observed loss on ignition It is smooth to the touch and sticky to the skin. It has no determined as directed in the preceding Loss on ignition, odor or a faint, characteristic odor. consumes not less than 140 mL and not more than 160 mL of It is practically insoluble in water and in ethanol (95). 0.1 mol/L hydrochloric acid VS. Identification (1) Mix 5.0 g of Magnesium Stearate with Assay (1) Silicon dioxide—Weigh accurately about 0.7 g 50 mL of peroxide-free diethyl ether, 20 mL of dilute nitric of Magnesium Silicate, add 10 mL of 0.5 mol/L sulfuric acid acid, and 20 mL of water in a round-bottom flask, and heat TS,evaporateonawaterbathtodryness,add25mLof to dissolve completely under a reflux condenser. After cool- water to the residue, and heat on a water bath for 15 minutes ing, transfer the contents of the flask to a separator, shake, with occasional stirring. Filter the supernatant liquid allow the layers to separate, and transfer the aqueous layer through filter paper for assay, add 25 mL of hot water to the to a flask. Extract the diethyl ether layer with two 4-mL por- residue, stir, and decant the supernatant liquid on the filter tions of water, and combine these extracts to the main aque- paper to filter. Wash the residue in the same manner with ous extract. After washing the combined aqueous extract two 25-mL portions of hot water, transfer the residue onto with 15 mL of peroxide-free diethyl ether, transfer to a the filter paper, and wash with hot water until the last wash- 50-mL volumetric flask, add water to make exactly 50 mL, ing does not respond to the Qualitative Tests <1.09> (1) for mix, and use this solution as the sample solution: the sample sulfate. Place the residue and the filter paper in a platinum solution responds to the Qualitative Tests <1.09> for magne- crucible, incinerate with strong heating, and ignite between sium. 7759Cand8259C for 30 minutes, then cool, and weigh the (2) The retention times of the peaks corresponding to residue as a (g). Moisten the residue with water, and add 6 methyl stearate and methyl palmitate in the chromatogram mL of hydrofluoric acid and 3 drops of sulfuric acid. of the sample solution correspond to those of methyl Evaporate to dryness, ignite for 5 minutes, cool, and weigh stearate and methyl palmitate in the chromatogram of the the residue as b (g). system suitability solution, as obtained in the Purity (5).

Content (z) of silicon dioxide (SiO2) Purity (1) Acidity or alkalinity—Heat 1.0 g of Magnesi- ＝ (a － b)/M × 100 um Stearate in 20 mL of freshly boiled and cooled water on a water bath for 1 minute while shaking, and filter after cool- M: Mass (g) of the sample ing. To 10 mL of the filtrate add 0.05 mL of bromothymol (2) Magnesium oxide—Weigh accurately about 0.3 g of blue TS, and add exactly 0.05 mL of 0.1 mol/L hydrochloric Magnesium Silicate, transfer to a 50-mL conical flask, add acid VS or 0.1 mol/L sodium hydroxide VS: the color of the 10 mL of 0.5 mol/L sulfuric acid VS, and heat on a water solution changes. bath for 15 minutes. Cool, transfer to a 100-mL volumetric (2) Chloride <1.03>—Perform the test with 10.0 mL of flask, wash the conical flask with water, add the washings to the sample solution obtained in Identification (1). Prepare the volumetric flask, dilute with water to 100 mL, and filter. the control solution with 1.40 mL of 0.02 mol/L hydrochlo- Pipet 50 mL of the filtrate, shake with 50 mL of water and 5 ricacidVS(notmorethan0.10z). mL of diluted 2,2?,2!-nitrilotrisethanol (1 in 2), add 2.0 mL (3) Sulfate <1.14>—Perform the test with 10.0 mL of the of ammonia TS and 10 mL of ammonia-ammonium chloride sample solution obtained in Identification (1). Prepare the buffer solution, pH 10.7, and titrate <2.50> with 0.05 mol/L control solution with 10.2 mL of 0.01 mol/L sulfuric acid disodium dihydrogen ethylenediamine tetraacetate VS (indi- VS (not more than 1.0z). cator: 40 mg of eriochrome black T-sodium chloride indi- (4) Heavy metals <1.07>—Heat 1.0 g of Magnesium cator). Stearate weakly first, then incinerate at about 500 ± 259C. After cooling, add 2 mL of hydrochloric acid, evaporate on Each mL of 0.05 mol/L disodium dihydrogen a water bath to dryness, add 20 mL of water and 2 mL of ethylenediamine tetraacetate VS dilute acetic acid to the residue, and heat for 2 minutes. ＝ 2.015 mg of MgO After cooling, filter this solution through a filter paper, (3) Ratio of percentage (z) of magnesium oxide (MgO) wash the filter paper with 15 mL of water, and combine the to silicon dioxide (SiO2)—Calculate the quotient from the washing with the filtrate. To the filtrate add water to make percentages obtained in (1) and (2). 50 mL, and perform the test with this solution as the test solution. Prepare the control solution as follows: evaporate 2 Containers and storage Containers—Well-closed contain- mL of hydrochloric acid on a water bath to dryness, add 2 ers. mL of dilute acetic acid, 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 20 ppm). (5) Relative content of stearic acid and palmitic Magnesium Stearate acid—Transfer exactly 0.10 g of Magnesium Stearate to a small conical flask fitted with a reflux condenser. Add 5.0 ステアリン酸マグネシウム mL of boron trifluoride-methanol TS, mix, and reflux for about 10 minutes to dissolve the solids. Add 4.0 mL of Magnesium Stearate consists chiefly magnesium heptane through the condenser, and reflux for about 10 salts of stearic acid (C18H36O2: 284.48) and palmitic minutes. After cooling, add 20 mL of saturated sodium acid (C16H32O2: 256.42). chloride solution, shake, and allow the layers to separate. It contains, when dried, not less than 4.0z and not Transfer the heptane layer through about 0.1 g of anhydrous more than 5.0z of magnesium (Mg: 24.31). sodium sulfate, previously washed with heptane, to another 1058 Magnesium Sulfate Hydrate / Official Monographs JP XVI flask. Transfer 1.0 mL of this solution to a 10-mL volumet- Loss on drying <2.41> Not more than 6.0z (2 g, 1059C, ric flask, dilute with heptane to volume, mix, and use this so- constant mass). lution as the sample solution. Perform the test with 1 mLof Microbial limit <4.05> The acceptance criteria of TAMC the sample solution as directed under Gas chromatography and TYMC are 103 CFU/g and 5 × 102 CFU/g, respectively. <2.02> according to the following conditions, and determine Salmonella and Escherichia coli are not observed. the area, A, of the methyl stearate peak and the total of the areas, B, of all of fatty acid ester peaks. Calculate the per- Assay Transfer about 0.5 g of previously dried Magnesium centage of stearic acid in the fatty acid fraction of Magne- Stearate, accurately weighed, to a 250-mL flask, add 50 mL sium Stearate by the following formula. of a mixture of 1-butanol and ethanol (99.5) (1:1), 5 mL of ammonia solution (28), 3 mL of ammonium chloride buffer Content (z) of stearic acid ＝ A/B × 100 solution, pH 10, 30.0 mL of 0.1 mol/L disodium dihydrogen Similarly, calculate the percentage of palmitic acid in ethylenediamine tetraacetate VS, and 1 to 2 drops of Magnesium Stearate. The methyl stearate peak, and the total eriochrome black T TS, and mix. Heat at 459Cto509Cto of the methyl stearate and methyl palmitate peaks are not make the solution clear, and after cooling, titrate <2.50> the less than 40z and not less than 90z of the total area of all excess disodium dihydrogen ethylenediamine tetraacetate fatty acid ester peaks, respectively, in the chromatogram. with 0.1 mol/L zinc sulfate VS until the solution changes Operating conditions— from blue to purple in color. Perform a blank determina- Detector: A hydrogen flame-ionization detector main- tion, and make any necessary correction. tained at a constant temperature of about 2609C. Each mL of 0.1 mol/L disodium dihydrogen Sample injection port: A splitless injection system main- ethylenediamine tetraacetate VS tained at a constant temperature of about 2209C. ＝ 2.431 mg of Mg Column: A fused silica capillary column 0.32 mm in inside diameter and 30 m in length, the inside coated with a 0.5-mm Containers and storage Containers—Tight containers. layer of polyethylene glycol 15000-diepoxide for gas chromatography. Column temperature: Maintain at 709C for 2 minutes Magnesium Sulfate Hydrate after injection, then program to increase the temperature at therateof59C per minute to 2409C and to maintain this 硫酸マグネシウム水和物 temperature for 5 minutes. Carrier gas: Helium. MgSO .7H O: 246.47 Flow rate: Adjust the flow rate so that the retention time 4 2 of methyl stearate is about 32 minutes. Magnesium Sulfate Hydrate, when ignited, contains Split ratio: Splitless. not less than 99.0z of magnesium sulfate (MgSO : Time span of measurement: About 1.5 time as long as the 4 120.37). retention time of methyl stearate beginning after the solvent peak. Description Magnesium Sulfate Hydrate occurs as color- System suitability— less or white crystals. It has a cooling, saline, bitter taste. Test for required detection: Place exactly 50 mg each of It is very soluble in water, and practically insoluble in stearic acid for gas chromatography and palmitic acid for ethanol (95). gas chromatography, each previously dried in a desiccator It dissolves in dilute hydrochloric acid. (silica gel) for 4 hours, in a small conical flask fitted with a Identification A solution of Magnesium Sulfate Hydrate reflux condenser. Add 5.0 mL of boron trifluoride-methanol (1 in 40) responds to the Qualitative Tests <1.09> for magne- TS, mix, and proceed in the same manner as directed for the sium salt and for sulfate. preparation of the sample solution, and use the solution so obtained as the solution for system suitability test. To ex- pH <2.54> Dissolve 1.0 g of Magnesium Sulfate Hydrate in actly 1 mL of the solution add heptane to make exactly 10 20 mL of water: the pH of this solution is between 5.0 and mL. Confirm that the peak area of methyl stearate obtained 8.2. from 1 mL of this solution is equivalent to 5 to 15z of that Purity (1) Clarity and color of solution—Dissolve 1.0 g from 1 mL of the solution for system suitability test. of Magnesium Sulfate Hydrate in 20 mL of water: the solu- System performance: When the procedure is run with 1 mL tion is clear and colorless. of the solution for system suitability test under the above op- (2) Chloride <1.03>—Perform the test with 1.0 g of Mag- erating conditions, methyl palmitate and methyl stearate are nesium Sulfate Hydrate. Prepare the control solution with eluted in this order, with the relative retention time of methyl 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than palmitate to methyl stearate being about 0.86, and with the 0.014z). resolution between these peaks being not less than 5. (3) Heavy metals <1.07>—Proceed with 2.0 g of Magnesi- System repeatability: When the test is repeated 6 times um Sulfate Hydrate according to Method 1, and perform the with the solution for system suitability test under the above test. Prepare the control solution with 2.0 mL of Standard operating conditions, the relative standard deviation of the Lead Solution (not more than 10 ppm). peak areas of methyl palmitate and methyl stearate are not (4) Zinc—Dissolve 2.0 g of Magnesium Sulfate Hydrate more than 6.0z, respectively, and the relative standard devi- in 20 mL of water, and add 1 mL of acetic acid and 5 drops ation of the ratios of the peak area of methyl palmitate to of potassium hexacyanoferrate (II) TS: no turbidity is pro- methyl stearate is not more than 1.0z. duced. JP XVI Official Monographs / Magnesium Sulfate Mixture 1059

(5) Calcium—Dissolve 1.0 g of Magnesium Sulfate Hy- form the test. drate in 5.0 mL of dilute hydrochloric acid, add water to Bacterial endotoxins <4.01> Less than 0.09 EU/mg. make 100 mL, and use this solution as the sample solution. Separately, dissolve 1.0 g of Magnesium Sulfate Hydrate in Extractable volume <6.05> It meets the requirement. 2.0 mL of Standard Calcium Solution and 5.0 mL of dilute Foreign insoluble matter <6.06> Perform the test according hydrochloric acid, add water to make exactly 100 mL, and to Method 1: it meets the requirement. use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed Insoluble particulate matter <6.07> It meets the require- under Atomic Absorption Spectrophotometry <2.23> accord- ment. ing to the following conditions, and determine the absor- Sterility <4.06> Perform the test according to the Mem- bances, A and A , of both solutions: A is not bigger than T S T brane filtration method: it meets the requirement. AS － AT (not more than 0.02z). Gas: Combustible gas—Acetylene or hydrogen. Assay Measure exactly a volume of Magnesium Sulfate Supporting gas—Air. Injection, equivalent to about 0.3 g of magnesium sulfate

Lamp: Calcium hollow-cathod lamp. hydrate (MgSO4.7H2O), and add water to make 75 mL. Wavelength: 422.7 nm. Then add 5 mL of ammonia-ammonium chloride buffer so- (6) Arsenic <1.11>—Prepare the test solution with 1.0 g lution, pH 10.7, and proceed as directed in the Assay under of Magnesium Sulfate Hydrate according to Method 1, and Magnesium Sulfate Hydrate. perform the test (not more than 2 ppm). Each mL of 0.05 mol/L disodium dihydrogen Loss on ignition <2.43> 45.0 – 52.0z (1 g, after drying at ethylenediamine tetraacetate VS

1059C for 2 hours, ignite at 4509C for 3 hours). ＝ 12.32 mg of MgSO4.7H2O Assay Weigh accurately about 0.6 g of Magnesium Sulfate Containers and storage Containers—Hermetic containers. Hydrate, previously ignited at 4509C for 3 hours after drying Plastic containers for aqueous injections may be used. at 1059C for 2 hours, and dissolve in 2 mL of dilute hydrochloric acid and water to make exactly 100 mL. Pipet 25 mL of this solution, add 50 mL of water and 5 mL of ammonia- Magnesium Sulfate Mixture ammonium chloride buffer solution, pH 10.7, and titrate <2.50> with 0.05 mol/L disodium dihydrogen ethylene- 硫酸マグネシウム水 diamine tetraacetate VS (indicator: 40 mg of eriochrome black T-sodium chloride indicator). Perform a blank deter- Magnesium Sulfate Mixture contains not less than mination, and make any necessary correction. 13.5 w/vz and not more than 16.5 w/vz of magne- Each mL of 0.05 mol/L disodium dihydrogen sium sulfate hydrate (MgSO4.7H2O: 246.47). ethylenediamine tetraacetate VS Method of preparation ＝ 6.018 mg of MgSO4 Magnesium Sulfate Hydrate 150 g Containers and storage Containers—Well-closed contain- Bitter Tincture 20 mL ers. Dilute Hydrochloric Acid 5 mL Purified Water or Purified Magnesium Sulfate Injection Water in Containers a sufficient quantity To make 1000 mL 硫酸マグネシウム注射液 Prepare before use, with the above ingredients. Description Magnesium Sulfate Mixture is a light yellowish Magnesium Sulfate Injection is an aqueous solution clear liquid. It has a bitter and acid taste. for injection. It contains not less than 95.0z and not more than Identification (1) Magnesium Sulfate Mixture responds 105.0z of the labeled amount of magnesium sulfate to the Qualitative Tests <1.09> for magnesium salt. hydrate (MgSO4.7H2O: 246.47). (2) Magnesium Sulfate Mixture responds to the Qualita- tive Tests <1.09> (2) for chloride. Method of preparation Prepare as directed under Injec- tions, with Magnesium Sulfate Hydrate. Assay Pipet 10 mL of Magnesium Sulfate Mixture, and add water to make exactly 100 mL. Pipet 10 mL of this solu- Description Magnesium Sulfate Injection is a clear, color- tion, add 50 mL of water and 5 mL of pH 10.7 ammonia- less liquid. ammonium chloride buffer solution, and titrate <2.50> with Identification Measure a volume of Magnesium Sulfate In- 0.05 mol/L disodium dihydrogen ethylenediamine tetraa- jection, equivalent to 0.5 g of Magnesium Sulfate Hydrate cetate VS (indicator: 40 mg of eriochrome black T-sodium according to the labeled amount, and add water to make 20 chloride indicator). mL: the solution responds to the Qualitative Tests <1.09> for Each mL of 0.05 mol/L disodium dihydrogen magnesium salt and for sulfate. ethylenediamine tetraacetate VS pH <2.54> 5.5 – 7.0 When the labeled concentration ＝ 12.32 mg of MgSO4.7H2O exceeds 5z, prepare a solution of 5z with water, and per- Containers and storage Containers—Tight containers. 1060 Maltose Hydrate / Official Monographs JP XVI

Maltose Hydrate in 10 mL of water, and add 1 drop of Maltose Hydrate iodine TS: a yellow color appears, and the color changes to a blue by adding 1 drop of starch TS. マルトース水和物 (7) Nitrogen—Weigh accurately about 2 g of Maltose Hydrate, and perform the test as directed under Nitrogen Determination <1.08> using 10 mL of sulfuric acid for the decomposition and 45 mL of a solution of sodium hydroxide (2 in 5) for the addition: the amount of nitrogen (N: 14.01) is not more than 0.01z. (8) Related substances—Dissolve 0.5 g of Maltose Hy- drate in 10 mL of water, and use this solution as the sample C H O .H O: 360.31 12 22 11 2 solution. Pipet 1 mL of the sample solution, add water to a-D-Glucopyranosyl-(1→4)-b-D-glucopyranose make exactly 100 mL, and use this solution as the standard monohydrate solution. Perform the test with exactly 20 mLeachofthe [6363-53-7] sample solution and standard solution as directed under Liq- uid Chromatography <2.01> according to the following oper- Maltose Hydrate, when dried, contains not less than ating conditions. Determine the peak areas from both solu- 98.0z of C H O .H O. 12 22 11 2 tions by the automatic integration method: the total area of Description Maltose Hydrate occurs as white crystals or the peaks which appear before the peak of maltose from the crystalline powder. sample solution is not larger than 1.5 times the peak area of It has a sweet taste. maltose from the standard solution, and the total area of the It is freely soluble in water, very slightly soluble in ethanol peaks which appear after the peak of maltose from the (95), and practically insoluble in diethyl ether. sample solution is not larger than 1/2 times the peak area of maltose from the standard solution. Identification (1) Dissolve 0.5 g of Maltose Hydrate in 5 Operating conditions— mL of water, add 5 mL of ammonia TS, and heat for 5 Detector, column, column temperature, mobile phase, minutes on a water bath: an orange color develops. flow rate, and selection of column: Proceed as directed in (2) Add 2 to 3 drops of a solution of Maltose Hydrate the operating conditions in the Assay. (1 in 50) to 5 mL of boiling Fehling TS: a red precipitate is Detection sensitivity: Adjust the sensitivity so that the formed. peak height of maltose obtained from 20 mL of the standard 20 Optical rotation <2.49> [a]D : ＋126 – ＋1319Weigh accu- solution is about 30 mm. rately about 10 g of Maltose Hydrate, previously dried, dis- Time span of measurement: About 2 times as long as the solve in 0.2 mL of ammonia TS and water to make exactly retention time of maltose. 100 mL, and determine the optical rotation of this solution Loss on drying <2.41> Not more than 0.5z (1 g, 809C, in a 100-mm cell. 4 hours). pH <2.54> The pH of a solution of Maltose Hydrate (1 in Residue on ignition <2.44> Not more than 0.1z (1 g). 10) is between 4.5 and 6.5. Assay Weigh accurately about 0.1 g each of Maltose Hy- Purity (1) Clarity and color of solution—Put 10 g of Mal- drate and Maltose RS, previously dried, dissolve in exactly tose Hydrate in 30 mL of water in a Nessler tube, warm at 10 mL each of the internal standard solution, and use these 609C in a water bath to dissolve, and after cooling, add solutions as the sample solution and the standard solution, water to make 50 mL: the solution is clear, and has no more respectively. Perform the test with 20 mLeachofthesample color than the following control solution. solution and standard solution as directed under Liquid Control solution: Add water to a mixture of 1.0 mL of Chromatography <2.01> according to the following operat- Cobalt (II) Chloride CS, 3.0 mL of Iron (III) Chloride CS ing conditions, and calculate the ratios, Q and Q ,ofthe and 2.0 mL of Copper (II) Sulfate CS to make 10.0 mL. To T S peak area of maltose to that of the internal standard. 1.0 mL of this solution add water to make 50 mL.

(2) Chloride <1.03>—Perform the test with 2.0 g of Mal- Amount (mg) of C12H22O11.H2O tose Hydrate. Prepare the control solution with 1.0 mL of ＝ MS × QT/QS 0.01 mol/L hydrochloric acid VS (not more than 0.018z). M : Amount (mg) of Maltose RS (3) Sulfate <1.14>—Perform the test with 2.0 g of Mal- S tose Hydrate. Prepare the control solution with 1.0 mL of Internal standard solution—A solution of ethylene glycol 0.005 mol/L sulfuric acid VS (not more than 0.024z). (1 in 50). (4) Heavy metals <1.07>—Proceed with 5.0 g of Maltose Operating conditions— Hydrate according to Method 1, and perform the test. Pre- Detector: A differential refractometer. pare the control solution with 2.0 mL of Standard Lead So- Column: A stainless steel column about 8 mm in inside lution (not more than 4 ppm). diameter and about 55 cm in length, packed with gel-type (5) Arsenic <1.11>—Dissolve 1.5 g of Maltose Hydrate in strong acid cation-exchange resin for liquid chromatography 5 mL of water, add 5 mL of dilute sulfuric acid and 1 mL of (degree of cross-linking: 8 z)(10mm in particle diameter). bromine TS, heat on a water bath for 5 minutes, then heat to Column temperature: A constant temperature of about concentrate to 5 mL, and use this solution as the test solu- 509C. tion after cooling. Perform the test (not more than 1.3 ppm). Mobile phase: Water. (6) Dextrin, soluble starch and sulfite—Dissolve 1.0 g of Flow rate: Adjust the flow rate so that the retention time JP XVI Official Monographs / Manidipine Hydrochloride 1061 of maltose is about 18 minutes. solution of Manidipine Hydrochloride in methanol (1 in Selection of column: Dissolve 0.25 g of maltose, 0.25 g of 100,000) as directed under Ultraviolet-visible Spectropho- glucose and 0.4 g of ethylene glycol in water to make 100 tometry <2.24>, and compare the spectrum with the Refer- mL. Proceed with 20 mL of this solution under the above ence Spectrum or the spectrum of a solution of Manidipine operating conditions, and calculate the resolution. Use a Hydrochloride RS prepared in the same manner as the sam- column giving elution of maltose, glucose and ethylene ple solution: both spectra exhibit similar intensities of ab- glycol in this order with the resolution of between the peaks sorption at the same wavelengths. of maltose and glucose being not less than 4. (2) Determine the infrared absorption spectrum of Manidipine Hydrochloride as directed in the potassium chlo- Containers and storage Containers—Tight containers. ride disc method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Manidipine Hydrochloride RS: both spectra Freeze-dried Mamushi Antivenom, exhibit similar intensities of absorption at the same wave Equine numbers. (3) Add 10 mL of water to 0.1 g of Manidipine Hydro- 乾燥まむしウマ抗毒素 chloride, shake vigorously, and filter. Add 1 drop of ammonia TS to 3 mL of the filtrate, allow to stand 5 minutes, and filter. The filtrate responds to the Qualitative Tests <1.09> (2) Freeze-dried Mamushi Antivenom, Equine, is a for chlorides. preparation for injection which is dissolved before use. It contains Agkistrodon Halys antivenom in im- Purity (1) Heavy metals <1.07>—Proceedwith1.0gof munoglobulin of horse origin. Manidipine Hydrochloride according to Method 2, and per- It conforms to the requirements of Freeze-dried form the test. Prepare the control solution with 1.0 mL of Mamushi Antivenom, Equine, in the Minimum Re- Standard Lead Solution (not more than 10 ppm). quirements for Biological Products. (2) Arsenic <1.11>—Prepare the test solution with 2.0 g of Manidipine Hydrochloride according to Method 4, and Description Freeze-dried Mamushi Antivenom, Equine, perform the test (not more than 1 ppm). becomes a colorless or light yellow-brown, clear liquid, or a (3) Related substances—Dissolve 20 mg of Manidipine slightly white-turbid liquid on addition of solvent. Hydrochloride in 200 mL of a mixture of water and acetonitrile (1:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mixture of water Manidipine Hydrochloride and acetonitrile (1:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex- マニジピン塩酸塩 actly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area from both solutions by the automatic integration method: the area of the peaks other than manidipine obtained from the sample solution is not larger than 1/5 times the manidipine peak area from the standard solution. Fur- thermore, the total of the areas of all peaks other than manidipine is not larger than 7/10 times the peak area of manidipine from the standard solution. C H N O .2HCl: 683.62 35 38 4 6 Operating conditions— 3-{2-[4-(Diphenylmethyl)piperazin-1-yl]ethyl} Detector, column, column temperature, mobile phase, and 5-methyl (4RS)-2,6-dimethyl-4-(3-nitrophenyl)- flow rate: Proceed as directed in the operating conditions in 1,4-dihydropyridine-3,5-dicarboxylate dihydrochloride the Assay. [126229-12-7] Time span of measurement: About 3.5 times as long as the retention time of manidipine, beginning after the solvent Manidipine Hydrochloride, when dried, contains peak. not less than 98.5z and not more than 101.0z of System suitability— C H N O .2HCl. 35 38 4 6 Test for required detectability: Pipet 10 mL of the stand- Description Manidipine Hydrochloride occurs as white to ard solution, add a mixture of water and acetonitrile (1:1) to pale yellow crystals or crystalline powder. make exactly 100 mL. Confirm that the peak area of mani- It is freely soluble in dimethylsulfoxide, sparingly soluble dipine obtained from 20 mL of this solution is equivalent to 8 in methanol, slightly soluble in ethanol (99.5), and practi- to 12z of that from 20 mL of the standard solution. cally insoluble in water. System performance: Dissolve 50 mg of Manidipine Hy- A solution of Manidipine Hydrochloride in dimethylsul- drochloride in a mixture of water and acetonitrile (1:1) to foxide (1 in 100) shows no optical rotation. make 50 mL. To 10 mL of this solution add 5 mL of a solu- Manidipine Hydrochloride turns slightly brown-yellowish tion of butyl benzoate in acetonitrile (7 in 5000) and the mix- white on exposure to light. ture of water and acetonitrile (1:1) to make 100 mL. When Melting point: about 2079C (with decomposition). the procedure is run with 20 mL of this solution under the above operating conditions, manidipine and butyl benzoate Identification (1) Determine the absorption spectrum of a 1062 Manidipine Hydrochloride Tablets / Official Monographs JP XVI are eluted in this order with the resolution between these peaks being not less than 5. Manidipine Hydrochloride Tablets System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat- マニジピン塩酸塩錠 ing conditions, the relative standard deviation of the peak area of manidipine is not more than 2.0z. Manidipine Hydrochloride Tablets contain not Loss on drying <2.41> Not more than 1.5z (1 g, 1059C, less than 92.0z and not more than 108.0z of 4 hours). the labeled amount of manidipine hydrochloride (C H N O .2HCl: 683.62). Residue on ignition <2.44> Not more than 0.2z (1 g). 35 38 4 6 Method of preparation Prepare as directed under Tablets, Assay Weigh accurately about 0.1 g of Manidipine Hydro- with Manidipine Hydrochloride. chloride, previously dried, anddissolveinamixtureofwater and acetonitrile (1:1) to make exactly 50 mL. Pipet 5 mL of Identification To a quantity of powdered Manidipine Hy- this solution, add exactly 5 mL of the internal standard solu- drochloride Tablets, equivalent to 10 mg of Manidipine Hy- tion, add the mixture of water and acetonitrile (1:1) to make drochloride according to the labeled amount, add 5 mL of 100 mL, and use this solution as the sample solution. Sepa- methanol, shake vigorously, centrifuge, and use the superna- rately, weigh accurately about 25 mg of Manidipine Hydro- tant liquid as the sample solution. Separately, dissolve 10 mg chloride RS, previously dried, and dissolve in the mixture of of Manidipine Hydrochloride RS in 5 mL of methanol, and water and acetonitrile (1:1) to make exactly 50 mL. Pipet 20 use this solution as the standard solution. Perform the test mL of this solution, add exactly 5 mL of the internal stand- with these solutions as directed under Thin-layer Chroma- ard solution, add the mixture of water and acetonitrile (1:1) tography <2.03>. Spot 5 mL each of the sample solution and to make 100 mL, and use this solution as the standard solu- standard solution on a plate of silica gel with fluorescent in- tion. Perform the test with 20 mL each of the sample solution dicator for thin-layer chromatography. Develop the plate and standard solution as directed under Liquid Chromatog- with a mixture of ethyl acetate and diethylamine (200:1) to a raphy <2.01> according to the following conditions, and cal- distance of about 10 cm, and air-dry the plate. Examine culate the ratios, QT and QS, of the peak area of manidipine under ultraviolet light (main wavelength: 254 nm): the prin- to that of the internal standard. cipal spot obtained from the sample solution and the spot obtained from the standard solution show the same Rf Amount (mg) of C H N O .2HCl 35 38 4 6 value. ＝ MS × QT/QS × 4 Uniformity of dosage units <6.02> Perform the test accord- M : Amount (mg) of Manidipine Hydrochloride RS S ing to the following method: it meets the requirement of the Internal standard solution—A solution of butyl benzoate in Content uniformity test. acetonitrile (7 in 5000). Conduct this procedure using light-resistant vessels. To 1 Operating conditions— tablet of Manidipine Hydrochloride Tablets, add exactly 1 Detector: An ultraviolet absorption photometer (wave- mL of the internal standard solution per 1 mg of manidipine length: 228 nm). hydrochloride (C35H38N4O6.2HCl), disintegrate by adding a Column: A stainless steel column 4.0 mm in inside diame- mixture of water and acetonitrile (1:1) to make V mL so that ter and 15 cm in length, packed with octadecylsilanized silica each mL contains about 0.1 mg of manidipine hydrochloride gel for liquid chromatography (5 mm in particle diameter). (C35H38N4O6.2HCl), shake vigorously for 10 minutes, and Column temperature: A constant temperature of about filter through a membrane filter with a pore size not 259C. exceeding 0.45 mm. Discard the first 1 mL of the filtrate, and Mobile phase: Dissolve 13.6 g of potassium dihydrogen use the subsequent filtrate as the sample solution. Then, phosphate in water to make 1000 mL, and adjust to pH 4.6 proceed as directed in the Assay. with diluted potassium hydroxide TS (1 in 10). To 490 mL of Amount (mg) of manidipine hydrochloride this solution add 510 mL of acetonitrile. (C H N O .2HCl) Flow rate: Adjust the flow rate so that the retention time 35 38 4 6 ＝ M × Q /Q × V/250 of manidipine is about 10 minutes. S T S

System suitability— MS: Amount (mg) of Manidipine Hydrochloride RS System performance: When the procedure is run with 20 Internal standard solution—A solution of butyl benzoate in mL of the standard solution under the above operating con- acetonitrile (7 in 10,000). ditions, manidipine and the internal standard are eluted in this order with the resolution between these peaks being not Dissolution <6.10> When the test is performed at 50 revolu- less than 5. tions per minute according to the Paddle method, using 900 System repeatability: When the test is repeated 6 times mL of 0.05 mol/L acetic acid-sodium acetate buffer solu- with 20 mL of the standard solution under the above operat- tion, pH 4.0, as the dissolution medium, the dissolution rate ing conditions, the relative standard deviation of the ratio of in 45 minutes of Manidipine Hydrochloride Tablets is not the peak area of manidipine to that of the internal standard less than 75z. is not more than 1.0z. Conduct this procedure using light-resistant vessels. Start the test with 1 tablet of Manidipine Hydrochloride Tablets, Containers and storage Containers—Tight containers. withdraw not less than 20 mL of the medium at the specified Storage—Light-resistant. minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the JP XVI Official Monographs / D-Mannitol 1063 first 10 mL of the filtrate, pipet V mL of the subsequent Then, proceed as directed in the Assay under Manidipine filtrate, and add the dissolution medium to make exactly Hydrochloride. V?mL so that each mL contains about 5.6 mg of manidipine Amount (mg) of manidipine hydrochloride hydrochloride (C H N O .2HCl) according to the labeled 35 38 4 6 (C H N O .2HCl) amount. Pipet 2 mL of this solution, add exactly 2 mL of 35 38 4 6 ＝ M × Q /Q × 2/5 methanol, and use this solution as the sample solution. Sepa- S T S rately, weigh accurately about 25 mg of Manidipine Hydro- MS: Amount (mg) of Manidipine Hydrochloride RS chloride RS, previously dried, dissolve in a mixture of water Internal standard solution—A solution of butyl benzoate in and acetonitrile (1:1) to make exactly 50 mL. Pipet 1 mL of acetonitrile (7 in 10,000). this solution, and add the dissolution medium to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 2 mL Containers and storage Containers—Tight containers. of methanol, and use this solution as the standard solution. Storage—Light-resistant. Perform the test with exactly 20 mLeachofthesamplesolu- tion and standard solution as directed under Liquid Chroma- tography <2.01> according to the following conditions, and D-Mannitol determine the manidipine peak areas, AT and AS, of both solutions. D-マンニトール Dissolution rate (z) with respect to the labeled amount

of manidipine hydrochloride (C35H38N4O6.2HCl) ＝ MS × AT/AS × V?/V × 1/C × 18

MS: Amount (mg) of Manidipine Hydrochloride RS C6H14O6: 182.17 C: Labeled amount (mg) of manidipine hydrochloride D-Mannitol

(C35H38N4O6.2HCl) in 1 tablet [69-65-8] Operating conditions— -Mannitol, when dried, contains not less than Detector: An ultraviolet absorption photometer (wave- D 98.0z of C H O . length: 228 nm). 6 14 6 Column: A stainless steel column 4.0 mm in inside diame- Description D-Mannitol occurs as white crystals or pow- ter and 15 cm in length, packed with octadecylsilanized silica der. It is odorless, and has a sweet taste with a cold sensa- gel for liquid chromatography (5 mm in particle diameter). tion. Column temperature: A constant temperature of about It is freely soluble in water, and practically insoluble in 259C. ethanol (95) and in diethyl ether. Mobile phase: A mixture of acetonitrile and a solution of It dissolves in sodium hydroxide TS. potassium dihydrogen phosphate (681 in 100,000) (3:2). Identification (1) To 5 drops of a saturated solution of D- Flow rate: Adjust the flow rate so that the retention time Mannitol add 1 mL of iron (III) chloride TS and 5 drops of a of manidipine is about 6 minutes. solution of sodium hydroxide (1 in 5): a yellow precipitate is System suitability— produced. Shake this solution vigorously: a clear solution is System performance: When the procedure is run with 20 produced. On addition of a solution of sodium hydroxide (1 mL of the standard solution under the above operating con- in 5), no precipitate is produced. ditions, the number of theoretical plates and the symmetry (2) Determine the infrared absorption spectrum of D- factor of the peak of manidipine are not less than 1500 and Mannitol as directed in the potassium bromide disk method not more than 1.5, respectively. under Infrared Spectrophotometry <2.25>, and compare the System repeatability: When the test is repeated 6 times spectrum with the Reference Spectrum: both spectra exhibit with 20 mL of the standard solution under the above operat- similar intensities of absorption at the same wave numbers. ing conditions, the relative standard deviation of the peak If any difference appears between the spectra, dissolve 1 g of area of manidipine is not more than 2.0z. D-Mannitol in 3 mL of warm water, then allow to stand at Assay Conduct this procedure using light-resistant vessels. 59C for 24 hours or until crystals appear, and filter. Wash Weigh accurately not less than 20 Manidipine Hydrochloride the crystals so obtained with a few amount of cold water, Tablets, and powder. Weigh accurately a portion of the dry at 1059C for 4 hours, and perform the test with the crys- powder, equivalent to about 10 mg of manidipine hydrochlo- tals. ride (C H N O .2HCl), add exactly 10 mL of the internal 35 38 4 6 Optical rotation <2.49> [a]20: ＋137 – ＋1459Weigh accu- standard solution, add a mixture of water and acetonitrile D rately about 1.0 g of D-Mannitol, previously dried, dissolve (1:1) to make 100 mL, shake vigorously for 10 minutes, and in 80 mL of a solution of hexaammonium heptamolybdate filter through a membrane filter with a pore size not tetrahydrate (1 in 20), and add diluted sulfuric acid (1 in 35) exceeding 0.45 mm. Discard the first 1 mL of the filtrate, and to make exactly 100 mL. Measure the optical rotation of this use the subsequent filtrate as the sample solution. Sepa- solution in a 100-mm cell. rately, weigh accurately about 25 mg of Manidipine Hydro- chloride RS, previously dried, and dissolve in the mixture of Melting point <2.60> 166 – 1699C water and acetonitrile (1:1) to make 50 mL. Pipet 20 mL of Purity (1) Clarity and color of solution—Dissolve 2.0 g this solution, add exactly 10 mL of the internal standard so- of D-Mannitol in 10 mL of water by warming: the solution is lution, add the mixture of water and acetonitrile (1:1) to clear and colorless. make 100 mL, and use this solution as the standard solution. 1064 D-Mannitol Injection / Official Monographs JP XVI

(2) Acidity—Dissolve 5.0 g of D-Mannitol in 50 mL of freshly boiled and cooled water, and add 1 drop of phenol- D-Mannitol Injection phthalein TS and 0.50 mL of 0.01 mol/L sodium hydroxide VS: a red color develops. D-Mannite Injection (3) Chloride <1.03>—Perform the test with 2.0 g of D- Mannitol. Prepare the control solution with 0.40 mL of 0.01 D-マンニトール注射液 mol/L hydrochloric acid VS (not more than 0.007z). (4) Sulfate <1.14>—Perform the test with 2.0 g of - D -Mannitol Injection is an aqueous solution for in- Mannitol. Prepare the control solution with 0.40 mL of D jection. 0.005 mol/L sulfuric acid VS (not more than 0.010z). It contains not less than 95.0z and not more than (5) Heavy metals <1.07>—Proceed with 5.0 g of D-Man- 105.0z of the labeled amount of D-mannitol nitol according to Method 1, and perform the test. Prepare (C H O : 182.17). the control solution with 2.5 mL of Standard Lead Solution 6 14 6 (not more than 5 ppm). Method of preparation Prepare as directed under Injec- (6) Nickel—Dissolve 0.5 g of D-Mannitol in 5 mL of tions, with D-Mannitol. No preservative is added. water, add 3 drops of dimethylglyoxime TS and 3 drops of Description D-Mannitol Injection is a clear, colorless liq- ammonia TS, and allow to stand for 5 minutes: no red color uid. It has a sweet taste. develops. It may precipitate crystals. (7) Arsenic <1.11>—Prepare the test solution with 1.5 g of D-Mannitol according to Method 1, and perform the test Identification Concentrate D-Mannitol Injection on a (not more than 1.3 ppm). waterbathtomakeasaturatedsolution.Proceedwith5 (8) Sugars—To 5.0 g of D-Mannitol add 15 mL of water drops of this solution as directed in the Identification (1) and 4.0 mL of dilute hydrochloric acid, and heat under a under D-Mannitol. reflux condenser in a water bath for 3 hours. After cooling, pH <2.54> 4.5–7.0 neutralize with sodium hydroxide TS (indicator: 2 drops of methyl orange TS), and add water to make 50 mL. Pipet 10 Bacterial endotoxins <4.01> Less than 0.50 EU/mL. mL of this solution into a flask, boil gently with 10 mL of Extractable volume <6.05> It meets the requirement. water and 40 mL of Fehling's TS for 3 minutes, and allow to stand to precipitate copper (I) oxide. Filter the supernatant Foreign insoluble matter <6.06> Perform the test according liquid through a glass filter (G4), wash the precipitate with to Method 1: it meets the requirement. hot water until the last washing no longer shows an alkaline Insoluble particulate matter <6.07> It meets the require- reaction, and filter the washings through the glass filter ment. described above. Dissolve the precipitate in 20 mL of iron (III) sulfate TS in the flask, filter through the glass filter Sterility <4.06> Perform the test according to the Mem- described above, and wash the filter with water. Combine brane filtration method: it meets the requirement. the washings and the filtrate, heat to 809C, and titrate <2.50> Assay Measure exactly a volume of D-Mannitol Injection, with 0.02 mol/L potassium permanganate VS: the consumed equivalent to about 5 g of D-mannitol (C H O ), and add volume is not more than 1.0 mL. 6 14 6 water to make exactly 250 mL. To exactly 10 mL of this so- Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, lution add water to make exactly 100 mL. Measure exactly 4 hours). 10 mL of this solution into an iodine flask, and proceed as directed in the Assay under D-Mannitol. Residue on ignition <2.44> Not more than 0.1z (1 g). Each mL of 0.1 mol/L sodium thiosulfate VS Assay Weigh accurately about 0.2 g of D-Mannitol, previ- ＝ 1.822 mg of C H O ously dried, and dissolve in water to make exactly 100 mL. 6 14 6 Pipet 10 mL of the solution into an iodine flask, add exactly Containers and storage Containers—Hermetic containers. 50 mL of potassium periodate TS, and heat for 15 minutes in Plastic containers for aqueous injections may be used. a water bath. After cooling, add 2.5 g of potassium iodide, stopper tightly, and shake well. Allow to stand for 5 minutes in a dark place, and titrate <2.50> with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Perform a blank determination. Each mL of 0.1 mol/L sodium thiosulfate VS

＝ 1.822 mg of C6H14O6 Containers and storage Containers—Tight containers. JP XVI Official Monographs / Meclofenoxate Hydrochloride 1065

Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, Maprotiline Hydrochloride 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). マプロチリン塩酸塩 Assay Weigh accurately about 0.25 g of Maprotiline Hy- drochloride, previously dried, dissolve in 180 mL of acetic acid (100), add 8 mL of a solution of bismuth nitrate pentahydrate in acetic acid (100) (1 in 50), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per- form a blank determination, and make any necessary correc-

C20H23N.HCl: 313.86 tion. 3-(9,10-Dihydro-9,10-ethanoanthracen-9-yl)- Each mL of 0.1 mol/L perchloric acid VS N-methylpropylamine monohydrochloride ＝ 31.39 mg of C H N.HCl [10347-81-6] 20 23 Containers and storage Containers—Well-closed contain- Maprotiline Hydrochloride, when dried, contains ers. not less than 99.0z of C20H23N.HCl. Description Maprotiline Hydrochloride occurs as a white crystalline powder. Freeze-dried Live Attenuated It is soluble in methanol and in acetic acid (100), sparingly Measles Vaccine soluble in ethanol (99.5), and slightly soluble in water. Melting point: about 2449C (with decomposition). 乾燥弱毒生麻しんワクチン Identification (1) Determine the absorption spectrum of a solution of Maprotiline Hydrochloride in methanol (1 in Freeze-dried Live Attenuated Measles Vaccine is a 10,000) as directed under Ultraviolet-visible Spectropho- preparation for injection which is dissolved before use. tometry <2.24>, and compare the spectrum with the Refer- It contains live attenuated measles virus. ence Spectrum: both spectra exhibit similar intensities of ab- It conforms to the requirements of Freeze-dried Live sorption at the same wavelengths. Attenuated Measles Vaccine in the Minimum Require- (2) Determine the infrared absorption spectrum of ments for Biological Products. Maprotiline Hydrochloride, previously dried, as directed in Description Freeze-dried Live Attenuated Measles Vaccine the potassium chloride disk method under Infrared Spectro- becomes a colorless, yellowish or reddish clear liquid on ad- photometry <2.25>, and compare the spectrum with the Ref- dition of solvent. erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, recrystallize Maprotiline Hydro- chloride with ethanol (99.5), filter, dry the crystals so ob- Meclofenoxate Hydrochloride tained, and perform the test with the crystals. メクロフェノキサート塩酸塩 (3) To 5 mL of a solution of Maprotiline Hydrochloride (1 in 200) add 2 mL of ammonia TS, heat on a water bath for 5 minutes, cool, and filter. Acidify the filtrate with dilute nitric acid: the solution responds to the Qualitative Tests <1.09> for chloride. Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of C H ClNO .HCl: 294.17 Maprotiline Hydrochloride according to Method 2, and per- 12 16 3 2-(Dimethylamino)ethyl (4-chlorophenoxy)acetate form the test. Prepare the control solution with 2.0 mL of monohydrochloride Standard Lead Solution (not more than 10 ppm). [3685-84-5] (2) Related substances—Dissolve 0.10 g of Maprotiline Hydrochloride in 5 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add Meclofenoxate Hydrochloride contains not less than 98.0 of C H ClNO .HCl, calculated on the anhy- methanol to make exactly 200 mL, and use this solution as z 12 16 3 the standard solution. Perform the test with these solutions drous basis. as directed under Thin-layer Chromatography <2.03>. Spot Description Meclofenoxate Hydrochloride occurs as white 10 mL each of the sample solution and standard solution on a crystals or crystalline powder. It has a faint, characteristic plate of silica gel with fluorescent indicator for thin-layer odor and a bitter taste. chromatography. Develop with a mixture of 2-butanol, It is freely soluble in water and in ethanol (95), sparingly diluted ammonia solution (28) (1 in 3) and ethyl acetate soluble in acetic anhydride, and practically insoluble in (14:5:4) to a distance of about 10 cm, and air-dry the plate. diethyl ether. Examine under ultraviolet light (main wavelength: 254 nm): The pH of a solution of Meclofenoxate Hydrochloride the number of the spot other than the principal spot from (1 in 20) is between 3.5 and 4.5. the sample solution is not more than 2 and they are not more Identification (1) To 10 mg of Meclofenoxate Hydrochlo- intense than the spot from the standard solution. 1066 Mecobalamin / Official Monographs JP XVI ride add 2 mL of ethanol (95), dissolve by warming if necessary, cool, add 2 drops of a saturated solution of hydroxyl- Mecobalamin ammonium chloride in ethanol (95) and 2 drops of a saturated solution of potassium hydroxide in ethanol (95), and heat メコバラミン in a water bath for 2 minutes. After cooling, render the solution slightly acidic with dilute hydrochloric acid, and add 3 drops of iron (III) chloride TS: a red-purple to dark purple color develops. (2) Dissolve 50 mg of Meclofenoxate Hydrochloride in 5 mL of water, and add 2 drops of Reinecke salt TS: a light red precipitate is formed. (3) Determine the absorption spectrum of a solution of Meclofenoxate Hydrochloride (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>,andcompare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (4) A solution of Meclofenoxate Hydrochloride (1 in 100) responds to the Qualitative Tests <1.09> for chloride. Melting point <2.60> 139–1439C Purity (1) Clarity and color of solution—Dissolve 0.5 g of Meclofenoxate Hydrochloride in 10 mL of water: the solution is clear and colorless. C63H91CoN13O14P: 1344.38 (2) Sulfate <1.14>—Perform the test with 1.0 g of Coa-[a-(5,6-Dimethyl-1H-benzoimidazol-1-yl)]-Cob- Meclofenoxate Hydrochloride. Prepare the control solution methylcobamide with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than [13422-55-4] 0.048z). (3) Heavy metals <1.07>—Proceed with 1.0 g of Meclo- Mecobalamin contains not less than 98.0z of fenoxate Hydrochloride according to Method 1, and per- C63H91CoN13O14P, calculated on the anhydrous basis. form the test. Prepare the control solution with 2.0 mL of Description Mecobalamin occurs as dark red crystals or Standard Lead Solution (not more than 20 ppm). crystalline powder. (4) Arsenic <1.11>—Prepare the test solution with 1.0 g It is sparingly soluble in water, slightly soluble in ethanol of Meclofenoxate Hydrochloride according to method 3, (99.5), and practically insoluble in acetonitrile. and perform the test (not more than 2 ppm). It is affected by light. (5) Organic acids—To 2.0 g of Meclofenoxate Hydro- chloride add 50 mL of diethyl ether, shake for 10 minutes, Identification (1) Conduct this procedure without ex- filter through a glass filter (G3), wash the residue with two posure to light, using light-resistant vessels. Determine the 5-mL portions of diethyl ether, and combine the washings absorption spectrum of a solution of Mecobalamin in hydro- with the filtrate. To this solution add 50 mL of neutralized chloric acid-potassium chloride buffer solution, pH 2.0 (1 in ethanol and 5 drops of phenolphthalein TS, and neutralize 20,000) as directed under Ultraviolet-visible Spectropho- with 0.1 mol/L sodium hydroxide VS: the volume of 0.1 tometry <2.24>, and compare the spectrum with the Refer- mol/L sodium hydroxide VS consumed is not more than ence Spectrum 1 or the spectrum of a solution of Mecobala- 0.54 mL. min RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the Water <2.48> Not more than 0.50z (1 g, volumetric titra- same wavelengths. Separately, determine the absorption tion, dirct titration). spectrum of a solution of Mecobalamin in phosphate buffer Residue on ignition <2.44> Not more than 0.1z (1 g). solution, pH 7.0 (1 in 20,000) as directed under Ultraviolet- visible Spectrophotometry <2.24>, and compare the spectrum Assay Weigh accurately about 0.4 g of Meclofenoxate Hy- with the Reference Spectrum 2 or the spectrum of a solution drochloride, dissolve in 70 mL of acetic anhydride, and of Mecobalamin RS prepared in the same manner as the titrate <2.50> with 0.1 mol/L perchloric acid VS until the sample solution: both spectra exhibit similar intensities of color of the solution changes from blue-green through yel- absorption at the same wavelengths. low-green to pale greenish yellow [indicator: 3 drops of a so- (2) Mix 1 mg of Mecobalamin with 50 mg of potassium lution of malachite green oxalate in acetic acid (100) (1 in bisulfate, and fuse by igniting. Cool, break up the mass with 100)]. Perform a blank determination, and make any neces- a glass rod, add 3 mL of water, and dissolve by boiling. Add sary correction. 1 drop of phenolphthalein TS, then add dropwise sodium Each mL of 0.1 mol/L perchloric acid VS hydroxide TS until a light red color just develops. Add 0.5 g

＝ 29.42 mg of C12H16ClNO3.HCl of sodium acetate, 0.5 mL of dilute acetic acid and 0.5 mL of a solution of disodium 1-nitroso-2-naphthol-3,6-disul- Containers and storage Containers—Tight containers. fonate (1 in 500): a red to orange-red color is immediately produced. Then add 0.5 mL of hydrochloric acid, and boil for 1 minute: the red color does not disappear. JP XVI Official Monographs / Medazepam 1067

Purity (1) Clarity and color of solution—Dissolve 20 mg 0.02 mol/L phosphate buffer solution, pH 3.5, then add of Mecobalamin in 10 mL of water: the solution is clear and 3.76 g of sodium 1-hexane sulfonate to dissolve. red color. Flow rate: Adjust the flow rate so that the retention time (2) Related substances—Perform the test with 10 mLof of mecobalamin is about 12 minutes. the sample solution obtained in the Assay as directed under System suitability— Liquid Chromatography <2.01> according to the following System performance: Dissolve 5 mg each of cyanocobala- conditions. Determine the peak area of mecobalamin and min and hydroxocobalamin acetate in the mobile phase to others of the sample solution by the automatic integration make 100 mL. When the procedure is run with 10 mLofthis method: each area of the peaks other than mecobalamin is solution under the above operating conditions, cyanocobala- not more than 0.5z of the peak area of mecobalamin, and min and hydroxocobalamin are eluted in this order with the the total area of the peaks other than mecobalamin is not resolution between these peaks being not less than 3. And more than 2.0z. when the procedure is run with 10 mL of the standard solu- Operating conditions— tion under the above operating conditions, the number of Detector, column, column temperature, mobile phase, and theoretical plates of the peak of mecobalamin is not less than flow rate: Proceed as directed in the operating conditions in 6000. the Assay. System repeatability: When the test is repeated 6 times Time span of measurement: About 2.5 times as long as the with 10 mL of the standard solution under the above operat- retention time of mecobalamin. ing conditions, the relative standard deviation of the peak System suitability— areas of mecobalamin is not more than 1.0z. Test for required detection: To exactly 1 mL of the sample Containers and storage Containers—Tight containers. solution add the mobile phase to make exactly 100 mL, and Storage—Light-resistant. use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, add the mobile phase to make exactly 10 mL. Confirm that the peak area of mecobalamin obtained from 10 mL of this solution is Medazepam equivalent to 7 to 13z of that from 10 mL of the solution for メダゼパム system suitability test. System performance: Proceed as directed in the system suitability in the Assay. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the peak areas of mecobalamin is not more than 3.0z.

Water <2.48> Not more than 12z (0.1 g, volumetric titra- C16H15ClN2:270.76 tion, direct titration). 7-Chloro-1-methyl-5-phenyl-2,3-dihydro-1H-1,4- benzodiazepine Assay Conduct this procedure without exposure to light, [2898-12-6] using light-resistant vessels. Weigh accurately about 50 mg of Mecobalamin and Mecobalamin RS (separately, deter- Medazepam, when dried, contains not less than mine the water <2.48> in the same manner as Mecobalamin), 98.5z and not more than 101.0z of C H ClN . dissolve each in the mobile phase to make exactly 50 mL, 16 15 2 and use these solutions as the sample solution and the stand- Description Medazepam occurs as white to light yellow ard solution, respectively. Perform the test with exactly 10 crystals or crystalline powder. mL of each of the sample solution and standard solution as It is freely soluble in methanol, in ethanol (99.5), in acetic directed under Liquid Chromatography <2.01> according to acid (100) and in diethyl ether, and practically insoluble in the following conditions, and determine the peak areas, AT water. and AS, of mecobalamin in each solution. It gradually turns yellow on exposure to light.

Amount (mg) of C63H91CoN13O14P Identification (1) Dissolve10mgofMedazepamin3mL ＝ MS × AT/AS of citric acid-acetic acid TS: a deep orange color develops. Heat in a water bath for 3 minutes: the color changes to dark M : Amount (mg) of Mecobalamin RS, calculated on the S red. anhydrous basis (2) Determine the absorption spectrum of a solution of Operating conditions— Medazepam in methanol (1 in 100,000) as directed under Ul- Detector: An ultraviolet absorption photometer (wave- traviolet-visible Spectrophotometry <2.24>, and compare the length: 266 nm). spectrum with the Reference Spectrum: both spectra exhibit Column: A stainless steel column 4.6 mm in inside diame- similar intensities of absorption at the same wavelengths. ter and 25 cm in length, packed with octadecylsilanized silica (3) Determine the infrared absorption spectrum of gel for liquid chromatography (5 mm in particle diameter). Medazepam as directed in the potassium bromide disk Column temperature: A constant temperature of about method under Infrared Spectrophotometry <2.25>,andcom- 409C. pare the spectrum with the Reference Spectrum: both spectra Mobile phase: To 200 mL of acetonitrile add 800 mL of exhibit similar intensities of absorption at the same wave 1068 Medicinal Carbon / Official Monographs JP XVI numbers. (4) Perform the test with Medazepam as directed under Medicinal Carbon Flame Coloration Test <1.04> (2): a green color appears. 薬用炭 Melting point <2.60> 101–1049C Purity (1) Clarity and color of solution—Dissolve 1.0 g Description Medicinal Carbon occurs as a black, odorless of Medazepam in 10 mL of methanol: the solution is clear and tasteless powder. and light yellow to yellow in color. Identification Place 0.5 g of Medicinal Carbon in a test (2) Chloride <1.03>—Dissolve 1.5 g of Medazepam in 50 tube, and heat by direct application of flame with the aid of mL of diethyl ether, add 46 mL of water and 4 mL of so- a current of air: it burns without any flame. Pass the evolved dium carbonate TS, shake, and collect the water layer. Wash gas through calcium hydroxide TS: a white turbidity is pro- the water layer with two 20-mL portions of diethyl ether, duced. and filter. To 20 mL of the filtrate add dilute nitric acid to neutralize, add 6 mL of dilute nitric acid and water to make Purity (1) Acidity or alkalinity—Boil 3.0 g of Medicinal 50 mL, and perform the test using this solution as the test so- Carbon with 60 mL of water for 5 minutes, allow to cool, lution. Prepare the control solution with 0.30 mL of 0.01 dilute to 60 mL with water, and filter: the filtrate is colorless mol/L hydrochloric acid VS (not more than 0.018z). and neutral. (3) Heavy metals <1.07>—Proceed with 1.0 g of (2) Chloride <1.03>—Take 4.0 mL of the filtrate ob- Medazepam according to Method 2, and perform the test. tained in (1) in a Nessler tube, add 6 mL of dilute nitric acid Prepare the control solution with 2.0 mL of Standard Lead and sufficient water to make 50 mL, and perform the test Solution (not more than 20 ppm). using this solution as the test solution. Prepare the control (4) Arsenic <1.11>—Prepare the test solution with 1.0 g solution with 0.80 mL of 0.01 mol/L hydrochloric acid VS of Medazepam according to Method 3, and perform the test (not more than 0.142z). (not more than 2 ppm). (3) Sulfate <1.14>—Take 5 mL of the filtrate obtained in (5) Related substances—Dissolve 0.25 g of Medazepam (1) in a Nessler tube, add 1 mL of dilute hydrochloric acid in 10 mL of methanol, and use this solution as the sample so- and sufficient water to make 50 mL, and perform the test lution. Pipet 1 mL of the sample solution, and add methanol using this solution as the test solution. Prepare the control to make exactly 20 mL. Pipet 2 mL of this solution, add solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not methanol to make exactly 50 mL, and use this solution as the more than 0.192z). standard solution. Perform the test with these solutions as (4) Sulfide—Boil 0.5 g of Medicinal Carbon with a mix- directed under Thin-layer Chromatography <2.03>. Spot 10 ture of 15 mL of dilute hydrochloric acid and 10 mL of mL each of the sample solution and standard solution on a water: lead (II) acetate paper does not become brown when plate of silica gel with fluorescent indicator for thin-layer held in the evolved gas within 5 minutes. chromatography. Develop the plate with a mixture of cyclo- (5) Cyanogen compounds—Place a mixture of 5 g of hexane, acetone and ammonia solution (28) (60:40:1) to a Medicinal Carbon, 2 g of L-tartaric acid and 50 mL of water distance of about 10 cm, and air-dry the plate. Examine in a distilling flask connected to a condenser provided with a under ultraviolet light (main wavelength: 254 nm): the spots tightly fitting adapter, the end of which dips below the sur- other than the principal spot from the sample solution are face of a mixture of 2 mL of sodium hydroxide TS and 10 not more intense than the spot from the standard solution. mL of water, contained in a small flask surrounded by ice. Heat the mixture in the distilling flask to boiling, and distil Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- to 25 mL. Dilute the distillate with water to 50 mL. To 25 um, 609C, 4 hours). mL of the diluted distillate add 1 mL of a solution of iron Residue on ignition <2.44> Not more than 0.1z (1 g). (II) sulfate heptahydrate (1 in 20), heat the mixture almost to boiling, cool, and filter. To the filtrate add 1 mL of hydro- Assay Weigh accurately about 0.4 g of Medazepam, previ- chloric acid and 0.5 mL of dilute iron (III) chloride TS: no ously dried, dissolve in 50 mL of acetic acid (100), and titrate blue color is produced. <2.50> with 0.1 mol/L perchloric acid VS (potentiometric (6) Acid soluble substances—To about 1 g of Medicinal titration). Perform a blank determination, and make any Carbon, accurately weighed, add 20 mL of water and 5 mL necessary correction. of hydrochloric acid, boil for 5 minutes, filter, wash the Each mL of 0.1 mol/L perchloric acid VS residue with 10 mL of hot water, and add the washings to the

＝ 27.08 mg of C16H15ClN2 filtrate. Add 5 drops of sulfuric acid to the filtrate, evaporate to dryness, and ignite the residue strongly: the mass of Containers and storage Containers—Tight containers. the residue is not more than 3.0z. Storage—Light-resistant. (7) Heavy metals <1.07>—Proceed with 0.5 g of Medici- nal Carbon according to Method 2, and perform the test. Prepare the control solution with 2.5 mL of Standard Lead Solution (not more than 50 ppm). (8) Zinc—Ignite 0.5 g of Medicinal Carbon to ash, add 5 mL of dilute nitric acid to the residue, boil gently for 5 minutes, filter, wash with 10 mL of water, and combine the washings and the filtrate. Add 3 mL of ammonia TS to the solution, filter again, wash with water, combine the washings and the filtrate, add another washing to make 25 mL, JP XVI Official Monographs / Mefenamic Acid 1069 add 1 drop of sodium sulfide TS, and allow to stand for 3 Purity (1) Acidity or alkalinity—Dissolve 5.0 g of Medici- minutes: the liquid produces no turbidity. nal Soap in 85 mL of neutralized ethanol by warming on a (9) Arsenic <1.11>—Prepare the test solution with 1.0 g water bath, filter while hot through absorbent cotton, and of Medicinal Carbon according to Method 3, and perform wash the filter and the residue with three 5-mL portions of the test (not more than 2 ppm). hot neutralized ethanol. Combine the filtrate and the washings, add hot neutralized ethanol to make exactly 100 mL, Loss on drying <2.41> Not more than 15.0z (1 g, 1059C, and perform the following tests quickly using this as the 4 hours). sample solution at 709C. Residue on ignition <2.44> Not more than 4.0z (1 g). (i) Add 3 drops of phenolphthalein TS and 0.20 mL of 0.1 mol/L sodium hydroxide VS to 40 mL of the sample so- Adsorptive power (1) Add 1.0 g of Medicinal Carbon, lution: a red color develops. previously dried, to 100 mL of water containing 120 mg of (ii) Add 3 drops of phenolphthalein TS and 0.20 mL of quinine sulfate, shake the mixture vigorously for 5 minutes, 0.05 mol/L sulfuric acid VS to 40 mL of the sample solu- filter immediately, and reject the first 20 mL of the filtrate. tion: no red color develops. Add 5 drops of iodine TS to 10 mL of the subsequent fil- (2) Heavy metals <1.07>—Proceed with 1.0 g of Medici- trate: no turbidity is produced. nal Soap according to Method 2, and perform the test. Pre- (2) Dissolve 250 mg of methylene blue trihydrate, exactly pare the control solution with 2.0 mL of Standard Lead So- weighed, in water to make exactly 250 mL. Measure two lution (not more than 20 ppm). 50-mL portions of this solution into each of two glass- (3) Ethanol-insoluble substances—Weigh accurately stoppered flasks. To one flask add exactly 250 mg of Medici- about 2 g of Medicinal Soap, dissolve by warming in 100 mL nal Carbon, previously dried, and shake vigorously for 5 of neutralized ethanol, filter the solution through a glass minutes. Filter the contents of each flask, rejecting the first filter (G4), wash the residue with hot neutralized ethanol, 20 mL of each filtrate. Pipet 25-mL portions of the remain- and dry at 1059C for 4 hours: the mass of the residue is not ing filtrate into two 250-mL volumetric flasks. To each volu- more than 1.0z. metric flask add 50 mL of a solution of sodium acetate trihy- (4) Water-insoluble substances—Wash thoroughly the drate (1 in 10), then add exactly 35 mL of 0.05 mol/L iodine dried substances obtained in (3) with 200 mL of water, and VS with swirling. Allow them to stand for 50 minutes, shak- dry at 1059C for 4 hours: the mass of the residue is not more ing vigorously from time to time. Dilute each mixture to ex- than 0.15z. actly 250 mL with water, allow to stand for 10 minutes, and (5) Alkali carbonates—To the washings obtained in (4) filter each solution at a temperature not exceeding 209C, add 3 drops of methyl orange TS and 2 mL of 0.05 mol/L rejecting the first 30 mL of each filtrate. Titrate <2.50> the sulfuric acid VS: a red color develops. excess iodine in a 100-mL aliquot of each filtrate with 0.1 mol/L sodium thiosulfate VS. The difference between the Loss on drying Not more than 5.0z in the case of the pow- two titrations is not less than 1.2 mL. der, and not more than 10.0z in the case of the granules. Weigh accurately about 0.5 g of Medicinal Soap in a tared Containers and storage Containers—Well-closed contain- beaker, add 10 g of sea sand (No. 1), previously dried at ers. 1059C for 1 hour, and again weigh the beaker. Add 10 mL of ethanol (95), evaporate on a water bath to dryness with Medicinal Soap thorough stirring, and dry at 1059C for 3 hours. Containers and storage Containers—Well-closed contain- 薬用石ケン ers.

Medicinal Soap is sodium salts of fatty acids. Mefenamic Acid Description Medicinal Soap occurs as white to light yellow powder or granules. It has a characteristic odor free from メフェナム酸 rancidity. Medicinal Soap is sparingly soluble in water, and slightly soluble in ethanol (95). A solution of Medicinal Soap (1 in 100) is alkaline. Fatty acid Dissolve 25 g of Medicinal Soap in 300 mL of hot water, add 60 mL of dilute sulfuric acid slowly, and warm in a water bath for 20 minutes. After cooling, filter off the precipitate, and wash with warm water until the washing C15H15NO2:241.29 no longer shows acidity to methyl orange TS. Transfer the 2-(2,3-Dimethylphenylamino)benzoic acid precipitate to a small beaker, and heat on a water bath to [61-68-7] complete separation of water and transparent fatty acids. Filter the fatty acid into a small beaker while warm, dry at Mefenamic Acid, when dried, contains not less than 1009C for 20 minutes, and perform the test with this mate- 99.0z of C15H15NO2. rial as directed under Fats and Fatty Oils <1.13>. The con- Description Mefenamic Acid occurs as a white to light yel- gealing point of the fatty acid is between 189Cand289C. low powder. It is odorless and tasteless at first, but leaves a The acid value is 185 – 205. The iodine value is 82 – 92. slightly bitter aftertaste. 1070 Mefloquine Hydrochloride / Official Monographs JP XVI

It is sparingly soluble in diethyl ether, slightly soluble in previously dried, and dissolve in 100 mL of ethanol (95), pre- methanol, in ethanol (95) and in chloroform, and practically viously neutralized to phenol red TS with 0.1 mol/L sodium insoluble in water. hydroxide VS, by warming gently. Cool, and titrate <2.50> It dissolves in sodium hydroxide TS. with 0.1 mol/L sodium hydroxide VS until the color of the Melting point: about 2259C (with decomposition). solution changes from yellow through yellow-red to red-purple (indicator: 2 to 3 drops of phenol red TS). Perform a Identification (1) Dissolve 10 mg of Mefenamic Acid in blank determination, and make any necessary correction. 1 mL of methanol by warming, cool, add 1 mL of a solution of p-nitrobenzene diazonium fluoroborate (1 in 1000) and Each mL of 0.1 mol/L sodium hydroxide VS

1 mL of sodium hydroxide TS, and mix thoroughly: an ＝ 24.13 mg of C15H15NO2 orange-red color is produced. Containers and storage Containers—Well-closed contain- (2) Dissolve 10 mg of Mefenamic Acid in 2 mL of sulfu- ers. ric acid, and heat: the solution shows a yellow color and a green fluorescence. (3) Dissolve 7 mg of Mefenamic Acid in a solution of hydrochloric acid in methanol (1 in 1000) to make 500 mL. De- Mefloquine Hydrochloride termine the absorption spectrum of the solution as directed メフロキン塩酸塩 under Ultraviolet-visible Spectrophotometry <2.24>,and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Purity (1) Chloride <1.03>—To 1.0 g of Mefenamic Acid add 20 mL of sodium hydroxide TS, and dissolve by warming. Cool, add 2 mL of acetic acid (100) and water to make 100 mL, and mix well. Remove the produced precipitate by filtration, discard the first 10 mL of the filtrate, and to sub- C H F N O.HCl: 414.77 sequent 25 mL of the filtrate add 6 mL of dilute nitric acid 17 16 6 2 (1RS)-[2,8-Bis(trifluoromethyl)quinolin-4-yl][(2SR)- and water to make 50 mL. Perform the test using this solu- piperidin-2-yl]methanol monohydrochloride tion as the test solution. Prepare the control solution as [51773-92-3] follows: to 0.50 mL of 0.01 mol/L hydrochloric acid VS add 5 mL of sodium hydroxide TS, 0.5 mL of acetic acid (100), 6 Mefloquine Hydrochloride, when dried, contains mL of nitric acid and water to make 50 mL (not more than not less than 99.0z and not more than 101.0z of 0.071z). C H F N O.HCl. (2) Heavy metals <1.07>—Proceed with 2.0 g of 17 16 6 2 Mefenamic Acid according to Method 2, and perform the Description Mefloquine Hydrochloride occurs as white test. Prepare the control solution with 2.0 mL of Standard crystals or a white crystalline powder. Lead Solution (not more than 10 ppm). It is freely soluble in methanol, soluble in ethanol (99.5), (3) Arsenic <1.11>—Prepare the test solution with 1.0 g and slightly soluble in water. of Mefenamic Acid according to Method 3, and perform the It dissolves in sulfuric acid. test (not more than 2 ppm). A solution of Mefloquine Hydrochloride in methanol (1 in (4) Related substances—Dissolve 0.10 g of Mefenamic 20) shows no optical rotation. Acid, in 5 mL of a mixture of chloroform and methanol Melting point: about 2609C (with decomposition). (3:1), and use this solution as the sample solution. Pipet 1 Identification (1) Dissolve 2 mg of Mefloquine Hydro- mL of the sample solution, add a mixture of chloroform and chloride in 1 mL of sulfuric acid: the solution shows a blue methanol (3:1) to make exactly 200 mL, pipet 10 mL of this fluorescence under ultraviolet light (main wavelength: 365 solution, add a mixture of chloroform and methanol (3:1) to nm). make exactly 50 mL, and use this solution as the standard (2) Determine the absorption spectrum of a solution of solution. Perform the test with these solutions as directed Mefloquine Hydrochloride in methanol (1 in 25,000) as di- under Thin-layer Chromatography <2.03>. Spot 25 mLeach rected under Ultraviolet-visible Spectrophotometry <2.24>, of the sample solution and standard solution on a plate of and compare the spectrum with the Reference Spectrum: silica gel with fluorescent indicator for thin-layer chromatog- both spectra exhibit similar intensities of absorption at the raphy. Develop the plate with a mixture of 2-methyl-1- same wavelengths. propanol and ammonia solution (28) (3:1) to a distance of (3) Determine the infrared absorption spectrum of about 10 cm, and air-dry the plate. Examine under ultravio- Mefloquine Hydrochloride, previously dried, as directed in let light (main wavelength: 254 nm): the spots other than the the potassium chloride disk method under Infrared Spectro- principal spot from the sample solution are not more intense photometry <2.25>, and compare the spectrum with the Ref- than the spot from the standard solution. erence Spectrum: both spectra exhibit similar intensities of Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- absorption at the same wave numbers. um, phosphorus (V) oxide, 4 hours). (4) To 5 mL of a solution of Mefloquine Hydrochloride (1 in 1000) add 1 mL of dilute nitric acid and 1 mL of silver Residue on ignition <2.44> Not more than 0.1z (1 g). nitrate TS: a white precipitate is formed, and the separated Assay Weigh accurately about 0.5 g of Mefenamic Acid, precipitate dissolves on the addition of an excess amount of JP XVI Official Monographs / Mefruside 1071 ammonia TS. ing conditions, the relative standard deviation of the peak area of mefloquine is not more than 2.0z. Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of (4) Residual solvent Being specified separately. Mefloquine Hydrochloride according to Method 2 using a quartz crucible, and perform the test. Prepare the control so- Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, lution with 2.0 mL of Standard Lead Solution (not more 2 hours). than 20 ppm). Residue on ignition <2.44> Not more than 0.1z (1 g, plati- (2) Arsenic <1.11>—To 1.0 g of Mefloquine Hydrochlo- num crucible). ride add 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10), burn the ethanol, gradually Assay Weigh accurately about 0.5 g of Mefloquine Hydro- heat, and incinerate by ignition at 8009C. If a carbonized chloride, previously dried, dissolve in 100 mL of a mixture residue still retains, moisten the residue with a little amount of acetic anhydride and acetic acid (100) (7:3), and titrate of nitric acid, and ignite again to incinerate. After cooling, <2.50> with 0.1 mol/L perchloric acid VS (potentiometric to the residue add 3 mL of hydrochloric acid, warm on a titration). Perform a blank determination in the same man- water bath to dissolve, and perform the test using this solu- ner, and make any necessary correction. tion as the test solution (not more than 2 ppm). Each mL of 0.1 mol/L perchloric acid VS (3) Related substances—Dissolve 50 mg of Mefloquine ＝ 41.48 mg of C H F N O.HCl Hydrochloride in 50 mL of the mobile phase, and use this 17 16 6 2 solution as the sample solution. Pipet 1 mL of the sample so- Containers and storage Containers—Well-closed contain- lution, and add the mobile phase to make exactly 50 mL. ers. Pipet 2 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mLeachofthesamplesolu- Mefruside tion and standard solution as directed under Liquid Chroma- tography <2.01> according to the following conditions, and メフルシド determine each peak area by the automatic integration method: the area of the peak other than mefloquine and the peak eluted first from the sample solution is not larger than the peak area of mefloquine from the standard solution, and the total area of the peaks other than the peak of mefloquine and the peak eluted first from the sample solution is not larger than 2.5 times the peak area of mefloquine from the standard solution. C13H19ClN2O5S2: 382.88 Operating conditions— 4-Chloro-N-methyl-N-[(2RS)-2-methyltetrahydrofuran-2- Detector: An ultraviolet absorption photometer (wave- ylmethyl]-3-sulfamoylbenzenesulfonamide length: 282 nm). [7195-27-9] Column: A stainless steel column 3.9 mm in inside diameter and 30 cm in length, packed with aminopropylsilanized Mefruside, when dried, contains not less than silica gel for liquid chromatography (10 mminparticlediam- 98.5z of C13H19ClN2O5S2. eter). Description Mefruside occurs as a white crystalline pow- Column temperature: A constant temperature of about der. 409C. It is very soluble in dimethylformamide, freely soluble in Mobile phase: A mixture of acetonitrile and diluted phos- acetone, soluble in methanol, sparingly soluble in ethanol phoric acid (1 in 14) (24:1). (95), and practically insoluble in water. Flow rate: Adjust the flow rate so that the retention time A solution of Mefruside in dimethylformamide (1 in 10) of mefloquine is about 10 minutes. has no optical rotation. Time span of measurement: About 3 times as long as the retention time of mefloquine. Identification (1) Determine the absorption spectrum of a System suitability— solution of Mefruside in methanol (1 in 40,000) as directed Test for required detectability: To exactly 10 mL of the under Ultraviolet-visible Spectrophotometry <2.24>,and standard solution add the mobile phase to make exactly 20 compare the spectrum with the Reference Spectrum: both mL. Confirm that the peak area of mefloquine obtained spectra exhibit similar intensities of absorption at the same with 10 mL of this solution is equivalent to 40 to 60z of that wavelengths. with 10 mL of the standard solution. (2) Determine the infrared absorption spectrum of System performance: Dissolve 10 mg of mefloquine hy- Mefruside, previously dried, as directed in the potassium drochloride and 5 mg of diprophylline in 50 mL of the mo- bromide disk method under Infrared Spectrophotometry bile phase. To 2 mL of this solution add the mobile phase to <2.25>, and compare the spectrum with the Reference Spec- make 20 mL. When the procedure is run with 10 mLofthis trum: both spectra exhibit similar intensities of absorption at solution under the above operating conditions, diprophylline the same wave numbers. and mefloquine are eluted in this order with the resolution (3) Perform the test with Mefruside as directed under between these peaks being not less than 5. Flame Coloration Test <1.04> (2): a green color appears. System repeatability: When the test is repeated 6 times Melting point <2.60> 149 – 1529C with 10 mL of the standard solution under the above operat- 1072 Mefruside Tablets / Official Monographs JP XVI

Purity (1) Heavy metals <1.07>—Dissolve 1.0 g of Mefru- equivalent to 0.01 g of Mefruside according to the labeled side in 30 mL of acetone, and add 2 mL of dilute acetic acid amount, shake with 70 mL of methanol strongly for 15 and water to make 50 mL. Perform the test using this solu- minutes, add methanol to make 100 mL, and filter. Deter- tion as the test solution. Prepare the control solution as mine the absorption spectrum of the filtrate as directed follows: to 2.0 mL of Standard Lead Solution add 30 mL of under Ultraviolet-visible Spectrophotometry <2.24>:itexhib- acetone, 2 mL of dilute acetic acid and water to make 50 mL its maxima between 274 nm and 278 nm, and between 283 (not more than 20 ppm). nm and 287 nm. (2) Arsenic <1.11>—Prepare the test solution with 1.0 g Uniformity of dosage units <6.02> Perform the test accord- of Mefruside according to Method 3, and perform the test ing to the following method: it meets the requirement of the (not more than 2 ppm). Content uniformity test. (3) Related substances—Dissolve 0.20 g of Mefruside in To 1 tablet of Mefruside Tablets add 40 mL of methanol, 10 mL of acetone, and use this solution as the sample solu- disintegrate the tablet using ultrasonic waves with occasional tion. Pipet 1 mL of the sample solution, add acetone to stirring, then further treat with ultrasonic waves for 10 make exactly 200 mL, and use this solution as the standard minutes, and add methanol to make exactly V mL of a solu- solution. Perform the test with these solutions as directed tion containing about 0.5 mg of mefruside (C H ClN O S ) under Thin-layer Chromatography <2.03>. Spot 10 mLeach 13 19 2 5 2 per mL. Centrifuge the solution, pipet 5 mL of the superna- of the sample solution and standard solution on a plate of tant liquid, add methanol to make exactly 20 mL, and use silica gel with fluorescent indicator for thin-layer chromatog- this solution as the sample solution. Then, proceed as di- raphy. Develop the plate with a mixture of chloroform and rected in the Assay. acetone (5:2) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 Amount (mg) of mefruside (C13H19ClN2O5S2) nm): the spots other than the principal spot from the sample ＝ MS × AT/AS × V/125 solution are not more intense than the spot from the stand- M : Amount (mg) of mefruside for assay ard solution. S Dissolution <6.10> When the test is performed at 50 revolu- Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, tions per minute according to the Paddle method, using 900 2 hours). mL of water as the dissolution medium, the dissolution rate Residue on ignition <2.44> Not more than 0.1z (1 g). in 45 minutes of Mefruside Tablets is not less than 85z. Start the test with 1 tablet of Mefruside Tablets, withdraw Assay Weigh accurately about 0.5 g of Mefruside, previ- not less than 20 mL of the medium at the specified minute ously dried, dissolve in 80 mL of N,N-dimethylformamide, after starting the test, and filter through a filter paper for and titrate <2.50> with 0.1 mol/L tetramethylammonium hy- quantitative analysis (5C). Discard the first 5 mL of the droxide VS (potentiometric titration). Separately, perform a filtrate, pipet V mL of the subsequent filtrate, add water to blank determination with a solution prepared by adding 13 make exactly V?mL so that each mL contains about 28 mg mL of water to 80 mL of N,N-dimethylformamide, and of mefruside (C H ClN O S ) according to the labeled make any necessary correction. 13 19 2 5 2 amount, and use this solution as the sample solution. Sepa- Each mL of 0.1 mol/L tetramethylammonium rately, weigh accurately about 70 mg of mefruside for assay, hydroxide VS previously dried at 1059C for 2 hours, dissolve in methanol

＝ 38.29 mg of C13H19ClN2O5S2 to make exactly 50 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the Containers and storage Containers—Well-closed contain- standard solution. Determine the absorbances, A and A , ers. T S of the sample solution and standard solution at 285 nm in a layer of 5 cm in length as directed under Ultraviolet-visible Mefruside Tablets Spectrophotometry <2.24>, using water as the blank. Dissolution rate (z) with respect to the labeled amount

メフルシド錠 of mefruside (C13H19ClN2O5S2) ＝ MS × AT/AS × V?/V × 1/C × 36

Mefruside Tablets contain not less than 95.0z and MS: Amount (mg) of mefruside for assay not more than 105.0z of the labeled amount of C: Labeled amount (mg) of mefruside (C13H19ClN2O5S2) mefruside (C13H19ClN2O5S2: 382.88). in 1 tablet Method of preparation Prepare as directed under Tablets, Assay Weigh accurately not less than 20 Mefruside with Mefruside. Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 65 mg of mefruside Identification (1) Weigh a quantity of powdered Mefru- (C H ClN O S ), shake with 70 mL of methanol for 15 side Tablets, equivalent to 0.3 g of Mefruside according to 13 19 2 5 2 minutes, then add methanol to make exactly 100 mL, and the labeled amount, shake with 15 mL of heated methanol filter. Discard the first 20 mL of the filtrate, take exactly 10 for 20 minutes, and filter. Add 25 mL of water to the fil- mL of the subsequent filtrate, add methanol to make exactly trate, and allow to stand while ice-cooling for 30 minutes. 50 mL, and use this solution as the sample solution. Sepa- Filter the white precipitate formed, wash with water, and dry rately, weigh accurately about 65 mg of mefruside for assay, at 1059C for 2 hours: the precipitate melts <2.60> between previously dried at 1059C for 2 hours, and dissolve in metha- 1499C and 1529C. nol to make exactly 100 mL. Pipet 10 mL of this solution, (2) Weigh a quantity of powdered Mefruside Tablets, JP XVI Official Monographs / Meglumine Iotalamate Injection 1073 add methanol to make exactly 50 mL, and use this solution water to make 50 mL. Perform the test using this solution as as the standard solution. Determine the absorbances, AT and the test solution. Prepare the control solution with 0.40 mL AS, of the sample solution and standard solution at 285 nm of 0.005 mol/L sulfuric acid VS (not more than 0.019z). as directed under Ultraviolet-visible Spectrophotometry (4) Heavy metals <1.07>—Proceed with 2.0 g of Meglu- <2.24>. mine according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution Amount (mg) of mefruside (C H ClN O S ) 13 19 2 5 2 (not more than 10 ppm). ＝ M × A /A S T S (5) Arsenic <1.11>—Prepare the test solution with 2.0 g

MS: Amount (mg) of mefruside for assay of Meglumine according to Method 3, and perform the test (not more than 1 ppm). Containers and storage Containers—Tight containers. (6) Reducing substances—To 5 mL of a solution of Meglumine (1 in 20) add 5 mL of Fehling's TS, and boil for Meglumine 2 minutes: no red-brown precipitate is produced. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, メグルミン 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.4 g of Meglumine, previously dried, dissolve in 25 mL of water, and titrate <2.50> with 0.1 mol/L hydrochloric acid VS (indicator: 2 drops of C H NO :195.21 7 17 5 methyl red TS). 1-Deoxy-1-methylamino-D-glucitol [6284-40-8] Each mL of 0.1 mol/L hydrochloric acid VS

＝ 19.52 mg of C7H17NO5 Meglumine, when dried, contains not less than Containers and storage Containers—Tight containers. 99.0z of C7H17NO5. Description Meglumine occurs as a white, crystalline powder. It is odorless, and has a slightly bitter taste. Meglumine Iotalamate Injection It is freely soluble in water, and slightly soluble in ethanol (95), and practically insoluble in diethyl ether. イオタラム酸メグルミン注射液 The pH of a solution of Meglumine (1 in 10) is between 11.0 and 12.0. Meglumine Iotalamate Injection is an aqueous solu- Identification (1) To 1 mL of a solution of Meglumine tion for injection. (1 in 10) add 1 mL of potassium 1,2-naphthoquinone-4- It contains not less than 95.0z and not more than sulfonate TS: a deep red color develops. 105.0z of the labeled amount of iotalamic acid (2) To 2 mL of a solution of Meglumine (1 in 10) add 1 (C11H9I3N2O4: 613.91). drop of methyl red TS, and add 0.5 mL of dilute sodium hy- Method of preparation droxide TS and 0.5 g of boric acid after neutralizing with 0.5 mol/L sulfuric acid TS: a deep red color develops. (1) (3) Dissolve 0.5 g of Meglumine in 1 mL of diluted hy- Iotalamic Acid 227.59 g drochloric acid (1 in 3), and add 10 mL of ethanol (99.5): a Meglumine 72.41 g white precipitate is produced. Then, rubbing the inside wall Water for Injection or Sterile Water of the container with a glass rod, cool with ice and produce for Injection in Containers a sufficient quantity more precipitate. Filter the precipitate by suction through a To make 1000 mL glass filter (G3), wash the precipitate with a small volume of ethanol (99.5), and dry at 1059C for 1 hour: the residue thus (2) obtained melts <2.60> between 1499Cand1529C. Iotalamic Acid 455 g

20 Meglumine 145 g Optical rotation <2.49> [a]D : －16.0 – －17.09(after dry- Water for Injection or Sterile Water ing, 1 g, water, 10 mL, 100 mm). for Injection in Containers a sufficient quantity Melting point <2.60> 128–1319C To make 1000 mL Purity (1) Clarity and color of solution—Dissolve 1.0 g Prepare as directed under Injections, with the above ingre- of Meglumine in 10 mL of water: the solution is clear and dients (1) or (2). colorless. (2) Chloride <1.03>—Dissolve 1.0 g of Meglumine in 30 Description Meglumine Iotalamate Injection is a clear, col- mL of water, and add 10 mL of dilute nitric acid and water orless to pale yellow, slightly viscous liquid. to make 50 mL. Perform the test using this solution as the It gradually changes in color by light. test solution. Prepare the control solution with 0.25 mL of Identification (1) To 1 mL of Meglumine Iotalamate In- 0.01 mol/L hydrochloric acid VS (not more than 0.009z). jection add 1 mL of potassium naphthoquinone sulfonate TS (3) Sulfate <1.14>—Dissolve 1.0 g of Meglumine in 30 and 0.2 mL of sodium hydroxide TS: a deep red color de- mL of water, and add 5 mL of dilute hydrochloric acid and velops. 1074 Meglumine Sodium Amidotrizoate Injection / Official Monographs JP XVI

(2) To a volume of Meglumine Iotalamate Injection, Operating conditions— equivalent to 1 g of Iotalamic Acid according to the labeled Detector: An ultraviolet absorption photometer (wave- amount, add 25 mL of water, and add 2.5 mL of dilute hy- length: 240 nm). drochloric acid while shaking: a white precipitate is pro- Column: A stainless steel column 4.6 mm in inside diame- duced. Filter the precipitate by suction through a glass filter ter and 15 cm in length, packed with octadecylsilanized silica (G4), wash the precipitate with two 10-mL portions of water, gel for liquid chromatography (5 mminparticlediameter). and dry at 1059C for 4 hours. Proceed with the precipitate so Column temperature: A constant temperature of about obtained as directed in the Identification (2) under Iotalamic 209C. Acid. Mobile phase: Dissolve 3.9 g of phosphoric acid and 2.8 mL of triethylamine in water to make 2000 mL. To this solu- Optical rotation <2.49> tion add 100 mL of acetonitrile. Method of preparation (1) a 20: －1.67 – －1.939(100 D Flow rate: Adjust the flow rate so that the retention time mm). of iotalamic acid is about 6 minutes. Method of preparation (2) a 20: －3.35 – －3.869(100 D System suitability— mm). System performance: When the procedure is run with 10 pH <2.54> 6.5–7.7 mL of the standard solution under the above operating conditions, iotalamic acid and the internal standard are eluted in Purity (1) Primary aromatic amines—To a volume of this order with the resolution between these peaks being not Meglumine Iotalamate Injection, equivalent to 0.20 g of less than 5. Iotalamic Acid according to the labeled amount, add 15 mL System repeatability: When the test is repeated 6 times of water, shake, add 4 mL of a solution of sodium nitrite with 10 mL of the standard solution under the above operat- (1 in 100) under ice-cooling, and proceed as directed in the ing conditions, the relative standard deviation of the ratios Purity (2) under Iotalamic Acid: the absorbance is not more of the peak area of iotalamic acid to that of the internal than 0.17. standard is not more than 1.0z. (2) Iodine and iodide—Take a volume of Meglumine Iotalamate Injection, equivalent to 1.5 g of Iotalamic Acid Containers and storage Containers—Hermetic containers, according to the labeled amount, and proceed as directed in and colored containers may be used. the Purity (2) under Sodium Iotalamate Injection. Storage—Light-resistant. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according Meglumine Sodium Amidotrizoate to Method 1: it meets the requirement. Injection Insoluble particulate matter <6.07> It meets the requirement. アミドトリゾ酸ナトリウムメグルミン注射液 Sterility <4.06> Perform the test according to the Mem- brane filtration method: it meets the requirement. Meglumine Sodium Amidotrizoate Injection is an aqueous solution for injection. Assay To an exactly measured volume of Meglumine Iotalamate Injection, equivalent to about 4 g of iotalamic It contains not less than 95.0z and not more than 105.0z of the labeled amount of amidotrizoic acid acid (C11H9I3N2O4), add water to make exactly 200 mL. Pipet 2 mL of this solution, add water to make exactly 200 (C11H9I3N2O4: 613.91). mL. To exactly 5 mL of this solution add exactly 5 mL of the Method of preparation internal standard solution, add the mobile phase to make (1) 100 mL, and use this solution as the sample solution. Sepa- Amidotrizoic Acid (anhydrous) 471.78 g rately, weigh accurately about 0.4 g of iotalamic acid for Sodium Hydroxide 5.03 g assay, previously dried at 1059C for 4 hours, dissolve in 100 Meglumine 125.46 g mL of water and 1 mL of sodium hydroxide TS, and add Water for Injection or Sterile Water water to make exactly 200 mL. Pipet 5 mL of this solution, for Injection in Containers a sufficient quantity add water to make exactly 50 mL. To exactly 5 mL of this solution add exactly 5 mL of the internal standard solution, To make 1000 mL add the mobile phase to make 100 mL, and use this solution (2) as the standard solution. Perform the test with 10 mLeachof Amidotorizoic Acid (anhydrous) 597.30 g the sample solution and standard solution as directed under Sodium Hydroxide 6.29 g Liquid Chromatography <2.01> according to the following Meglumine 159.24 g conditions, and calculate the ratios, Q and Q , of the peak T S Water for Injection or Sterile Water area of iotalamic acid to that of the internal standard. for Injection in Containers a sufficient quantity Amount (mg) of iotalamic acid (C11H9I3N2O4) To make 1000 mL ＝ MS × QT/QS × 10 Prepare as directed under Injections, with the above ingre- MS: Amount (mg) of iotalamic acid for assay dients (1) or (2). Internal standard solution—A solution of L-tryptophan in Description Meglumine Sodium Amidotrizoate Injection is the mobile phase (3 in 2500). a clear, colorless to pale yellow, slightly viscous liquid. JP XVI Official Monographs / Meglumine Sodium Iodamide Injection 1075

It gradually changes in color by light. weigh accurately about 0.25 g of amidotrizoic acid for assay (previously determine the loss on drying <2.41> in the same Identification (1) To a volume of Meglumine Sodium manner as Amidotrizoic Acid), dissolve in a solution of Amidotrizoate Injection, equivalent to 1 g of Amidotrizoic meglumine (3 in 1000) to make exactly 100 mL, then pipet 2 Acid according to the labeled amount, add 25 mL of water, mL of this solution, add exactly 10 mL of the internal stand- and add 2.5 mL of dilute hydrochloric acid with stirring: a ard solution and water to make 100 mL, and use this solu- white precipitate is produced. Filter the precipitate by suction as the standard solution. Perform the test with 5 mL tion through a glass filter (G4), wash with two 10-mL por- each of the sample solution and standard solution as directed tions of water, and dry at 1059C for 1 hour. Proceed with under Liquid Chromatography <2.01> according to the fol- the precipitate so obtained as directed in the Identification lowing conditions, and calculate the ratios, Q and Q ,of (2) under Amidotrizoic Acid. T S the peak area of amidotrizoic acid to that of the internal (2) To 1 mL of Meglumine Sodium Amidotrizoate Injec- standard. tion add 1 mL of potassium 1,2-naphthoquinone-4-sulfonate

TS and 0.2 mL of sodium hydroxide TS: a deep red color de- Amount (mg) of amidotrizoic acid (C11H9I3N2O4) velops. ＝ MS × QT/QS × 2 (3) Meglumine Sodium Amidotrizoate Injection re- M : Amount (mg) of amidotrizoic acid for assay, calcu- sponds to the Qualitative Tests <1.09> (1) for sodium salt. S lated on the dried basis Optical rotation <2.49> Internal standard solution—Dissolve 0.06 g of acetrizoic Method of preparation (1) a 20: －2.91 – －3.369(100 D acid in a solution of meglumine (3 in 1000) to make 100 mL. mm). Operating conditions— Method of preparation (2) a 20: －3.69 – －4.279(100 D Detector: An ultraviolet absorption photometer (wave- mm). length: 254 nm). pH <2.54> 6.0–7.7 Column: A stainless steel column 4.6 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica Purity (1) Primary aromatic amines—To a volume of gel for liquid chromatography (5 mminparticlediameter). Meglumine Sodium Amidotrizoate Injection, equivalent to Column temperature: A constant temperature of about 0.20 g of Amidotrizoic Acid according to the labeled 259C. amount, add 6 mL of water, mix, add 4 mL of a solution of Mobile phase: Dissolve 1.7 g of tetrabutylammonium sodium nitrite (1 in 100) and 10 mL of 1 mol/L hydrochloric phosphate and 7.0 g of dipotassium hydrogenphosphate in acid TS, and shake. Proceed as directed in the Purity (2) 750 mL of water, adjust the pH to 7.0 with diluted phos- under Amidotrizoic Acid: the absorbance is not more than phoric acid (1 in 10), add water to make 800 mL, then add 0.19. 210 mL of acetonitrile, and mix. (2) Iodine and iodide—To a volume of Meglumine So- Flow rate: Adjust the flow rate so that the retention time dium Amidotrizoate Injection, equivalent to 0.25 g of of amidotrizoic acid is about 5 minutes. Amidotrizoic Acid according to the labeled amount, add System suitability— water to make 20 mL, add 5 mL of dilute nitric acid, shake System performance: When the procedure is run with 5 mL well, and filter by suction through a glass filter (G4). Add 5 of the standard solution under the above operating condi- mL of chloroform to the filtrate, and shake vigorously: no tions, amidotrizoic acid and the internal standard are eluted color develops in the chloroform layer. Then add 1 mL of in this order with the resolution between these peaks being hydrogen peroxide (30), and shake vigorously: the chlo- not less than 6. roform layer has no more color than the following control System repeatability: When the test is repeated 6 times solution. with 5 mL of the standard solution under the above operating Control solution: Dissolve 0.10 g of potassium iodide in conditions, the relative standard deviation of the ratios of water to make 100 mL. Add 20 mL of water to 0.10 mL of the peak area of amidotrizoic acid to that of the internal this solution, add 5 mL of dilute nitric acid, 5 mL of chlo- standard is not more than 1.0z. roform and 1 mL of hydrogen peroxide (30), and shake vigorously. Containers and storage Containers—Hermetic containers, and colored containers may be used. Extractable volume <6.05> It meets the requirement. Storage—Light-resistant. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require- Meglumine Sodium Iodamide ment. Injection Sterility <4.06> Perform the test according to the Mem- ヨーダミドナトリウムメグルミン注射液 brane filtration method: it meets the requirement. Assay To an exactly measured volume of Meglumine So- Meglumine Sodium Iodamide Injection is an aque- dium Amidotrizoate Injection, equivalent to about 0.5 g of ous solution for injection. amidotrizoic acid (C11H9I3N2O4), add water to make exactly 200 mL. Pipet 2 mL of this solution, add exactly 10 mL of It contains not less than 59.7 w/vz and not more than 65.9 w/v of iodamide (C H I N O : 627.94). the internal standard solution and water to make 100 mL, z 12 11 3 2 4 and use this solution as the sample solution. Separately, 1076 Melphalan / Official Monographs JP XVI

Method of preparation tion with isotonic sodium chloride solution so as to contain 0.30 mL of Meglumine Sodium Iodamide Injection per mL Iodamide 627.9 g according to the labeled amount, and perform the test: it Sodium Hydroxide 6.0 g meets the requirements. Meglumine 165.9 g Water for Injection or Sterile Water Assay To an exactly measured 8 mL of Meglumine Sodium for Injection in Containers a sufficient quantity Iodamide Injection add sodium hydroxide TS to make ex- To make 1000 mL actly 100 mL, and use this solution as the sample solution. Pipet 10 mL of the sample solution into a saponification Prepare as directed under Injections, with the above ingre- flask, add 30 mL of sodium hydroxide TS and 1 g of zinc dients. powder, and proceed as directed in the Assay under Io- Description Meglumine Sodium Iodamide Injection is a damide. clear, colorless to pale yellow, slightly viscous liquid. Each mL of 0.1 mol/L silver nitrate VS

It gradually changes in color by light. ＝ 20.93 mg of C12H11I3N2O4 Identification (1) To 2 mL of Meglumine Sodium Io- Containers and storage Containers—Hermetic containers, damide Injection add 25 mL of water, and add 3 mL of and colored containers may be used. dilute hydrochloric acid with thorough stirring: a white pre- Storage—Light-resistant. cipitate is formed. Filter the precipitate by suction through a glass filter (G3), and wash with two 10-mL portions of water. Transfer the precipitate to a suitable flask, add 100 Melphalan mL of water, dissolve by heating, and gently boil until the volume becomes about 30 mL. After cooling, collect the メルファラン separated crystals by filtration, dry at 1059C for 1 hour, and proceed as directed in the Identification (1) and (2) under Iodamide. (2) Determine the infrared absorption spectrum of the dried crystals obtained in (1) as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of about C13H18Cl2N2O2: 305.20 3390 cm－1, 1369 cm－1, 1296 cm－1, 1210 cm－1 and 1194 4-Bis(2-chloroethyl)amino-L-phenylalanine cm－1. [148-82-3] (3) To 1 mL of Meglumine Sodium Iodamide Injection add 1 mL of potassium 1,2-naphthoquinone-4-sulfonate TS Melphalan contains not less than 93.0z of and 0.2 mL of sodium hydroxide TS: a deep red color is pro- C13H18Cl2N2O2,calculatedonthedriedbasis. duced. Description Melphalan occurs as a white, to light yellowish (4) Meglumine Sodium Iodamide Injection responds to white, crystalline powder. the Qualitative Tests <1.09> (1) for sodium salt. It is slightly soluble in water, in methanol and in ethanol 20 (95), and practically insoluble in diethyl ether. Optical rotation <2.49> a D : －3.84 – －4.429(100 mm). It dissolves in dilute hydrochloric acid and in dilute so- pH <2.54> 6.5–7.5 dium hydroxide TS. It is gradually colored by light. Purity (1) Primary aromatic amines—Mix 0.30 mL of 20 Meglumine Sodium Iodamide Injection and 6 mL of water, Optical rotation [a]D : about －329(0.5 g, calculated on then add 4 mL of a solution of sodium nitrite (1 in 100) and the dried basis, methanol, 100 mL, 100 mm). 10 mL of 1 mol/L hydrochloric acid TS, shake well, and Identification (1) To 20 mg of Melphalan add 50 mL of proceed as directed in the Purity (2) under Iodamide: the ab- methanol, dissolve by warming, add 1 mL of a solution of 4- sorbance is not more than 0.22. (4-nitrobenzyl)pyridine in acetone (1 in 20), and evaporate (2) Iodine and iodide—To 0.40 mL of Meglumine So- on a water bath to dryness. Dissolve the residue in 1 mL of dium Iodamide Injection add water to make 20 mL, then warmed methanol and add 2 drops of ammonia solution add 5 mL of dilute nitric acid, shake well, filter by suction (28): a purple color develops. through a glass filter (G3). To the filtrate add 5 mL of chlo- (2) Dissolve 0.1 g of Melphalan in 10 mL of dilute so- roform, and shake vigorously: no color develops in the chlo- dium hydroxide TS, and heat on a water bath for 10 roform layer. Then add 1 mL of a strong hydrogen peroxide minutes. After cooling, add dilute nitric acid to acidify, and solution, and shake vigorously: the chloroform layer has no filter: the filtrate responds to the Qualitative Tests <1.09> for more color than the control solution. chloride. Control solution: Dissolve 0.10 g of potassium iodide in (3) Determine the absorption spectrum of a solution of water to make 100 mL. To a 0.10-mL portion of this solu- Melphalan in methanol (1 in 100,000) as directed under Ul- tion add 20 mL of water, 5 mL of dilute nitric acid, 5 mL of traviolet-visible Spectrophotometry <2.24>, and conpare the chloroform and 1 mL of strong hydrogen peroxide solution, spectrum with the Reference Spectrum: both spectra exhibit and shake vigorously. similar intensities of absorption at the same wavelengths. Extractable volume <6.05> It meets the requirement. Purity (1) Ionisable chloride—Weigh accurately about Pyrogen <4.04> Dilute Meglumine Sodium Iodamide Injec- 0.5 g of Melphalan, dissolve in 80 mL of diluted nitric acid JP XVI Official Monographs / Menatetrenone 1077

(1 in 40), stir for 2 minutes, and titrate <2.50> with 0.1 mol/L ence Spectrum or the spectrum of Menatetrenone RS: both silver nitrate VS (potentiometric titration): the consumed spectra exhibit similar intensities of absorption at the same volume is not more than 1.0 mL to 0.50 g of Melphalan. wave numbers. (2) Heavy metals <1.07>—Proceed with 1.0 g of Melpha- Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of lan according to Method 4, and perform the test. Prepare the Menatetrenone according to Method 4, and perform the test. control solution with 2.0 mL of Standard Lead Solution (not Prepare the control solution with 2.0 mL of Standard Lead more than 20 ppm). Solution (not more than 20 ppm). (3) Arsenic <1.11>—Prepare the test solution with 1.0 g (2) Menadione—To 0.20 g of Menatetrenone add 5 mL of Melphalan according to Method 3, and perform the test of diluted ethanol (1 in 2), shake well, and filter. To 0.5 mL (not more than 2 ppm). of the filtrate add 1 drop of a solution of 3-methyl-1-phenyl- Loss on drying <2.41> Not more than 7.0z (1 g, in vacuum 5-pyrazorone in ethanol (99.5) (1 in 20) and 1 drop of ammo- at a pressure not exceeding 0.67 kPa, 1059C, 2 hours). nia water (28), and allow to stand for 2 hours: no blue-purple color develops. Residue on ignition <2.44> Not more than 0.3z (1 g). (3) cis Isomer—Dissolve 0.10 g of Menatetrenone in 10 Assay Weigh accurately about 0.25 g of Melphalan, add 20 mL of hexane, and use this solution as the sample solution. mL of a solution of potassium hydroxide (1 in 5), and heat Pipet 1 mL of the sample solution, add hexane to make ex- under a reflux condenser on a water bath for 2 hours. After actly 50 mL, and use this solution as the standard solution. cooling, add 75 mL of water and 5 mL of nitric acid, cool, Perform the test with these solutions as directed under Thin- and titrate <2.50> with 0.1 mol/L silver nitrate VS (potentio- layer Chromatography <2.03>. Spot 10 mLeachofthesample metric titration). Make any necessary correction by using the solution and standard solution on a plate of silica gel with results obtained in the Purity (1). fluorescent indicator for thin-layer chromatography. De- velop the chromatogram with a mixture of hexane and Each mL of 0.1 mol/L silver nitrate VS dibutyl ether (17:3) to a distance of about 12 cm, and air-dry ＝ 15.26 mg of C H Cl N O 13 18 2 2 2 the plate. Examine under ultraviolet light (main wavelength: Containers and storage Containers—Tight containers. 254 nm): the spot corresponding to relative Rfvalue1.1 Storage—Light-resistant. regarding to the principal spot from the sample solution is not more intense than the spot from the standard solution. (4) Related substances—Conduct this procedure without Menatetrenone exposure to daylight, using a light-resistant vessel. Dissolve 0.10 g of Menatetrenone in 100 mL of ethanol (99.5), and メナテトレノン use this solution as the sample solution. Pipet 1 mL of the sample solution, add ethanol (99.5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of these solutions by the automatic integration method: the total area of peaks other than the peak of C H O :444.65 31 40 2 menatetrenone from the sample solution is not larger than 2-Methyl-3-[(2E,6E,10E)-3,7,11,15-tetramethylhexadeca- the peak area of menatetrenone from the standard solution. 2,6,10,14-tetraen-1-yl]-1,4-naphthoquinone Operating conditions— [863-61-6] Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in Menatetrenone contains not less than 98.0z of the Assay. C H O , calculated on the dehydrated basis. 31 40 2 Time span of measurement: About 6 times as long as the Description Menatetrenone occurs as yellow, crystals, crys- retention time of menatetrenone beginning after the solvent talline powder, waxy mass or oily material. peak. It is very soluble in hexane, soluble in ethanol (99.5), spar- System suitability— ingly soluble in 2-propanol, slightly soluble in methanol, and Test for required detection: To exactly 5 mL of the stand- practically insoluble in water. ard solution add ethanol (99.5) to make exactly 50 mL. Con- It decomposes and the color becomes more intense by firm that the peak area of menatetrenone obtained from light. 20 mL of this solution is equivalent to 7 to 13z of that from Melting point: about 379C. 20 mL of the standard solution. System performance: Proceed as directed in the system Identification (1) Dissolve 0.1 g of Menatetrenone in 5 suitability in the Assay. mL of ethanol (99.5) by warming, cool, and add 1 mL of a System repeatability: When the test is repeated 6 times solution of potassium hydroxide in ethanol (95) (1 in 10): a with 20 mL of the standard solution under the above operat- blue color develops, and upon standing it changes from blue- ing conditions, the relative standard deviation of the peak purple to red-brown through red-purple. areas of menatetrenone is not more than 1.0z. (2) Determine the infrared absorption spectrum of Menatetrenone, after melting by warming if necessary, as di- Water <2.48> Not more than 0.5z (0.5 g, volumetric titra- rected in the liquid film method under Infrared Spectropho- tion, direct titration). tometry <2.25>, and compare the spectrum with the Refer- 1078 dl-Menthol / Official Monographs JP XVI

Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Conduct this procedure without exposure to day- dl-Menthol light, using a light-resistant vessel. Weigh accurately about dl-メントール 0.1 g each of Menatetrenone and Menatetrenone RS (separately, determine the water <2.48> inthesamemanneras Menatetrenone), dissolve each in 50 mL of 2-propanol, and add ethanol (99.5) to make exactly 100 mL. Pipet 10 mL of these solutions, and add ethanol (99.5) to make exactly 100 mL. Pipet 2 mL each of these solutions, add exactly 4 mL each of the internal standard solution, and use these solu- C10H20O: 156.27 tions as the sample solution and standard solution. Perform (1RS,2SR,5RS)-5-Methyl-2-(1-methylethyl)cyclohexanol the test with 20 mL each of the sample solution and standard [89-78-1] solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the dl-Menthol contains not less than 98.0z of ratios, QT and QS, of the peak area of menatetrenone to that C10H20O. of the internal standard. Description dl-Menthol occurs as colorless crystals. It has a

Amount (mg) of C31H40O2 ＝ MS × QT/QS characteristic and refreshing odor and a burning taste, fol- lowedbyacooltaste. M : Amount (mg) of Menatetrenone RS, calculated on the S It is very soluble in ethanol (95) and in diethyl ether, and dehydrated basis very slightly soluble in water. Internal standard solution—A solution of phytonadione in It sublimes gradually at room temperature. 2-propanol (1 in 20,000). Identification (1) Triturate dl-Menthol with an equal Operating conditions— amount of camphor, chloral hydrate or thymol: the mixture Detector: An ultraviolet absorption photometer (wave- liquefies. length: 270 nm). (2) Shake 1 g of dl-Menthol with 20 mL of sulfuric acid: Column: A stainless steel column 4.6 mm in inside diame- the mixture becomes turbid with a yellow-red color. Allow to ter and 15 cm in length, packed with octadecylsilanized silica stand for 3 hours: a clear, oily layer possesses no aroma of gel for liquid chromatography (5 mm in particle diameter). menthol is separated. Column temperature: A constant temperature of about 409C. Congealing point <2.42> 27–289C Mobile phase: Methanol. Optical rotation <2.49> [a]20: －2.0 – ＋2.09(2.5 g, ethanol Flow rate: Adjust the flow rate so that the retention time D (95), 25 mL, 100 mm). of menatetrenone is about 7 minutes. System suitability— Purity (1) Non-volatile residue—Volatilize 2.0 g of dl- System performance: When the procedure is run with 20 Menthol on a water bath, and dry the residue at 1059Cfor2 mL of the standard solution under the above operating con- hours: the residue weighs not more than 1.0 mg. ditions, menatetrenone and the internal standard are eluted (2) Thymol—Add 0.20 g of dl-Menthol to a cold mixture in this order with the resolution between these peaks being of 2 mL of acetic acid (100), 6 drops of sulfuric acid and 2 not less than 4. drops of nitric acid: no green to blue-green color immedi- System repeatability: When the test is repeated 6 times ately develops. with 20 mL of the standard solution under the above operat- (3) Nitromethane or nitroethane—To 0.5 g of dl-Men- ing conditions, the relative standard deviation of the ratios thol placed in a flask add 2 mL of a solution of sodium hy- of the peak area of menatetrenone to that of the internal droxide (1 in 2) and 1 mL of strong hydrogen peroxide, con- standard is not more than 1.0z. nect a reflux condenser to the flask, and boil the mixture gently for 10 minutes. After cooling, add water to make ex- Containers and storage Containers—Tight containers. actly 20 mL, and filter. Take 1 mL of the filtrate in a Nessler Storage—Light-resistant. tube, add water to make 10 mL, neutralize with dilute hydrochloric acid, then add 1 mL of dilute hydrochloric acid, and cool. To the mixture add 1 mL of a solution of sulfanilic acid (1 in 100), allow to stand for 2 minutes, and then add 1 mL of a solution of N,N-diethyl-N?-1-naphthylethylenedia- mine oxalate (1 in 1000) and water to make 25 mL: no red- purple color immediately develops. Assay Weigh accurately about 2 g of dl-Menthol, add exactly 20 mL of a mixture of dehydrated pyridine and acetic anhydride (8:1), connect a reflux condenser, and heat on a water bath for 2 hours. Wash down the condenser with 20 mL of water, and titrate <2.50> with 1 mol/L sodium hydroxide VS (indicator: 5 drops of phenolphthalein TS). Per- form a blank determination, and make any necessary correction. JP XVI Official Monographs / Mepenzolate Bromide 1079

Each mL of 1 mol/L sodium hydroxide VS water, and titrate <2.50> with 1 mol/L sodium hydroxide VS

＝ 156.3 mg of C10H20O (indicator: 5 drops of phenolphthalein TS). Perform a blank determination and make any necessary correction. Containers and storage Containers—Tight containers. Storage—In a cold place. Each mL of 1 mol/L sodium hydroxide VS

＝ 156.3 mg of C10H20O Containers and storage Containers—Tight containers. l-Menthol Storage—In a cold place. l-メントール Mepenzolate Bromide

メペンゾラート臭化物

C10H20O: 156.27 (1R,2S,5R)-5-Methyl-2-(1-methylethyl)cyclohexanol [2216-51-5]

l-Menthol contains not less than 98.0z of C10H20O. C H BrNO : 420.34 Description l-Menthol occurs as colorless crystals. It has a 21 26 3 (3RS)-3-[(Hydroxy)(diphenyl)acetoxy]-1,1- characteristic and refreshing odor and a burning taste, fol- dimethylpiperidinium bromide lowedbyacooltaste. [76-90-4] It is very soluble in ethanol (95) and in diethyl ether, and very slightly soluble in water. Mepenzolate Bromide, when dried, contains not less It sublimes gradually at room temperature. than 98.5z of C21H26BrNO3. Identification (1) Triturate l-Menthol with an equal Description Mepenzolate Bromide is white to pale yellow amount of camphor, chloral hydrate or thymol: the mixture crystals or crystalline powder. It is odorless, and has a bitter liquefies. taste. (2) Shake 1 g of l-Menthol with 20 mL of sulfuric acid: It is very soluble in formic acid, freely soluble in metha- the mixture becomes turbid with a yellow-red color. Allow to nol, soluble in hot water, slightly soluble in water and in stand for 3 hours: a clear, oily layer which possesses no aro- ethanol (95), very slightly soluble in acetic anhydride, and ma of menthol is separated. practically insolubleindiethylether. 20 Optical rotation <2.49> [a]D : －45.0 – －51.09 (2.5 g, Melting point: about 2309C (with decomposition). ethanol (95), 25 mL, 100 mm). Identification (1) To 30 mg of Mepenzolate Bromide add Melting point <2.60> 42–449C 10 drops of sulfuric acid: a red color develops. (2) Dissolve 10 mg of Mepenzolate Bromide in 20 mL of Purity (1) Non-volatile residue—Volatilize 2.0 g of l- water and 5 mL of dilute hydrochloric acid, and to 5 mL of Menthol on a water bath, and dry the residue at 1059Cfor this solution add 1 mL of Dragendorff's TS: an orange pre- 2 hours: the residue weighs not more than 1.0 mg. cipitate is produced. (2) Thymol—Add 0.20 g of l-Menthol to a cold mixture (3) Determine the absorption spectrum of a solution of of 2 mL of acetic acid (100), 6 drops of sulfuric acid and 2 Mepenzolate Bromide in 0.01 mol/L hydrochloric acid TS (1 drops of nitric acid: no green to blue-green color immedi- in 2000) as directed under Ultraviolet-visible Spectropho- ately develops. tometry <2.24>, and compare the spectrum with the Refer- (3) Nitromethane or nitroethane—To 0.5 g of l-Menthol ence Spectrum: both spectra exhibit similar intensities of ab- placed in a flask add 2 mL of sodium hydroxide solution (1 sorption at the same wavelengths. in 2) and 1 mL of strong hydrogen peroxide, connect a reflux (4) Dissolve 0.5 g of Mepenzolate Bromide in 50 mL of condenser to the flask, and boil the mixture gently for 10 water and 3 mL of nitric acid by heating. This solution re- minutes. After cooling, add water to make exactly 20 mL, sponds to the Qualitative Tests <1.09> for Bromide. and filter. Take 1 mL of the filtrate in a Nessler tube, add water to make 10 mL, neutralize with dilute hydrochloric Purity (1) Heavy Metals <1.07>—Proceed with 1.0 g of acid, add another 1 mL of dilute hydrochloric acid, and Mepenzolate Bromide according to Method 2, and perform cool. To the mixture add 1 mL of a solution of sulfanilic the test. Prepare the control solution with 2.0 mL of Stand- acid (1 in 100), allow to stand for 2 minutes, and then add 1 ard Lead Solution (not less than 20 ppm). mL of a solution of N,N-diethyl-N?-1-naphthylethylenedia- (2) Arsenic <1.11>—Prepare the test solution with 1.0 g mine oxalate (1 in 1000) and water to make 25 mL: no red- of Mepenzolate Bromide according to Method 3, and per- purple color immediately develops. form the test (not more than 2 ppm). (3) Related substances—Dissolve 0.40 g of Mepenzolate Assay Weigh accurately about 2 g of l-Menthol, add ex- Bromide in exactly measured 10 mL of methanol, and use actly 20 mL of a mixture of dehydrated pyridine and acetic this solution as the sample solution. Pipet 1 mL of the sam- anhydride (8:1), connect a reflux condenser, and heat on a ple solution, add methanol to make exactly 200 mL, and use water bath for 2 hours. Wash the condenser with 20 mL of 1080 Mepitiostane / Official Monographs JP XVI this solution as the standard solution (1). Separately, dis- acetone, slightly soluble in methanol and in ethanol (99.5), solve 40 mg of benzophenone in methanol to make exactly and practically insoluble in water. 100 mL. Pipet 2 mL of this solution, add methanol to make It is hydrolyzed in moist air. exactly 10 mL, and use this solution as the standard solution Identification (1) Dissolve 1 mg of Mepitiostane in 1 mL (2). Perform the test with these solutions as directed under of methanol, and add 0.5 mL of palladium (II) chloride TS: Thin-layer Chromatography <2.03>. Spot 10 mLeachofthe an orange precipitate is formed. To this suspension add 1 sample solution, standard solutions (1) and (2) on a plate of mL of water and 2 mL of chloroform, shake well, and allow silica gel with fluorecent indicator for thin-layer chromatog- to stand: an orange color develops in the chloroform layer. raphy. Develop the plate with a mixture of 1-butanol, meth- (2) Dissolve 0.1 g of Mepitiostane in 2 mL of diethylene anol, water and acetic acid (100) (3:3:2:1) to a distance of glycol dimethyl ether, shake with 1 mL of 1 mol/L hydro- about 10 cm, and air-dry the plate and then at 809Cfor30 chloric acid TS, and filter. To the filtrate add 1.5 mL of 2,4- minutes. Examine under ultraviolet light (main wavelength: dinitrophenylhydrazine-diethylene glycol dimethyl ether TS 254 nm): the spots other than either the principal spot or the and 1.5 mL of diluted ethanol (95) (2 in 3): an orange-yellow spot corresponding to benzophenone from the sample solu- precipitate is formed. Filter the precipitate, recrystallize tion are not more intense than the spot from standard solu- from ethanol (99.5), and dry in a desiccator (in vacuum, tion (1), and the spot corresponding to benzophenone from phosphorus (V) oxide) for 4 hours: the crystals melt <2.60> the sample solution is not more intense than the spot from between 1449C and 1499C. standard solution (2). Spray evenly Dragendorff's TS on the (3) Determine the infrared absorption spectrum of plate: the spots other than the principal spot from the sample Mepitiostane as directed in the potassium bromide disk solution are not more intense than the spot from standard method under Infrared Spectrophotometry <2.25>,andcom- solution (1). pare the spectrum with the Reference Spectrum: both spectra Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, exhibit similar intensities of absorption at the same wave 4 hours). numbers.

20 Residue on ignition <2.44> Not more than 0.1z (1 g). Optical rotation <2.49> [a]D : ＋20 – ＋239 (0.1 g, chloroform, 10 mL, 100 mm). Assay Weigh accurately about 0.35 g of Mepenzolate Bromide, previously dried, dissolve in 2 mL of formic acid, Purity (1) Clarity and color of solution—Dissolve 0.10 g add 60 mL of acetic anhydride, and titrate <2.50> with 0.1 of Mepitiostane in 4 mL of petroleum ether: the solution is mol/L perchloric acid VS (potentiometric titration). Per- clear and colorless to pale yellow. form a blank determination, and make any necessary correc- (2) Heavy metals <1.07>—Proceedwith1.0gof tion. Mepitiostane according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Each mL of 0.1 mol/L perchloric acid VS Solution (not more than 20 ppm). ＝ 42.03 mg of C H BrNO 21 26 3 (3) Related substances—Dissolve 20 mg of Mepitiostane Containers and storage Containers—Tight containers. in exactly 5 mL of a mixture of acetone and triethylamine (1000:1), and use this solution as the sample solution. Sepa- rately, dissolve 10 mg of Epitiostanol RS in a mixture of Mepitiostane acetone and triethylamine (1000:1) to make exactly 10 mL. Pipet 1 mL and 3 mL of this solution, to each add a mixture メピチオスタン of acetone and triethylamine (1000:1) to make exactly 25 mL, and use these solutions as the standard solution (1) and the standard solution (2), respectively. Perform the test with these solutions as directed under Thin-layer Chromatogra- phy <2.03>. Spot 5 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane and acetone (3:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly diluted sulfuric acid (1 in 5) on the plate, heat between 1209Cand1309C for 5 minutes, and examine under ultravi- C H O S: 404.65 25 40 2 olet light (main wavelength: 365 nm): the spots other than 2a,3a-Epithio-17b-(1-methoxycyclopentyloxy)-5a-androstane the principal spot from the sample solution showing the [21362-69-6] same Rf value as the standard solutions are not more intense than the spot from the standard solution (2), and the remain- Mepitiostane contains not less than 96.0z and not ing spots other than the principal spot are not more intense more than 102.0z of C H O S, calculated on the an- 25 40 2 than the spot from the standard solution (1). hydrous basis. Water <2.48> Not more than 0.7z (0.3 g, volumetric titra- Description Mepitiostane occurs as white to pale yellow tion, back titration). crystals or crystalline powder. It is freely soluble in triethylamine, in chloroform, in Residue on ignition <2.44> Not more than 0.1z (0.5 g). diethyl ether and in cyclohexane, soluble in diethylene glycol Assay Weigh accurately about 0.3 g of Mepitiostane, and dimethyl ether and in petroleum ether, sparingly soluble in dissolve in cyclohexane to make exactly 10 mL. Pipet 2 mL JP XVI Official Monographs / Mepivacaine Hydrochloride 1081 of this solution, add 10 mL of ethanol (99.5), mix with exactly 2 mL each of 0.01 mol/L hydrochloric acid TS and the Mepivacaine Hydrochloride internal standard solution, add ethanol (99.5) to make 20 mL, allow to stand at ordinary temperature for 30 minutes, メピバカイン塩酸塩 and use this solution as the sample solution. Separately, weigh accurately about 45 mg of Epitiostanol RS, dissolve in exactly 2 mL of the internal standard solution, add ethanol (99.5) to make 20 mL, and use this solution as the standard solution. Perform the test with 10 mLeachofthesample solution and standard solution as directed under Liquid C H N O.HCl: 282.81 Chromatography <2.01> according to the following condi- 15 22 2 (2RS)-N-(2,6-Dimethylphenyl)-1-methylpiperidine-2- tions, and calculate the ratios, Q and Q , of the peak area T S carboxamide monohydrochloride of epitiostanol to that of the internal standard, respectively. [1722-62-9]

Amount (mg) of C25H40O2S ＝ MS × QT/QS × 5 × 1.320 Mepivacaine Hydrochloride, when dried, contains not less than 98.5z and not more than 101.0z of M : Amount (mg) of Epitiostanol RS, calculated on the S C H N O.HCl. anhydrous basis 15 22 2 Description Mepivacaine Hydrochloride occurs as white Internal standard solution—A solution of n-octylbenzene in crystals or crystalline powder. ethanol (99.5) (1 in 300). It is freely soluble in water and in methanol, soluble in Operating conditions— acetic acid (100), and sparingly soluble in ethanol (99.5). Detector: An ultraviolet absorption photometer (wave- A solution of Mepivacaine Hydrochloride (1 in 10) shows length: 265 nm). no optical rotation. Column: A stainless steel column 4.0 mm in inside diame- Melting point: about 2569C (with decomposition). ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mminparticlediameter). Identification (1) Determine the absorption spectrum of a Column temperature: A constant temperature of about solution of Mepivacaine Hydrochloride (1 in 2500) as di- 259C. rected under Ultraviolet-visible Spectrophotometry <2.24>, Mobile phase: A mixture of methanol and water (20:3). and compare the spectrum with the Reference Spectrum: Flow rate: Adjust the flow rate so that the retention time both spectra exhibit similar intensities of absorption at the of epitiostanol is about 6 minutes. same wavelengths. System suitability— (2) Determine the infrared absorption spectrum of System performance: When the procedure is run with 10 Mepivacaine Hydrochloride as directed in the potassium mL of the standard solution under the above operating con- chloride disk method under Infrared Spectrophotometry ditions, epitiostanol and the internal standard are eluted in <2.25>, and compare the spectrum with the Reference Spec- this order with the resolution between these peaks being not trum: both spectra exhibit similar intensities of absorption at less than 4. the same wave numbers. System repeatability: When the test is repeated 6 times (3) A solution of Mepivacaine Hydrochloride (1 in 50) with 10 mL of the standard solution under the above operat- responds to the Qualitative Tests <1.09š > for chloride. ing conditions, the relative standard deviation of the ratios pH <2.54> Dissolve 0.2 g of Mepivacaine Hydrochloride in of the peak area of epitiostanol to that of the internal stand- 10 mL of water: the pH of this solution is between 4.0 and ard is not more than 1.0z. 5.0. Containers and storage Containers—Hermetic containers. Purity (1) Clarity and color of solution—Dissolve 1.0 g Storage—Light-resistant, under Nitrogen atmosphere, and of Mepivacaine Hydrochloride in 10 mL of water: the solu- in a cold place. tion is clear and colorless. (2) Sulfate <1.14>—Perform the test with 0.5 g of Mepivacaine Hydrochloride. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.038z). (3) Heavy metals <1.07>—Proceedwith2.0gof Mepivacaine Hydrochloride according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (4) Related substances—Dissolve 0.10 g of Mepivacaine Hydrochloride in 5 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add methanol to make exactly 20 mL. Pipet 2 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard 1082 Mepivacaine Hydrochloride Injection / Official Monographs JP XVI solution on a plate of silica gel for thin-layer chromatogra- Assay Pipet a volume of Mepivacaine Hydrochloride Injec- phy. Develop the plate with a mixture of diethyl ether, meth- tion, equivalent to about 40 mg of mepivacaine hydrochlo- anol and ammonia solution (28) (100:5:1) to a distance of ride (C15H22N2O.HCl), add exactly 4 mL of the internal about 10 cm, and air-dry the plate. Spray evenly bismuth ni- standard solution and 0.001 mol/L hydrochloric acid TS to trate-potassium iodide TS on the plate: the spots other than make 20 mL, and use this solution as the sample solution. the principal spot from the sample solution are not more in- Separately, weigh accurately about 40 mg of mepivacaine hy- tense than the spot from the standard solution. drochloride for assay, previously dried at 1059C for 3 hours, dissolve in 0.001 mol/L hydrochloric acid TS, add exactly 4 Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, mL of the internal standard solution and 0.001 mol/L hy- 3 hours). drochloride TS to make 20 mL, and use this solution as the Residue on ignition <2.44> Not more than 0.1z (1 g). standard solution. Perform the test with 5 mLeachofthe sample solution and standard solution as directed under Liq- Assay Weigh accurately about 0.4 g of Mepivacaine Hy- uid Chromatography <2.01> according to the following con- drochloride, previously dried, dissolve in 10 mL of acetic ditions, and calculate the ratios, Q and Q , of the peak area acid (100) and add 70 mL of acetic anhydride. Titrate <2.50> T S of mepivacaine to that of the internal standard. with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination, and make any necessary Amount (mg) of mepivacaine hydrochloride correction. (C15H22N2O.HCl) ＝ M × Q /Q Each mL of 0.1 mol/L perchloric acid VS S T S

＝ 28.28 mg of C15H22N2O.HCl MS: Amount (mg) of mepivacaine hydrochloride for assay Containers and storage Containers—Tight containers. Internal standard solution—A solution of benzophenone in methanol (1 in 4000). Operating conditions— Mepivacaine Hydrochloride Detector: An ultraviolet absorption photometer (wavelength: 254 nm). Injection Column: A stainless steel column about 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized メピバカイン塩酸塩注射液 silica gel for liquid chromatography (10 mminparticlediameter). Mepivacaine Hydrochloride Injection is an aqueous Column temperature: A constant temperature of about solution for injection. 259C. It contains not less than 95.0z and not more than Mobile phase: Dissolve 2.88 g of sodium lauryl sulfate in 105.0z of the labeled amount of mepivacaine hydro- 1000 mL of a mixture of 0.02 mol/L phosphate buffer solu- chloride (C15H22N2O.HCl: 282.81). tion, pH 3.0, and acetonitrile (11:9). Flow rate: Adjust the flow rate so that the retention time Method of preparation Prepare as directed under Injec- of mepivacaine is about 6 minutes. tions, with Mepivacaine Hydrochloride. System suitability— Description Mepivacaine Hydrochloride Injection is a System performance: When the procedure is run with 5 mL clear, colorless liquid. of the standard solution under the above operating conditions, mepivacaine and the internal standard are eluted in Identification To a volume of Mepivacaine Hydrochloride this order with the resolution between these peaks being not Injection, equivalent to 20 mg of Mepivacaine Hydrochlo- less than 6. ride according to the labeled amount, add 1 mL of sodium System repeatability: When the test is repeated 6 times hydrochloride TS, and extract with 20 mL of hexane. To 8 with 5 mL of the standard solution under the above operating mL of the hexane extract add 20 mL of 1 mol/L hydrochlo- conditions, the relative standard deviation of the ratio of the ric acid TS, shake vigorously, and determine the absorption peak area of mepivacaine to that of the internal standard is spectrum of the water layer separated as directed under Ul- not more than 1.0z. traviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 261 nm and 265 nm, and between 270 nm and Containers and storage Containers—Hermetic containers. 273 nm. pH Being specified separately. Bacterial endotoxins <4.01> Less than 0.6 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the requirement. Sterility <4.06> Perform the test according to the Mem- brane filtration method: it meets the requirement. JP XVI Official Monographs / Mercaptopurine Hydrate 1083

Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- Mequitazine um, phosphorus (V) oxide, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). メキタジン Assay Weigh accurately about 0.25 g of Mequitazine, dissolve in 50 mL of acetic acid (100), titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per- form a blank determination, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS

＝ 32.25 mg of C20H22N2S Containers and storage Containers—Tight containers. C H N S: 322.47 20 22 2 Storage—Light-resistant. 10-[(3RS)-1-Azabicyclo[2.2.2]oct-3-ylmethyl]-10H- phenothiazine [29216-28-2] Mercaptopurine Hydrate Mequitazine, when dried, contains not less than メルカプトプリン水和物 98.5z of C20H22N2S. Description Mequitazine occurs as white crystals or crystalline powder. It is freely soluble in methanol and in acetic acid (100), soluble in ethanol (95), and practically insoluble in water. It is gradually colored by light. C H N S.H O: 170.19 A solution of Mequitazine in methanol (1 in 50) shows no 5 4 4 2 1,7-Dihydro-6H-purine-6-thione monohydrate optical rotation. [6112-76-1] Identification (1) Determine the absorption spectrum of a solution of Mequitazine in ethanol (95) (1 in 250,000) as di- Mercaptopurine Hydrate contains not less than rected under Ultraviolet-visible Spectrophotometry <2.24>, 98.0z of mercaptopurine (C5H4N4S: 152.18), calcu- and compare the spectrum with the Reference Spectrum: lated on the anhydrous basis. both spectra exhibit similar intensities of absorption at the Description Mercaptopurine Hydrate occurs as light yellow same wavelengths. to yellow crystals or crystalline powder. It is odorless. (2) Determine the infrared absorption spectrum of Me- It is practically insoluble in water, in acetone and in quitazine, previously dried, as directed in the potassium bro- diethyl ether. mide disk method under Infrared Spectrophotometry <2.25>, It dissolves in sodium hydroxide TS and in ammonia TS. and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the Identification (1) Dissolve 0.6 g of Mercaptopurine Hy- same wave numbers. drate in 6 mL of sodium hydroxide solution (3 in 100), and add slowly 0.5 mL of iodomethane with vigorous stirring. Melting point <2.60> 146–1509C Stir well for 10 minutes, cool in an ice bath, and adjust the Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of pH with acetic acid (31) to about 5. Collect the separated Mequitazine according to Method 2, and perform the test. crystals by filtration, recrystallize from water, and dry at Prepare the control solution with 2.0 mL of Standard Lead 1209C for 30 minutes: the crystals melt <2.60> between Solution (not more than 20 ppm). 2189Cand2229C (with decomposition). (2) Related substances—Conduct this procedure without (2) Determine the absorption spectrum of a solution of exposure to light, using light-resistant vessels. Dissolve 50 Mercaptopurine Hydrate in 0.1 mol/L hydrochloric acid TS mg of Mequitazine in 5 mL of methanol, and use this solu- (1 in 200,000) as directed under Ultraviolet-visible Spectro- tion as the sample solution. Pipet 1 mL of the sample solu- photometry <2.24>, and compare the spectrum with the Ref- tion, add methanol to make exactly 50 mL, then pipet 5 mL erence Spectrum: both spectra exhibit similar intensities of of this solution, add methanol to make exactly 50 mL, and absorption at the same wavelengths. use this solution as the standard solution. Perform the test Purity (1) Clarity of solution—Dissolve 0.20 g of Mer- with these solutions as directed under Thin-layer Chroma- captopurine Hydrate in 10 mL of ammonia TS: the solution tography <2.03>. Spot 5 mL each of the sample solution and is clear. standard solution on a plate of silica gel with fluorescent in- (2) Sulfate <1.14>—Dissolve 50 mg of Mercaptopurine dicator for thin-layer chromatography. Develop with a mix- Hydrate in 10 mL of dilute hydrochloric acid, add 5 drops of ture of ethyl acetate, methanol and diethylamine (7:2:2) to a barium chloride TS, and allow to stand for 5 minutes: no distance of about 10 cm, and air-dry the plate. Examine turbidity is produced. under ultraviolet light (main wavelength: 254 nm): the num- (3) Heavy metals <1.07>—Proceed with 1.0 g of Mercap- ber of the spot other than the principal spot from the sample topurine Hydrate according to Method 2, and perform the solution is not more than 3 and they are not more intense test. Prepare the control solution with 2.0 mL of Standard than the spot from the standard solution. Lead Solution (not more than 20 ppm). 1084 Mercurochrome / Official Monographs JP XVI

(4) Hypoxanthine—Dissolve 50 mg of Mercaptopurine Hydrate in exactly 10 mL of a solution of ammonia solution Mercurochrome (28) in methanol (1 in 10), and use this solution as the sample solution. Separately, dissolve 5.0 mg of hypoxanthine in a Merbromin solution of ammonia solution (28) in methanol (1 in 10) to make exactly 100 mL, and use this solution as the standard マーキュロクロム solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mLeach Mercurochrome is a sodium salt of a mixture of of the sample solution and standard solution on a plate of brominated and mercurized fluoresceins. silica gel with fluorescent indicator for thin-layer chromatog- When dried, it contains not less than 18.0z and not raphy. Develop the plate with a mixture of methanol, chlo- more than 22.4z of bromine (Br: 79.90), and not less roform, n-butyl formate and ammonia solution (28) than 22.4z and not more than 26.7z of mercury (Hg: (8:6:4:1) to a distance of about 10 cm, and air-dry the plate. 200.59). Examine under ultraviolet light (main wavelength: 254 nm): the spot from the sample solution observed at the same place Description Mercurochrome occurs as blue-green to green- as that from the standard solution, is not larger and not ish red-brown scales or granules. It is odorless. more intense than that from the standard solution. It is freely soluble in water, but sometimes leaves a small (5) Phosphorus—Take 0.20 g of Mercaptopurine Hy- amount of insoluble matter. It is practically insoluble in drate in a crucible, add 2 mL of diluted sulfuric acid (3 in 7), ethanol (95) and in diethyl ether. then heat gently, slowly adding dropwise several 0.5-mL Identification (1) A solution of Mercurochrome (1 in portions of nitric acid, until the liquid becomes colorless. 2000) shows a red color and a yellow-green fluorescence. Continue to heat until most of the liquid has evaporated, (2) To 5 mL of a solution of Mercurochrome (1 in 250) cool, and dissolve the residue in 10 mL of water. Transfer add 3 drops of dilute sulfuric acid: a reddish orange precipi- the solution to a 25-mL volumetric flask, wash the crucible tate is produced. with two 4-mL portions of water, combine the washings with (3) Heat 0.1 g of Mercurochrome with small crystals of the solution in the volumetric flask, and use this solution as iodine in a test tube: red crystals are sublimed on the upper the sample solution. Separately, dissolve 0.4396 g of potas- part of the tube. If yellow crystals are produced, scratch with sium dihydrogenphosphate in water to make exactly 200 mL. a glass rod: the color of the crystals changes to red. To 2.0 mL of this solution add water to make exactly 100 (4) Place 0.1 g of Mercurochrome in a porcelain cruci- mL. Transfer 2.0 mL of this solution to a 25-mL volumetric ble, add 1 mL of a solution of sodium hydroxide (1 in 6), flask, add 16 mL of water, and use this solution as the stand- evaporate to dryness with stirring, and ignite. Dissolve the ard solution. To the sample solution and standard solution residue in 5 mL of water, acidify with hydrochloric acid, and add 1 mL of diluted sulfuric acid (3 in 7), 0.5 mL of nitric shake with 3 drops of chlorine TS and 2 mL of chloroform: acid, 0.75 mL of hexaammonium heptamolybdate TS, 1 mL a yellowish brown color develops in the chloroform layer. of 1-amino-2-naphthol-4-sulfonic acid TS and water to make 25 mL, and allow to stand for 5 minutes. Perform the test Purity (1) Dyestuff—Dissolve 0.40 g of Mercurochrome with these solutions as directed under Ultraviolet-visible in 20 mL of water, add 3 mL of dilute sulfuric acid, and Spectrophotometry <2.24>, using water as the blank: the ab- filter: the filtrate has no more color than Matching Fluid C. sorbance of the subsequent solution of the sample solution (2) Soluble halides—Dissolve 5.0 g of Mercurochrome in at 750 nm is not larger than that of the subsequent solution 80 mL of water, add 10 mL of dilute nitric acid and water to of the standard solution. make 100 mL, shake, and filter. Transfer 40 mL of the filtrate to a Nessler tube, add 6 mL of dilute nitric acid and Water <2.48> 10.0 – 12.0z (0.2 g, volumetric titration, water to make 50 mL, then add 1 mL of silver nitrate TS, back titration). mix well, and allow to stand for 5 minutes protected from Residue on ignition <2.44> Not more than 0.1z (1 g). direct sunlight: no turbidity is produced, or even if produced, it is not more than that of the following control solu- Assay Weigh accurately about 0.25 g of Mercaptopurine tion. Hydrate, dissolve in 90 mL of N,N-dimethylformamide, and Control solution: To 0.25 mL of 0.01 mol/L hydrochloric titrate <2.50> with 0.1 mol/L tetramethylammonium hydrox- acid VS add 6 mL of dilute nitric acid and water to make 50 ide VS (potentiometric titration). Perform a blank determi- mL, then add 1 mL of silver nitrate TS, and proceed as di- nation with a mixture of 90 mL of N,N-dimethylformamide rected above. and 15 mL of water, and make any necessary correction. (3) Soluble mercury salts—To 5 mL of the filtrate ob- Each mL of 0.1 mol/L tetramethylammonium tained in (1) add 5 mL of water, and use this solution as the hydroxide VS sample solution. Dissolve 40 mg of mercury (II) chloride,

＝ 15.22 mg of C5H4N4S weighed accurately, in water to make 1000 mL, and add 3 mL of dilute sulfuric acid to 20 mL of this solution. To 5 mL Containers and storage Containers—Well-closed contain- of the solution add 5 mL of water, and use this as the control ers. solution. Add 1 drop each of sodium sulfide TS to these solutions, and compare: the sample solution has no more color than the control solution. (4) Insoluble mercury compounds—Dissolve 2.5 g of Mercurochrome in 50 mL of water, allow to stand for 24 hours, centrifuge, and wash the precipitate with small por- JP XVI Official Monographs / Meropenem Hydrate 1085 tions of water until the last washing becomes colorless. of water, and add 3 drops of dilute sulfuric acid: a red- Transfer the precipitate to a glass-stoppered flask, add ex- orange precipitate is produced. actly 5 mL of 0.05 mol/L iodine VS, allow to stand for 1 (3) Evaporate 5 mL of Mercurochrome Solution to dry- hour with frequent agitation, add 4.3 mL of 0.1 mol/L so- ness, and proceed with the residue as directed in the Identifi- dium thiosulfate VS dropwise with shaking, and add 1 mL of cation (3) under Mercurochrome. starch TS: a blue color develops. (4) To 5 mL of Mercurochrome Solution add 1 mL of a solution of sodium hydroxide (1 in 6), and proceed as di- Loss on drying <2.41> Not more than 5.0z (1 g, 1059C, rected in the Identification (4) under Mercurochrome. 5 hours). Purity Dyestuff—To 20 mL of Mercurochrome Solution Assay (1) Mercury—Weigh accurately about 0.6 g of add 3 mL of dilute sulfuric acid, and filter: the filtrate has Mercurochrome, previously powdered and dried, transfer to no more color than Matching Fluid C. an iodine flask, dissolve in 50 mL of water, add 8 mL of acetic acid (31), 20 mL of chloroform and exactly 30 mL of 0.05 Assay Transfer exactly measured 30 mL of Mercuro- mol/L iodine VS, stopper tightly, and allow to stand for 1 chrome Solution to an iodine flask, dilute with 20 mL of hour with frequent, vigorous shaking. Titrate <2.50> the ex- water, add 8 mL of acetic acid (31) and 20 mL of chlo- cess iodine with 0.1 mol/L sodium thiosulfate VS with roform, and proceed as directed in the Assay (1) under vigorous shaking (indicator: 1 mL of starch TS). Perform a Mercurochrome. blank determination, and make any necessary correction. Each mL of 0.05 mol/L iodine VS ＝ 10.03 mg of Hg Each mL of 0.05 mol/L iodine VS ＝ 10.03 mg of Hg Containers and storage Containers—Tight containers. (2) Bromine—Weigh accurately about 0.5 g of Mer- Storage—Light-resistant. curochrome, previously powdered and dried, in a porcelain crucible, add 2 g of potassium nitrate, 3 g of potassium carbonate and 3 g of anhydrous sodium carbonate, mix well, Meropenem Hydrate cover the surface of the mixture with 3 g of a mixture of equal amounts of potassium carbonate and anhydrous so- メロペネム水和物 dium carbonate, and ignite almost to fusion. Cool, dissolve the ignited mixture in 80 mL of warm water, acidify with nitric acid, and add exactly 25 mL of 0.1 mol/L silver nitrate VS. Shake well, and titrate <2.50> the excess silver nitrate with 0.1 mol/L ammonium thiocyanate VS (indicator: 2 mL of ammonium iron (III) sulfate TS). Perform a blank determination and make any necessary correction.

Each mL of 0.1 mol/L silver nitrate VS ＝ 7.990 mg of Br C17H25N3O5S.3H2O: 437.51 (4R,5S,6S)-3-[(3S,5S)-5-(Dimethylcarbamoyl)pyrrolidin- Containers and storage Containers—Tight containers. 3-ylsulfanyl]-6-[(1R)-1-hydroxyethyl]-4-methyl-7-oxo-1- Storage—Light-resistant. azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid trihydrate [119478-56-7]

Mercurochrome Solution Meropenem Hydrate contains not less than 980 mg (potency) and not more than 1010 mg (potency) per Merbromin Solution mg, calculated on the anhydrous basis. The potency of Meropenem Hydrate is expressed as mass (potency) of マーキュロクロム液 meropenem (C17H25N3O5S: 383.46). Description Meropenem Hydrate occurs as a white to light Mercurochrome Solution contains not less than yellow crystalline powder. 0.42 w/vz and not more than 0.56 w/vz of mercury It is sparingly soluble in water, and practically insoluble in (Hg: 200.59). ethanol (95) and in diethyl ether. Method of preparation Identification (1) Dissolve 10 mg of Meropenem Hydrate Mercurochrome 20 g in 2 mL of water, add 3 mL of hydroxylammonium chloride- Purified Water or Purified ethanol TS, allow to stand for 5 minutes, add 1 mL of acidic Water in Containers a sufficient quantity ammonium iron (III) sulfate TS, and shake: a red-brown To make 1000 mL color develops. (2) Determine the absorption spectra of solutions of Prepare by mixing the above ingredients. Meropenem Hydrate and Meropenem RS (3 in 100,000) as Description Mercurochrome Solution is a dark red liquid. directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectra: both spectra exhibit similar intensi- Identification (1) To 1 mL of Mercurochrome Solution ties of absorption at the same wavelengths. add 40 mL of water: the resulting solution shows a red color (3) Determine the infrared absorption spectra of and a yellow-green fluorescence. Meropenem Hydrate and Meropenem RS as directed in the (2) Dilute 1 mL of Mercurochrome Solution with 4 mL potassium bromide disk method under Infrared Spectropho- 1086 Meropenem Hydrate / Official Monographs JP XVI tometry <2.25>, and compare the spectra: both spectra ex- equivalent to 16 to 24z of that from 10 mL of the standard hibit similar intensities of absorption at the same wave num- solution. bers. System performance: Warm the sample solution at 609C for 30 minutes. When the procedure is run with 10 mLofthe Optical rotation <2.49> [a]20: －17 – －219(0.22 g calcu- D sample solution under the above operating conditions, the lated as the anhydrous basis, water, 50 mL, 100 mm). ring-opened meropenem, meropenem and the dimer are pH <2.54> Dissolve 0.2 g of Meropenem Hydrate in 20 mL eluted in this order, and the resolution between the peaks of of water: the pH of the solution is between 4.0 and 6.0. the ring-opened meropenem and meropenem is not less than 1.5. Purity (1) Clarity and color of solution—Dissolve 0.5 g System repeatability: When the test is repeated 6 times of Meropenem Hydrate in 10 mL of sodium hydrogen car- with 10 mL of the standard solution under the above operat- bonate TS: the solution is clear and has no more color than ing conditions, the relative standard deviation of the peak the following control solution. area of meropenem is not more than 1.5z. Control solution: To a mixture of 0.3 mL of Cobalt (II) Chloride CS and 1.2 mL of Iron (III) Chloride CS add 18.5 Water <2.48> Not less than 11.4z and not more than mLofdilutedhydrochloricacid(1in40). 13.4z (0.35 g, volumetric titration, direct titration). (2) Heavy metals <1.07>—Proceed with 2.0 g of Residue on ignition <2.44> Not more than 0.1z (1 g). Meropenem Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand- Assay Weigh accurately an amount of Meropenem Hy- ard Lead Solution (not more than 10 ppm). drate and Meropenem RS, equivalent to about 50 mg (po- (3) Related substances—Dissolve 50 mg of Meropenem tency), add exactly 10 mL of the internal standard solution Hydrate in 10 mL of triethylamine-phosphate buffer solu- to dissolve, add triethylamine-phosphate buffer solution, pH tion, pH 5.0, and use this solution as the sample solution. 5.0 to make 100 mL, and use these solutions as the sample Prepare the sample solution before use. Pipet 1 mL of the solution and the standard solution, respectively. Perform the sample solution, and add triethylamine-phosphate buffer so- test with 5 mL of the sample solution and standard solution lution, pH 5.0 to make exactly 100 mL. Pipet 3 mL of this as directed under Liquid Chromatography <2.01> according solution, add triethylamine-phosphate buffer solution, pH to the following conditions, and calculate the ratios, QT and 5.0 to make exactly 10 mL, and use this solution as the QS, of the peak area of meropenem to that of the internal standard solution. Perform the test with exactly 10 mLeach standard. of the sample solution and standard solution as directed Amount [mg (potency)] of meropenem (C H N O S) under Liquid Chromatography <2.01> according to the fol- 17 25 3 5 ＝ M × Q /Q × 1000 lowing conditions, and determine each peak area by the au- S T S tomatic integration method: the peak area of ring-opened MS: Amount [mg (potency)] of Meropenem RS meropenem, having the relative retention time about 0.5 to Internal standard solution—A solution of benzyl alcohol in meropenem, and the peak area of the dimmer, having the triethylamine-phosphate buffer solution, pH 5.0 (1 in 300). relative retention time about 2.2 to meropenem, obtained Operating conditions— from the sample solution are not larger than the peak area of Detector: An ultraviolet absorption photometer (wave- meropenem from the standard solution, the area of the peak length: 220 nm). other than meropenem and the peaks mentioned above from Column: A stainless steel column 6.0 mm in inside diame- the sample solution is not larger than 1/3 times the peak area ter and 15 cm in length, packed with octadecylsilanized silica of meropenem from the standard solution, and the total area gel for liquid chromatography (5 mminparticlediameter). of the peaks other than meropenem from the sample solu- Column temperature: A constant temperature of about tion is not larger than 3 times the peak area of meropenem 259C. from the standard solution. Mobile phase: A mixture of triethylamine-phosphate Operating conditions— buffer solution, pH 5.0 and methanol (5:1). Detector: An ultraviolet absorption photometer (wave- Flow rate: Adjust the flow rate so that the retention time length: 220 nm). of meropenem is about 7 minutes. Column: A stainless steel column 6.0 mm in inside diame- System suitability— ter and 15 cm in length, packed with octadecylsilanized silica System performance: When the procedure is run with 5 mL gel for liquid chromatography (5 mm in particle diameter). of the standard solution under the above operating condi- Column temperature: A constant temperature of about tions, meropenem and the internal standard are eluted in this 409C. order with the resolution between these peaks being not less Mobile phase: A mixture of triethylamine-phosphate than 20. buffer solution, pH 5.0 and acetonitrile (100:7). System repeatability: When the test is repeated 6 times Flow rate: Adjust the flow rate so that the retention time with 5 mL of the standard solution under the above operating of meropenem is about 6 minutes. conditions, the relative standard deviation of the ratios of Time span of measurement: About 7 times as long as the the peak area of meropenem to that of the internal standard retention time of meropenem. is not more than 1.0z. System suitability— Test for required detectability: Pipet 5 mL of the standard Containers and storage Containers—Tight containers. solution, and add triethylamine-phosphate buffer solution, pH 5.0 to make exactly 25 mL. Confirm that the peak area of meropenem obtained from 10 mL of this solution is JP XVI Official Monographs / Mestranol 1087

ard solution. Perform the test with 5 mLeachofthesample Meropenem for Injection solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-

注射用メロペネム tions, and calculate the ratios, QT and QS, of the peak area of meropenem to that of the internal standard.

Meropenem for Injection is a preparation for injec- Amount [mg (potency)] of meropenem (C17H25N3O5S) tion, which is dissolved before use. ＝ MS × QT/QS It contains not less than 93.0z and not more M : Amount [mg (potency)] of Meropenem RS than 107.0z of the labeled potency of meropenem S (C17H25N3O5S: 383.46). Internal standard solution—A solution of benzyl alcohol in triethylamine-phosphate buffer solution, pH 5.0 (1 in 300). Method of preparation Prepare as directed under Injec- Operating conditions— tions, with Meropenem Hydrate. Proceed as directed in the operating conditions in the Description Meropenem for Injection occurs as a white to Assay under Meropenem Hydrate. light yellow crystalline powder. System suitability— System performance: Proceed as directed in the system Identification Determine the infrared absorption spectrum suitability in the Assay under Meropenem Hydrate. of Meropenem for Injection as directed in the potassium System repeatability: When the test is repeated 6 times bromide disk method under Infrared Spectrophotometry with 5 mL of the standard solution under the above operating <2.25>: it exhibits absorption at the wave numbers of about conditions, the relative standard deviation of the ratio of the 3410 cm－1, 1750 cm－1, 1655 cm－1, 1583 cm－1 and 1391 peak area of meropenem to that of the internal standard is cm－1. not more than 1.0z. pH <2.54> Dissolve an amount of Meropenem for Injec- Containers and storage Containers—Hermetic containers. tion, equivalent to 0.25 g (potency) of Meropenem Hydrate Plastic containers for aqueous injections may be used. according to the labeled amount, in 5 mL of water: the pH of the solution is between 7.3 and 8.3. Purity (1) Clarity and color of solution—Dissolve an Mestranol amount of Meropenem for Injection, equivalent to 1.0 g (potency) of Meropenem Hydrate according to the labeled メストラノール amount, in 20 mL of water: the solution is clear and is not more intensely colored than the following matching fluid. Matching fluid: To a mixture of 0.3 mL of Cobalt (II) Chloride CS and 1.2 mL of Iron (III) Chloride CS add 18.5 mLofdilutedhydrochloricacid(1in40). (2) Related substances—Being specified separately.

Loss on drying <2.41> 9.5–12.0z (0.1 g, reduced pressure C21H26O2:310.43 not exceeding 0.67 kPa, 609C, 3 hours). 3-Methoxy-19-nor-17a-pregna-1,3,5(10)-trien-20-yn-17-ol [72-33-3] Bacterial endotoxins <4.01> Less than 0.12 EU/mg (potency). Mestranol, when dried, contains not less than Uniformity of dosage units <6.02> It meets the requirement 97.0z and not more than 102.0z of C21H26O2. of the Mass variation test. Description Mestranol occurs as a white to pale yellowish Foreign insoluble matter <6.06> Perform the test according white, crystalline powder. It is odorless. to Method 2: it meets the requirement. It is freely soluble in chloroform, soluble in 1,4-dioxane, sparingly soluble in ethanol (99.5) and in diethyl ether, and Insoluble particulate matter <6.07> It meets the require- practically insoluble in water. ment. Identification (1) Dissolve 2 mg of Mestranol in 1 mL of Sterility <4.06> Perform the test according to the Mem- a mixture of sulfuric acid and ethanol (99.5) (2:1): a red- brane filtration method: it meets the requirement. purple color develops with a yellow-green fluorescence. Assay Weigh accurately the mass on the contents of not (2) Determine the absorption spectrum of a solution of less than 10 containers of Meropenem for Injection. Weigh Mestranol in ethanol (99.5) (1 in 10,000) as directed under accurately an amount of the contents, equivalent to about Ultraviolet-visible Spectrophotometry <2.24>,andcompare 50 mg (potency) of Meropenem Hydrate, dissolve in exactly the spectrum with the Reference Spectrum or the spectrum 10 mL of the internal standard solution, add triethylamine- of a solution of Mestranol RS prepared in the same manner phosphate buffer solution, pH 5.0, to make 100 mL, and use as the sample solution: both spectra exhibit similar intensi- this solution as the sample solution. Separately, weigh accu- ties of absorption at the same wavelengths. rately an amount of Meropenem RS, equivalent to about 50 (3) Determine the infrared absorption spectrum of Mes- mg (potency), dissolve in exactly 10 mL of the internal stand- tranol, previously dried, as directed in the potassium bro- ard solution, add triethylamine-phosphate buffer solution, mide disk method under Infrared Spectrophotometry <2.25>, pH 5.0, to make 100 mL, and use this solution as the stand- and compare the spectrum with the Reference Spectrum or 1088 Metenolone Acetate / Official Monographs JP XVI the spectrum of previously dried Mestranol RS: both spectra exhibit similar intensities of absorption at the same wave Metenolone Acetate numbers.

20 メテノロン酢酸エステル Optical rotation <2.49> [a]D : ＋2–＋89 (after drying, 0.2 g, 1,4-dioxane, 10 mL, 100 mm). Melting point <2.60> 148–1549C Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Mestranol according to Method 2, and perform the test. Pre- pare the control solution with 2.0 mL of Standard Lead So- lution (not more than 20 ppm). (2) Arsenic <1.11>—Prepare the test solution with 1.0 g of Mestranol according to Method 3, and perform the test C22H32O3:344.49 (not more than 2 ppm). 1-Methyl-3-oxo-5a-androst-1-en-17b-yl acetate (3) Related substances—Dissolve 0.10 g of Mestranol in [434-05-9] 20 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add chloroform to Metenolone Acetate, when dried, contains not less make exactly 200 mL, and use this solution as the standard than 97.0z and not more than 103.0z of C22H32O3. solution. Perform the test with these solutions as directed Description Metenolone Acetate occurs as a white to pale under Thin-layer Chromatography <2.03>. Spot 10 mLeach yellowish white, crystalline powder. It is odorless. of the sample solution and standard solution on a plate of It is freely soluble in acetone, in 1,4-dioxane and in chlo- silica gel for thin-layer chromatography. Develop the plate roform, soluble in methanol and in ethanol (95), sparingly with a mixture of chloroform and ethanol (99.5) (29:1) to a soluble in diethyl ether and in sesame oil, slightly soluble in distance of about 10 cm, and air-dry the plate. Spray evenly hexane and in petroleum ether, and practically insoluble in diluted sulfuric acid (1 in 5) on the plate, and heat at 1059C water. for 15 minutes: the spots other than the principal spot from the sample solution are not more intense than the spot from Identification (1) Dissolve 1 mg of Metenolone Acetate in the standard solution. 5 mL of a mixture of ethanol (95) and sulfuric acid (1:1), and heat for 30 minutes in a water bath: a red-brown color de- Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, velops. 3 hours). (2) To 10 mg of Metenolone Acetate add 0.5 mL of Residue on ignition <2.44> Not more than 0.1z (0.5 g). dilute sodium hydroxide-ethanol TS, and heat for 1 minute on a water bath. After cooling, add 0.5 mL of diluted sulfu- Assay Weigh accurately about 10 mg each of Mestranol ric acid (1 in 2), and boil gently for 1 minute: the odor of and Mestranol RS, previously dried, dissolve in ethanol ethyl acetate is perceptible. (99.5) to make exactly 100 mL, and use these solutions as the (3) Dissolve 50 mg of Metenolone Acetate in 3 mL of sample solution and the standard solution, respectively. De- methanol, add 0.3 mL of a solution of potassium carbonate termine the absorbances, A and A , of the sample solution T S (1 in 6), and boil for 2 hours under a reflux condenser. After and the standard solution at 279 nm as directed under Ultra- cooling, add this solution gradually to 50 mL of cold water, violet-visible Spectrophotometry <2.24>. and stir for 15 minutes. Filter the precipitate so obtained by

Amount (mg) of C21H26O2 ＝ MS × AT/AS suction through a glass filter (G4), wash with 10 mL of water, and dry at 1059C for 1 hour: it melts <2.60> between M : Amount (mg) of Mestranol RS S 1579Cand1619C. Containers and storage Containers—Tight containers. (4) Determine the infrared absorption spectrum of Storage—Light-resistant. Metenolone Acetate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho- tometry <2.25>, and compare the spectrum with the Refer- ence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.

20 Optical rotation <2.49> [a]D : ＋39 – ＋429(after drying, 0.2 g, chloroform, 10 mL, 100 mm). Melting point <2.60> 141 – 1449C Purity (1) Clarity and color of solution—Dissolve 0.50 g of Metenolone Acetate in 10 mL of 1,4-dioxane: the solution is clear and colorless to pale yellow. (2) Heavy metals <1.07>—Proceed with 2.0 g of Meteno- lone Acetate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (3) Related substances—Dissolve 35 mg of Metenolone Acetate in 20 mL of chloroform, and use this solution as the JP XVI Official Monographs / Metenolone Enanthate Injection 1089 sample solution. Pipet 1 mL of the sample solution, dilute minutes. Filter the produced precipitate by suction through a with chloroform to exactly 250 mL, and use this solution as glass filter (G4), wash with water until the washings become the standard solution. Perform the test with these solutions neutral, and dry at 1059C for 1 hour: it melts <2.60> between as directed under Thin-layer Chromatography <2.03>. Spot 1569Cand1629C. 10 mL each of the sample solution and standard solution on a Optical rotation <2.49> [a]20: ＋39 – ＋439(after drying, plate of silica gel with fluorescent indicator for thin-layer D 0.2 g, chloroform, 10 mL, 100 mm). chromatography. Develop the plate with a mixture of ethyl acetate and cyclohexane (1:1) to a distance of about 12 cm, Melting point <2.60> 67–729C and air-dry the plate. Examine under ultraviolet light (main Purity (1) Clarity and color of solution—Dissolve 0.5 g wavelength: 254 nm): the spots other than the principal spot of Metenolone Enanthate in 10 mL of 1,4-dioxane: the solu- from the sample solution are not more intense than the spot tion is clear and colorless. from the standard solution. (2) Heavy metals <1.07>—Proceed with 2.0 g of Meteno- Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, lone Enanthate according to Method 2, and perform the test. 3 hours). Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Residue on ignition <2.44> Not more than 0.1z (0.5 g). (3) Related substances—Dissolve 20 mg of Metenolone Assay Weigh accurately about 10 mg of Metenolone Ace- Enanthate in exactly 10 mL of chloroform, and use this solu- tate, previously dried, and dissolve in methanol to make ex- tion as the sample solution. Perform the test with the sample actly 100 mL. Pipet 5 mL of this solution, and dilute with solution as directed under Thin-layer Chromatography methanol to exactly 50 mL. Determine the absorbance A of <2.03>. Spot 10 mL of the sample solution on a plate of silica this solution at the wavelength of maximum absorption at gel with fluorescent indicator for thin-layer chromatogra- about 242 nm as directed under Ultraviolet-visible Spectro- phy. Develop the plate with a mixture of ethyl acetate and photometry <2.24>. cyclohexane (1:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: Amount (mg) of C H O ＝ A/391 × 10,000 22 32 3 254 nm): any spot other than the principal spot does not Containers and storage Containers—Tight containers. appear. Storage—Light-resistant. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacuum, phosphorus (V) oxide, 4 hours). Metenolone Enanthate Residue on ignition <2.44> Not more than 0.1z (0.5 g). Assay Weigh accurately about 0.1 g of Metenolone Enan- メテノロンエナント酸エステル thate, previously dried, and dissolve in methanol to make exactly 100 mL. Pipet 10 mL of this solution, and dilute with methanol to make exactly 100 mL. Pipet 10 mL of this solution, and dilute again with methanol to make exactly 100 mL. Determine the absorbance, A, of this solution at the wavelength of maximum absorption at about 242 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>.

Amount (mg) of C27H42O3 ＝ A/325 × 100,000

C27H42O3:414.62 Containers and storage Containers—Tight containers. 1-Methyl-3-oxo-5a-androst-1-en-17b-yl heptanoate Storage—Light-resistant. [303-42-4]

Metenolone Enanthate, when dried, contains not Metenolone Enanthate Injection less than 97.0z and not more than 103.0z of C27H42O3. メテノロンエナント酸エステル注射液 Description Metenolone Enanthate occurs as white crystals or crystalline powder. It is odorless. Metenolone Enanthate Injection is an oily solution It is very soluble in ethanol (95), in acetone, in 1,4-dioxane for injection. and in chloroform, freely soluble in methanol, in ethyl ace- It contains not less than 90.0z and not more than tate, in diethyl ether, in cyclohexane, in petroleum ether and 110.0z of the labeled amount of metenolone enan- in toluene, soluble in sesame oil, and practically insoluble in thate (C27H42O3: 414.62). water. Method of preparation Prepare as directed under Injec- Identification (1) Heat 1 mg of Metenolone Enanthate tions, with Metenolone Enanthate. with 5 mL of a mixture of ethanol (95) and sulfuric acid (1:1) Description Metenolone Enanthate Injection is a clear, on a water bath for 30 minutes: a red-brown color develops. pale yellow, oily liquid. (2) Dissolve 0.05 g of Metenolone Enanthate in 3 mL of methanol, add 0.3 mL of a solution of potassium carbonate Identification (1) Measure a volume of Metenolone (1 in 6), boil under a reflux condenser for 2 hours, cool, add Enanthate Injection, equivalent to 0.1 g of Metenolone slowly this solution to 50 mL of cold water, and stir for 15 Enanthate according to the labeled amount, add 20 mL of 1090 Metformin Hydrochloride / Official Monographs JP XVI petroleum ether, and extract with three 20-mL portions of diluted acetic acid (100) (5 in 7). Combine the extracts, wash Metformin Hydrochloride with 20 mL of petroleum ether, add 300 mL of cold water while cooling in an ice bath, and stir sufficiently. Filter the メトホルミン塩酸塩 produced precipitate by suction through a glass filter (G4), wash with water until the last washing becomes neutral, and dry in a desiccator (in vacuum, phosphorus (V) oxide) for 6 hours. With this sample, proceed as directed in the Identifi- cation (1) under Metenolone Enanthate.

(2) Measure a volume of Metenolone Enanthate Injec- C4H11N5.HCl: 165.62 tion, equivalent to 10 mg of Metenolone Enanthate accord- 1,1-Dimethylbiguanide monohydrochloride ing to the labeled amount, dissolve in 10 mL of chloroform, [1115-70-4] and use this solution as the sample solution. Separately dissolve 10 mg of metenolone enanthate in 10 mL of chlo- Metformin Hydrochloride, when dried, contains roform, and use this solution as the standard solution. Per- not less than 98.5z and not more than 101.0z of form the test with these solutions as directed under Thin- C4H11N5.HCl. layer Chromatography <2.03>. Spot 10 mLeachofthesample Description Metformin Hydrochloride occurs as white solution and standard solution on a plate of silica gel with crystals or crystalline powder. fluorescent indicator for thin-layer chromatography. De- It is freely soluble in water, sparingly soluble in acetic acid velop the plate with toluene to a distance of about 15 cm, (100), and slightly soluble in ethanol (99.5). and air-dry the plate. Again develop this plate with a mixture Melting point: about 2219C (with decomposition). of ethyl acetate and cyclohexane (1:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light Identification (1) Determine the absorption spectrum of a (main wavelength: 254 nm): the principal spot from the sam- solution of Metformin Hydrochloride (1 in 100,000) as di- ple solution and the spot from the standard solution show rected under Ultraviolet-visible Spectrophotometry <2.24>, the same Rfvalue. and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the Extractable volume <6.05> It meets the requirement. same wavelengths. Assay To an exactly measured volume of Metenolone (2) Determine the infrared absorption spectrum of Met- Enanthate Injection, equivalent to about 0.1 g of meteno- formin Hydrochloride as directed in the potassium chloride lone enanthate (C27H42O3), add chloroform to make exactly disk method under Infrared Spectrophotometry <2.25>,and 100 mL. Pipet 5 mL of this solution, add chloroform to compare the spectrum with the Reference Spectrum: both make exactly 50 mL, and use this solution as the sample so- spectra exhibit similar intensities of absorption at the same lution. Weigh accurately about 0.1 g of metenolone enan- wave numbers. thate for assay, previously dried in a desiccator (in vacuum, (3) A solution of Metformin Hydrochloride (1 in 50) phosphorus (V) oxide) for 4 hours, and prepare the standard responds to the Qualitative Tests <1.09> for chloride. solution in the same manner as directed for the preparation Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of of the sample solution. Pipet 3 mL each of the sample solu- Metformin Hydrochloride according to Method 1, and per- tion and standard solution, and treat each solution as form the test. Prepare the control solution with 2.0 mL of follows: add 10 mL of isoniazid TS, exactly measured, add Standard Lead Solution (not more than 10 ppm). methanol to make exactly 20 mL, and allow to stand for 60 (2) Related substances—Dissolve 2.5 g of Metformin minutes. Determine the absorbances, A and A ,ofthe T S Hydrochloride in 10 mL of water, and use this solution as solutions from the sample solution and standard solution, the sample solution. Pipet 1 mL of the sample solution, and respectively, at 384 nm as directed under Ultraviolet-visible add water to make exactly 50 mL. Pipet 1 mL of this solu- Spectrophotometry <2.24>, using a solution obtained by tion, add water to make exactly 10 mL, and use this solution proceeding with 3 mL of chloroform as the blank. as the standard solution (1). Pipet 5 mL of the standard solu-

Amount (mg) of metenolone enanthate (C27H42O3) tion (1), add water to make exactly 10 mL, and use this solu- ＝ MS × AT/AS tion as the standard solution (2). Separately, to 0.10 g of 1- cyanoguanidine add water to make exactly 50 mL. Pipet 1 M : Amount (mg) of metenolone enanthate for assay S mL of this solution, add water to make exactly 20 mL, and Containers and storage Containers—Hermetic containers. use this solution as the standard solution (3). Perform the Storage—Light-resistant. test with these solutions as directed under Thin-layer Chro- matography <2.03>. Spot 10 mL each of the sample solution and standard solutions (1), (2) and (3) on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of 4-methyl-2-pentanone, 2-methoxyethanol, water and acetic acid (100) (30:20:5:3) to a distance of about 10 cm, air-dry the plate, then dry at 1059C for 10 minutes. Spray evenly sodium pentacyanonitrosylferrate (III)-potassium hexacyanoferrate (III) TS on the plate: the spot other than the principal spot with the sample solution is not more intense than the spot with the standard solution (1), the num- JP XVI Official Monographs / Methamphetamine Hydrochloride 1091 ber of them showing more intense than the spot with the 3 mL of this solution, add exactly 3 mL of the internal stand- standard solution (2) is not more than two, and the spot with ard solution and the mixture of water and acetonitrile (3:2) the sample solution appeared at the position corresponding to make 50 mL, and use this solution as the standard solu- to the spot with the standard solution (3) is not more intense tion. Perform the test with 5 mL each of the sample solution than the spot with the standard solution (3). and standard solution as directed under Liquid Chromatog- raphy <2.01> according to the following conditions, and cal- Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, culate the ratios, Q and Q , of the peak area of metformin 3 hours). T S to that of the internal standard. Residue on ignition <2.44> Not more than 0.1z (1 g). Amount (mg) of metformin hydrochloride (C4H11N5.HCl) Assay Weigh accurately about 0.1 g of Metformin Hydro- ＝ MS × QT/QS chloride, previously dried, dissolve in 40 mL of acetic acid M : Amount (mg) of metformin hydrochloride for assay (100), add 40 mL of acetic anhydride, and titrate <2.50> with S 0.05 mol/L perchloric acid VS (potentiometric titration). Internal standard solution—Dissolve 0.3 g of isobutyl par- Perform a blank determination in the same manner, and ahydroxybenzoate in 100 mL of the mixture of water and make any necessary correction. acetonitrile (3:2). Operating conditions— Each mL of 0.05 mol/L perchloric acid VS Detector: An ultraviolet absorption photometer (wave- ＝ 4.141 mg of C H N .HCl 4 11 5 length: 235 nm). Containers and storage Containers—Tight containers. Column: A stainless steel column 4.6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticlediameter). Metformin Hydrochloride Tablets Column temperature: A constant temperature of about 409C. メトホルミン塩酸塩錠 Mobile phase: Dissolve 0.8 g of sodium lauryl sulfate in 620 mL of diluted phosphoric acid (1 in 2500), and add 380 mL of acetonitrile. Metformin Hydrochloride Tablets contain not less Flow rate: Adjust the flow rate so that the retention time than 95.0z and not more than 105.0z of the labeled of metformin is about 10 minutes. amount of metformin hydrochloride (C H N .HCl: 4 11 5 System suitability— 165.62). System performance: When the procedure is run with 5 mL Method of preparation Prepare as directed under Tablets, of the standard solution under the above operating condi- with Metformin Hydrochloride. tions, metformin and the internal standard are eluted in this order with the resolution between these peaks being not less Identification Shake an amount of powdered Metformin than 6. Hydrochloride Tablets, equivalent to 250 mg of Metformin System repeatability: When the test is repeated 6 times Hydrochloride according to the labeled amount, with 25 mL with 5 mL of the standard solution under the above operating of 2-propanol, and filter. Evaporate the filtrate under conditions, the relative standard deviation of the ratio of the reduced pressure in a water bath at 409C, and determine the peak area of metformin to that of the internal standard is infrared absorption spectrum of the residue as directed in the not more than 1.0z. potassium chloride disk method under Infrared Spectropho- tometry <2.25>: it exhibits absorption at the wave numbers of Containers and storage Containers—Well-closed contain- about 3370 cm－1, 3160 cm－1,1627cm－1,1569cm－1 and ers. 1419 cm－1. Uniformity of dosage unit <6.02> It meets the requirement of the Mass variation test. Methamphetamine Hydrochloride Dissolution Being specified separately. メタンフェタミン塩酸塩 Assay Weigh accurately the mass of not less than 20 Met- formin Hydrochloride Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.15 g of metformin hydrochloride (C4H11N5.HCl), add 70 mL of a mixture of water and acetonitrile (3:2), shake for 10 minutes, C10H15N.HCl: 185.69 add the mixture of water and acetonitrile (3:2) to make ex- (2S)-N-Methyl-1-phenylpropan-2-amine actly 100 mL, and filter through a membrane filter with a monohydrochloride pore size of not more than 0.45 mm. Discard the first 10 mL [51-57-0] of the filtrate, pipet 3 mL of the subsequent filtrate, add exactly 3 mL of the internal standard solution and the mixture Methamphetamine Hydrochloride, when dried, con- of water and acetonitrile (3:2) to make 50 mL, and use this tains not less than 98.5z of C10H15N.HCl. solution as the sample solution. Separately, weigh accurately Description Methamphetamine Hydrochloride occurs as about 0.15 g of metformin hydrochloride for assay, previ- colorless crystals or a white, crystalline powder. It is odor- ously dried at 1059C for 3 hours, and dissolve in the mixture less. of water and acetonitrile (3:2) to make exactly 100 mL. Pipet It is freely soluble in water, in ethanol (95) and in chlo- 1092 L-Methionine / Official Monographs JP XVI roform, and practically insoluble in diethyl ether. Description L-Methionine occurs as white crystals or crys- The pH of a solution of Methamphetamine Hydrochloride talline powder. It has a characteristic odor. (1 in 10) is between 5.0 and 6.0. It is freely soluble in formic acid, soluble in water, and very slightly soluble in ethanol (95). Identification (1) To 5 mL of a solution of Metham- It dissolves in dilute hydrochloric acid. phetamine Hydrochloride (1 in 100) add 0.5 mL of hydrogen hexachloroplatinate (IV) TS: an orange-yellow, crystalline Identification Determine the infrared absorption spectrum precipitate is produced. of L-Methionine, previously dried, as directed in the potas- (2) To 5 mL of a solution of Methamphetamine Hydro- sium bromide disk method under Infrared Spectrophotome- chloride (1 in 100) add 0.5 mL of iodine TS: a brown precipi- try <2.25>, and compare the spectrum with the Reference tate is produced. Spectrum: both spectra exhibit similar intensities of absorp- (3) To 5 mL of a solution of Methamphetamine Hydro- tion at the same wave numbers. chloride (1 in 100) add 0.5 mL of 2,4,6-trinitrophenol TS: a Optical rotation <2.49> [a]20: ＋21.0 – ＋25.09(after dry- yellow, crystalline precipitate is produced. D ing, 0.5 g, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm). (4) A solution of Methamphetamine Hydrochloride (1 in 20) responds to the Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 0.5 g of L-Methionine in 20 mL of

20 water: the pH of this solution is between 5.2 and 6.2. Optical rotation <2.49> [a]D : ＋16 – ＋199(after drying, 0.2 g, water, 10 mL, 100 mm). Purity (1) Clarity and color of solution—Dissolve 0.5 g of L-Methionine in 20 mL of water: the solution is clear and Melting point <2.60> 171–1759C colorless. Purity (1) Acidity or alkalinity—Dissolve 2.0 g of (2) Chloride <1.03>—Dissolve 0.5 g of L-Methionine in Methamphetamine Hydrochloride in 40 mL of freshly boiled 20 mL of water, and add 6 mL of dilute nitric acid and water and cooled water, add 2 drops of methyl red TS, and use this to make 40 mL. Perform the test using this solution as the solution as the sample solution. test solution. Prepare the control solution with 0.30 mL of (i) To 20 mL of the sample solution add 0.20 mL of 0.01 0.01 mol/L hydrochloric acid VS, 6 mL of dilute nitric acid mol/L sulfuric acid VS: a red color develops. and water to make 40 mL. In this test, to the test solution (ii) To 20 mL of the sample solution add 0.20 mL of 0.02 and the control solution add 10 mL each of silver nitrate TS mol/L sodium hydroxide VS: a yellow color develops. (not more than 0.021z). (2) Sulfate <1.14>—Dissolve 0.05 g of Methamphetamine (3) Sulfate <1.14>—Perform the test with 0.6 g of L- Hydrochloride in 40 mL of water, add 1 mL of dilute hydro- Methionine. Prepare the control solution with 0.35 mL of chloric acid and 1 mL of barium chloride TS, and allow to 0.005 mol/L sulfuric acid VS (not more than 0.028z). stand for 10 minutes: the solution remains unchanged. (4) Ammonium <1.02>—Perform the test with 0.25 g of L-Methionine. Prepare the control solution with 5.0 mL of Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, Standard Ammonium Solution (not more than 0.02z). 2 hours). (5) Heavy metals <1.07>—Dissolve 1.0 g of L-Methionine Residue on ignition <2.44> Not more than 0.1z (1 g). in 40 mL of water and 2 mL of dilute acetic acid, dissolve by warming, cool, and add water to make 50 mL. Perform the Assay Weigh accurately about 0.4 g of Methamphetamine test using this solution as the test solution. Prepare the con- Hydrochloride, previously dried, and dissolve in 50 mL of a trol solution as follows: to 2.0 mL of Standard Lead Solu- mixture of acetic anhydride and acetic acid (100) (7:3). tion add 2 mL of dilute acetic acid and water to make 50 mL Titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio- (not more than 20 ppm). metric titration). Perform a blank determination, and make (6) Arsenic <1.11>—Transfer 1.0 g of L-Methionine to a any necessary correction. 100-mL decomposition flask, add 5 mL of nitric acid and 2 Each mL of 0.1 mol/L perchloric acid VS mL of sulfuric acid, put a small funnel on the mouth of the

＝ 18.57 mg of C10H15N.HCl flask, and heat carefully until white fumes are evolved. After cooling, add two 2-mL portions of nitric acid, heat, add Containers and storage Containers—Tight containers. 2-mL portions of hydrogen peroxide (30) several times, and Storage—Light-resistant. heat until the solution becomes colorless or pale yellow. After cooling, add 2 mL of saturated ammonium oxalate monohydrate solution, and heat again until white fumes are L-Methionine evolved. After cooling, add water to make 5 mL, and perform the test with this solution as the test solution (not more L-メチオニン than 2 ppm). (7) Related substances—Dissolve 0.10 g of L-Methionine in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add water to

C5H11NO2S: 149.21 make exactly 50 mL. Pipet 5 mL of this solution, add water (2S)-2-Amino-4-(methylsulfanyl)butanoic acid to make exactly 20 mL, and use this solution as the standard [63-68-3] solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mLeachof L-Methionine, when dried, contains not less than the sample solution and standard solution on a plate of silica 98.5z of C5H11NO2S. gel for thin-layer chromatography. After air-drying, immediately develop the plate with a mixture of 1-butanol, water JP XVI Official Monographs / Methotrexate Capsules 1093 and acetic acid (100) (3:1:1) to a distance of about 10 cm, intensities of absorption at the same wave numbers. and dry the plate at 809C for 30 minutes. Spray evenly a so- Water <2.48> Take 5 mL of pyridine for water determina- lution of ninhydrin in acetone (1 in 50) on the plate, and heat tion and 20 mL of methanol for Karl Fischer method in a at 809C for 5 minutes: the spots other than the principal spot dried titration flask, and titrate with water determination TS from the sample solution are not more intense than the spot until the end point. Weigh accurately about 0.2 g of from the standard solution. Methotrexate, immediately place in the titration flask, and Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, add a known excess volume of Karl Fischer TS. Mix well for 3 hours). 30 minutes, and perform the test: the water content is not more than 12.0z. Residue on ignition <2.44> Not more than 0.1z (1 g). Residue on ignition <2.44> Not more than 0.1z (0.5 g). Assay Weigh accurately about 0.15 g of L-Methionine, previously dried, and dissolve in 3 mL of formic acid, add 50 Assay Weigh accurately about 25 mg each of Methotrexate mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L and Methotrexate RS, dissolve in the mobile phase to make perchloric acid VS (potentiometric titration). Perform a exactly 250 mL, and use these solutions as the sample solu- blank determination, and make any necessary correction. tion and standard solution. Perform the test with exactly 10 mL each of these solutions as directed under Liquid Chroma- Each mL of 0.1 mol/L perchloric acid VS tography <2.01> according to the following conditions, and ＝ 14.92 mg of C5H11NO2S determine the peak areas, AT and AS, of methotrexate in Containers and storage Containers—Tight containers. each solution.

Amount (mg) of C20H22N8O5 ＝ MS × AT/AS

Methotrexate MS: Amount (mg) of Methotrexate RS, calculated on the anhydrous basis メトトレキサート Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 302 nm). Column: A stainless steel column 4.6 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. C H N O : 454.44 20 22 8 5 Mobile phase: A mixture of disodium hydrogen phos- N-{4-[(2,4-Diaminopteridin- phate-citric acid buffer solution, pH 6.0 and acetonitrile 6-ylmethyl)(methyl)amino]benzoyl}-L-glutamic acid (89:11). [59-05-2] Flow rate: Adjust the flow rate so that the retention time of methotrexate is about 8 minutes. Methotrexate is a mixture of 4-amino-10-methylfolic System suitability— acid and closely related compounds. System performance: Dissolve 10 mg each of Metho- It contains not less than 94.0z and not more than trexate and folic acid in 100 mL of the mobile phase. When 102.0z of C H N O , calculated on the anhydrous 20 22 8 5 the procedure is run with 10 mL of this solution under the basis. above operating conditions, folic acid and methotrexate are Description Methotrexate occurs as a yellow-brown, crys- eluted in this order with the resolution between these peaks talline powder. being not less than 8. It is slightly soluble in pyridine, and practically insoluble System repeatability: When the test is repeated 6 times in water, in acetonitrile, in ethanol (95) and in diethyl ether. with 10 mL of the standard solution under the above operat- It dissolves in dilute sodium hydroxide TS and in dilute so- ing conditions, the relative standard deviation of the peak dium carbonate TS. area of methotrexate is not more than 1.0z. It is gradually affected by light. Containers and storage Containers—Tight containers. Identification (1) Dissolve 1 mg of Methotrexate in 100 Storage—Light-resistant. mL of 0.1 mol/L hydrochloric acid TS. Determine the absorption spectrum of this solution as directed under Ultra- violet-visible Spectrophotometry <2.24>, and compare the Methotrexate Capsules spectrum with the Reference Spectrum or the spectrum of a solution of Methotrexate RS prepared in the same manner as メトトレキサートカプセル the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Methotrexate Capsules contain not less than 95.0z (2) Determine the infrared absorption spectrum of and not more than 105.0z of the labeled amount of Methotrexate as directed in the potassium bromide disk methotrexate (C H N O : 454.44). method under Infrared Spectrophotometry <2.25>,and 20 22 8 5 compare the spectrum with the Reference Spectrum or the Method of preparation Prepare as directed under Cap- spectrum of Methotrexate RS: both spectra exhibit similar sules, with Methotrexate. 1094 Methotrexate Capsules / Official Monographs JP XVI

Identification To an amount of the content of trate, add water to make exactly V?mL so that each mL con-

Methotrexate Capsules, equivalent to 2 mg of Methotrexate tains about 2.2 mg of methotrexate (C20H22N8O5) according according to the labeled amount, add 100 mL of 0.1 mol/L to the labeled amount, and use this solution as the sample hydrochloric acid TS, shake, and filter. To 10 mL of the fil- solution. Separately, weigh accurately about 10 mg of trate add 0.1 mol/L hydrochloric acid TS to make 20 mL, Methotrexate RS (separately determine the water <2.48> in and determine the absorption spectrum of this solution as di- the same manner as Methotrexate), and dissolve in the mo- rected under Ultraviolet-visible Spectrophotometry <2.24>:it bile phase to make exactly 100 mL. Pipet 2 mL of this solu- exhibits maxima between 240 nm and 244 nm and between tion, add water to make exactly 100 mL, and use this solu- 304 nm and 308 nm. tion as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as Uniformity of dosage units <6.02> Perform the test accord- directed under Liquid Chromatography <2.01> according to ing to the following method: it meets the requirement of the the following conditions, and determine the peak areas, A Content uniformity test. T and A , of methotrexate of both solutions. To the content of 1 capsule of Methotrexate Capsules add S 3V/5 mL of the mobile phase, agitate with the aid of ultra- Dissolution rate (z) with respect to the labeled sonic waves for 15 minutes, then shake for 25 minutes, and amount of methotrexate (C20H22N8O5) add the mobile phase to make exactly V mL so that each mL ＝ MS × AT/AS × V?/V × 1/C × 18 contains about 20 mg of methotrexate (C H N O ). Centri- 20 22 8 5 M : Amount (mg) of Methotrexate RS, calculated on the fuge this solution, pipet 2 mL of the supernatant liquid, add S anhydrous basis exactly 2 mL of the internal standard solution, then add the C: Labeled amount (mg) of methotrexate (C H N O )in mobile phase to make 20 mL, and use this solution as the 20 22 8 5 1capsule sample solution. Separately, weigh accurately about 10 mg of Methotrexate RS (separately determine the water <2.48> in Operating conditions— the same manner as Methotrexate), and dissolve in the mo- Proceed as directed in the operating conditions in the bile phase to make exactly 100 mL. Pipet 10 mL of this solu- Assay. tion, and add the mobile phase to make exactly 50 mL. Pipet System suitability— 2 mL of this solution, add exactly 2 mL of the internal stand- System performance: When the procedure is run with 50 ard solution, then add the mobile phase to make 20 mL, and mL of the standard solution under the above operating con- use this solution as the standard solution. Perform the test ditions, the number of theoretical plates and the symmetry with 20 mL each of the sample solution and standard solution factor of the peak of methotrexate are not less than 3500 and as directed under Liquid Chromatography <2.01> according not more than 1.5, respectively. to the following conditions, and calculate the ratios, QT and System repeatability: When the test is repeated 6 times QS, of the peak area of methotrexate to that of the internal with 50 mL of the standard solution under the above operat- standard. ing conditions, the relative standard deviation of the peak area of methotrexate is not more than 1.0z. Amount (mg) of methotrexate (C20H22N8O5) ＝ MS × QT/QS × V/500 Assay Accurately weigh the mass of not less than 20 Methotrexate Capsules, take out all of the content, and ac- M : Amount (mg) of Methotrexate RS, calculated on the S curately weigh the mass of the empty capsules. Powder the anhydrous basis content, weigh accurately a portion of the powder, equiva-

Internal standard solution—A solution of 4-nitrophenol in lent to about 10 mg of methotrexate (C20H22N8O5), add 60 methanol (1 in 10,000). mL of the mobile phase, shake for 25 minutes, and add the Operating conditions— mobile phase to make exactly 100 mL. Centrifuge this solu- Proceed as directed in the operating conditions in the tion, pipet 2 mL of the supernatant liquid, add exactly 2 mL Assay. of the internal standard solution, then add the mobile phase System suitability— to make 20 mL, and use this solution as the sample solution. System performance: Proceed as directed in the system Separately, weigh accurately about 10 mg of Methotrexate suitability in the Assay. RS (separately determine the water <2.48> in the same man- System repeatability: When the test is repeated 6 times ner as Methotrexate), and dissolve in the mobile phase to with 20 mL of the standard solution under the above operat- make exactly 100 mL. Pipet 2 mL of this solution, add ex- ing conditions, the relative standard deviation of the ratio of actly 2 mL of the internal standard solution, then add the the peak area of methotrexate to that of the internal stand- mobile phase to make 20 mL, and use this solution as the ard is not more than 1.0z. standard solution. Perform the test with 20 mLeachofthe sample solution and standard solution as directed under Liq- Dissolution <6.10> When the test is performed at 50 revolu- uid Chromatography <2.01>, and calculate the ratios, Q and tions per minute according to the Paddle method using a T Q , of the peak area of methotrexate to that of the internal sinker, using 900 mL of water as the dissolution medium, the S standard. dissolution rate in 30 minutes of Methotrexate Capsules is not less than 85z. Amount (mg) of methotrexate (C20H22N8O5) Start the test with 1 capsule of Methotrexate Capsules, ＝ MS × QT/QS withdraw not less than 20 mL of the medium at the specified M : Amount (mg) of Methotrexate RS, calculated on the minute after starting the test, and filter through a membrane S anhydrous basis filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet V mL of the subsequent fil- Internal standard solution—A solution of 4-nitrophenol in JP XVI Official Monographs / Methoxsalen 1095 methanol (1 in 10,000). of a solution of Methoxsalen RSpreparedinthesameman- Operating conditions— ner as the sample solution: both spectra exhibit similar inten- Detector: An ultraviolet absorption photometer (wave- sities of absorption at the same wavelengths. length: 302 nm). Melting point <2.60> 145 – 1499C Column: A stainless steel column 4.6 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of gel for liquid chromatography (5 mm in particle diameter). Methoxsalen according to Method 4, and perform the test. Column temperature: A constant temperature of about Prepare the control solution with 2.0 mL of Standard Lead 259C. Solution (not more than 20 ppm). Mobile phase: To 250 mL of 0.2 mol/L potassium dihy- (2) Arsenic <1.11>—Prepare the test solution with 1.0 g drogen phosphate TS add 28.5 mL of 0.2 mol/L sodium hy- of Methoxsalen according to Method 3, and perform the test droxide TS and water to make 1000 mL. To 890 mL of this (not more than 2 ppm). solution add 110 mL of acetonitrile. (3) Related substances—Dissolve 50 mg of Methoxsalen Flow rate: Adjust the flow rate so that the retention time in 10 mL of chloroform, and use this solution as the sample of methotrexate is about 6 minutes. solution. Pipet 2 mL of the sample solution, add chloroform System suitability— to make exactly 50 mL. Pipet 1 mL of this solution, add System performance: Dissolve 10 mg each of methotrexate chloroform to make exactly 10 mL, and use this solution as and folic acid in 100 mL of the mobile phase. To 2 mL of the standard solution. Perform the test with these solutions this solution add the mobile phase to make 20 mL. When the as directed under Thin-layer Chromatography <2.03>. Spot 5 procedure is run with 20 mL of this solution under the above mL each of the sample solution and standard solution on a operating conditions, folic acid and methotrexate are eluted plate of silica gel with fluorescent indicator for thin-layer in this order with the resolution between these peaks being chromatography. Develop the plate with a mixture of chlo- not less than 8. roform, hexane and ethyl acetate (40:10:3) to a distance of System repeatability: When the test is repeated 6 times about 10 cm, and air-dry the plate. Examine under ultravio- with 20 mL of the standard solution under the above operat- let light (main wavelength: 254 nm): the spots other than the ing conditions, the relative standard deviation of the ratio of principal spot from the sample solution are not more intense the peak area of methotrexate to that of the internal stand- than the spot from the standard solution. ard is not more than 1.0z. Water <2.48> Not more than 0.5z (1 g, volumetric titra- Containers and storage Containers—Tight containers. tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g). Methoxsalen Assay Weigh accurately about 50 mg each of Methoxsalen andMethoxsalenRS,anddissolveeachinethanol(95)to メトキサレン make exactly 100 mL. Pipet 2 mL each of these solutions, and dilute each with ethanol (95) to make exactly 25 mL. Pipet 10 mL each of these solutions, and dilute each again with ethanol (95) to make exactly 50 mL, and use these solutions as the sample solution and the standard solution, re-

spectively. Determine the absorbances, AT and AS,ofthe sample solution and the standard solution at 300 nm as di- C H O : 216.19 12 8 4 rected under Ultraviolet-visible Spectrophotometry <2.24>. 9-Methoxy-7H-furo[3,2-g]chromen-7-one

[298-81-7] Amount (mg) of C12H8O4 ＝ MS × AT/AS M : Amount (mg) of Methoxsalen RS, calculated on the Methoxsalen contains not less than 98.0z and not S anhydrous basis more than 102.0z of C12H8O4,calculatedonthe anhydrous basis. Containers and storage Containers—Well-closed containers. Description Methoxsalen occurs as white to pale yellow Storage—Light-resistant. crystals or crystalline powder. It is odorless and tasteless. It is freely soluble in chloroform, slightly soluble in methanol, in ethanol (95) and in diethyl ether, and practically insoluble in water. Identification (1) To 10 mg of Methoxsalen add 5 mL of dilute nitric acid, and heat: a yellow color develops. Make this solution alkaline with a solution of sodium hydroxide (2 in 5): the color changes to red-brown. (2) To 10 mg of Methoxsalen add 5 mL of sulfuric acid, and shake: a yellow color develops. (3) Determine the absorption spectrum of a solution of Methoxsalen in ethanol (95) (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>,andcompare the spectrum with the Reference Spectrum or the spectrum 1096 Methylbenactyzium Bromide / Official Monographs JP XVI

ture of acetic anhydride and acetic acid (100) (4:1). Titrate Methylbenactyzium Bromide <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination, and make any メチルベナクチジウム臭化物 necessary correction. Each mL of 0.1 mol/L perchloric acid VS

＝ 42.24 mg of C21H28BrNO3 Containers and storage Containers—Tight containers.

Methylcellulose C H BrNO :422.36 21 28 3 Cellulose, methyl ether N,N-Diethyl-2-[(hydroxyl)(diphenyl)acetoxy]-N- methylethylaminium bromide メチルセルロース [3166-62-9]

Methylbenactyzium Bromide, when dried, contains [9004-67-5] not less than 99.0z of C21H28BrNO3. This monograph is harmonized with the European Phar- Description Methylbenactyzium Bromide occurs as white macopoeia and the U.S. Pharmacopeia. The parts of the text crystals or crystalline powder. It is odorless, and has an ex- that are not harmonized are marked with symbols ( ). tremely bitter taste.  It is freely soluble in water and in acetic acid (100), soluble Methylcellulose is a methyl ether of cellulose. in ethanol (95), slightly soluble in acetic anhydride, and It contains not less than 26.0z and not more than practically insoluble in diethyl ether. 33.0z of methoxy group (-OCH : 31.03), calculated The pH of a solution of Methylbenactyzium Bromide (1 in 3 on the dried basis. 50) is between 5.0 and 6.0. The viscosity of Methylcellulose is shown in mil- Identification (1) Shake 0.5 mL of a solution of Methyl- lipascal second (mPa･s). benactyzium Bromide (1 in 100) with 5 mL of phosphate Description Methylcellulose occurs as a white to yellow- buffer solution, pH 7.0, 2 to 3 drops of bromothymol blue ish white, powder or granules. TS and 5 mL of chloroform: a yellow color develops in the It is practically insoluble in ethanol (99.5). chloroform layer. Methylcellulose swells, when water is added, and forms a (2) To about 1 g of Methylbenactyzium Bromide add 5 clear or slightly turbid, viscous liquid. mL of water and 10 mL of sodium hydroxide TS, allow to  stand for 5 minutes, add 5 mL of dilute hydrochloric acid, Identification (1) Disperse evenly 1.0 g of Methylcellulose collect the precipitate, wash well with water, recrystallize over the surface of 100 mL of water in a beaker, while gently from a mixture of water and ethanol (95) (10:3), and dry at tapping the top of the container, if necessary, and allow the 1059C for 1 hour: the crystals melt <2.60> between 1459C beaker to stand: it aggregates on the surface of water. and 1509C. Continue the heating up to about 2009C: a red (2) Add 1.0 g of Methylcellulose to 100 mL of hot water, color develops. and stir: it becomes a suspension. Cool the suspension to (3) Add 2 mL of dilute nitric acid to 5 mL of a solution 59C, and stir: the resulting liquidisaclearoraslightly of Methylbenactyzium Bromide (1 in 10): the solution re- cloudy, viscous fluid. sponds to the Qualitative Tests <1.09> (1) for bromide. (3) To 0.1 mL of the viscous fluid obtained in (2) add 9 mL of diluted sulfuric acid (9 in 10), stir, heat in a water Melting point <2.60> 168–1729C bath for exactly 3 minutes, and immediately cool in ice Purity (1) Clarity and color of solution—Dissolve 1.0 g water.Add carefully 0.6 mL of ninhydrin TS, stir, and allow of Methylbenactyzium Bromide in 10 mL of water: the solu- to stand at 259C: the solution shows a light red color, and it tion is clear and colorless. does not change to purple color within 100 minutes. (2) Sulfate <1.14>—Perform the test with 0.5 g of (4) Pour and spread out 2 to 3 mL of the viscous fluid Methylbenactyzium Bromide. Prepare the control solution obtained in (2) onto a glass plate, and allow the water to with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more evaporate: a transparent film results. than 0.038z). (5) Pipet 50 mL of water, add exactly 50 mL of the vis- (3) Heavy metals <1.07>—Proceed with 2.0 g of Methyl- cous fluid obtained in (2), and warm to rise the temperature benactyzium Bromide according to Method 2, and perform at a rate of 2 to 59C per minute while stirring: the tempera- the test. Prepare the control solution with 2.0 mL of Stand- ture, when a white turbidity of the solution starts to increase, ard Lead Solution (not more than 10 ppm). is not less than 509C. Loss on drying <2.41> Not more than 0.5z (2 g, 1059C, Viscosity <2.53> 2 hours). (i) Method I: Apply to Methylcellulose having a labeled viscosityoflessthan600mPa･s. Put exactly an amount of Residue on ignition <2.44> Not more than 0.1z (1 g). Methylcellulose, equivalent to 4.000 g on the dried basis, in a Assay Weigh accurately about 0.5 g of Methylbenactyzium tared, wide-mouth bottle, add hot water to make 200.0 g, Bromide, previously dried, and dissolve in 80 mL of a mix- stopper the bottle, stir by mechanical means at 350- to 450- JP XVI Official Monographs / Methylcellulose 1097 revolutions per minute for 10 to 20 minutes to get a homoge- Solution in a 100-mL kjeldahl flask, add 18 mL of the mix- neous dispersion. If necessary,takeoffthesampleattached ture of nitric acid and sulfuric acid (5:4) and an amount of on the walls of the bottle, put them in the dispersed solution, nitric acid equal to that used for preparation of the test solu- and dissolve by continuing the stirring in a water bath not tion, and heat until white fumes are evolved. After cooling, exceeding 59C for 20 to 40 minutes. Add cooled water, if add 10 mL of water. In the case where hydrogen peroxide necessary, to make 200.0 g, and use this solution as the sam- (30) is added for the preparation of the test solution, add the ple solution. Centrifuge the solution if necessary to expel any same amount of hydrogen peroxide (30), then proceed in the entrapped air bubbles. Perform the test with the sample so- same manner for preparation of the test solution, and use so lution at 20 ± 0.19C as directed in Method I under Viscosity obtained solution as the control solution. Adjust the test Determination: not less than 80z and not more than 120z solution and the control solution to pH 3.0 to 4.0 with of the labeled viscosity. ammonia solution (28), and add water to make 40 mL, re- (ii) Method II: Apply to Methylcellulose having a labeled spectively. To these solutions add 1.2 mL of thioacetamide- viscosity of not less than 600 mPa･s. Put exactly an amount alkaline glycerin TS, 2 mL of acetate buffer solution, pH 3.5 of Methylcellulose, equivalent to 10.00 g on the dried basis, and water to make 50 mL, separately. After allowing to in a tared, wide-mouth bottle, add hot water to make 500.0 stand for 5 minutes, observe vertically both tubes on a white g, stopper the bottle, and prepare the sample solution in the background: the color obtained with the test solution is not same manner as directed in Method I. Perform the test with more intense than that with the control solution (not more the sample solution at 20 ± 0.19CasdirectedinMethodII than 20 ppm). (2) under Viscosity Determination, using a single cylinder- Loss on drying <2.41> Not more than 5.0z (1 g, 1059C, type rotational viscometer, according to the following oper- 1 hour). ating conditions: not less than 75z and not more than 140z of the labeled viscosity. Residue on ignition <2.44> Not more than 1.5z (1 g). Operating conditions— Assay (i) Apparatus—Reaction bottle: A 5-mL pressure- Apparatus: Brookfield type viscometer LV model. tight glass vial, having 20 mm in outside diameter and 50 Rotor No., rotation frequency, and conversion factor: mm in height, the neck 20 mm in outside diameter and 13 According to the following table, depending on the labeled mm in inside diameter, equipped with a septum of butyl- viscosity. rubber processed the surface with fluoroplastics, which can be fixed tightly to vial with aluminum cap, or equivalent. Retation Labeled viscosity Rotor Conversion frequency Heater: A square-shaped aluminum block, having holes 20 (mPa･s) No. factor /min mm in diameter and 32 mm in depth, adopted to the reaction Not less than 600 and less than 1400 3 60 20 bottle. Capable of stirring the content of the reaction bottle 〃 1400 〃 3500 3 12 100 by means of magnetic stirrer or of reciprocal shaker about 〃 3500 〃 9500 4 60 100 100 times per minute. 〃 9500 〃99,500 4 6 1000 (ii) Procedure—Weigh accurately about 65 mg of 〃99,500 4 3 2000 Methylcellulose, transfer to the reaction bottle, add 0.06 to 0.10 g of adipic acid, 2.0 mL of the internal standard solu- Procedure of apparatus: Read value after 2 minutes of tion and 2.0 mL of hydroiodic acid, stopper the bottle imme- rotation, and stop the rotation for 2 minutes. Repeat this diately, and weigh accurately. Stir or shake for 60 minutes procedure more two times, and average three observed while heating so that the temperature of the bottle content is values. 130 ± 29C. In the case when the stirrer or shaker is not pH <2.54> Allow the sample solution obtained in the Vis- available, heat for 30 minutes with repeated shaking at 5- cosity to stand at 20 ± 29C for 5 minutes: the pH of the so- minute intervals by hand, and continue heating for an addi- lution thus obtained is between 5.0 and 8.0. tional 30 minutes. Allow the bottle to cool, and again weigh accurately. If the mass loss is less than 0.50z or there is no Purity Heavy metals—Put 1.0 g of Methylcellulose in a evidence of a leak, use the upper layer of the mixture as the 100-mL Kjeldahl flask, add a sufficient amount of a mixture sample solution. Separately, put 0.06 to 0.10 g of adipic acid of nitric acid and sulfuric acid (5:4) to wet the sample, and in a reaction bottle, 2.0 mL of the internal standard solution heat gently. Repeat this procedure until to use totally 18 mL and 2.0 mL of hydroiodic acid, stopper the bottle immedi- of the mixture of nitric acid and sulfuric acid. Then boil ately, and weigh accurately. Add 45 mL of iodomethane for gently until the solution changes to black. After cooling, add assay through the septum using micro-syringe, weigh accu- 2 mL of nitric acid, and heat until the solution changes to rately, stir thoroughly, and use the upper layer of the mix- black. Repeat this procedure until the solution no longer ture as the standard solution. Perform the test with 1 to 2 mL changes to black, and heat strongly until dense white fumes each of the sample solution and standard solution as directed are evolved. After cooling, add 5 mL of water, boil gently under Gas Chromatography <2.02> according to the follow- until dense white fumes are evolved, then heat until the ing conditions, and calculate the ratios, Q and Q ,ofthe volume of the solution becomes to 2 to 3 mL. After cooling, T S peak area of iodomethane to that of the internal standard. if the solution reveals yellow color by addition of 5 mL of water, add 1 mL of hydrogen peroxide (30), and heat until Content (z) of methoxy group (CH3O) the volume of the solution becomes to 2 to 3 mL. After cool- ＝ MS/M × QT/QS × 21.86 ing, dilute the solution with 2 to 3 mL of water, transfer to a M : Amount (mg) of iodomethane in the standard solu- Nessler tube, add water to make 25 mL, and use this solution S tion as the test solution. Separately, put 2.0 mL of Standard Lead M: Amount (mg) of sample, calculated on the dried basis 1098 Methyldopa Hydrate / Official Monographs JP XVI

Internal standard solution—A solution of n-octane in o- spectrum of Methyldopa RS: both spectra exhibit similar in- xylene (3 in 100). tensities of absorption at the same wave numbers. Operating conditions— Optical rotation <2.49> [a]20: －25 – －289(calculated on Detector: A thermal conductivity detector or hydrogen D the anhydrous basis, 1 g, aluminum (III) chloride TS, 20 flame-ionization detector. mL, 100 mm). Column:Aglasscolumn3–4mmininsidediameterand 1.8–3minlength,packedwithsiliceousearthforgaschro- Purity (1) Acidity—Shake 1.0 g of Methyldopa Hydrate matography, 125 to 150 mm in diameter, coated with methyl with 100 mL of freshly boiled and cooled water, and add silicone polymer at the ratio of 10 – 20z. 0.20 mL of 0.1 mol/L sodium hydroxide VS and 2 drops of Column temperature: A constant temperature of about methyl red TS: a yellow color develops. 1009C. (2) Chloride <1.03>—Perform the test with 0.5 g of Carrier gas: Helium for thermal conductivity detector, or Methyldopa Hydrate. Prepare the control solution with Helium or Nitrogen for hydrogen, flame-ionization detector. 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than Flow rate: Adjust the flow rate so that the retention time 0.028z). of the internal standard is about 10 minutes. (3) Heavy metals <1.07>—Proceed with 2.0 g of Methyl- System suitability— dopa Hydrate according to Method 2, and perform the test. System performance: When the procedure is run with 1 – 2 Prepare the control solution with 2.0 mL of Standard Lead mL of the standard solution under the above operating con- Solution (not more than 10 ppm). ditions, iodomethane and the internal standard are eluted in (4) Arsenic <1.11>—Prepare the test solution with 1.0 g this order, with complete separation of these peaks. of Methyldopa Hydrate in 5 mL of dilute hydrochloric acid, and perform the test (not more than 2 ppm). Containers and storage Containers—Well-closed contain- (5) 3-O-Methylmethyldopa—Dissolve 0.10 g of Methyl- ers.  dopa Hydrate in methanol to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve 5 mg of 3-O-methylmethyldopa for thin-layer chromatography Methyldopa Hydrate in methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions メチルドパ水和物 as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (13:5:3) to a distance of about 10 cm, and air-dry the plate. Spray evenly 4-nitroaniline-sodium nitrite TS on the 1 C10H13NO4.1 2 H2O: 238.24 plate, and air-dry the plate, then spray evenly a solution of (2S)-2-Amino-3-(3,4-dihydroxyphenyl)-2-methylpropanoic sodium carbonate decahydrate (1 in 4) on the plate: the spot acid sesquihydrate from the sample solution corresponding to that from the [41372-08-1] standard solution is not more intense than the spot from the standard solution. Methyldopa Hydrate contains not less than 98.0z Water <2.48> 10.0 – 13.0z (0.2 g, volumetric titration, of methyldopa (C H NO : 211.21), calculated on the 10 13 4 direct titration). anhydrous basis. Residue on ignition <2.44> Not more than 0.1z (1 g). Description Methyldopa Hydrate occurs as a white to pale grayish white, crystalline powder. Assay Weigh accurately about 0.3 g of Methyldopa Hy- It is slightly soluble in water, in methanol and in acetic drate, dissolve in 80 mL of acetic acid (100), and titrate acid (100), very slightly soluble in ethanol (95), and practi- <2.50> with 0.1 mol/L perchloric acid VS until the color of cally insoluble in diethyl ether. the solution changes from purple through blue to blue-green It dissolves in dilute hydrochloric acid. (indicator: 2 to 3 drops of crystal violet TS). Perform a blank determination, and make any necessary correction. Identification (1) To 10 mg of Methyldopa Hydrate add 3 drops of ninhydrin TS, and heat in a water bath for 3 Each mL of 0.1 mol/L perchloric acid VS minutes: a purple color develops. ＝ 21.12 mg of C10H13NO4 (2) Determine the absorption spectrum of a solution of Containers and storage Containers—Well-closed contain- Methyldopa Hydrate in 0.1 mol/L hydrochloric acid TS (1 in ers. 25,000) as directed under Ultraviolet-visible Spectropho- Storage—Light-resistant. tometry <2.44>, and compare the spectrum with the Refer- ence Spectrum or the spectrum of a solution of Methyldopa RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (3) Determine the infrared absorption spectrum of Methyldopa Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>,and compare the spectrum with the Reference Spectrum or the JP XVI Official Monographs / Methyldopa Tablets 1099

Start the test with 1 tablet of Methyldopa Tablets, with- Methyldopa Tablets draw not less than 30 mL of the medium at the specified minute after starting the test, and filter through a membrane メチルドパ錠 filter with a pore size not exceeding 0.8 mm. Discard the first 10 mL of the filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V?mL so that each mL contains Methyldopa Tablets contain not less than 90.0z about 25 mg of methyldopa (C H NO ) according to the and not more than 110.0z of the labeled amount of 10 13 4 labeled amount, and use this solution as the sample solution. methyldopa (C H NO : 211.21). 10 13 4 Separately, weigh accurately about 56 mg of methyldopa for Method of preparation Prepare as directed under Tablets, assay (separately deternine the loss on drying <2.41> at 1259C with Methyldopa Hydrate. for 2 hours), and dissolve in water to make exactly 200 mL. Pipet 10 mL of this solution, add water to make exactly 100 Identification (1) To a quantity of powdered Methyldopa mL, and use this solution as the standard solution. Deter- Tablets, equivalent to 0.1 g of Methyldopa Hydrate accord- mine the absorbances, A and A , of the sample solution ing to the labeled amount, add 10 mL of water, and heat in a T S and the standard solution at 280 nm as directed under Ultra- water bath for 5 minutes with occasional shaking. After violet-visible Spectrophotometry <2.24>. cooling, centrifuge for 5 minutes at 2000 rotations per minute, apply 1 drop of the supernatant solution to a filter Dissolution rate (z) with respect to the labeled amount paper, and dry with warm air. Place 1 drop of ninhydrin TS of methyldopa (C10H13NO4) over the spot, and heat for 5 minutes at 1009C: a purple ＝ MS × AT/AS × V?/V × 1/C × 45 color develops. M : Amount (mg) of methyldopa for assay, calculated on (2) To 0.5 mL of the supernatant liquid obtained in the S the dried basis Identification (1) add 2 mL of 0.05 mol/L sulfuric acid TS, 2 C: Labeled amount (mg) of methyldopa (C H NO )in1 mL of iron (II) tartrate TS and 4 drops of ammonia TS, and 10 13 4 tablet shake well: a deep purple color develops. (3) To 0.7 mL of the supernatant liquid obtained in the Assay Weigh accurately and powder not less than 20 Identification (1) add 0.1 mol/L hydrochloric acid TS to Methyldopa Tablets. Weigh accurately a portion of the pow- make 20 mL. To 10 mL of this solution add 0.1 mol/L hy- der, equivalent to about 0.1 g of methyldopa (C10H13NO4), drochloric acid TS to make 100 mL, and determine the ab- add 50 mL of 0.05 mol/L sulfuric acid TS, shake thoroughly sorption spectrum of the solution as directed under Ultravio- for 15 minutes, add 0.05 mol/L sulfuric acid TS to make ex- let-visible Spectrophotometry <2.24>: it exhibits a maximum actly 100 mL, and filter through a dry filter paper. Discard between 277 nm and 283 nm. the first 20 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about Uniformity of dosage units <6.02> Perform the test accord- 0.11 g of Methyldopa RS (previously dry at 1259Cfor2 ing to the following method: it meets the requirement of the hours, and determine the loss on drying <2.41>), dissolve in Content uniformity test. 0.05 mol/L sulfuric acid TS to make exactly 100 mL, and To 1 tablet of Methyldopa Tablets add 50 mL of 0.05 use this solution as the standard solution. Pipet 5 mL each of mol/L sulfuric acid TS, shake for 15 minutes, then add 0.05 the sample solution and standard solution, add exactly 5 mL mol/L sulfuric acid TS to make exactly 100 mL, and filter. of iron (II) tartrate TS, and add ammonia-ammonium Discard the first 20 mL of the filtrate, pipet V mL of the acetate buffer solution, pH 8.5, to make exactly 100 mL. subsequent filtrate equivalent to about 5 mg of methyldopa Perform the test with these solutions as directed under Ultra- (C H NO ), add exactly 5 mL of iron (II) tartrate TS, then 10 13 4 violet-visible Spectrophotometry <2.24>, using a solution add ammonia-ammonium acetate buffer solution, pH 8.5, to prepared with 5 mL of 0.05 mol/L sulfuric acid TS in the make exactly 100 mL, and use this solution as the sample so- same manner, as the blank. Determine the absorbances, A lution. Separately, weigh accurately about 0.11 g of Methyl- T and A , of the subsequent solutions of the sample solution dopa RS (separately determine the loss on drying <2.41> at S and the standard solution at 520 nm, respectively. 1259C for 2 hours), and dissolve in 0.05 mol/L sulfuric acid

TS to make exactly 100 mL. Pipet 5 mL of this solution, add Amount (mg) of methyldopa (C10H13NO4) exactly 5 mL of iron (II) tartrate TS, then add ammonia- ＝ MS × AT/AS ammonium acetate buffer solution, pH 8.5, to make exactly M : amount (mg) of Methyldopa RS, calculated on the 100 mL, and use this solution as the standard solution. De- S dried basis termine the absorbances at 520 nm, AT and AS,ofthesample solution and standard solution as directed under Ultravi- Containers and storage Containers—Well-closed contain- olet-visible Spectrophotometry <2.24>. ers.

Amount (mg) of methyldopa (C10H13NO4) ＝ MS × AT/AS × 5/V

MS: Amount (mg) of Methyldopa RS, calculated on the dried basis Dissolution <6.10> When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 60 minutes of Methyldopa Tablets is not less than 75z. 1100 dl-Methylephedrine Hydrochloride / Official Monographs JP XVI

standard solution. dl-Methylephedrine Hydrochloride Operating conditions— Detector: An ultraviolet absorption photometer (wave- dl-メチルエフェドリン塩酸塩 length: 257 nm). Column: A stainless steel column 4.6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticlediameter). Column temperature: A constant temperature of about 409C.

C11H17NO.HCl: 215.72 Mobile phase: Dissolve 13.6 g of potassium dihydrogen (1RS,2SR)-2-Dimethylamino-1-phenylpropan-1-ol phosphate and 3 g of sodium 1-heptane sulfonate in 1000 mL monohydrochloride of water, and adjust the pH to 2.5 with phosphoric acid. To [18760-80-0] 900 mL of this solution add 200 mL of acetonitrile. Flow rate: Adjust the flow rate so that the retention time dl-Methylephedrine Hydrochloride, when dried, of methylephedrine is about 10 minutes. contains not less than 99.0z and not more than Time span of measurement: About 2 times as long as the 101.0z of C11H17NO.HCl. retention time of methylephedrine beginning after the solvent peak. Description dl-Methylephedrine Hydrochloride occurs as System suitability— colorlesscrystalsorawhite,crystallinepowder. Test for required detectability: To exactly 2 mL of the It is freely soluble in water, sparingly soluble in ethanol standard solution add water to make exactly 20 mL. Con- (99.5), slightly soluble in acetic acid (100), and practically firm that the peak area of methylephedrine obtained from insoluble in acetic anhydride. 20 mL of this solution is equivalent to 7 to 13z of that from A solution of dl-Methylephedrine Hydrochloride (1 in 20) 20 mL of the standard solution. shows no optical rotation. System performance: Dissolve 50 mg of dl-Methylephe- Identification (1) Determine the absorption spectrum of a drine Hydrochloride and 0.4 mg of methyl parahydroxyben- solution of dl-Methylephedrine Hydrochloride (1 in 2000) as zoate in 50 mL of water. When the procedure is run with 20 directed under Ultraviolet-visible Spectrophotometry <2.24>, mL of this solution under the above operating conditions, and compare the spectrum with the Reference Spectrum: methylephedrine and methyl parahydroxybenzoate are both spectra exhibit similar intensities of absorption at the eluted in this order with the resolution between these peaks same wavelengths. being not less than 3. (2) Determine the infrared absorption spectrum of dl- System repeatability: When the test is repeated 6 times Methylephedrine Hydrochloride, previously dried, as di- with 20 mL of the standard solution under the above operat- rected in the potassium chloride disk method under Infrared ing conditions, the relative standard deviation of the peak Spectrophotometry <2.25>, and compare the spectrum with area of methylephedrine is not more than 2.0z. the Reference Spectrum: both spectra exhibit similar intensi- Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, ties of absorption at the same wave numbers. 3 hours). (3) A solution of dl-Methylephedrine Hydrochloride (1 in 10) responds to the Qualitative Tests <1.09> for chloride. Residue on ignition <2.44> Not more than 0.1z (1 g). pH <2.54> The pH of a solution prepared by dissolving Assay Weigh accurately about 0.4 g of dl-Methylephedrine 1.0 g of dl-Methylephedrine Hydrochloride in 20 mL of Hydrochloride, previously dried, dissolve in 80 mL of a mix- water is between 4.5 and 6.0. ture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio- Melting point <2.60> 207–2119C metric titration). Perform a blank determination, and make Purity (1) Clarity and color of solution—Dissolve 1.0 g any necessary correction. of dl-Methylephedrine Hydrochloride in 10 mL of water: the Each mL of 0.1 mol/L perchloric acid VS solution is clear and colorless. ＝ 21.57 mg of C H NO.HCl (2) Heavy metals <1.07>—Proceed with 1.0 g of dl- 11 17 Methylephedrine Hydrochloride according to Method 4, and Containers and storage Containers—Well-closed contain- perform the test. Prepare the control solution with 1.0 mL of ers. Standard Lead Solution (not more than 10 ppm). Storage—Light-resistant. (3) Related substances—Dissolve 50 mg of dl-Methyl- ephedrine Hydrochloride in 20 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the total area of the peaks other than the peak of methylephedrine is not larger than the peak area of methylephedrine from the JP XVI Official Monographs / Methylergometrine Maleate 1101

of water, and adjust the pH to 2.5 with phosphoric acid. To 10 dl-Methylephedrine 900 mL of this solution add 200 mL of acetonitrile. Flow rate: Adjust the flow rate so that the retention time Hydrochloride Powder of methylephedrine is about 10 minutes. System suitability— dl-Methylephedrine Hydrochloride Powder System performance: When the procedure is run with 20 mL of the standard solution under the above operating con- dl-メチルエフェドリン塩酸塩散 10 ditions, methylephedrine and the internal standard are eluted in this order with the resolution between these peaks being 10z dl-Methylephedrine Hydrochloride Powder not less than 3. contains not less than 9.3z and not more than 10.7z System repeatability: When the test is repeated 6 times of dl-methylephedrine hydrochloride (C11H17NO.HCl: with 20 mL of the standard solution under the above operat- 215.72). ing conditions, the relative standard deviation of the ratio of the peak area of methylephedrine to that of the internal Method of preparation standard is not more than 1.0z. dl-Methylephedrine Hydrochloride 100 g Containers and storage Containers—Well-closed contain- Starch, Lactose Hydrate or ers. their mixture a sufficient quantity Storage—Light-resistant. To make 1000 g Prepare as directed under Granules or Powders, with the above ingredients. Methylergometrine Maleate Identification Determine the absorption spectrum of a so- メチルエルゴメトリンマレイン酸塩 lution of 10z dl-Methylephedrine Hydrochloride Powder (1 in 200) as directed under Ultraviolet-visible Spectrophotome- try <2.24>: it exhibits maxima between 250 nm and 253 nm, between 255 nm and 259 nm, and between 261 nm and 264 nm. Assay Weigh accurately about 0.5 g of 10z dl-Methyl- ephedrine Hydrochloride Powder, add exactly 4 mL of the internal standard solution and 25 mL of water, shake vigorously for 20 minutes to dissolve, add water to make 50 mL, filter through a membrane filter with pore size of 0.45 mm, if C20H25N3O2.C4H4O4: 455.50 necessary, discard the first 10 mL of the filtrate, and use the (8S)-N-[(1S)-1-(Hydroxymethyl)propyl]-6-methyl-9,10- subsequent filtrate as the sample solution. Separately, weigh didehydroergoline-8-carboxamide monomaleate accurately about 50 mg of dl-methylephedrine hydrochloride [7054-07-1] for assay, previously dried at 1059C for 3 hours, add exactly 4 mL of the internal standard solution and water to make 50 Methylergometrine Maleate, when dried, contains mL, and use this solution as the standard solution. Perform not less than 95.0z and not more than 105.0z of the test with 20 mL each of the sample solution and standard C20H25N3O2.C4H4O4. solution as directed under Liquid Chromatography <2.01> Description Methylergometrine Maleate occurs as a white according to the following conditions, and calculate the to pale yellow, crystalline powder. It is odorless. ratios of the peak area, QT and QS, of methylephedrine to that of the internal standard. It is slightly soluble in water, in methanol and in ethanol (95), and practically insoluble in diethyl ether. Amount (mg) of dl-methylephedrine hydrochloride It gradually changes to yellow by light. (C11H17NO.HCl) ＝ MS × QT/QS Melting point: about 1909C (with decomposition).

MS: Amount (mg) of dl-methylephedrine hydrochloride Identification (1) A solution of Methylergometrine Male- for assay ate (1 in 200) shows a blue fluorescence. Internal standard solution—A solution of methyl parahy- (2) The colored solution obtained in the Assay develops a deep blue in color. Determine the absorption spectrum of droxybenzoate in acetonitrile (1 in 10,000). Operating conditions— the colored solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with Detector: An ultraviolet absorption photometer (wave- the Reference Spectrum or the spectrum of a solution of length: 257 nm). Column: A stainless steel column 4.6 mm in inside diame- Methylergometrine Maleate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensi- ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). ties of absorption at the same wavelengths. (3) To 5 mL of a solution of Methylergometrine Maleate Column temperature: A constant temperature of about (1 in 500) add 1 drop of potassium permanganate TS: the red 409C. Mobile phase: Dissolve 13.6 g of potassium dihydrogen color of the test solution fades immediately. 20 phosphate and 3 g of sodium 1-heptane sulfonate in 1000 mL Optical rotation <2.49> [a]D : ＋44 – ＋509(after drying, 1102 Methylergometrine Maleate Tablets / Official Monographs JP XVI

0.1 g, water, 20 mL, 100 mm). trophotometry <2.24>: it exhibits maxima between 543 nm and 547 nm and between 620 nm and 630 nm. Purity Related substances—Conduct this procedure without exposure to daylight, using light-resistant vessels. Dis- Uniformity of dosage unit <6.02> Perform the test accord- solve 8 mg of Methylergometrine Maleate in 2 mL of a mixing to the following method: it meets the requirement of the ture of ethanol (95) and ammonia solution (28) (9:1), and use Content uniformity test. this solution as the sample solution. Pipet 1 mL of the sam- Transfer 1 tablet of Methylergometrine Maleate Tablets ple solution, add a mixture of ethanol (95) and ammonia so- to a brown glass-stoppered centrifuge tube, add 10 mL of lution (28) (9:1) to make exactly 100 mL, and use this solu- water, shake for 10 minutes vigorously, and disintegrate the tion as the standard solution. Perform the test immediately tablet. Add 3 g of sodium chloride and 2 mL of ammonia so- with these solutions as directed under Thin-layer Chroma- lution (28), add exactly 25 mL of chloroform, and after tography <2.03>. Spot 10 mL each of the sample solution and vigorous shaking for 10 minutes, centrifuge for 5 minutes. standard solution on a plate of silica gel with fluorescent in- Discard the water layer, take the chloroform extracts, dicator for thin-layer chromatography, and immediately de- add chloroform to make exactly V mL of a solution velop the plate with a mixture of chloroform, methanol and containing about 5 mg of methylergometrine maleate water (75:25:3) to a distance of about 10 cm, and air-dry the (C20H25N3O2.C4H4O4) per mL, and use this solution as the plate. Examine under ultraviolet light (main wavelength: 365 sample solution. Separately, weigh accurately about 1.25 mg nm): the spots other than the principal spot from the sample of Methylergometrine Maleate RS, previously dried in a solution are not more intense than the spot from the stand- desiccator ( in vacuum, phosphorus (V) oxide) for 4 hours, ard solution. and dissolve in water to make exactly 100 mL. Pipet 10 mL of this solution into a brown glass-stoppered centrifuge tube, Loss on drying <2.41> Not more than 2.0z (0.2 g, in vacu- and add 3 g of sodium chloride and 2 mL of ammonia solu- um, phosphorus (V) oxide, 4 hours). tion (28). Add exactly 25 mL of chloroform, shake vigor- Assay Weigh accurately about 10 mg each of Methyler- ously for 10 minutes, and centrifuge for 5 minutes. Discard gometrine Maleate and Methylergometrine Maleate RS, pre- the water layer, and use the chloroform layer as the standard viously dried, add water to make exactly 250 mL, and use solution. Pipet 20 mL each of the sample solution and the these solutions as the sample solution and the standard solu- standard solution separately into brown glass-stoppered tion. Pipet 2 mL each of the sample solution and the stand- centrifuge tubes, add immediately exactly 10 mL of dilute ard solution separately into brown glassstoppered test tubes, 4-dimethylaminobenzaldehyde-iron (III) chloride TS, and add exactly 4 mL each of 4-dimethylaminobenzaldehyde- shake for 5 minutes vigorously. Centrifuge these solutions iron (III) chloride TS while ice cooling, warm for 10 minutes for 5 minutes, take the water layers, and allow them to stand at 459C, and allow to stand for 20 minutes at room tempera- for 1 hour. Perform the test with these solutions as directed ture. Perform the test with these solutions as directed under under Ultraviolet-visible Spectrophotometry <2.24>,using Ultraviolet-visible Spectrophotometry <2.24>, using a solu- dilute 4-dimethylaminobenzaldehyde-iron (III) chloride TS tion, prepared with 2.0 mL of water in the same manner, as as the blank. Determine the absorbances, AT and AS,ofthe the blank. Determine the absorbances, AT and AS,ofthe subsequent solutions of the sample solution and standard subsequent solutions of the sample solution and the standard solution at 545 nm, respectively. solution at 545 nm, respectively. Amount (mg) of methylergometrine maleate

Amount (mg) of C20H25N3O2.C4H4O4 (C20H25N3O2.C4H4O4) ＝ MS × AT/AS ＝ MS × AT/AS × V/250

MS: Amount (mg) of Methylergometrine Maleate RS MS: Amount (mg) of Methylergometrine Maleate RS Containers and storage Containers—Tight containers. Dissolution <6.10> When the test is performed at 100 revo- Storage—Light-resistant. lutions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Methylergometrine Maleate Tablets is Methylergometrine Maleate Tablets not less than 70z. Start the test with 1 tablet of Methylergometrine Maleate メチルエルゴメトリンマレイン酸塩錠 Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Dis- Methylergometrine Maleate Tablets contain not card the first 10 mL of the filtrate, to exactly V mL of the less than 90.0z and not more than 110.0z of subsequent filtrate add water to make exactly V?mL so that the labeled amount of methylergometrine maleate each mL contains about 0.13 mg of methylergometrine male- (C20H25N3O2.C4H4O4: 455.50). ate (C20H25N3O2.C4H4O4) according to the labeled amount, Method of preparation Prepare as directed under Tablets, and use this solution as the sample solution. Separately, with Methylergometrine maleate. weigh accurately about 25 mg of Methylergometrine Maleate RS, previously dried in a desiccator for 4 hours (in vacuum, Identification (1) The sample solution obtained in the phosphorus (V) oxide), and dissolve in water to make exactly Assay shows a blue fluorescence. 100 mL. Pipet 5 mL of this solution, add water to make ex- (2) The colored solution obtained in the Assay shows a actly 100 mL, then pipet 1 mL of this solution, add water to deep blue color. Determine the absorption spectrum of the make exactly 100 mL, and use this solution as the standard colored solution as directed under Ultraviolet-visible Spec- solution. Determine immediately the intensities of the JP XVI Official Monographs / Methyl Parahydroxybenzoate 1103

 fluorescence, FT and FS, of the sample solution and standard that are not harmonized are marked with symbols ( ). solution at 338 nm as the excitation wavelength and at 427 nm as the fluorescence wavelength as directed under Fluoro- Methyl Parahydroxybenzoate, when dried, contains metry <2.22>. not less than 98.0z and not more than 102.0z of C H O . Dissolution rate (z) with respect to the labeled amount 8 8 3  of methylergometrine maleate (C20H25N3O2.C4H4O4) Description Methyl Parahydroxybenzoate, occurs as col- ＝ MS × FT/FS × V?/V × 1/C × 9/20 orless crystals or a white, crystalline powder. It is freely soluble in ethanol (95) and in acetone, and M : Amount (mg) of Methylergometrine Maleate RS S slightly soluble in water. C: Labeled amount (mg) of methylergometrine maleate 

(C20H25N3O2.C4H4O4)in1tablet Identification (1) The melting point <2.60> of Methyl Parahydroxybenzoate is between 1259C and 1289C. Assay Weigh accurately and powder not less than 20 (2) Determine the infrared absorption spectrum of Methylergometrine Maleate Tablets. Weigh accurately a Methyl Parahydroxybenzoate as directed in the potassium portion of the powder, equivalent to about 0.3 mg of bromide disk method under Infrared Spectrophotometry methylergometrine maleate (C H N O .C H O ), transfer 20 25 3 2 4 4 4 <2.25>, and compare the spectrum with the Reference Spec- to a brown separator, add 15 mL of sodium hydrogen car- trum: both spectra exhibit similar intensities of absorption at bonate solution (1 in 20), and extract with four 20-mL por- the same wave numbers. tions of chloroform. Filter each portion of the chloroform  extracts through a pledget of absorbent cotton, previously Purity (1) Clarity and color of solution—Dissolve 1.0 g moistened with chloroform, into another dried, brown sepa- of Methyl Parahydroxybenzoate in 10 mL of ethanol (95): rator, combine all the extracts, and use this extract as the the solution is clear and not more intensely colored than the sample solution. Separately, weigh accurately about 10 mg following control solution. of Methylergometrine Maleate RS, previously dried in a Control solution: To 5.0 mL of Cobalt (II) Chloride CS, desiccator (silica gel) for 4 hours, dissolve in water, and add 12.0 mL of Iron (III) Chloride CS and 2.0 mL of Copper (II) water to make exactly 100 mL. Pipet 3 mL of this solution, Sulfate CS add water to make 1000 mL. and transfer to a brown separator, proceed in the same man- (2) Acidity—Dissolve 0.20 g of Methyl Parahydroxyben- ner as the preparation of the sample solution, and use this zoate in 5 mL of ethanol (95), add 5 mL of freshly boiled extract as the standard solution. To each total volume of the and cooled water and 0.1 mL of bromocresol green-sodium sample solution and the standard solution add exactly 25 mL hydroxide-ethanol TS, then add 0.1 mL of 0.1 mol/L so- each of dilute p-dimethylaminobenzadehyde-ferric chloride dium hydroxide VS: the solution shows a blue color. TS, and after vigorous shaking for 5 minutes, allow to stand (3) Heavy metals <1.07>—Dissolve 1.0 g of Methyl Par- for 30 minutes. Draw off the water layer, centrifuge, and ahydroxybenzoate in 25 mL of acetone, add 2 mL of dilute allow to stand for 1 hour. Perform the test with these solu- acetic acid and water to make 50 mL, and perform the test tions as directed under Ultraviolet-visible Spectrophotome- using this solution as the test solution. Prepare the control try <2.24>, using dilute 4-dimethylaminobenzaldehyde-ferric solution as follows: to 2.0 mL of Standard Lead Solution chloride TS as the blank. Determine the absorbances, AT add 25 mL of acetone, 2 mL of dilute acetic acid, and water and AS, of the subsequent solutions of the sample solution to make 50 mL (not more than 20 ppm). and the standard solution at 545 nm, respectively. (4) Related substances—Dissolve 0.10 g of Methyl Par- ahydroxybenzoate in 10 mL of acetone, and use this solution Amount (mg) of methylergometrine maleate as the sample solution. Pipet 0.5 mL of the sample solution, (C H N O .C H O ) 20 25 3 2 4 4 4 add acetone to make exactly 100 mL, and use this solution as ＝ M × A /A × 3/100 S T S the standard solution. Perform the test with these solutions

MS: Amount (mg) of Methylergometrine Maleate RS as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a Containers and storage Containers—Well-closed contain- plate of silica gel with fluorescent indicator for thin-layer ers. chromatography. Develop the plate with a mixture of metha- Storage—Light-resistant. nol, water and acetic acid (100) (70:30:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot other than the Methyl Parahydroxybenzoate principal spot with the sample solution is not more intense than the spot obtained with the standard solution. パラオキシ安息香酸メチル Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 1.0 g of Methyl Parahy- droxybenzoate, add exactly 20 mL of 1 mol/L sodium hydroxide VS, heat at about 709C for 1 hour, and immediately cool in ice. Titrate <2.50> the excess sodium hydroxide with C H O : 152.15 8 8 3 0.5 mol/L sulfuric acid VS up to the second equivalent point Methyl 4-hydroxybenzoate (potentiometric titration). Perform a blank determination. [98-76-3] Each mL of 1 mol/L sodium hydroxide VS This monograph is harmonized with the European Phar- ＝ 152.1 mg of C H O macopoeia and the U.S. Pharmacopeia. The parts of the text 8 8 3 1104 Methylprednisolone / Official Monographs JP XVI

Containers and storage Containers—Well-closed contain- Residue on ignition <2.44> Not more than 0.2z (0.2 g). ers.  Assay Weigh accurately about 10 mg of Methylpredniso- lone, previously dried, and dissolve in methanol to make exactly 100 mL. To exactly 5 mL of this solution add methanol Methylprednisolone to make exactly 50 mL, and determine the absorbance A at the wavelength of maximum absorption at about 243 nm as メチルプレドニゾロン directed under Ultraviolet-visible Spectrophotometry <2.24>.

Amount (mg) of C22H30O5 ＝ A/400 × 10,000 Containers and storage Containers—Tight containers.

Methylprednisolone Succinate C H O :374.47 22 30 5 メチルプレドニゾロンコハク酸エステル 11b,17,21-Trihydroxy-6a-methylpregna-1,4-diene- 3,20-dione [83-43-2]

Methylprednisolone, when dried, contains not less than 96.0z and not more than 104.0z of C22H30O5. Description Methylprednisolone occurs as a white, crystalline powder. It is odorless. It is sparingly soluble in methanol and in 1,4-dioxane, slightly soluble in ethanol (95) and in chloroform, and prac- C26H34O8:474.54 tically insoluble in water and in diethyl ether. 11b,17,21-Trihydroxy-6a-methylpregna-1,4-diene- Melting point: 232 – 2409C (with decomposition). 3,20-dione 21-(hydrogen succinate) [2921-57-5] Identification (1) Add 2 mL of sulfuric acid to 2 mg of Methylprednisolone: a deep red color develops with no Methylprednisolone Succinate, when dried, contains fluorescence. Then add 10 mL of water to this solution: the not less than 97.0z and not more than 103.0z of color fades, and a gray, flocculent precipitate is produced. C H O . (2) Dissolve 10 mg of Methylprednisolone in 1 mL of 26 34 8 methanol, add 1 mL of Fehling's TS, and heat: a red precipi- Description Methylprednisolone Succinate occurs as a tate is produced. white, crystals or crystalline powder. (3) Determine the absorption spectrum of a solution of It is soluble in methanol, sparingly soluble in ethanol (95), Methylprednisolone in methanol (1 in 100,000) as directed and practically insoluble in water. under Ultraviolet-visible Spectrophotometry <2.24>,and Melting point: about 2359C (with decomposition). compare the spectrum with the Reference Spectrum: both Identification (1) Determine the absorption spectrum of a spectra exhibit similar intensities of absorption at the same solution of Methylprednisolone Succinate in methanol (1 in wavelengths. 50,000) as directed under Ultraviolet-visible Spectropho- 20 Optical rotation <2.49> [a]D : ＋79 – ＋869(after drying, tometry <2.24>, and compare the spectrum with the Refer- 0.1 g, 1,4-dioxane, 10 mL, 100 mm). ence Spectrum or the spectrum of a solution of Methylpred- nisolone Succinate RS prepared in the same manner as the Purity Related substances—Dissolve 50 mg of Methylpred- sample solution: both spectra exhibit similar intensities of nisolone in 5 mL of a mixture of chloroform and methanol absorption at the same wavelengths. (9:1), and use this solution as the sample solution. Pipet 1 (2) Determine the infrared absorption spectrum of mL of the sample solution, add a mixture of chloroform and Methylprednisolone Succinate, previously dried, as directed methanol (9:1) to make exactly 200 mL, and use this solution in the potassium bromide disk method under Infrared Spec- as the standard solution. Perform the test with these solu- trophotometry <2.25>, and compare the spectrum with the tions as directed under Thin-layer Chromatography <2.03>. Reference Spectrum or the spectrum of previously dried Spot 10 mL each of the sample solution and standard solu- Methylprednisolone Succinate RS: both spectra exhibit simi- tion on a plate of silica gel for thin-layer chromatography. lar intensities of absorption at the same wave numbers. In Develop the plate with a mixture of dichloromethane, diethyl case when some differences are found between the spectra, ether, methanol and water (385:75:40:6) to a distance of repeat the test with residues obtained by dissolving these sub- about 12 cm, and air-dry the plate. Then heat at 1059Cfor stances in ethanol (95), evaporating to dryness, and drying. 10 minutes, cool, and spray evenly alkaline blue tetrazolium 25 TS on the plate: the spots other than the principal spot from Optical rotation <2.49> [a]D : ＋99 – ＋1039(after drying, the sample solution are not more intense than the spot from 0.2 g, ethanol (95), 20 mL, 100 mm). the standard solution. Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C, Methylprednisolone Succinate according to Method 4, and 3 hours). perform the test. Prepare the control solution with 1.0 mL of JP XVI Official Monographs / Methylrosanilinium Chloride 1105

Standard Lead Solution (not more than 10 ppm). droxybenzoate in a mixture of 0.05 mol/L phosphate buffer (2) Arsenic <1.11>—Prepare the test solution with 2.0 g solution, pH 3.5 and acetonitrile (1:1) (3 in 20,000). of Methylprednisolone Succinate according to Method 3, Operating conditions— and perform the test (not more than 1 ppm). Detector: An ultraviolet absorption photometer (wave- (3) Related substances—Dissolve 15 mg of Methylpred- length: 254 nm). nisolone Succinate in 5 mL of methanol, add a mixture of Column: A stainless steel column 4.6 mm in inside diame- 0.05 mol/L phosphate buffer solution, pH 3.5 and aceto- ter and 25 cm in length, packed with octadecylsilanized silica nitrile (1:1) to make 50 mL, and use this solution as the sam- gel for liquid chromatography (5 mminparticlediameter). ple solution. Pipet 1 mL of the sample solution, add the mix- Column temperature: A constant temperature of about ture of 0.05 mol/L phosphate buffer solution, pH 3.5 and 259C. acetonitrile (1:1) to make exactly 100 mL, and use this solu- Mobile phase: To 1000 mL of 0.05 mol/L potassium dihy- tion as the standard solution. Perform the test with exactly drogen phosphate TS add a suitable amount of 0.05 mol/L 5 mL each of the sample solution and standard solution as disodium hydrogen phosphate TS to make a solution having directed under Liquid Chromatography <2.01> according to pH 5.5. To 640 mL of this solution add 360 mL of aceto- the following conditions, and determine each peak area by nitrile. the automatic integration method: the area of the peaks Flow rate: Adjust the flow rate so that the retention time other than the peak of methylprednisolone succinate from of methylprednisolone succinate is about 6 minutes. sample solution is not larger than 1/2 times the peak area of System suitability— methylprednisolone succinate from the standard solution, System performance: When the procedure is run with 5 mL and the total area of the peaks other than the peak of of the standard solution under the above operating condi- methylprednisolone succinate is not larger than the peak area tions, methylprednisolone succinate and the internal stand- of methylprednisolone succinate from the standard solution. ard are eluted in this order with the resolution between these Operating conditions— peaks being not less than 6. Detector, column, column temperature, mobile phase, and System repeatability: When the test is repeated 6 times flow rate: Proceed as directed in the operating conditions in with 5 mL of the standard solution under the above operating the Assay. conditions, the relative standard deviation of the ratio of the Time span of measurement: About 3 times as long as the peak area of methylprednisolone succinate to that of the in- retention time of methylprednisolone succinate. ternal standard is not more than 1.0z. System suitability— Containers and storage Containers—Tight containers. Test for required detectability: Pipet 1 mL of the standard solution, and add the mixture of 0.05 mol/L phosphate buffer solution, pH 3.5 and acetonitrile (1:1) to make exactly 10 mL. Confirm that the peak area of methylprednisolone Methylrosanilinium Chloride succinate obtained from 5 mL of this solution is equivalent to Crystal Violet 7to13z of that from 5 mL of the standard solution. System performance: Proceed as directed in the System メチルロザニリン塩化物 suitability in the Assay. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating C25H30ClN3:407.98 conditions, the relative standard deviation of the peak area of methylprednisolone succinate is not more than 1.0z. Methylrosanilinium Chloride is hexamethylpararosaniline chloride, and is usually admixed with pen- Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, tamethylpararosaniline chloride and tetramethylpara- 3 hours). rosaniline chloride. Residue on ignition <2.44> Not more than 0.2z (0.5 g). It contains not less than 96.0z of methylrosanilinium chloride [as hexamethylpararosaniline chloride Assay Weigh accurately about 15 mg each of Methylpred- (C H ClN )], calculated on the dried basis. nisolone Succinate and Methylprednisolone Succinate RS, 25 30 3 previously dried, dissolve separately in 5 mL of methanol, Description Methylrosanilinium Chloride occurs as green and add the mixture of 0.05 mol/L phosphate buffer solu- fragments having a metallic luster or a dark green powder. It tion, pH 3.5 and acetonitrile (1:1) to make exactly 50 mL. is odorless or has a slight odor. Pipet 5 mL each of these solutions, add exactly 5 mL of the It is soluble in ethanol (95), sparingly soluble in water, and internal standard solution, and use these solutions as the practically insolubleindiethylether. sample solution and the standard solution, respectively. Per- Identification (1) To 1 mL of sulfuric acid add 1 mg of form the test with 5 mL each of the sample solution and Methylrosanilinium Chloride: it dissolves, and shows an standard solution as directed under Liquid Chromatography orange to red-brown color. To this solution add water drop- <2.01> according to the following conditions, and calculate wise: the color of the solution changes from brown through the ratios, Q and Q , of the peak area of methylpredniso- T S green to blue. lone succinate to that of the internal standard. (2) Dissolve 0.02 g of Methylrosanilinium Chloride in 10

Amount (mg) of C26H34O8 ＝ MS × QT/QS mL of water, add 5 drops of hydrochloric acid, and use this solution as the sample solution. To 5 mL of the sample solu- M : Amount (mg) of Methylprednisolone Succinate RS S tion add tannic acid TS dropwise: an intense blue precipitate Internal standard solution—A solution of ethyl parahy- is formed. 1106 Methyl Salicylate / Official Monographs JP XVI

(3) To 5 mL of the sample solution obtained in (2) add 0.5 g of zinc powder, and shake: the solution is decolorized. Methyl Salicylate Place 1 drop of this solution on filter paper, and apply 1 drop of ammonia TS adjacent to it: a blue color is produced サリチル酸メチル at the zone of contact of the both solutions. Purity (1) Ethanol-insoluble substances—Weigh accurately about 1 g of Methylrosanilinium Chloride, previously dried at 1059C for 4 hours, heat with 50 mL of ethanol (95) under a reflux condenser for 15 minutes in a water bath, and filter the mixture through a tared glass filter (G4). Wash the C8H8O3:152.15 residue on the filter with warm ethanol (95) until the last Methyl 2-hydroxybenzoate washing does not show a purple color, and dry at 1059Cfor [119-36-8] 2 hours: the mass of the residue is not more than 1.0z. (2) Heavy metals <1.07>—Proceed with 1.0 g of Methyl- Methyl Salicylate contains not less than 98.0z of rosanilinium Chloride according to Method 2, and perform C8H8O3. the test. Prepare the control solution with 3.0 mL of Stand- Description Methyl Salicylate is a colorless to pale yellow ard Lead Solution (not more than 30 ppm). liquid. It has a strong, characteristic odor. (3) Zinc—To 0.10 g of Methylrosanilinium Chloride add It is miscible with ethanol (95) and with diethyl ether. 0.1 mL of sulfuric acid, and incinerate by ignition. After It is very slightly soluble in water. cooling, boil with 5 mL of dilute hydrochloric acid, 0.5 mL Specific gravity d 20: 1.182 – 1.192 of dilute nitric acid and 4 mL of water, add 5 mL of ammo- 20 Boiling point: 219 – 2249C nia TS, boil again, and filter. To the filtrate add 2 to 3 drops of sodium sulfide TS: no turbidity is produced. Identification Shake 1 drop of Methyl Salicylate thor- (4) Arsenic <1.11>—Prepare the test solution with 0.40 g oughly with 5 mL of water for 1 minute, and add 1 drop of of Methylrosanilinium Chloride, according to Method 3, iron (III) chloride TS: a purple color develops. and perform the test (not more than 5 ppm). Purity (1) Acidity—Shake 5.0 mL of Methyl Salicylate Loss on drying <2.41> Not more than 7.5z (1 g, 1059C, thoroughly with 25 mL of freshly boiled and cooled water 4 hours). and 1.0 mL of 0.1 mol/L sodium hydroxide VS for 1 minute, add 2 drops of phenol red TS, and titrate <2.50> with Residue on ignition <2.44> Not more than 1.5z (0.5 g). 0.1 mol/L hydrochloric acid VS until the red color disap- Assay Transfer about 0.4 g of Methylrosanilinium Chlo- pears: not more than 0.45 mL of 0.1 mol/L sodium hydrox- ride, accurately weighed, to a wide-mouthed, conical flask, ide VS is consumed. add 25 mL of water and 10 mL of hydrochloric acid, dis- (2) Heavy metals—Shake 10.0 mL of Methyl Salicylate solve, and add exactly 50 mL of 0.1 mol/L titanium (III) thoroughly with 10 mL of water, add 1 drop of hydrochloric chloride VS while passing a stream of carbon dioxide acid, and saturate with hydrogen sulfide by passing it through the flask. Heat to boil, and boil gently for 15 through the mixture: neither the oily layer nor the aqueous minutes, swirling the liquid frequently. Cool while passing a layer shows a dark color. stream of carbon dioxide through the flask, titrate <2.50> the Assay Weigh accurately about 2 g of Methyl Salicylate, excess titanium (III) chloride with 0.05 mol/L ammonium add an exactly measured 50 mL of 0.5 mol/L potassium hy- iron (III) sulfate VS until a faint, red color is produced (indi- droxide-ethanol VS, and heat on a water bath for 2 hours cator: 5 mL of ammonium thiocyanate TS). Perform a blank under a reflux condenser. Cool, and titrate <2.50> the excess determination. potassium hydroxide with 0.5 mol/L hydrochloric acid VS Each mL of 0.1 mol/L titanium (III) chloride VS (indicator: 3 drops of phenolphthalein TS). Perform a blank

＝ 20.40 mg of C25H30ClN3 determination. Containers and storage Containers—Tight containers. Each mL of 0.5 mol/L potassium hydroxide-ethanol VS

＝ 76.08 mg of C8H8O3 Containers and storage Containers—Tight containers. JP XVI Official Monographs / Methyltestosterone 1107

Identification (1) Determine the absorption spectrum of a Compound Methyl Salicylate Spirit solution of Methyltestosterone in ethanol (95) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry 複方サリチル酸メチル精 <2.24>, and compare the spectrum with the Reference Spec- trum or the spectrum of a solution of Methyltestosterone RS prepared in the same manner as the sample solution: both Method of preparation spectra exhibit similar intensities of absorption at the same Methyl Salicylate 40 mL wavelengths. Capsicum Tincture 100 mL (2) Determine the infrared absorption spectrum of d-ordl-Camphor 50 g Methyltestosterone, previously dried, as directed in the po- Ethanol a sufficient quantity tassium bromide disk method under Infrared Spectropho- To make 1000 mL tometry <2.25>, and compare the spectrum with the Refer- ence Spectrum or the spectrum of dried Methyltestosterone Prepare as directed under Medicated Spirits, with the RS: both spectra exhibit similar intensities of absorption at above ingredients. the same wave numbers.

20 Description Compound Methyl Salicylate Spirit is a red- Optical rotation <2.49> [a]D : ＋79 – ＋859(after drying, dish yellow liquid, having a characteristic odor and a burn- 0.1 g, ethanol (95), 10 mL, 100 mm). ing taste. Melting point <2.60> 163 – 1689C Identification (1) Shake 1 mL of Compound Methyl Salicylate Spirit with 5 mL of dilute ethanol, and add 1 drop Purity Related substances—Dissolve 40 mg of Methyltesto- of iron (III) chloride TS: a purple color is produced (methyl sterone in 2 mL of ethanol (95), and use this solution as the salicylate). sample solution. Pipet 1 mL of the sample solution, add (2) Shake thoroughly 0.5 mL of Compound Methyl ethanol (95) to make exactly 100 mL, and use this solution as Salicylate Spirit with 10 mL of chloroform, and use this so- the standard solution. Perform the test with these solutions lution as the sample solution. Dissolve 40 mg of methyl as directed under Thin-layer Chromatography <2.03>. Spot salicylate in 10 mL of chloroform, and use this solution as 10 mL each of the sample solution and standard solution on a the standard solution. Perform the test with these solutions plate of silica gel with fluorescent indicator for thin-layer as directed under Thin-layer Chromatography <2.03>. Spot 5 chromatography. Develop the plate with a mixture of chlo- mL each of the sample solution and standard solution on the roform and diethylamine (19:1) to a distance of about 15 cm, plate of silica gel with fluorescent indicator for thin-layer and air-dry the plate. Examine under ultraviolet light (main chromatography. Develop the plate with a mixture of hexane wavelength: 254 nm): the spots other than the principal spot and chloroform (4:1) to a distance of about 10 cm, air-dry from the sample solution are not more intense than the spot the plate, and examine under ultraviolet light (main wave- from the standard solution. length: 254 nm): the spots from the sample solution and the Loss on drying <2.41> Not more than 1.0z (0.5 g, in vacu- standard solution show the same Rf value. Spray evenly iron um, phosphorus (V) oxide, 10 hours). (III) chloride TS upon the plate: the spot from the standard solution and the corresponding spot from the sample solu- Residue on ignition <2.44> Not more than 0.1z (0.5 g). tion reveal a purple color. Assay Weigh accurately about 20 mg each of Methyltesto- Containers and storage Containers—Tight containers. sterone and Methyltestosterone RS, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 10 hours, dissolve each in methanol to make exactly 200 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal Methyltestosterone standard solution, add methanol to make 50 mL, and use メチルテストステロン these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate

the ratios, QT and QS, of the peak area of methyltestosterone to that of the internal standard.

Amount (mg) of C20H30O2 ＝ MS × QT/QS

C20H30O2:302.45 MS: Amount (mg) of Methyltestosterone RS 17b-Hydroxy-17a-methylandrost-4-en-3-one [58-18-4] Internal standard solution—A solution of propyl parahydroxybenzoate in methanol (1 in 10,000). Methyltestosterone, when dried, contains not less Operating conditions— than 98.0z and not more than 102.0z of C H O . Detector: An ultraviolet absorption photometer (wave- 20 30 2 length: 241 nm). Description Methyltestosterone occurs as white to pale yel- Column: A stainless steel column 6 mm in inside diameter low, crystals or crystalline powder. and 15 cm in length, packed with octadecylsilanized silica gel It is freely soluble in methanol and in ethanol (95), and for liquid chromatography (5 mm in particle diameter). practically insoluble in water. Column temperature: A constant temperature of about 1108 Methyltestosterone Tablets / Official Monographs JP XVI

359C. absorbances, AT and AS, of the sample solution and the Mobile phase: A mixture of acetonitrile and water (11:9). standard solution at the wavelength of maximum absorption Flow rate: Adjust the flow rate so that the retention time at about 241 nm, respectively, as directed under Ultraviolet- of methyltestosterone is about 10 minutes. visible Spectrophotometry <2.25>. System suitability— Amount (mg) of methyltestosterone (C H O ) System performance: When the procedure is run with 10 20 30 2 ＝ M × A /A × V?/V × 1/10 mL of the standard solution under the above operating con- S T S ditions, the internal standard and methyltestosterone are MS: Amount (mg) of Methyltestosterone RS eluted in this order with the resolution between these peaks Dissolution <6.10> When the test is performed at 100 revo- being not less than 9. lutions per minute according to the Paddle method, using System repeatability: When the test is repeated 6 times 900 mL of a solution prepared by dissolving 1 g of polysor- with 10 mL of the standard solution under the above operat- bate 80 in water to make 5 L as the dissolution medium, the ing conditions, the relative standard deviation of the ratios dissolution rate in 30 minutes of a 10-mg tablet is not less of the peak area of methyltestosterone to that of the internal than 75z and that in 60 minutes of a 25-mg tablet is not less standard is not more than 1.0z. than 70z. Containers and storage Containers—Tight containers. Start the test with 1 tablet of Methyltestosterone Tablets, Storage—Light-resistant. withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the Methyltestosterone Tablets first 10 mL of the filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V?mL so メチルテストステロン錠 that each mL contains about 11 mg of methyltestosterone

(C20H30O2) according to the labeled amount, and use this solution as the sample solution. Separately, weigh accurately Methyltestosterone Tablets contain not less than about 22 mg of Methyltestosterone RS, previously dried in 90.0z and not more than 110.0z of the labeled vacuum using phosphorus (V) oxide as a desiccant for 10 amount of methyltestosterone (C H O : 302.45). 20 30 2 hours, and dissolve in ethanol (99.5) to make exactly 100 Method of preparation Prepare as directed under Tablets, mL. Pipet 5 mL of this solution, add the dissolution medium with Methyltestosterone. to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, A and A ,at249 Identification To a portion of powdered Methyltestoster- T S nm of the sample solution and standard solution as directed one Tablets, equivalent to 10 mg of Methyltestosterone ac- under Ultraviolet-visible Spectrophotometry <2.24>,using cording to the labeled amount, add 50 mL of acetone, shake the dissolution medium as the blank. for 30 minutes, and filter. Evaporate the filtrate to dryness, dissolve the residue in 10 mL of acetone, and use this solu- Dissolution rate (z) with respect to the labeled amount tion as the sample solution. Separately, dissolve 10 mg of of methyltestosterone (C20H30O2) Methyltestosterone RS in 10 mL of acetone, and use this so- ＝ MS × AT/AS × V?/V × 1/C × 45 lution as the standard solution. Perform the test with these M : Amount (mg) of Methyltestosterone RS solutions as directed under Thin-layer Chromatography S C: Labeled amount (mg) of methyltestosterone (C H O ) <2.03>. Spot 10 mL each of the sample solution and standard 20 30 2 in 1 tablet solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform and Assay Weigh accurately the mass of not less than 20 ethanol (95) (9:1) to a distance of about 12 cm, and air-dry Methyltestosterone Tablets, and powder. Weigh accurately a the plate. Spray evenly dilute sulfuric acid on the plate, and portion of the powder, equivalent to about 25 mg of methyl- heat at 1109C for 10 minutes: the spot from the sample solu- testosterone (C20H30O2), add about 70 mL of methanol, tion and the standard solution show the same Rfvalue. shake for 30 minutes, and add methanol to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 5 mL of the in- Uniformity of dosage units <6.02> Perform the test accord- ternal standard solution and methanol to make 50 mL, filter ing to the following method: it meets the requirement of the through a membrane filter (not exceeding 0.45 mm in pore Content uniformity test. size), and use the filtrate as the sample solution. Separately, To 1 tablet of Methyltestosterone Tablets add 5 mL of weigh accurately about 20 mg of Methyltestosterone RS, water to disintegrate, add 50 mL of methanol, and shake for previously dried in a desiccator (in vacuum, phosphorus (V) 30 minutes. Add methanol to make exactly 100 mL, and cen- oxide) for 10 hours, dissolve in methanol to make exactly trifuge. Measure exactly V mL of the supernatant liquid, add 200 mL. Pipet 5 mL of this solution, add exactly 5 mL of the methanol to make exactly V?mL of a solution containing internal standard solution, add methanol to make 50 mL, about 10 mg of methyltestosterone (C H O )perml,and 20 30 2 and use this solution as the standard solution. Perform the use this solution as the sample solution. Separately, weigh test with 10 mL each of the sample solution and standard so- accurately about 10 mg of Methyltestosterone RS, previously lution as directed under Liquid Chromatography <2.01> ac- dried in a desiccator (in vacuum, phosphorus (V) oxide) for cording to the following conditions, and calculate the ratios, 10 hours, and dissolve in 5 mL of water and 50 mL of meth- Q and Q , of the peak area of methyltestosterone to that of anol, then add methanol to make exactly 100 mL. Pipet 5 T S the internal standard. mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Determine the JP XVI Official Monographs / Meticrane 1109

Amount (mg) of methyltestosterone (C20H30O2) spectra exhibit similar intensities of absorption at the same ＝ MS × QT/QS × 5/4 wave numbers.

MS: Amount (mg) of Methyltestosterone RS Purity (1) Ammonium <1.02>—Perform the test with 0.10 g of Meticrane. Prepare the control solution with Internal standard solution—A solution of propyl parahy- 3.0 mL of Standard Ammonium Solution (not more than droxybenzoate in methanol (1 in 10,000). 0.03z). Operating conditions— (2) Heavy metals <1.07>—Proceed with 1.0 g of Meti- Detector: An ultraviolet absorption photometer (wave- crane according to Method 2, and perform the test. Prepare length: 241 nm). the control solution with 2.0 mL of Standard Lead Solution Column: A stainless steel column 6 mm in inside diameter (not more than 20 ppm). and 15 cm in length, packed with octadecylsilanized silica gel (3) Arsenic <1.11>—Prepare the test solution with 1.0 g for liquid chromatography (5 mminparticlediameter). of Meticrane according to Method 3, and perform the test Column temperature: A constant temperature of about (not more than 2 ppm). 359C. (4) Related substances—Dissolve 50 mg of Meticrane in Mobile phase: A mixture of acetonitrile and water (11:9). 50 mL of acetonitrile. To 5 mL of this solution add the mo- Flow rate: Adjust the flow rate so that the retention time bile phase to make 25 mL, and use this solution as the sam- of methyltestosterone is about 10 minutes. ple solution. Pipet 1 mL of the sample solution, add the mo- System suitability— bile phase to make exactly 100 mL, and use this solution as System performance: When the procedure is run with 10 the standard solution. Perform the test with exactly 10 mL mL of the standard solution under the above operating con- each of the sample solution and standard solution as directed ditions, the internal standard and methyltestosterone are under Liquid Chromatography <2.01> according to the fol- eluted in this order with the resolution between these peaks lowing conditions, and determine each peak area of both so- being not less than 9. lutions by the automatic integration method: the total area System repeatability: When the test is repeated 6 times of the peaks other than meticrane from the sample solution with 10 mL of the standard solution under the above operat- is not larger than the peak area of meticrane from the standing conditions, the relative standard deviation of the ratio of ard solution. the peak area of methyltestosterone to that of the internal Operating conditions 1— standard is not more than 1.0z. Detector: An ultraviolet absorption photometer (wave- Containers and storage Containers—Tight containers. length: 230 nm). Column: A stainless steel column 4.6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica Meticrane gel for liquid chromatography (5 mminparticlediameter). Column temperature: A constant temperature of about メチクラン 409C. Mobile phase: A mixture of water and acetonitrile (17:3). Flow rate: Adjust the flow rate so that the retention time of meticrane is about 7 minutes. Time span of measurement: About 4 times as long as the retention time of meticrane beginning after the solvent peak. System suitability 1— C H NO S : 275.34 10 13 4 2 Test for required detection: To exactly 2 mL of the stand- 6-Methylthiochromane-7-sulfonamide 1,1-dioxide ard solution add the mobile phase to make exactly 20 mL. [1084-65-7] Confirm that the peak area of meticrane obtained from 10 mL of this solution is equivalent to 7 to 13z of that from 10 Meticrane, when dried, contains not less than 98.0z mL of the standard solution. of C H NO S . 10 13 4 2 System performance: Dissolve 10 mg each of Meticrane Description Meticrane occurs as white, crystals or crystal- and caffeine in 100 mL of acetonitrile. To exactly 2 mL of line powder. It is odorless and has a slight bitter taste. this solution add the mobile phase to make exactly 10 mL. It is freely soluble in dimethylformamide, slightly soluble When the procedure is run with 10 mL of this solution under in acetonitrile and in methanol, very slightly soluble in the above operating conditions 1, caffeine and meticrane are ethanol (95), and practically insoluble in water. eluted in this order with the resolution between these peaks Melting point: about 2349C (with decomposition). being not less than 10. System repeatability: When the test is repeated 6 times Identification (1) Determine the absorption spectrum of a with 10 mL of the standard solution under the above operat- solution of Meticrane in methanol (3 in 10,000) as directed ing conditions 1, the relative standard deviation of the peak under Ultraviolet-visible Spectrophotometry <2.24>,and area of meticrane is not more than 2.0z. compare the spectrum with the Reference Spectrum: both Operating conditions 2— spectra exhibit similar intensities of absorption at the same Detector, column, and column temperature: Proceed as wavelengths. directed in the operating conditions 1. (2) Determine the infrared absorption spectrum of Meti- Mobile phase: A mixture of water and acetonitrile (1:1). crane, previously dried, as directed in the potassium bromide Flow rate: Adjust the flow rate so that the retention time disk method under Infrared Spectrophotometry <2.25>,and of meticrane is about 2 minutes. compare the spectrum with the Reference Spectrum: both 1110 Metildigoxin / Official Monographs JP XVI

Time span of measurement: About 10 times as long as the retention time of meticrane beginning after the solvent peak. Metildigoxin System suitability 2— Test for required detection: To exactly 2 mL of the stand- メチルジゴキシン ard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of meticrane obtained from 10 mL of this solution is equivalent to 7 to 13z of that from 10 mL of the standard solution. System performance: Dissolve 20 mg each of Meticrane and methyl parahydroxybenzoate in 100 mL of acetonitrile. To exactly 2 mL of this solution add the mobile phase to make exactly 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions 2, meticrane and methyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operating conditions 2, the relative standard deviation of the peak area of meticrane is not more than 2.0z. C H O . 1 C H O: 824.00 Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 42 66 14 2 3 6 3b-[2,6-Dideoxy-4-O-methyl-b-D-ribo-hexopyranosyl- 4 hours). (1→4)-2,6-dideoxy-b-D-ribo-hexopyranosyl-(1→4)- Residue on ignition <2.44> Not more than 0.1z (1 g). 2,6-dideoxy-b-D-ribo-hexopyranosyloxy]-12b,14- dihydroxy-5b-card-20(22)-enolide—acetone (2/ ) Assay Weigh accurately about 0.5 g of Meticrane, previ- 1 [30685-43-9, acetone-free] ously dried, dissolve in 50 mL of dimethylformamide, add 5 mL of water, and titrate <2.50> with 0.1 mol/L potassium Metildigoxin contains not less than 96.0z and not hydroxide-ethanol VS (potentiometric titration). Perform a 1 more than 103.0z of C H O . C H O, calculated on blank determination, and make any necessary correction. 42 66 14 2 3 6 the anhydrous basis. Each mL of 0.1 mol/L potassium hydroxide-ethanol VS Description Metildigoxin occurs as a white to light yellow- ＝ 27.54 mg of C H NO S 10 13 4 2 ish white, crystalline powder. Containers and storage Containers—Well-closed contain- It is freely soluble in N,N-dimethylformamide, in pyridine ers. and in acetic acid (100), soluble in chloroform, sparingly soluble in methanol, slightly soluble in ethanol (95) and in acetone, very slightly soluble in water, and practically insoluble in diethyl ether. Identification (1) Dissolve 2 mg of Metildigoxin in 2 mL of acetic acid (100), shake well with 1 drop of iron (III) chloride TS, and add gently 2 mL of sulfuric acid to divide into two layers: a brown color develops at the interface, and a deep blue color gradually develops in the acetic acid layer. (2) Dissolve 2 mg of Metildigoxin in 2 mL of 1,3-dinitrobenzene TS, add 2 mL of a solution of tetramethylammonium hydroxide in ethanol (95) (1 in 200), and shake: a purple color gradually develops, and changes to blue-purple. (3) Determine the absorption spectrum of a solution of Metildigoxin in methanol (1 in 50,000) as directed under Ul- traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Metildigoxin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (4) Determine the infrared absorption spectrum of Metil- digoxin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Metildigoxin RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve Metildigoxin and Metildigoxin RS in acetone, respectively, then evaporate the acetone to dryness, and repeat the test on the residues. JP XVI Official Monographs / Metoclopramide 1111

20 Optical rotation <2.49> [a]546.1: ＋22.0 – ＋25.59(1 g, cal- Metildigoxin RS (separately, determine the water <2.48> in culated on the anhydrous basis, pyridine, 10 mL, 100 mm). the same mammer as Metildigoxin), and dissolve each in methanol to make exactly 50 mL. Pipet 5 mL each of the so- Purity (1) Arsenic <1.11>—Prepare the test solution with lutions, add methanol to each to make exactly 100 mL, and 0.5 g of Metildigoxin according to Method 3, and perform use these solutions as the sample solution and the standard the test (not more than 4 ppm). solution, respectively. Pipet 5 mL each of the sample solu- (2) Related substances—Dissolve 10 mg of Metildigoxin tion and standard solution, add 15 mL of 2,4,6-trinitro- in 10 mL of chloroform, and use this solution as the sample phenol-ethanol TS and 2 mL of sodium hydroxide TS to solution. Pipet 1 mL of the sample solution, add chloroform each, shake well, add methanol to make exactly 25 mL, and to make exactly 50 mL, and use this solution as the standard allow to stand at 20 ± 0.59C for 20 minutes. Perform the solution. Perform the test with these solutions as directed test with these solutions as directed under Ultraviolet-visible under Thin-layer Chromatography <2.03>. Spot 20 mLeach Spectrophotometry <2.24> using a solution prepared by mix- of the sample solution and standard solution on a plate of ing 15 mL of 2,4,6-trinitrophenol-ethanol TS and 2 mL of silica gel for thin-layer chromatography. Develop the plate sodium hydroxide TS and adding methanol to make exactly with a mixture of 2-butanone and chloroform (3:1) to a dis- 25 mL as the blank. Determine the maximum absorbances, tance of about 15 cm, and air-dry the plate. Spray evenly A and A , of the subsequent solutions obtained from the dilute sulfuric acid on the plate, and heat at 1109Cfor10 T S sample solution and the standard solution, respectively, by minutes: the spots other than the principal spot from the measuring every 5 minutes, at 495 nm. sample solution are not more intense than the spot from the 1 standard solution. Amount (mg) of C42H66O14. 2 C3H6O ＝ M × A /A Acetone Weigh accurately about 0.1 g of Metildigoxin, dis- S T S solve in exactly 2 mL of the internal standard solution, add MS: Amount (mg) of Metildigoxin RS, calculated on the N,N-dimethylformamide to make 10 mL, and use this solu- anhydrous basis tion as the sample solution. Separately, weigh accurately Containers and storage Containers—Tight containers. about 0.4 g of acetone in a 50-mL volumetric flask containing about 10 mL of N,N-dimethylformamide, and add N,N- dimethylformamide to make 50 mL. Pipet 5 mL of this solution, add exactly 20 mL of the internal standard solution, Metoclopramide then add N,N-dimethylformamide to make 100 mL, and use メトクロプラミド this solution as the standard solution. Perform the test with 1 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02>, and calculate the ratios, QT and QS, of the peak area of acetone to that of the internal standard: the amount of acetone is between 2.0z and 5.0z.

Amount (z) of acetone ＝ MS/MT × QT/QS C H ClN O : 299.80 M : Amount (g) of acetone 14 22 3 2 S 4-Amino-5-chloro-N-[2-(diethylamino)ethyl]-2- M : amount (g) of the sample T methoxybenzamide Internal standard solution—A solution of t-butanol in N,N- [364-62-5] dimethylformamide (1 in 2000). Operating conditions— Metoclopramide, when dried, contains not less than Detector: A hydrogen flame-ionization detector. 99.0z of C14H22ClN3O2. Column: A glass column about 2 mm in inside diameter Description Metoclopramide occurs as white crystals or a and 1 to 2 m in length, packed with porous ethylvinylben- crystalline powder, and is odorless. zene-divinylbenzene copolymer for gas chromatography (150 It is freely soluble in acetic acid (100), soluble in methanol to 180 mm in particle diameter). and in chloroform, sparingly soluble in acetic anhydride, in Column temperature: A constant temperature between ethanol (95) and in acetone, very slightly soluble in diethyl 1709C and 2309C. ether, and practically insoluble in water. Carrier gas: Nitrogen. It dissolves in dilute hydrochloric acid. Flow rate: Adjust the flow rate so that the retention time of acetone is about 2 minutes. Identification (1) Dissolve 10 mg of Metoclopramide in 1 Selection of column: Proceed with 1 mL of the standard mL of dilute hydrochloric acid and 4 mL of water: the solu- solution under the above operating conditions, and calculate tion responds to the Qualitative Tests <1.09> for Primary the resolution. Use a column giving elution of acetone and t- Aromatic Amines. butanol in this order with the resolution between these peaks (2) Dissolve 10 mg of Metoclopramide in 5 mL of dilute being not less than 2.0. hydrochloric acid and 20 mL of water, and to 5 mL of this solution add 1 mL of Dragendorff's TS: a reddish orange Water <2.48> Not more than 3.0z (0.3 g, volumetric titra- precipitate is produced. tion, direct titration). (3) Dissolve 0.1 g of Metoclopramide in 1 mL of 1 Residue on ignition <2.44> Not more than 0.1z (0.5 g). mol/LhydrochloricacidTS,anddilutewithwatertomake 100 mL. To 1 mL of the solution add water to make 100 mL, Assay Weigh accurately 0.1 g each of Metildigoxin and 1112 Metoclopramide Tablets / Official Monographs JP XVI determine the absorption spectrum of the solution as di- according to the labeled amount, add 15 mL of 0.5 mol/L rected under Ultraviolet-visible Spectrophotometry <2.24>, hydrochloric acid TS, and heat in a water bath at 709Cfor and compare the spectrum with the Reference Spectrum: 15 minutes while frequent shaking. After cooling, centrifuge both spectra exhibit similar intensities of absorption at the for 10 minutes, and to 5 mL of the supernatant liquid add 1 same wavelengths. mL of 4-dimethylaminobenzaldehyde-hydrochloric acid TS: a yellow color develops. Melting point <2.60> 146–1499C (2) Determine the absorption spectrum of the sample so- Purity (1) Clarity and color of solution—Dissolve 1.0 g lution obtained in the Assay as directed under Ultraviolet- of Metoclopramide in 10 mL of 1 mol/L hydrochloric acid visible Spectrophotometry <2.24>: it exhibits maxima be- TS: the solution is clear and colorless. tween 270 nm and 274 nm, and between 306 nm and 310 nm. (2) Heavy metals <1.07>—Proceed with 1.0 g of Metoclo- Uniformity of dosage units <6.02> Perform the test accord- pramide as directed under Method 2, and perform the test. ing to the following method: it meets the requirement of the Prepare the control solution with 2.0 mL of Standard Lead Content uniformity test. Solution (not more than 20 ppm). To 1 tablet of Metoclopramide Tablets add 10 mL of 0.1 (3) Arsenic <1.11>—Dissolve 1.0 g of Metoclopramide in mol/L hydrochloric acid TS, disperse the particles with the 5 mL of 1 mol/L hydrochloric acid TS, and use this solution aid of ultrasonic waves, then add 0.1 mol/L hydrochloric as the sample solution. Perform the test (not more than 2 acid TS to make exactly 25 mL, and centrifuge for 10 ppm). minutes. Pipet 4 mL of the supernatant liquid, add 0.1 (4) Related substances—Dissolve 0.10 g of Metoclo- mol/L hydrochloric acid TS to make exactly V mL of a solu- pramide in 10 mL of methanol, and use this solution as the tion so that each mL contains about 12 mgofmetoclo- sample solution. Dilute 1 mL of the sample solution, exactly pramide (C H ClN O ), and use this solution as the sample measured, with methanol to make exactly 200 mL, and use 14 22 3 2 solution. Separately, weigh accurately about 80 mg of this solution as the standard solution. Perform the test with metoclopramide for assay, previously dried at 1059Cfor3 these solutions as directed under Thin-layer Chromatogra- hours, and dissolve in 0.1 mol/L hydrochloric acid TS to phy <2.03>. Spot 10 mL each of the sample solution and make exactly 500 mL. Pipet 4 mL of this solution, add 0.1 standard solution on a plate of silica gel with fluorescent in- mol/LhydrochloricacidTStomakeexactly50mL,anduse dicator for thin-layer chromatography. Develop the plate this solution as the standard solution. Determine the absor- with a mixture of 1-butanol and ammonia solution (28) bances, A and A , of the sample solution and standard so- (19:1) to a distance of about 10 cm. Dry the plate, first in air T S lution at 308 nm as directed under Ultraviolet-visible Spec- and then at 809C for 30 minutes. Examine under ultraviolet trophotometry <2.24>. light (main wavelength: 254 nm): the spots other than the principal spot from the sample solution are not more intense Amount (mg) of metoclopramide (C14H22ClN3O2) than the spot from the standard solution. ＝ MS × AT/AS × V/1000

Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, MS: Amount (mg) of metoclopramide for assay 3 hours). Dissolution Being specified separately. Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately not less than 20 Metoclopramide Assay Dissolve about 0.4 g of Metoclopramide, previously Tablets, and powder. Weigh accurately a portion of the dried and accurately weighed, in 50 mL of acetic acid (100), powder, equivalent to about 75 mg of metoclopramide add 5 mL of acetic anhydride, and warm for 5 minutes. (C14H22ClN3O2), add 300 mL of 0.1 mol/L hydrochloric acid Allow to cool, and titrate <2.50> with 0.1 mol/L perchloric TS, shake for 1 hour, and add 0.1 mol/L hydrochloric acid acid VS (indicator: 2 drops of crystal violet TS). Perform the TS to make exactly 500 mL. Centrifuge for 10 minutes, pipet blank determination, and make any necessary correction. 4 mL of the supernatant liquid, add 0.1 mol/hydrochloric acid TS to make exactly 50 mL, and use this solution as the Each mL of 0.1 mol/L perchloric acid VS sample solution. Separately, weigh accurately about 80 mg ＝ 29.98 mg of C H ClN O 14 22 3 2 of metoclopramide for assay, previously dried at 1059Cfor3 Containers and storage Containers—Well-closed contain- hours, and dissolve in 0.1 mol/L hydrochloric acid TS to ers. make exactly 500 mL. Pipet 4 mL of this solution, add 0.1 mol/hydrochloric acid TS to make exactly 50 mL, and use this solution as the standard solution. Determine the absor- Metoclopramide Tablets bances, AT and AS, of the sample solution and standard solution at 308 nm as directed under Ultraviolet-visible Spec- メトクロプラミド錠 trophotometry <2.24>.

Amount (mg) of metoclopramide (C14H22ClN3O2) Metoclopramide Tablets contain not less than ＝ MS × AT/AS 95.0z and not more than 105.0z of the labeled MS: Amount (mg) of metoclopramide for assay amount of metoclopramide (C14H22ClN3O2: 299.80). Containers and storage Containers—Tight containers. Method of preparation Prepare as directed under Tablets, with Metoclopramide. Identification (1) To a quantity of powdered Metoclo- pramide Tablets, equivalent to 50 mg of Metoclopramide JP XVI Official Monographs / Metoprolol Tartrate Tablets 1113

tate and methanol (4:1), to a distance of about 12 cm, and Metoprolol Tartrate air-dry the plate. Allow to stand the plate in an iodine vapors until the spot with the standard solution appears obviously: メトプロロール酒石酸塩 the spot other than the principal spot and other than the spot on the original point with the sample solution is not more than three spots, and they are not more intense than the spot with the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum, 609C, 4 hours). (C H NO )・C H O : 684.81 15 25 3 2 4 6 6 Residue on ignition <2.44> Not more than 0.1z (1 g). (2RS)-1-[4-(2-Methoxyethyl)phenoxy]-3- [(1-methylethyl)amino]propan-2-ol hemi-(2R,3R)-tartrate Assay Weigh accurately about 0.5 g of Metoprolol Tar- [56392-17-7] trate, previously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (poten- Metoprolol Tartrate, when dried, contains not tiometric titration). Perform a blank determination in the less than 99.0z and not more than 101.0z of same manner, and make any necessary correction. (C H NO ) .C H O . 15 25 3 2 4 6 6 Each mL of 0.1 mol/L perchloric acid VS

Description Metoprolol Tartrate occurs as a white crystal- ＝ 34.24 mg of (C15H25NO3)2.C4H6O6 line powder. Containers and storage Containers—Well-closed contain- It is very soluble in water, and freely soluble in methanol, ers. in ethanol (95) and in acetic acid (100). 20 Optical rotation [a]D : ＋7.0 – ＋10.09(after drying, 1 g, water, 50 mL, 100 mm). Metoprolol Tartrate Tablets Identification (1) Determine the absorption spectrum of a solution of Metoprolol Tartrate (1 in 10,000) as directed メトプロロール酒石酸塩錠 under Ultraviolet-visible Spectrophotometry <2.24>,and compare the spectrum with the Reference Spectrum: both Metoprolol Tartrate Tablets contain not less than spectra exhibit similar intensities of absorption at the same 93.0z and not more than 107.0z of the labeled wavelengths. amount of metoprolol tartrate ((C H NO ) .C H O : (2) Determine the infrared absorption spectrum of 15 25 3 2 4 6 6 684.81). Metoprolol Tartrate, previously dried, as directed in the paste method under Infrared Spectrophotometry <2.25>,and Method of preparation Prepare as directed under Tablets, compare the spectrum with the Reference Spectrum: both with Metoprolol Tartrate. spectra exhibit similar intensities of absorption at the same Identification To an amount of powdered Metoprolol Tar- wave numbers. If any difference appears between the spec- trate Tablets, equivalent to 10 mg of Metoprolol Tartrate ac- tra, recrystallize Metoprolol Tartrate from a solution in ace- cording to the labeled amount, add 100 mL of ethanol (95), tone (23 in 1000), filter and dry the crystals, and perform the shake for 15 minutes, and filter. Determine the absorption test with the crystals. spectrum of the filtrate as directed under Ultraviolet-visible (3) A solution of Metoprolol Tartrate (1 in 5) responds Spectrophotometry <2.24>: it exhibits maxima between 274 to the Qualitative Tests <1.09> (1) for tartrate. nm and 278 nm and between 281 nm and 285 nm. pH <2.54> The pH of a solution obtained by dissolving Uniformity of dosage units <6.02> Perform the test accord- 1.0 g of Metoprolol Tartrate in 10 mL of water is between ing to the following method: it meets the requirement of the 6.0 and 7.0. Content uniformity test. Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of To 1 tablet of Metoprolol Tartrate Tablets add 1 mL of Metoprolol Tartrate according to Method 1, and perform water for every 10 mg of Metoprolol Tartrate, shake for 20 the test. Prepare the control solution with 2.0 mL of Stand- minutes, then add 75 mL of ethanol (95), shake for 15 ard Lead Solution (not more than 10 ppm). minutes, add ethanol (95) to make exactly 100 mL, and cen- (2) Related substances—Dissolve 0.10 g of Metoprolol trifuge. Pipet V mL of the supernatant liquid, add ethanol Tartrate in 5 mL of methanol, and use this solution as the (95) to make exactly V? so that each mL contains about 0.1 sample solution. Pipet 1 mL of the sample solution, and add mg of metoprolol tartrate ((C15H25NO3)2.C4H6O6), and use methanol to make exactly 100 mL. Pipet 2 mL of this solu- this solution as the sample solution. Separately, weigh accu- tion, add methanol to make exactly 10 mL, and use this solu- rately about 50 mg of metoprolol tartrate for assay, previ- tion as the standard solution. Perform the test with these so- ously dried in vacuum at 609C for 4 hours, dissolve in 5 mL lutions as directed under Thin-layer Chromatography <2.03>. of water, and add ethanol (95) to make exactly 100 mL. Spot 10 mL each of the sample solution and standard solu- Pipet 10 mL of this solution, add ethanol (95) to make ex- tion on a plate of silica gel for thin-layer chromatography. actly 50 mL, and use this solution as the standard solution.

After saturating the plate with the atmosphere by allowing to Determine the absorbances, AT and AS, of the sample solu- stand in a developing vessel, which contains the developing tion and standard solution at 276 nm as directed under Ul- solvent and a glass vessel containing ammonia water (28), traviolet-visible Spectrophotometry <2.24>, using ethanol develop with the developing solvent, a mixture of ethyl ace- (95) as the blank. 1114 Metronidazole / Official Monographs JP XVI

Amount (mg) of metoprolol tartrate ((C15H25NO3)2.C4H6O6) solution and standard solution as directed under Liquid ＝ MS × AT/AS × V?/V × 1/5 Chromatography <2.01> according to the following conditions, and calculate the ratios, Q and Q , of the peak area M : Amount (mg) of metoprolol tartrate for assay T S S of metoprolol to that of the internal standard. Dissolution <6.10> When the test is performed at 50 revolu- Amount (mg) of metoprolol tartrate ((C H NO ) .C H O ) tions per minute according to the Paddle method, using 900 15 25 3 2 4 6 6 ＝ M × Q /Q mL of water as the dissolution medium, the dissolution rate S T S in 30 minutes of Metoprolol Tartrate Tablets is not less than MS: Amount (mg) of metoprolol tartrate for assay 80z. Internal standard solution—A solution of ethyl parahy- Start the test with 1 tablet of Metoprolol Tartrate Tablets, droxybenzoate in the mixture of ethanol (99.5) and 1 mol/L withdraw not less than 20 mL of the medium at the specified hydrochloric acid TS (100:1) (1 in 500). minute after starting the test, and filter through a membrane Operating conditions— filter with a pore size not exceeding 0.5 mm. Discard the first Detector: An ultraviolet absorption photometer (wave- 10 mL of the filtrate, pipet V mL of the subsequent filtrate, length: 274 nm). add water to make exactly V?mL so that each mL contains Column: A stainless steel column 4.6 mm in inside diame- about 22 mg of metoprolol tartrate ((C H NO ) .C H O ) 15 25 3 2 4 6 6 ter and 15 cm in length, packed with octadecylsilanized silica according to the labeled amount, and use this solution as the gel for liquid chromatography (5 mminparticlediameter). sample solution. Separately, weigh accurately about 56 mg Column temperature: A constant temperature of about of metoprolol tartrate for assay, previously dried in vacuum 259C. at 609C for 4 hours, and dissolve in water to make exactly Mobile phase: Dissolve 14.0 g of sodium perchlorate 200 mL. Pipet 8 mL of this solution, add water to make ex- monohydrate in 1000 mL of water, and adjust to pH 3.2 actly 100 mL, and use this solution as the standard solution. with diluted perchloric acid (17 in 2000). To 750 mL of this Perform the test with exactly 50 mLeachofthesamplesolu- solution add 250 mL of acetonitrile. tion and standard solution as directed under Liquid Chroma- Flow rate: Adjust the flow rate so that the retention time tography <2.01> according to the following conditions, and of metoprolol is about 8 minutes. determine the peak areas, A and A , of metoprolol. T S System suitability— Dissolution rate (z) with respect to the labeled amount System performance: When the procedure is run with 10

of metoprolol tartrate ((C15H25NO3)2.C4H6O6) mL of the standard solution under the above operating con- ＝ MS × AT/AS × V?/V × 1/C × 36 ditions, metoprolol and the internal standard are eluted in this order with the resolution between these peaks being not M : Amount (mg) of metoprolol tartrate for assay S less than 5. C: Labeled amount (mg) of metoprolol tartrate System repeatability: When the test is repeated 6 times ((C H NO ) .C H O )in1tablet 15 25 3 2 4 6 6 with 10 mL of the standard solution under the above operat- Operating conditions— ing conditions, the relative standard deviation of the ratio of Proceed as directed in the Assay. the peak area of metoprolol to that of the internal standard System suitability— is not more than 1.0z. System performance: When the procedure is run with 50 Containers and storage Containers—Well-closed contain- mL of the standard solution under the above operating con- ers. ditions, the number of theoretical plates and the symmetry factor of the peak of metoprolol are not less than 2000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times Metronidazole with 50 mL of the standard solution under the above operat- メトロニダゾール ing conditions, the relative standard deviation of the peak area of metoprolol is not more than 2.0z. Assay Weigh accurately the mass of not less than 20 Metoprolol Tartrate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.12 g of metoprolol tartrate ((C15H25NO3)2.C4H6O6), add 60 mL of a C6H9N3O3: 171.15 mixture of ethanol (99.5) and 1 mol/L hydrochloric acid TS 2-(2-Methyl-5-nitro-1H-imidazol-1-yl)ethanol (100:1) and exactly 10 mL of the internal standard solution, [443-48-1] shake for 15 minutes, and add the mixture of ethanol (99.5) and 1 mol/L hydrochloric acid TS (100:1) to make 100 mL. Metronidazole, when dried, contains not less than Centrifuge, and use the supernatant liquid as the sample so- 99.0z and not more than 101.0z of C6H9N3O3. lution. Separately, weigh accurately about 0.12 g of meto- Description Metronidazole occurs as white to pale yellow- prolol tartrate for assay, previously dried in vacuum at 609C ish white crystals or crystalline powder. for 4 hours, dissolve in 60 mL of the mixture of ethanol It is freely soluble in acetic acid (100), sparingly soluble in (99.5) and 1 mol/L hydrochloric acid TS (100:1), add exactly ethanol (99.5) and in acetone, and slightly soluble in water. 10 mL of the internal standard solution, then add the mix- It dissolves in dilute hydrochloric acid. ture of ethanol (99.5) and 1 mol/L hydrochloric acid TS It is colored to yellow-brown by light. (100:1) to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mLeachofthesample Identification (1) Determine the absorption spectrum of a JP XVI Official Monographs / Metronidazole Tablets 1115 solution of Metronidazole in 0.1 mol/L hydrochloric acid TS zole Tablets, equivalent to 0.1 g of Metronidazole according (1 in 100,000) as directed under Ultraviolet-visible Spectro- to the labeled amount, add 100 mL of 0.1 mol/L hydrochlo- photometry <2.24>, and compare the spectrum with the Ref- ric acid TS, and allow to stand for 30 minutes with occa- erence Spectrum: both spectra exhibit similar intensities of sional stirring. Then, shake vigorously, and centrifuge a part absorption at the same wavelengths. of this solution. To 1 mL of the supernatant liquid add 0.1 (2) Determine the infrared absorption spectrum of mol/LhydrochloricacidTStomake100mL.Determinethe Metronidazole as directed in the potassium bromide disk absorption spectrum of this solution as directed under Ultra- method under Infrared Spectrophotometry <2.25>,andcom- violet-visible Spectrophotometry <2.24>: it exhibits a maxi- pare the spectrum with the Reference Spectrum: both spectra mum between 275 nm and 279 nm. exhibit similar intensities of absorption at the same wave (2) Shake vigorously a quantity of powdered Metronida- numbers. zole Tablets, equivalent to 0.20 g of Metronidazole according to the labeled amount, with 20 mL of acetone for 10 Melting point <2.60> 159–1639C. minutes, centrifuge, and use the supernatant liquid as the Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of sample solution. Separately, dissolve 0.10 g of metronida- Metronidazole according to Method 2, and perform the test. zole in 10 mL of acetone, and use this solution as the stand- Prepare the control solution with 2.0 mL of Standard Lead ard solution. Perform the test with these solutions as di- Solution (not more than 20 ppm). rected under Thin-layer Chromatography <2.03>. Spot 5 mL (2) 2-Methyl-5-nitroimidazol—Dissolve 0.10 g of Metro- each of the sample solution and standard solution on a plate nidazole in acetone to make exactly10mL,andusethissolu- of silica gel with fluorescent indicator for thin-layer chroma- tion as the sample solution. Separately, dissolve 20 mg of 2- tography, develop the plate immediately with a mixture of methyl-5-nitroimidazole for thin-layer chromatography in acetone, water and ethyl acetate (8:1:1) to a distance of acetone to make exactly 20 mL, then pipet 5 mL of this solu- about 10 cm, and air-dry the plate. Examine under ultravio- tion, add acetone to make exactly 100 mL, and use this solu- let light (main wavelength: 254 nm): the Rf value of the prin- tion as the standard solution. Perform the test with these so- cipal spots obtained from the sample solution and the stand- lutions as directed under Thin-layer Chromatography <2.03>. ard solution is the same. Spot 20 mL each of the sample solution and standard solu- Uniformity of dosage units <6.02> Perform the test accord- tion on a plate of silica gel with fluorescent indicator for ing to the following method: it meets the requirement of the thin-layer chromatography. Immediately develop the plate Content uniformity test. with a mixture of acetone, water and ethyl acetate (8:1:1) to To 1 tablet of Metronidazole Tablets add 25 mL of a mix- a distance of about 15 cm, and air-dry the plate. Examine ture of water and methanol (1:1), shake vigorously for 25 under ultraviolet light (main wavelength: 254 nm): the spot minutes, and add the mixture of water and methanol (1:1) to from the sample solution corresponding to the spot from the make exactly 50 mL. Pipet 5 mL of this solution, and add a standard solution is not more intense than the spot from the mixture of water and methanol (4:1) to make exactly 100 standard solution. mL. Filter the solution through a membrane filter with pore Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- size of 0.45 mm, discard the first 3 mL of the filtrate, and use um, silica gel, 24 hours). the subsequent filtrate as the sample solution. Hereinafter, proceed as directed in the Assay. Residue on ignition <2.44> Not more than 0.1z (1 g). Amount (mg) of metronidazole (C H N O ) Assay Weigh accurately about 0.2 g of Metronidazole, pre- 6 9 3 3 ＝ M × A /A × 10 viously dried, and dissolve in 30 mL of acetic acid (100). S T S

Titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: MS: Amount (mg) of metronidazole for assay 0.5 mL of p-naphtholbenzein TS) until the color of the solu- Dissolution <6.10> When the test is performed at 50 revolu- tion changes from orange-yellow to green. Perform a blank tions per minute according to the Paddle method, using 900 determination, and make any necessary correction. mL of water as the dissolution medium, the dissolution rate Each mL of 0.1 mol/L perchloric acid VS in 90 minutes of Metronidazole Tablets is not less than 70z.

＝ 17.12 mg of C6H9N3O3 Start the test with 1 tablet of Metronidazole Tablets, withdraw not less than 20 mL of the medium at the specified Containers and storage Containers—Tight containers. minute after starting the test, and filter through a membrane Storage—Light-resistant. filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet V mL of the subsequent fil- Metronidazole Tablets trate, add water to make exactly V?mL so that each mL contains about 11 mg of metronidazole (C6H9N3O3) according to the labeled amount, and use this solution as the sample メトロニダゾール錠 solution. Separately, weigh accurately about 22 mg of metronidazole for assay, previously dried in vacuum with Metronidazole Tablets contain not less than 93.0z silica gel for 24 hours, and dissolve in water to make exactly and not more than 107.0z of the labeled amount of 100 mL. Pipet 5 mL of this solution, add water to make ex- metronidazole (C6H9N3O3: 171.15). actly 100 mL, and use this solution as the standard solution. Determine the absorbances, A and A ,at320nmofthe Method of preparation Prepare as directed under Tablets, T S sample solution and standard solution as directed under Ul- with Metronidazole. traviolet-visible Spectrophotometry <2.24>. Identification (1) To an amount of powdered Metronida- 1116 Metyrapone / Official Monographs JP XVI

Dissolution rate (z) with respect to the labeled amount of metronidazole (C6H9N3O3) Metyrapone ＝ MS × AT/AS × V?/V × 1/C × 45 メチラポン MS: Amount (mg) of metronidazole for assay C: Labeled amount (mg) of metronidazole (C6H9N3O3)in 1tablet Assay Weigh accurately the mass of not less than 20 Metronidazole Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.25 g of metronida- C H N O: 226.27 zole (C H N O ),add25mLofamixtureofwaterandmeth- 14 14 2 6 9 3 3 2-Methyl-1,2-di(pyridin-3-yl)propan-1-one anol (1:1), shake vigorously for 10 minutes, and add the mix- [54-36-4] ture of water and methanol (1:1) to make exactly 50 mL. Pipet 5 mL of this solution, and add a mixture of water and Metyrapone, when dried, contains not less than methanol (4:1) to make exactly 100 mL. Filter this solution 98.0z of C H N O. through a membrane filter with pore size of 0.45 mm, discard 14 14 2 the first 3 mL of the filtrate, and use the subsequent filtrate Description Metyrapone occurs as a white to pale yellow, as the sample solution. Separately, weigh accurately about crystalline powder. It has a characteristic odor and a bitter 25 mg of metronidazole for assay, previously dried in vacu- taste. um on silica gel for 24 hours, dissolve in the mixture of water It is very soluble in methanol, in ethanol (95), in acetic an- and methanol (4:1) to make exactly 100 mL, and use this so- hydride, in chloroform, in diethyl ether and in nitrobenzene, lution as the standard solution. Perform the test with exactly and sparingly soluble in water. 10 mL each of the sample solution and standard solution as It dissolves in 0.5 mol/L sulfuric acid TS. directed under Liquid Chromatography <2.01> according to Identification (1) Mix 5 mg of Metyrapone with 10 mg of the following conditions, and determine the peak areas, A T 1-chloro-2,4-dinitrobenzene, melt by gently heating for 5 to and A , of metronidazole. S 6 seconds, cool, and add 4 mL of potassium hydroxide-

Amount (mg) of metronidazole (C6H9N3O3) ethanol TS: a dark red color develops. ＝ MS × AT/AS × 10 (2) Determine the absorption spectrum of a solution of Metyrapone in 0.5 mol/L sulfuric acid TS (1 in 100,000) as M : Amount (mg) of metronidazole for assay S directed under Ultraviolet-visible Spectrophotometry <2.24>, Operating conditions— and compare the spectrum with the Reference Spectrum: Detector: An ultraviolet absorption photometer (wave- both spectra exhibit similar intensities of absorption at the length: 320 nm). same wavelengths. Column: A stainless steel column 4.6 mm in inside diame- Melting point <2.60> 50–549C. ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Purity (1) Clarity and color of solution—Dissolve 0.5 g Column temperature: A constant temperature of about of Metyrapone in 5 mL of methanol: the solution is clear 259C. and colorless to pale yellow. Mobile phase: A mixture of water and methanol (4:1). (2) Heavy metals <1.07>—Proceed with 2.0 g of Metyra- Flow rate: Adjust the flow rate so that the retention time pone according to Method 2, and perform the test. Prepare of metronidazole is about 5 minutes. the control solution with 2.0 mL of Standard Lead Solution System suitability— (not more than 10 ppm). System performance: When the procedure is run with 10 (3) Arsenic <1.11>—Prepare the test solution with 1.0 g mL of the standard solution under the above operating con- of Metyrapone, according to Method 3, and perform the test ditions, the number of theoretical plates and the symmetry (not more than 2 ppm). factor of the peak of metronidazole are not less than 3000 (4) Related substances—Dissolve 0.25 g of Metyrapone and not more than 1.5, respectively. in 5 mL of methanol, and use this solution as the sample so- System repeatability: When the test is repeated 6 times lution. Pipet 1 mL of the sample solution, and add methanol with 10 mL of the standard solution under the above operat- to make exactly 50 mL. Pipet 5 mL of this solution, add ing conditions, the relative standard deviation of the peak methanol to make exactly 50 mL, and use this solution as the area of metronidazole is not more than 1.0z. standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 Containers and storage Containers—Tight containers. mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform and methanol (15:1) to a distance of about 10 cm, and air-dry the plate for about 15 minutes. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum, silica gel, 24 hours). JP XVI Official Monographs / Mexiletine Hydrochloride 1117

Residue on ignition <2.44> Not more than 0.1z (1 g). Purity (1) Clarity and color of solution—Dissolve 1.0 g of Mexiletine Hydrochloride in 10 mL of water: the solution Assay Weigh accurately about 0.2 g of Metyrapone, previ- is clear and colorless. ously dried, dissolve in 10 mL of nitrobenzene and 40 mL of (2) Heavy Metals <1.07>—Proceed with 2.0 g of Mexile- acetic anhydride, and titrate <2.50> with 0.1 mol/L perchlo- tine Hydrochloride according to Method 1, and perform the ric acid VS (potentiometric titration). Perform a blank deter- test. Prepare the control solution with 2.0 mL of Standard mination, and make any necessary correction. Lead Solution (not more than 10 ppm). Each mL of 0.1 mol/L perchloric acid VS (3) Related substances—Dissolve 20 mg of Mexiletine

＝ 11.31 mg of C14H14N2O Hydrochloride in 20 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so- Containers and storage Containers—Tight containers. lution, add the mobile phase to make exactly 250 mL, and Storage—Light-resistant. use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> Mexiletine Hydrochloride according to the following conditions. Determine each peak area of both solutions by the automatic integration method: メキシレチン塩酸塩 each peak area of the peaks other than the peak of mexiletine from the sample solution is not larger than the peak area of mexiletine from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, flow rate, and selection of column: Proceed as directed in

C11H17NO.HCl: 215.72 the operating conditions in the Assay. (1RS)-2-(2,6-Dimethylphenoxy)-1-methylethylamine Detection sensitivity: Adjust the detection sensitivity so monohydrochloride that the peak height of mexiletine obtained from 20 mLof [5370-01-4] the standard solution is between 5 mm and 10 mm. Time span of measurement: About 3 times as long as the Mexiletine Hydrochloride, when dried, contains retention time of mexiletine beginning after peaks of the sol- not less than 98.0z and not more than 102.0z of vent. C H NO.HCl. 11 17 Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, Description Mexiletine Hydrochloride occurs as a white 3 hours). powder. Residue on ignition <2.44> Not more than 0.1z (1 g). It is freely soluble in water and in ethanol (95), slightly soluble in acetonitrile, and practically insoluble in diethyl Assay Weigh accurately about 20 mg each of Mexiletine ether. Hydrochloride and Mexiletine Hydrochloride RS, each pre- A solution of Mexiletine Hydrochloride (1 in 20) shows no viously dried, and dissolve each in the mobile phase to make optical rotation. exactly 20 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard solution, then add the Identification (1) Determine the absorption spectrum of a mobile phase to make 100 mL, and use these solutions as the solution of Mexiletine Hydrochloride in 0.01 mol/L hydro- sample solution and the standard solution, respectively. Per- chloric acid TS (1 in 2000) as directed under Ultraviolet- form the test with 20 mL each of the sample solution and visible Spectrophotometry <2.24>, and compare the spectrum standard solution as directed under Liquid Chromatography with the Reference Spectrum or the spectrum of a solution of <2.01> according to the following conditions, and calculate Mexiletine Hydrochloride RS prepared in the same manner the ratios, Q and Q , of the peak area of mexiletine to that as the sample solution: both spectra exhibit similar intensi- T S of the internal standard, respectively. ties of absorption at the same wavelengths.

(2) Determine the infrared absorption spectrum of Amount (mg) of C11H17NO.HCl Mexiletine Hydrochloride, previously dried, as directed in ＝ MS × QT/QS the potassium bromide disk method under Infrared Spec- M : Amount (mg) of Mexiletine Hydrochloride RS trophotometry <2.25>, and compare the spectrum with the S Reference Spectrum or the spectrum of dried Mexiletine Internal standard solution—A solution of phenetylamine hy- Hydrochloride RS: both spectra exhibit similar intensities of drochloride in the mobile phase (3 in 5000). absorption at the same wave numbers. If any difference Operating conditions— appears between the spectra, recrystallize Mexiletine Hydro- Detector: An ultraviolet absorption photometer (wave- chloride from ethanol (95), filter, dry the crystals, and repeat length: 210 nm). the test on the crystals. Column: A stainless steel column about 4 mm in inside di- (3) A solution of Mexiletine Hydrochloride (1 in 100) ameter and about 15 cm in length, packed with octylsilanized responds to the Qualitative Tests <1.09> (2) for chloride. silica gel for liquid chromatography (about 7 mminparticle diameter). pH <2.54> Dissolve 1.0 g of Mexiletine Hydrochloride in 10 Column temperature: A constant temperature of about mL of water: the pH of this solution is between 3.8 and 5.8. 309C. Melting point <2.60> 200–2049C. Mobile phase: Dissolve 2.5 g of sodium lauryl sulfate and 3 g of sodium dihydrogenphosphate dihydrate in 600 mL of 1118 Miconazole / Official Monographs JP XVI water, and add 420 mL of acetonitrile. tion. Pipet 1 mL of the sample solution, add methanol to Flow rate: Adjust the flow rate so that the retention time make exactly 20 mL. Pipet 1 mL of this solution, add metha- of mexiletine is about 6 minutes. nol to make exactly 20 mL, and use this solution as the Selection of column: Proceed with 20 mL of the standard standard solution. Perform the test with these solutions as solution under the above conditions, and calculate the reso- directed under Thin-layer Chromatography <2.03>. Spot 50 lution. Use a column giving elution of the internal standard mL each of the sample solution and standard solution on a and mexiletine in this order with the resolution between these plate of silica gel for thin-layer chromatography. Develop peaks being not less than 9. the plate with a mixture of hexane, chloroform, methanol and ammonia solution (28) (60:30:10:1) to a distance of Containers and storage Containers—Tight containers. about 12 cm, and air-dry the plate. Allow the plate to stand Storage—Light-resistant. in iodine vapor for 20 minutes: the spots other than the principal spot from the sample solution are not more intense Miconazole than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- ミコナゾール um, silica gel, 60z, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.3 g of Miconazole, previously dried, dissolve in 40 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 3 drops of p-naphtholbenzein TS) until the color of the solution changes from light yellow-brown to light yellow-green. Per- form a blank determination, and make any necessary correction. C H Cl N O: 416.13 18 14 4 2 Each mL of 0.1 mol/L perchloric acid VS 1-[(2RS)-2-(2,4-Dichlorobenzyloxy)-2-(2,4- ＝ 41.61 mg of C H Cl N O dichlorophenyl)ethyl]-1H-imidazole 18 14 4 2 [22916-47-8] Containers and storage Containers—Tight containers.

Miconazole, when dried, contains not less than 98.5z of C18H14Cl4N2O. Miconazole Nitrate Description Miconazole occurs as a white to pale yellowish ミコナゾール硝酸塩 white, crystalline powder. It is freely soluble in methanol, in ethanol (95) and in acetic acid (100), soluble in diethyl ether, and practically insoluble in water. A solution of Miconazole in methanol (1 in 20) shows no optical rotation. Identification (1) Determine the absorption spectrum of a solution of Miconazole in methanol (1 in 2500) as directed under Ultraviolet-visible Spectrophotometry <2.24>,and compare the spectrum with the Reference Spectrum: both C18H14Cl4N2O.HNO3:479.14 spectra exhibit similar intensities of absorption at the same 1-[(2RS)-2-(2,4-Dichlorobenzyloxy)-2-(2,4- wavelengths. dichlorophenyl)ethyl]-1H-imidazole mononitrate (2) Determine the infrared absorption spectrum of [22832-87-7] Miconazole, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry Miconazole Nitrate, when dried, contains not less <2.25>, and compare the spectrum with the Reference Spec- than 98.5z of C18H14Cl4N2O.HNO3. tum: both spectra exhibit similar intensities of absorption at Description Miconazole Nitrate occurs as a white crystal- the same wave numbers. line powder. Melting point <2.60> 84–879C. It is freely soluble in N,N-dimethylformamide, sparingly soluble in methanol, slightly soluble in ethanol (95), in ace- Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of tone and in acetic acid (100), and very slightly soluble in Miconazole according to Method 2, and perform the test. water and in diethyl ether. Prepare the control solution with 1.0 mL of Standard Lead Melting point: about 1809C (with decomposition). Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 1.0 g Identification (1) To 2 mL of a solution of Miconazole of Miconazole according to Method 3, and perform the test Nitrate in methanol (1 in 100) add 2 mL of Reinecke salt TS: (not more than 2 ppm). a light red precipitate is formed. (3) Related substances—Dissolve 0.10 g of Miconazole in (2) Determine the absorption spectrum of a solution of 10 mL of methanol, and use this solution as the sample solu- Miconazole Nitrate in methanol (1 in 2500) as directed under JP XVI Official Monographs / Micronomicin Sulfate 1119

Ultraviolet-visible Spectrophotometry <2.24>,andcompare the spectrum with the Reference Spectrum: both spectra Micronomicin Sulfate exhibit similar intensities of absorption at the same wavelengths. ミクロノマイシン硫酸塩 (3) Perform the test with a solution of Miconazole Nitrate in methanol (1 in 100) as directed under Flame Coloration Test <1.04> (2): a green color appears. (4) A solution of Miconazole Nitrate in methanol (1 in 100) responds to the Qualitative Tests <1.09> for nitrate. Purity (1) Clarity and color of solution—Dissolve 1.0 g of Miconazole Nitrate in 100 mL of methanol: the solution is clear and colorless. (2) Chloride <1.03>—Dissolve 0.10 g of Miconazole Nitrate in 6 mL of dilute nitric acid and N,N-dimethylformamide to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.25 mL of 0.01 mol/L hydrochloric acid VS add 6 mL of dilute nitric acid and N,N-dimethylformamide to make 50 mL (not more than 0.09z). (3) Heavy metals <1.07>—Proceed with 1.0 g of Micona- zole Nitrate according to Method 2, and perform the test. (C20H41N5O7)2.5H2SO4: 1417.53 Prepare the control solution with 1.0 mL of Standard Lead 2-Amino-2,3,4,6-tetradeoxy-6-methylamino-a-D- Solution (not more than 10 ppm). erythro-hexopyranosyl-(1→4)-[3-deoxy-4-C-methyl-3- (4) Arsenic <1.11>—Prepare the test solution with 1.0 g methylamino-b-L-arabinopyranosyl-(1→6)]-2-deoxy-D- of Miconazole Nitrate according to Method 3, and perform streptamine hemipentasulfate the test (not more than 2 ppm). [52093-21-7, Micronomicin] (5) Related substances—Dissolve 0.10 g of Miconazole Nitrate in 10 mL of methanol, and use this solution as the Micronomicin Sulfate is the sulfate of an amino- sample solution. Pipet 1 mL of the sample solution, add glycoside substance having antibacterial activity pro- methanol to make exactly 20 mL, pipet 1 mL of this solu- duced by the growth of Micromonospora sagamiensis. tion, add methanol to make exactly 20 mL, and use this solu- It contains not less than 590 mg (potency) and not tion as the standard solution. Perform the test with these so- more than 660 mg (potency) per mg, calculated on lutions as directed under Thin-layer Chromatography <2.03>. the anhydrous basis. The potency of Micronomicin Spot 50 mL each of the sample solution and standard solu- Sulfate is expressed as mass (potency) of micronomicin tion on a plate of silica gel for thin-layer chromatography. (C20H41N5O7: 463.57). Develop the plate with a mixture of n-hexane, chloroform, Description Micronomicin Sulfate occurs as a white to methanol and ammonia solution (28) (60:30:10:1) to a dis- light yellowish white powder. tance of about 12 cm, and air-dry the plate. Allow the plate It is very soluble in water, sparingly soluble in ethylene in iodine vapor for 20 minutes: the spots other than the prin- glycol, and practically insoluble in methanol and in ethanol cipal spot from the sample solution are not more intense (99.5). than the spot from the standard solution. It is hygroscopic. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- Identification (1) Dissolve 50 mg each of Micronomicin um, silica gel, 609C, 3 hours). Sulfate and Micronomicin Sulfate RS in 10 mL of water, and Residue on ignition <2.44> Not more than 0.1z (1 g). use these solutions as the sample solution and the standard solution. Perform the test with these solutions as directed Assay Weigh accurately about 0.35 g of Miconazole under Thin-layer Chromatography <2.03>. Spot 5 mLofthe Nitrate, previously dried, dissolve in 50 mL of acetic acid sample solution and standard solution on a plate of silica gel (100) by warming, cool, and titrate <2.50> with 0.1 mol/L for thin-layer chromatography. Develop the plate with a perchloric acid VS (potentiometric titration). Perform a mixture of ethanol (99.5), 1-buthanol and ammonia solution blank determination, and make any necessary correction. (28) (10:8:7) to a distance of about 10 cm, and air-dry the Each mL of 0.1 mol/L perchloric acid VS plate. Spray evenly a solution of ninhydrin in a mixture of

＝ 47.91 mg of C18H14Cl4N2O.HNO3 acetone and pyridine (25:1) (1 in 500), and heat at 1009Cfor 10 minutes: the spots obtained from the sample solution and Containers and storage Containers—Tight containers. the standard solution are red-purple to red-brown and their Storage—Light-resistant. Rf values are the same. (2) To 5 mL of a solution of Micronomicin Sulfate (1 in 100) add 1 mL of barium chloride TS: a white precipitate is formed, and it does not dissolve by addition of dilute nitric acid.

20 Optical rotation <2.49> [a]D : ＋110 – ＋1309(0.25 g calculated on the anhydrous basis, water, 25 mL, 100 mm). 1120 Midecamycin / Official Monographs JP XVI pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Micronomicin Sulfate in 10 mL of water is between Midecamycin 3.5 and 5.5. ミデカマイシン Purity (1) Clarity and color of solution—Dissolve 1.5 g of Micronomicin Sulfate in 10 mL of water: the solution is clear and colorless to pale yellow. (2) Heavy metals <1.07>—Proceed with 1.0 g of Micronomicin Sulfate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand- ard Lead Solution (not more than 20 ppm). (3) Related substances—Dissolve 0.40 g of Micronomi- cin Sulfate in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the C41H67NO15: 813.97 plate with a mixture of ethanol (99.5), 1-buthanol and am- (3R,4R,5S,6R,8R,9R,10E,12E,15R)- monia solution (28) (10:8:7) to a distance of about 10 cm, 5-[2,6-Dideoxy-3-C-methyl-4-O-propanoyl-a-L-ribo- and air-dry the plate. Spray evenly a solution of ninhydrin in hexopyranosyl-(1→4)-3,6-dideoxy-3-dimethylamino-b-D- a mixture of acetone and pyridine (25:1) (1 in 500), and heat glucopyranosyloxy]-6-formylmethyl-9-hydroxy-4-methoxy- at 1009C for 10 minutes: the spot other than the principal 8-methyl-3-propanoyloxyhexadeca-10,12-dien-15-olide spot obtained from the sample solution is not more intense [35457-80-8] than the spot from the standard solution. Midecamycin is a macrolide substance having anti- Water <2.48> Not more than 10.0z (0.2 g, volumetric bacterial activity produced by the growth of Strep- titration, back titration). Use a mixture of methanol for tomyces mycarofaciens. water determination and ethylene glycol for water determi- It contains not less than 950 mg (potency) and not nation (1:1) instead of methanol for water determination. more than 1020 mg (potency) per mg, calculated on the Assay Perform the test according to the Cylinder-plate dried basis. The potency of Midecamycin is expressed method as directed under Microbial Assay for Antibiotics as mass (potency) of midecamycin (C41H67NO15). <4.02> according to the following conditions. Description Midecamycin occurs as a white crystalline (i) Test organism—Bacillus subtilis ATCC 6633 powder. (ii) Culture medium—Use the medium i in 1) Medium It is very soluble in methanol, freely soluble in ethanol for test organism [5] under (1) Agar media for seed and base (95), and very slightly soluble in water. layer. (iii) Standard solutions—Weigh accurately an amount of Identification (1) Determine the absorption spectrum of a Micronomicin Sulfate RS, equivalent to about 20 mg (po- solution of Midecamycin in methanol (1 in 50,000) as di- tency), dissolve in 0.1 mol/L phosphate buffer solution for rected under Ultraviolet-visible Spectrophotometry <2.24>, antibiotics, pH 8.0 to make exactly 20 mL, and use this solu- and compare the spectrum with the Reference Spectrum or tion as the standard stock solution. Keep the standard stock the spectrum of a solution of Midecamycin RS prepared in solutionat5–159C, and use within 30 days. Take exactly a the same manner as the sample solution: both spectra exhibit suitable amount of the standard stock solution before use, similar intensities of absorption at the same wavelengths. add 0.1 mol/L phosphate buffer solution for antibiotics, pH (2) Determine the infrared absorption spectrum of 8.0 to make solutions so that each mL contains 2 mg (po- Midecamycin as directed in the potassium bromide disk tency) and 0.5 mg (potency), and use these solutions as the method under the Infrared Spectrophotometry <2.25>,and high concentration standard solution and the low concentra- compare the spectrum with the Reference Spectrum or the tion standard solution, respectively. spectrum of Midecamycin RS: both spectra exhibit similar (iv) Sample solutions—Weigh accurately an amount of intensities of absorption at the same wave numbers. Micronomicin Sulfate, equivalent to about 20 mg (potency), Melting point <2.60> 153 – 1589C. and dissolve in 0.1 mol/L phosphate buffer solution for antibiotics, pH 8.0 to make exactly 20 mL. Take exactly a suit- Purity Heavy metals <1.07>—Proceed with 1.0 g of able amount of this solution, add 0.1 mol/L phosphate Midecamycin according to Method 2, and perform the test. buffer solution for antibiotics, pH 8.0 to make solutions so Prepare the control solution with 3.0 mL of Standard Lead that each mL contains 2 mg (potency) and 0.5 mg (potency), Solution (not more than 30 ppm). and use these solutions as the high concentration sample Loss on drying <2.41> Not more than 2.0z (1.0 g, in vacu- solution and the low concentration sample solution, respec- um not exceeding 0.67 kPa, 609C, 3 hours). tively. Residue on ignition <2.44> Not more than 0.2z (1 g). Containers and storage Containers—Tight containers. Assay Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics JP XVI Official Monographs / Midecamycin Acetate 1121

<4.02> according to the following conditions. (C45H71NO17). (i) Test organism—Bacillus subtilis ATCC 6633 Description Midecamycin Acetate occurs as white, crystals (ii) Culture medium—Use the medium i in 1) Medium or crystalline powder. for test organism [5] under (1) Agar media for seed and base It is sparingly soluble in methanol, slightly soluble in layer. ethanol (95), and practically insoluble in water. (iii) Standard solutions—Weigh accurately an amount of Midecamycin RS, previously dried, equivalent to about 20 Identification (1) Determine the absorption spectrum of a mg (potency), dissolve in 10 mL of methanol, add water to solution of Midecamycin Acetate in methanol (1 in 50,000) make exactly 50 mL, and use this solution as the standard as directed under Ultraviolet-visible Spectrophotometry stock solution. Keep the standard stock solution at 59Cor <2.24>, and compare the spectrum with the Reference Spec- below and use within 7 days. Take exactly a suitable amount trum or the spectrum of a solution of Midecamycin Acetate of the standard stock solution before use, add 0.1 mol/L RS prepared in the same manner as the sample solution: phosphate buffer solution, pH 8.0 to make solutions so that both spectra exhibit similar intensities of absorption at the each mL contains 20 mg (potency) and 5 mg (potency), and same wavelengths. use these solutions as the high concentration standard solu- (2) Determine the infrared absorption spectrum of tion and the low concentration standard solution, respec- Midecamycin Acetate, previously dried, as directed in the tively. potassium bromide disk method under Infrared Spectropho- (iv) Sample solutions—Weigh accurately an amount of tometry <2.25>, and compare the spectrum with the Refer- Midecamycin, previously dried, equivalent to about 20 mg ence Spectrum or spectrum of dried Midecamycin Acetate (potency), dissolve in 10 mL of methanol, and add water to RS: both spectra exhibit similar intensities of absorption at make exactly 50 mL. Take exactly a suitable amount of the the same wave numbers. solution, add 0.1 mol/L phosphate buffer solution, pH 8.0 Purity Heavy metals <1.07>—Proceed with 1.0 g of to make solutions so that each mL contains 20 mg (potency) Midecamycin Acetate according to Method 2, and perform and 5 mg (potency), and use these solutions as the high con- the test. Prepare the control solution with 2.0 mL of Stand- centration sample solution and the low concentration sample ard Lead Solution (not more than 20 ppm). solution, respectively. Loss on drying <2.41> Not more than 2.0z (1 g, in vacuum Containers and storage Containers—Tight containers. not exceeding 0.67 kPa, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g). Midecamycin Acetate Assay Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics ミデカマイシン酢酸エステル <4.02> according to the following conditions. (i) Test organism—Micrococcus luteus ATCC 9341 (ii) Culture medium—Use the medium i in 3) Medium for other organisms under (1) Agar media for seed and base layer. (iii) Standard solutions—Weigh accurately an amount of Midecamycin Acetate RS, previously dried, equivalent to about 25 mg (potency), and dissolve in methanol to make exactly 50 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 5 – 159Cand use within 7 days. Take exactly a suitable amount of the standard stock solution before use, add 0.1 mol/L phosphate buffer solution, pH 4.5 to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high concentration standard solution and the low concentration standard solution, respectively. (iv) Sample solutions—Weigh accurately an amount of

C45H71NO17: 898.04 Midecamycin Acetate, previously dried, equivalent to about (3R,4S,5S,6R,8R,9R,10E,12E,15R)-9-Acetoxy-5-[3-O- 25 mg (potency), and dissolve in methanol to make exactly acetyl-2,6-dideoxy-3-C-methyl-4-O-propanoyl-a-L- 50 mL. Take exactly a suitable amount of the solution, add ribo-hexopyranosyl-(1→4)-3,6-dideoxy-3-dimethylamino- 0.1 mol/L phosphate buffer solution, pH 4.5 to make solu- b-D-glucopyranosyloxy]-6-formylmethyl-4-methoxy-8- tions so that each mL contains 20 mg (potency) and 5 mg (po- methyl-3-propioyloxyhexadeca-10,12-dien-15-olide tency), and use these solutions as the high concentration [55881-07-7] sample solution and the low concentration sample solution, respectively. Midecamycin Acetate is a derivative of midecamy- Containers and storage Containers—Tight containers. cin. It contains not less than 950 mg (potency) and not more than 1010 mg (potency) per mg, calculated on the dried basis. The potency of Midecamycin Acetate is expressed as mass of midecamycin acetate 1122 Migrenin / Official Monographs JP XVI

nal standard solution, dissolve in chloroform to make 10 Migrenin mL, and use this solution as the sample solution. Separately, weigh accurately about 90 mg of Caffeine RS, previously ミグレニン dried at 809C for 4 hours, add exactly 5 mL of the internal standard solution, dissolve in chloroform to make 10 mL, and use this solution as the standard solution. Perform the Migrenin is composed of 90 parts of antipyrine, 9 test with 1 mL each of the sample solution and standard solu- parts of caffeine, and 1 part of citric acid in mass. tion as directed under Gas Chromatography <2.02> accord- Migrenin, when dried, contains not less than 87.0z ing to the following conditions, and calculate the ratios, Q and not more than 93.0z of antipyrine (C H N O: T 11 12 2 and Q , of the peak area of caffeine to that of the internal 188.23) and not less than 8.6z and not more than S standard. 9.5z of caffeine (C8H10N4O2: 194.19). Amount (mg) of caffeine (C H N O ) ＝ M × Q /Q Description Migrenin occurs as a white powder or crystal- 8 10 4 2 S T S line powder. It is odorless and has a bitter taste. MS: Amount (mg) of Caffeine RS It is very soluble in water, freely soluble in ethanol (95) Internal standard solution—A solution of ethenzamide in and in chloroform, and slightly soluble in diethyl ether. chloroform (1 in 50). The pH of a solution of Migrenin (1 in 10) is between 3.0 Operating conditions— and 4.0. Detector: A hydrogen flame-ionization detector. It is affected by moisture and light. Column: A glass column 2.6 mm in inside diameter and Identification (1) To 5 mL of a solution of Migrenin (1 in 210 cm in length, packed with siliceous earth for gas chroma- 100) add 2 drops of sodium nitrite TS and 1 mL of dilute sul- tography (180 to 250 mm in particle diameter) coated with furic acid: a deep green color develops. 50z phenyl-methyl silicon polymer for gas chromatography (2) To 5 mL of a solution of Migrenin (1 in 50) add 1 at the ratio of 15z. drop of hydrochloric acid and 0.2 mL of formaldehyde solu- Column temperature: A constant temperature of about tion, heat in a water bath for 30 minutes, add an excess of 2109C. ammonia TS, and filter. Acidify the filtrate with hydrochlo- Carrier gas: Nitrogen. ric acid, shake with 3 mL of chloroform, and separate the Flow rate: Adjust the flow rate so that the retention time chloroform layer. Evaporate the chloroform solution on a of ethenzamide is about 4 minutes. water bath, add 10 drops of hydrogen peroxide TS and 1 System suitability— drop of hydrochloric acid to the residue, and evaporate on a System performance: Dissolve 0.9 g of antipyrine and water bath to dryness: the residue shows a yellow-red color. 0.09 g of caffeine in 10 mL of chloroform. When the proce- Invert the residue over a vessel containing 3 drops of ammo- dure is run with 1 mL of this solution under the above operat- nia TS: a red-purple color develops, disappearing on the ad- ing conditions, caffeine and antipyrine are eluted in this dition of 2 to 3 drops of sodium hydroxide TS. order with the resolution between these peaks being not less (3) A solution of Migrenin (1 in 10) responds to the than 1.5. Qualitative Tests <1.09> for citrate. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating Melting point <2.60> 104–1109C. conditions, the relative standard deviation of the ratios of Purity (1) Clarity and color of solution—Dissolve 1.0 g the peak area of caffeine to that of the internal standard is of Migrenin in 40 mL of water: the solution is clear and col- not more than 1.0z. orless to pale yellow. Containers and storage Containers—Tight containers. (2) Heavy metals <1.07>—Proceed with 1.0 g of Migrenin Storage—Light-resistant. according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay (1) Antipyrine—Weigh accurately about 0.25 g of Migrenin, previously dried in an iodine flask, dissolve in 25 mL of sodium acetate TS, add exactly 30 mL of 0.05 mol/L iodine VS, and allow to stand for 20 minutes with occasional shaking. Add 15 mL of chloroform to dissolve the precipitate so obtained, and titrate <2.50> the excess iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 3 mL of starch TS). Perform a blank determination. Each mL of 0.05 mol/L iodine VS

＝ 9.411 mg of C11H12N2O (2) Caffeine—To about 1 g of Migrenin, previously dried and accurately weighed, add exactly 5 mL of the inter- JP XVI Official Monographs / Minocycline Hydrochloride 1123

(3) Related substances—Dissolve 50 mg of Minocycline Minocycline Hydrochloride Hydrochloride in 100 mL of the mobile phase, and use this solution as the sample solution. Perform the test immedi- ミノサイクリン塩酸塩 ately after the preparation of the sample solution with 20 mL of the sample solution as directed under Liquid Chromatog- raphy <2.01> according to the following conditions, and determine each peak area by the automatic integration method. Calculate the amount of each peak area by the area percentage method: the amount of epiminocycline is not more than 1.2z, the amount of each peak other than minocycline and epiminocycline is not more than 1.0z, and the total area of the peaks other than minocycline and epiminocycline is not C H N O .HCl: 493.94 23 27 3 7 more than 2.0z. (4S,4aS,5aR,12aS)-4,7-Bis(dimethylamino)- Operating conditions— 3,10,12,12a-tetrahydroxy-1,11-dioxo- Detector, column, column temperature, and mobile phase: 1,4,4a,5,5a,6,11,12a-octahydrotetracene-2-carboxamide Proceed as directed in the operating conditions in the Assay. monohydrochloride Flow rate: Adjust the flow rate so that the retention time [13614-98-7] of minocycline is about 12 minutes. The retention time of epiminocycline is about 10 minutes under this condition. Minocycline Hydrochloride is the hydrochloride of Time span of measurement: About 2.5 times as long as the a derivative of tetracycline. retention time of minocycline beginning after the solvent It contains not less than 890 mg (potency) and not peak. more than 950 mg (potency) per mg, calculated on the System suitability— anhydrous basis. The potency of Minocycline Hydro- Test for required detection: To exactly 2 mL of the sample chloride is expressed as mass (potency) of minocycline solution add the mobile phase to make exactly 100 mL, and (C H N O : 457.48). 23 27 3 7 use this solution as the solution for system suitability test. Description Minocycline Hydrochloride occurs as a yellow Pipet 5 mL of the solution for system suitability test, and crystalline powder. add the mobile phase to make exactly 100 mL. Confirm that It is freely soluble in N,N-dimethylformamide, soluble in the peak area of minocycline obtained from 20 mLofthisso- methanol, sparingly soluble in water, and slightly soluble in lution is equivalent to 3.5 to 6.5z of that from 20 mLofthe ethanol (95). solution for system suitability test. System performance: Proceed as directed in the system Identification (1) Determine the absorption spectrum of a suitability in the Assay. solution of Minocycline Hydrochloride in a solution of hy- System repeatability: When the test is repeated 6 times drochloric acid in methanol (19 in 20,000) (1 in 62,500) as di- with 20 mL of the solution for system suitability test under rected under Ultraviolet-visible Spectrophotometry <2.24>, the above operating conditions, the relative standard devia- and compare the spectrum with the Reference Spectrum or tion of the peak area of minocycline is not more than 2.0z. the spectrum of a solution of Minocycline Hydrochloride RS prepared in the same manner as the sample solution: both Water <2.48> Not less than 4.3z and not more than 8.0z spectra exhibit similar intensities of absorption at the same (0.3 g, volumetric titration, direct titration). wavelengths. Residue on ignition <2.44> Not more than 0.5z (1 g). (2) Determine the infrared absorption spectrum of Minocycline Hydrochloride as directed in the potassium Assay Weigh accurately an amount of Minocycline Hydro- chloride disk method under Infrared Spectrophotometry chloride and Minocycline Hydrochloride RS, equivalent to <2.25>, and compare the spectrum with the Reference Spec- about 50 mg (potency), dissolve each in the mobile phase to trum or the spectrum of Minocycline Hydrochloride RS: make exactly 100 mL, and use these solutions as the sample both spectra exhibit similar intensities of absorption at the solution and the standard solution. Perform the test with same wave numbers. exactly 20 mL each of the sample solution and standard solu- (3) A solution of Minocycline Hydrochloride (1 in 100) tion as directed under Liquid Chromatography <2.01> ac- responds to the Qualitative Tests <1.09> (2) for chloride. cording to the following conditions, and determine the peak areas, A and A , of minocycline of these solutions. pH <2.54> Dissolve 1.0 g of Minocycline Hydrochloride in T S

100 mL of water: the pH of the solution is between 3.5 and Amount [mg (potency)] of minocycline (C23H27N3O7) 4.5. ＝ MS × AT/AS × 1000

Purity (1) A solution of Minocycline Hydrochloride (1 in MS: Amount [mg (potency)] of Minocycline Hydrochlo- 100) is clear, and when the test is performed within 1 hour ride RS after preparation of this solution, the absorbance of the so- Operating conditions— lution at 560 nm, determined as directed under Ultraviolet- Detector: An ultraviolet absorption photometer (wave- visible Spectrophotometry <2.24>, is not more than 0.06. length: 280 nm). (2) Heavy metals <1.07>—Proceed with 0.5 g of Mino- Column: A stainless steel column 4.6 mm in inside diame- cycline Hydrochloride according to Method 2, and perform ter and 15 cm in length, packed with octylsilanized silica gel the test. Prepare the control solution with 2.5 mL of Stand- for liquid chromatography (5 mm in particle diameter). ard Lead Solution (not more than 50 ppm). Column temperature: A constant temperature of about 1124 Minocycline Hydrochloride for Injection / Official Monographs JP XVI

259C. the following conditions, and determine each peak area by Mobile phase: Adjust to pH 6.5 of a mixture of a solution the automatic integration method. Calculate the amounts of of ammonium oxalate monohydrate (7 in 250), N,N- each peak by the area percentage method: the amount of dimethylformamide and 0.1 mol/L disodium dihydrogen epiminocycline, having the relative retention time of about ethylenediamine tetraacetate TS (11:5:4) with tetrabutylam- 0.83 with respect to minocycline, is not more than 6.0z. monium hydroxide TS. Operating conditions— Flow rate: Adjust the flow rate so that the retention time Detector, column, column temperature, mobile phase and of minocycline is about 12 minutes. flow rate: Proceed as directed in the operating conditions in System suitability— the Assay. System performance: Dissolve 50 mg of Minocycline Hy- Time span of measurement: About 2.5 times as long as the drochloride in 25 mL of water. Heat 5 mL of this solution on retention time of minocycline, beginning after the solvent a water bath for 60 minutes, then add water to make 25 mL. peak. When the procedure is run with 20 mL of this solution under System suitability— the above operating conditions, epiminocycline and minocy- Test for required detectability: Pipet 2 mL of the standard cline are eluted in this order with the resolution between solution obtained in the Assay, add the mobile phase to these peaks being not less than 2.0. make exactly 100 mL, and use this solution as the solution System repeatability: When the test is repeated 6 times for system suitability test. Pipet 5 mL of the solution for sys- with 20 mL of the standard solution under the above operat- tem suitability test, add the mobile phase to make exactly ing conditions, the relative standard deviation of peak areas 100 mL. Confirm that the peak area of minocycline obtained of minocycline is not more than 1.0z. from 20 mL of this solution is equivalent to 3.5 to 6.5z of that from 20 mL of the solution for system suitability test. Containers and storage Containers—Tight containers. System performance: Proceed as directed in the system Storage—Light-resistant. suitability in the Assay. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under Minocycline Hydrochloride for the above operating conditions, the relative standard devia- Injection tion of the peak area of minocycline is not more than 2.0z. Water <2.48> Weigh accurately the mass of the content of 注射用ミノサイクリン塩酸塩 one container of Minocycline Hydrochloride for Injection, dissolve in exactly 2 mL of methanol for water determina- Minocycline Hydrochloride for Injection is a prepa- tion, and perform the test with exactly 1 mL of this solution ration for injection, which is dissolved before use. as directed in the Volumetric titration (back titration): not It contains not less than 90.0z and not more more than 3.0z. than 110.0z of the labeled amount of minocycline Bacterial endotoxins <4.01> Less than 1.25 EU/mg (po- (C23H27N3O7: 457.48). tency). Method of preparation Prepare as directed under Injec- Uniformity of dosage units <6.02> It meets the requirement tions, with Minocycline Hydrochloride. of the Mass variation test. Description Minocycline Hydrochloride for Injection oc- Foreign insoluble matter <6.06> Perform the test according curs as a yellow to yellow-brown powder or flakes. to Method 2: it meets the requirement. Identification Dissolve 4 mg of Minocycline Hydrochloride Insoluble particulate matter <6.07> It meets the require- for Injection in 250 mL of a solution of hydrochloric acid in ment. methanol (19 in 20,000). Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro- Sterility <4.06> Perform the test according to the Mem- photometry <2.24>: it exhibits maxima between 221 nm and brane filtration method: it meets the requirement. 225 nm, between 261 nm and 265 nm, and between 354 nm Assay Weigh accurately the mass of the contents of not less and 358 nm. than 10 containers of Minocycline Hydrochloride for Injec- pH <2.54> The pH of a solution, prepared by dissolving an tion. Weigh accurately an amount of the contents, equiva- amount of Minocycline Hydrochloride for Injection, equivalent to about 0.1 g (potency) of Minocycline Hydrochloride, lent to 0.1 g (potency) of Minocycline Hydrochloride accord- dissolve in the mobile phase to make exactly 100 mL. Pipet ing to the labeled amount, in 10 mL of water is 2.0 to 3.5. 25 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the sample solution. Sepa- Purity Related substances—Conduct this procedure rapidly rately, weigh accurately an amount of Minocycline Hydro- after the preparation of the sample solution. Take an chloride RS, equivalent to about 25 mg (potency), dissolve in amount of Minocycline Hydrochloride for Injection, equiva- the mobile phase to make exactly 50 mL, and use this solu- lent to 0.1 g (potency) of Minocycline Hydrochloride accord- tion as the standard solution. Then, proceed as directed in ing to the labeled amount, dissolve in the mobile phase to the Assay under Minocycline Hydrochloride. make 100 mL. To 25 mL of this solution, add the mobile phase to make 50 mL, and use this solution as the sample so- Amount [mg (potency)] of minocycline (C23H27N3O7) lution. Perform the test with 20 mL of the sample solution as ＝ MS × AT/AS × 4 directed under Liquid Chromatography <2.01> according to MS: Amount [mg (potency)] of Minocycline Hydrochlo- JP XVI Official Monographs / Minocycline Hydrochloride Tablets 1125

ride RS Water <2.48> Not more than 12.0z (0.5 g of powdered Minocycline Hydrochloride Tablets, volumetric titration, Containers and storage Containers—Hermetic containers. back titration). Uniformity of dosage units <6.02> Perform the test accord- Minocycline Hydrochloride Tablets ing to the following method: it meets the requirement of the Content uniformity test. ミノサイクリン塩酸塩錠 To 1 tablet of Minocycline Hydrochloride Tablets add 60 mL of the mobile phase, treat with ultrasonic waves for 15 minutes, and add the mobile phase to make exactly V mL so Minocycline Hydrochloride Tablets contain not less that each mL contains about 0.5 mg (potency) of Mino- than 90.0z and not more than 110.0z of the labeled cycline Hydrochloride. Centrifuge this solution, and use the potency of Minocycline (C H N O : 457.48). 23 27 3 7 supernatant liquid as the sample solution. Then, proceed as Method of preparation Prepare as directed under Tablets, directed in the Assay. with Minocycline Hydrochloride. Amount [mg (potency)] of minocycline (C23H27N3O7) Identification To a quantity of powdered Minocycline Hy- ＝ MS × AT/AS × V/50 drochloride Tablets, equivalent to 10 mg (potency) of Mino- M : Amount [mg (potency)] of Minocycline Hydrochlo- cycline Hydrochloride according to the labeled amount, add S ride RS 625 mL of a solution of hydrochloric acid in methanol (19 in 20,000), shake well, and filter. Determine the absorption Dissolution <6.10> When the test is performed at 50 revolu- spectrum of the filtrate as directed under Ultraviolet-visible tions per minute according to the Paddle method using 900 Spectrophotometry <2.24>: it exhibits maxima between 221 mL of water as the dissolution medium, the dissolution rate nm and 225 nm, between 261 nm and 265 nm, and between in 30 minutes of Minocycline Hydrochloride Tablets is not 354 nm and 358 nm. less than 85z. Start the test with 1 tablet of Minocycline Hydrochloride Purity Related substances—Conduct this procedure rapidly Tablets, withdraw not less than 20 mL of the medium at the after preparation of the sample solution. Powder not less specified minute after starting the test, and filter through a than 5 Minocycline Hydrochloride Tablets. Weigh accurately membrane filter with a pore size not exceeding 0.45 mm. Dis- a portion of the powder, equivalent to 50 mg (potency) card the first 10 mL of the filtrate, pipet V mL of the subse- of Minocycline Hydrochloride according to the labeled quent filtrate, add water to make exactly V?mL so that each amount, add 60 mL of the mobile phase, shake vigorously, mL contains about 9 mg (potency) of Minocycline Hydro- and add the mobile phase to make 100 mL. Centrifuge this chloride according to the labeled amount, and use this solu- solution, and use the supernatant liquid as the sample solution as the sample solution. Separately, weigh accurately an tion. Perform the test with 20 mL of the sample solution as amount of Minocycline Hydrochloride RS, equivalent to directed under Liquid Chromatography <2.01> according to about 30 mg (potency), and dissolve in water to make exactly the following conditions. Determine each peak area by the 100 mL. Pipet 4 mL of this solution, add water to make ex- automatic integration method. Calculate the amounts of actly 100 mL, and use this solution as the standard solution. these peaks by the area percentage method: the amount of Perform the test with the sample solution and standard solu- the peak of epiminocycline, having the relative retention tion as directed under Ultraviolet-visible Spectrophotometry time of about 0.83 with respect to minocycline, is not more <2.24>, and determine the absorbances, A and A ,at348 than 2.0z. T S nm. Operating conditions— Detector, column, column temperature, mobile phase and Dissolution rate (z) with respect to the labeled amount flow rate: Proceed as directed in the operating conditions in of minocycline (C23H27N3O7) the Assay. ＝ MS × AT/AS × V?/V × 1/C × 36 Time span of measurement: About 2.5 times as long as the M : Amount [mg (potency)] of Minocycline Hydrochlo- retention time of minocycline, beginning after the solvent S ride RS peak. C: Labeled amount [mg (potency)] of minocycline System suitability— (C H N O )in1tablet Test for required detectability: To 2 mL of the sample so- 23 27 3 7 lution add the mobile phase to make 100 mL, and use this Assay To a number of Minocycline Hydrochloride Tablets, solution as the solution for system suitability test. Pipet 5 equivalent to about 1 g (potency) of Minocycline Hydrochlo- mL of the solution for system suitability test, and add the ride, add 120 mL of the mobile phase, treat with ultrasonic mobile phase to make exactly 100 mL. Confirm that the waves for 15 minutes, and add the mobile phase to make ex- peak area of minocycline obtained from 20 mL of this solu- actly 200 mL. Centrifuge this solution, pipet 5 mL of the su- tion is equivalent to 3.5 to 6.5z of that of minocycline from pernatant liquid, add the mobile phase to make exactly 50 20 mL of the solution for system suitability test. mL, and use this solution as the sample solution. Separately, System performance: Proceed as directed in the system weigh accurately an amount of Minocycline Hydrochloride suitability in the Assay. RS, equivalent to about 25 mg (potency), dissolve in the System repeatability: When the test is repeated 6 times mobile phase to make exactly 50 mL, and use this solution as with 20 mL of the solution for system suitability test under the standard solution. Then, proceed as directed in the Assay the above operating conditions, the relative standard devia- under Minocycline Hydrochloride. tion of the peak area of minocycline is not more than 2.0z. 1126 Mitomycin C / Official Monographs JP XVI

Amount [mg (potency)] of minocycline (C23H27N3O7) the peak other than mitomycin C obtained from the sample ＝ MS × AT/AS × 40 solution is not larger than the peak area of mitomycin C from the standard solution, and the total area of the peaks M : Amount [mg (potency)] of Minocycline Hydrochlo- S other than mitomycin C from the sample solution is not ride RS larger than 3 times the peak area of mitomycin C from the Containers and storage Containers—Tight containers. standard solution. Storage—Light-resistant. Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 254 nm). Mitomycin C Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel マイトマイシン C for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase A: To 20 mL of 0.5 mol/L ammonium acetate TS add water to make 1000 mL. To 800 mL of this solution add 200 mL of methanol. Mobile phase B: To 20 mL of 0.5 mol/L ammonium acetate TS add water to make 1000 mL. To this solution add 1000 mL of methanol. C H N O : 334.33 15 18 4 5 Flowing of the mobile phase: Control the gradient by mix- (1aS,8S,8aR,8bS)-6-Amino-4,7-dioxo-8a-methoxy- ing the mobile phases A and B as directed in the following 5-methyl-1,1a,2,8,8a,8b- table. hexahydroazirino[2?,3?:3,4]pyrrolo[1,2-a]indol- 8-ylmethyl carbamate [50-07-7] Time after injection Mobile phase A Mobile phase B of sample (min) (volz) (volz)

Mitomycin C is a substance having antitumor 0–10 100 0 activity produced by the growth of Streptomyces 10–30 100→ 00→ 100 caespitosus. 30–45 0 100 It contains not less than 970 mg (potency) and not more than 1030 mg (potency) per mg, calculated on the dried basis. The potency of Mitomycin C is expressed Flow rate: About 1.0 mL per minute. Time span of measurement: About 2 times as long as the as mass (potency) of mitomycin C (C15H18N4O5). retention time of mitomycin C beginning after the solvent Description Mitomycin C occurs as blue-purple, crystals or peak. crystalline powder. System suitability— It is freely soluble in N,N-dimethylacetamide, slightly Test for required detection: Pipet 10 mL of the standard soluble in water and in methanol, and very slightly soluble in solution, and add methanol to make exactly 100 mL. Con- ethanol (99.5). firm that the peak area of mitomycin C obtained from 10 mL Identification (1) Determine the absorption spectrum of a of this solution is equivalent to 7 to 13z of that from 10 mL solution of Mitomycin C (1 in 100,000) as directed under Ul- of the standard solution. traviolet-visible Spectrophotometry <2.24>, and compare the System performance: Dissolve 25 mg of Mitomycin C and spectrum with the Reference Spectrum or the spectrum of a 40 mg of 3-ethoxy-4-hydroxybenzaldehyde in 50 mL of solution of Mitomycin C RS prepared in the same manner as methanol. When the procedure is run with 10 mLofthisso- the sample solution: both spectra exhibit similar intensities lution under the above operating conditions, mitomycin C of absorption at the same wavelengths. and 3-ethoxy-4-hydroxybenzaldehyde are eluted in this order (2) Determine the infrared absorption spectrum of with the resolution between these peaks being not less than Mitomycin C as directed in the potassium bromide disk 15. method under Infrared Spectrophotometry <2.25>,and System repeatability: When the test is repeated 3 times compare the spectrum with the Reference Spectrum or the with 10 mL of the standard solution under the above operat- spectrum of Mitomycin C RS: both spectra exhibit similar ing conditions, the relative standard deviation of the peak intensities of absorption at the same wave numbers. area of mitomycin C is not more than 3.0z. Purity Related substances—Conduct this procedure rapidly Loss on drying <2.41> Not more than 1.0z (0.1 g, reduced after the sample and the standard solutions are prepared. pressure not exceeding 0.67 kPa, 609C, 3 hours). Dissolve 50 mg of Mitomycin C in 10 mL of methanol, and Assay Weigh accurately an amount of Mitomycin C and use this solution as the sample solution. Pipet 1 mL of the Mitomycin C RS, equivalent to about 25 mg (potency), sample solution, add methanol to make exactly 100 mL, and dissolve each in N,N-dimethylacetamide to make exactly use this solution as the standard solution. Perform the test 50 mL, and use these solutions as the sample solution and with exactly 10 mL each of the sample solution and standard standard solution. Perform the test with exactly 10 mLeach solution as directed under Liquid Chromatography <2.01> of the sample solution and standard solution as directed according to the following conditions, and determine each under Liquid Chromatography <2.01> according to the fol- peak area by the automatic integration method: each area of JP XVI Official Monographs / Mizoribine 1127 lowing conditions, and determine the peak areas, AT and AS, um not exceeding 0.67 kPa, phosphorus (V) oxide, 609C, of mitomycin C. 3 hours).

Amount [mg (potency)] of C15H18N4O5 Bacterial endotoxins <4.01> Less than 10 EU/mg (potency). ＝ M × A /A × 1000 S T S Uniformity of dosage units <6.02> Perform the test accord-

MS: Amount [mg (potency)] of Mitomycin C RS ing to the following method: it meets the requirement of the Content uniformity test. Operating conditions— To 1 container of Mitomycin C for Injection add exactly Detector: An ultraviolet absorption photometer (wave- V mL of N,N-dimethylacetamide so that each mL contains length: 365 nm). about 0.5 mg (potency) of Mitomycin C, shake, centrifuge, Column: A stainless steel column 4 mm in inside diameter and use the supernatant liquid as the sample solution. Sepa- and 30 cm in length, packed with phenylated silica gel for rately, weigh accurately about 25 mg (potency) of Mitomycin liquid chromatography (10 mm in particle diameter). CRS,addN,N-dimethylacetamide to make exactly 50 mL, Column temperature: A constant temperature of about and use this solution as the standard solution. Then, proceed 259C. as directed in the Assay under Mitomycin C. Mobile phase: To 40 mL of 0.5 mol/L ammonium acetate

TS add 5 mL of diluted acetic acid (100) (1 in 20) and water Amount [mg (potency)] of mitomycin C (C15H18N4O5) to make 1000 mL. To 600 mL of this solution add 200 mL of ＝ MS × AT/AS × V/50 methanol. M : Amount [mg (potency)] of Mitomycin C RS Flow rate: Adjust the flow rate so that the retention time S of mitomycin C is about 7 minutes. Foreign insoluble matter <6.06> Perform the test according System suitability— to Method 2: it meets the requirement. System performance: Dissolve about 25 mg of Mitomycin Insoluble particulate matter <6.07> It meets the require- C RS and about 0.375 g of 3-ethoxy-4-hydroxybenzaldehyde ment. in 50 mL of N,N-dimethylacetamide. When the procedure is run with 10 mL of this solution under the above operating Sterility <4.06> Perform the test according to the Mem- conditions, mitomycin C and 3-ethoxy-4-hydroxybenzalde- brane filtration method: it meets the requirement. hyde are eluted in this order with the resolution between Assay Weigh accurately the mass of the contents of not less these peaks being not less than 3. than 10 containers of Mitomycin C for Injection. Weigh ac- System repeatability: When the test is repeated 6 times curately an amount of the contents, equivalent to about 10 with 10 mL of the standard solution under the above operat- mg (potency) of Mitomycin C, add exactly 20 mL of N,N- ing conditions, the relative standard deviation of the peak dimethylacetamide, shake, centrifuge, and use the superna- area of mitomycin C is not more than 1.0z. tant liquid as the sample solution. Separately, weigh accu- Containers and storage Containers—Tight containers. rately an amount of Mitomycin C RS, equivalent to about 25 mg (potency), dissolve in N,N-dimethylacetamide to make exactly 50 mL, and use this solution as the standard solution. Mitomycin C for Injection Then, proceed as directed in the Assay under Mitomycin C. Amount [mg (potency)] of mitomycin C (C H N O ) 注射用マイトマイシン C 15 18 4 5 ＝ MS × AT/AS × 2/5 M : Amount [mg (potency)] of Mitomycin C RS Mitomycin C for Injection is a preparation for in- S jection, which is dissolved before use. Containers and storage Containers—Hermetic containers. It contains not less than 90.0z and not more than 110.0z of the labeled amount of mitomycin C (C15H18N4O5: 334.33). Mizoribine Method of preparation Prepare as directed under Injec- ミゾリビン tions, with Mitomycin C. Description Mitomycin C for Injection occurs as a blue- purple powder. Identification Dissolve an amount of Mitomycin C for In- jection, equivalent to 2 mg (potency) of Mitomycin C according to the labeled amount, in 200 mL of water, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>:itexhib- C H N O :259.22 its maxima between 216 nm and 220 nm, and between 362 9 13 3 6 5-Hydroxy-1-b-D-ribofuranosyl-1H-imidazole-4-carboxamide nm and 366 nm. [50924-49-7] pH <2.54> The pH of a solution, prepared by dissolving 0.25 g of Mitomycin C for Injection in 20 mL of water, is 5.5 Mizoribine contains not less than 98.0z and not to 8.5. more than 102.0z of C9H13N3O6,calculatedonthe anhydrous basis. Loss on drying <2.41> Not more than 1.0z (0.4 g, in vacu- 1128 Mizoribine Tablets / Official Monographs JP XVI

Description Mizoribine occurs as a white to yellowish white tion, direct titration). crystalline powder. Residue on ignition <2.44> Not more than 0.1z (1 g). It is freely soluble in water, and practically insoluble in methanol and in ethanol (99.5). Assay Weigh accurately about 0.1 g of Mizoribine, and dissolve in the mobile phase to make exactly 50 mL. Pipet 5 mL Identification (1) Determine the absorption spectrum of a of this solution, add the mobile phase to make exactly 50 solution of Mizoribine (1 in 100,000) as directed under Ultra- mL, and use this solution as the sample solution. Separately, violet-visible Spectrophotometry <2.24>, and compare the weigh accurately about 10 mg of Mizoribine RS (separately spectrum with the Reference Spectrum or the spectrum of a determine the water <2.48> using the same manner as Mizori- solution of Mizoribine RS prepared in the same manner as bine), dissolve in the mobile phase to make exactly 50 mL, the sample solution: both spectra exhibit similar intensities and use this solution as the standard solution. Perform the of absorption at the same wavelengths. test with exactly 5 mL each of the sample solution and stand- (2) Determine the infrared absorption spectrum of ard solution as directed under Liquid Chromatography Mizoribine as directed in the potassium bromide disk <2.01> according to the following conditions, and determine method under Infrared Spectrophotometry <2.25>,andcom- the peak areas of mizoribine, A and A , of both solutions. pare the spectrum with the Reference Spectrum or the spec- T S trum of Mizoribine RS: both spectra exhibit similar intensi- Amount (mg) of C9H13N3O6 ＝ MS × AT/AS × 10 ties of absorption at the same wave numbers. MS: Amount (mg) of Mizoribine RS, calculated on the an- 20 Optical rotation <2.49> [a]D : －25 – －279(0.5 g calculated hydrous basis on the anhydrous basis, water, 25 mL, 100 mm). Operating conditions— Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Detector: An ultraviolet absorption photometer (wave- Mizoribine according to Method 1, and perform the test. length: 279 nm). Prepare the control solution with 2.0 mL of Standard Lead Column: A stainless steel column 4.6 mm in inside diame- Solution (not more than 20 ppm). ter and 25 cm in length, packed with octadecylsilanized silica (2) Related substances—Dissolve 0.10 g of Mizoribine in gel for liquid chromatography (5 mminparticlediameter). the mobile phase to make 50 mL, and use this solution as the Column temperature: A constant temperature of about sample solution. Pipet 5 mL of the sample solution, and add 259C. the mobile phase to make exactly 50 mL. Pipet 1 mL of this Mobile phase: Diluted phosphoric acid (1 in 1500). solution, add the mobile phase to make exactly 100 mL, and Flow rate: Adjust the flow rate so that the retention time use this solution as the standard solution. Perform the test of mizoribine is about 9 minutes. with exactly 5 mL each of the sample solution and standard System suitability— solution as directed under Liquid Chromatography <2.01> System performance: When the procedure is run with 5 mL according to the following conditions. Determine each peak of the standard solution under the above operating condi- area of both solutions by the automatic integration method: tions, the number of theoretical plates and the symmetry fac- the areas of the peaks other than mizoribine obtained from tor of the peak of mizoribine are not less than 10,000 and the sample solution are not larger than the mizoribine peak not more than 1.4, respectively. area from the standard solution. System repeatability: When the test is repeated 6 times Operating conditions— with 5 mL of the standard solution under the above operating Column, column temperature, mobile phase, and flow conditions, the relative standard deviation of the peak area rate: Proceed as directed in the operating conditions in the of mizoribine is not more than 1.0z. Assay. Containers and storage Containers—Tight containers. Detector: An ultraviolet absorption photometer (wavelength: 220 nm). Time span of measurement: About 3 times as long as the retention time of mizoribine, beginning after the solvent Mizoribine Tablets peak. ミゾリビン錠 System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 5 mL. Mizoribine Tablets contain not less than 93.0z and Confirm that the peak area of mizoribine obtained from 5 not more than 107.0z of the labeled amount of mL of this solution is equivalent to 14 to 26z of that from 5 mizoribine (C9H13N3O6: 259.22). mL of the standard solution. Method of preparation Prepare as directed under Tablets, System performance: When the procedure is run with 5 mL with Mizoribine. of the standard solution under the above operating conditions, the number of theoretical plates and the symmetry fac- Identification To a quantity of powdered Mizoribine tor of the peak of mizoribine are not less than 10,000 and Tablets, equivalent to 0.1 g of Mizoribine according to the not more than 1.4, respectively. labeled amount, add 5 mL of water, shake, filter, and use System repeatability: When the test is repeated 6 times the filtrate as the sample solution. Separately, dissolve 20 mg with 5 mL of the standard solution under the above operating of Mizoribine RS in 1 mL of water, and use this solution as conditions, the relative standard deviation of the peak area the standard solution. Perform the test with the sample solu- of mizoribine is not more than 2.0z. tion and standard solution as directed under Thin-Layer Chromatography <2.03>. Spot 1 mL each of the sample solu- Water <2.48> Not more than 0.5z (0.5 g, volumetric titra- JP XVI Official Monographs / Mizoribine Tablets 1129 tion and standard solution on a plate of silica gel for thin- about 25 mg of Mizoribine RS (separately determine the layer chromatography. Then develop the plate with a mix- water <2.48> in the same manner as Mizoribine), and dissolve ture of methanol, ammonia solution (28) and 1-propanol in water to make exactly 100 mL. Pipet 2 mL of the solution, (2:1:1) to a distance of about 10 cm, and air-dry the plate. add water to make exactly 100 mL, and use this solution as

Allow the plate to stand in iodine vapor: the principal spot the standard solution. Determine the absorbances, AT and from the sample solution and the spot from the standard so- AS, at 279 nm of the sample solution and standad solution as lution show a red-brown color and the same Rf value. directed under Ultraviolet-visible Spectrophotometry <2.24>.

Purity Related substances—To a quantity of powdered Amount of mizoribine (C9H13N3O6) Mizoribine Tablets, equivalent to 0.10 g of Mizoribine ac- ＝ MS × AT/AS × V?/V × 1/50 cording to the labeled amount, add 30 mL of the mobile M : Amount (mg) of Mizoribine RS, calculated on the phase, shake, then add the mobile phase to make 50 mL. S anhydrous basis Filter the solution through a membrane filter with a pore size not exceeding 0.5 mm and use the filtrate as the sample solu- Dissolution <6.10> When the test is performed at 50 revolu- tion. Pipet 2 mL of the sample solution, add the mobile tions per minute according to the Paddle method, using 900 phase to make exactly 20 mL. Pipet 1 mL of the solution, mL of water as the dissolution medium, the dissolution rate add the mobile phase to make exactly 20 mL, and use this so- in 45 minutes of Mizoribine Tablets is not less than 80z. lution as the standard solution. Perform the test with exactly Start the test with 1 tablet of Mizoribine Tablets, with- 5 mL each of the sample solution and standard solution as di- draw not less than 20 mL of the medium at the specified rected under Liquid Chromatography <2.01> according to minute after starting the test, and filter through a membrane the following conditions. Determine each peak area of each filter with a pore size not exceeding 0.5 mm. Discard not less solution by the automatic integration method: the area of than 10 mL of the first filtrate, pipet V mL of the subsequent the peak, having the relative retention time of about 0.3 with filtrate, add water to make exactly V?mL so that each mL respect to mizoribine, obtained from the sample solution is contains about 14 mg of mizoribine (C9H13N3O6) according not larger than the peak area of mizoribine from the stand- to the labeled amount, and use this solution as the sample ard solution, and the area of the peak other than mizoribine solution. Separately, weigh accurately about 28 mg of and other than the peak mentioned above is not larger than Mizoribine RS (separately determine the water <2.48> in the 2/5 times the peak area of mizoribine from the standard so- same manner as Mizoribine), and dissolve in water to make lution. exactly 100 mL. Pipet 1 mL of this solution, add water to Operating conditions— make exactly 20 mL, and use this solution as the standard

Column, column temperature, mobile phase, and flow solution. Determine the absorbances, AT and AS,at279nm rate: Proceed as directed in the operating conditions in the of the sample solution and standard solution as directed Assay under Mizoribine. under Ultraviolet-visible Spectrophotometry <2.24>. Detector: An ultraviolet absorption photometer (wave- Dissolution rate (z) with respect to the labeled amount length: 220 nm). of mizoribine (C H N O ) Time span of measurement: About 3 times as long as the 9 13 3 6 ＝ M × A /A × V?/V × 1/C × 45 retention time of mizoribine, beginning after the solvent S T S peak. MS: Amount (mg) of Mizoribine RS, calculated on the System suitability— anhydrous basis

Test for required detectability: To exactly 1 mL of the C: Labeled amount (mg) of mizoribine (C9H13N3O6)in1 standard solution add the mobile phase to make exactly 5 tablet mL. Confirm that the peak area of mizoribine obtained Assay Weigh accurately not less than 20 Mizoribine from 5 mL of this solution is equivalent to 14 to 26z of that Tablets, and powder. Weigh accurately a portion of from 5 mL of the standard solution. the powder, equivalent to about 25 mg of mizoribine System performance: When the procedure is run with 5 mL (C H N O ), add 50 mL of water and shake, then add water of the standard solution under the above operating condi- 9 13 3 6 to make exactly 100 mL. Filter the solution, discard not less tions, the number of theoretical plates and the symmetry fac- than 10 mL of the first filtrate, pipet 2 mL of the subsequent tor of the peak of mizoribine are not less than 10,000 and filtrate, add water to make exactly 100 mL, and use this so- not more than 1.4, respectively. lution as the sample solution. Separately, weigh accurately System repeatability: When the test is repeated 6 times about 25 mg of Mizoribine RS (separately determine the with 5 mL of the standard solution under the above operating water <2.48> in the same manner as Mizoribine), and dissolve conditions, the relative standard deviation of the peak area in water to make exactly 100 mL. Pipet 2 mL of the solution, of mizoribine is not more than 2.0z. add water to make exactly 100 mL, and use this solution as

Uniformity of dosage units <6.02> Perform the test accord- the standard solution. Determine the absorbances, AT and ing to the following method: it meets the requirement of the AS, at 279 nm of the sample solution and standard solution Content uniformity test. as directed under Ultraviolet-visible Spectrophotometry To 1 tablet of Mizoribine Tablets add 50 mL of water, <2.24>. shake until the tablet is disintegrated, and add water to make Amount (mg) of mizoribine (C H N O ) ＝ M × A /A exactly 100 mL. Filter the solution, discard not less than 10 9 13 3 6 S T S mL of the first filtrate, pipet V mL of the subsequent fil- MS: Amount (mg) of Mizoribine RS, calculated on the trate, add water to make exactly V?mL so that each mL con- anhydrous basis tains about 5 mg of mizoribine (C H N O ), and use this so- 9 13 3 6 Containers and storage Containers—Tight containers. lution as the sample solution. Separately, weigh accurately 1130 Morphine and Atropine Injection / Official Monographs JP XVI

the sample solution and standard solution as directed under Morphine and Atropine Injection Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak モルヒネ・アトロピン注射液 area of morphine to that of the internal standard. Amount (mg) of morphine hydrochloride hydrate

Morphine and Atropine Injection is an aqueous so- (C17H19NO3.HCl.3H2O) lution for injection. ＝ MS × QT/QS × 1.168 It contains not less than 0.91 w/vz and not M : Amount (mg) of morphine hydrochloride for assay, more than 1.09 w/vz of morphine hydrochloride S calculated on the anhydrous basis hydrate (C17H19NO3.HCl.3H2O: 375.84), and not less than 0.027 w/vz and not more than 0.033 w/vz of Internal standard solution—A solution of etilefrine hydro- atropine sulfate hydrate [(C17H23NO3)2.H2SO4.H2O: chloride (1 in 500). 694.83]. Operating conditions— Detector: An ultraviolet absorption photometer (wave- Method of preparation length: 285 nm). Morphine Hydrochloride Hydrate 10 g Column: A stainless steel column 4.6 mm in inside diame- Atropine Sulfate Hydrate 0.3 g ter and 15 cm in length, packed with octadecylsilanized silica Water for Injection or Sterile Water gel for liquid chromatography (5 mminparticlediameter). for Injection in Containers a significant quantity Column temperature: A constant temperature of about To make 1000 mL 409C. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in Prepare as directed under Injections, with the above ingre- 500 mL of diluted phosphoric acid (1 in 1000), and adjust dients. the pH with sodium hydroxide TS to 3.0. To 240 mL of this Description Morphine and Atropine Injection is a clear, solution add 70 mL of tetrahydrofuran, and mix. colorless liquid. Flow rate: Adjust the flow rate so that the retention time It is gradually colored by light. of morphine is about 10 minutes. pH:2.5–5.0 System suitability— System performance: When the procedure is run with 20 Identification To 2 mL of Morphine and Atropine Injec- mL of the standard solution under the above operating con- tion add 2 mL of ammonia TS, and extract with 10 mL of ditions, morphine and the internal standard are eluted in this diethyl ether. Filter the extract with a filter paper, evaporate order with the resolution between these peaks being not less the filtrate on a water bath to dryness, dissolve the residue in than 3. 1 mL of ethanol (99.5), and use this solution as the sample System repeatability: When the test is repeated 6 times solution. Separately, dissolve 0.1 g of morphine hydrochlo- with 20 mL of the standard solution under the above operat- ride in 10 mL of water, perform with 2 mL of this solution ing conditions, the relative standard deviation of the ratios the same procedure as used for preparation of the sample so- of the peak area of morphine to that of the internal standard lution, and use the solution so obtained as the standard solu- is not more than 1.0z. tion (1). Separately, dissolve 3 mg of atropine sulfate in 10 (2) Atropine sulfate hydrate—Pipet 2 mL of Morphine mL of water, perform with 2 mL of this solution the same and Atropine Injection, add exactly 2 mL of the internal procedure as used for preparation of the sample solution, standard solution, and use this solution as the sample solu- and use the solution so obtained as the standard solution (2). tion. Separately, weigh accurately about 15 mg of Atropine Perform the test with these solutions as directed under Thin- Sulfate RS (separately determine the loss on drying <2.41> layer Chromatography <2.03>. Spot 10 mLeachofthesample under the same conditions as Atropine Sulfate Hydrate), and solution and standard solutions (1) and (2) on a plate of dissolve in water to make exactly 50 mL. Pipet 2 mL of this silica gel for thin-layer chromatography. Develop the plate solution, add exactly 2 mL of the internal standard solution, with a mixture of methanol and ammonia solution (28) and use this solution as the standard solution. Perform the (200:3) to a distance of about 10 cm, and air-dry the plate. test with 20 mL each of the sample solution and standard so- Spray evenly Dragendorff's TS on the plate: the two spots lution as directed under Liquid Chromatography <2.01> ac- obtained from the sample solution show the same color tone cording to the following conditions, and calculate the ratios, and the same Rf value with either spot of orange color ob- QT and QS, of the peak areas of atropine to that of the inter- tained from the standard solution (1) or the standard solu- nal standard. tion (2) (morphine and atropine). Amount (mg) of atropine sulfate hydrate 6.05 Extractable volume < > It meets the requirement. [(C17H23NO3)2.H2SO4.H2O] Assay (1) Morphine hydrochloride hydrate—Pipet 2 mL ＝ MS × QT/QS × 1/25 × 1.027 of Morphine and Atropine Injection, add exactly 10 mL of MS: Amount (mg) of Atropine Sulfate RS, calculated on the internal standard solution, then add water to make 50 the dried basis mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of morphine hydrochloride for Internal standard solution—A solution of etilefrine hydro- assay, add exactly 10 mL of the internal standard solution to chloride (1 in 12,500). dissolve, then add water to make 50 mL, and use this solu- Operating conditions— tion as the standard solution. Perform the test with 20 mLof Column, column temperature, and mobile phase: Proceed as directed in the operating conditions in the Assay (1). JP XVI Official Monographs / Morphine Hydrochloride Injection 1131

Detector: An ultraviolet absorption photometer (wave- trum: both spectra exhibit similar intensities of absorption at length: 225 nm). the same wave numbers. Flow rate: Adjust the flow rate so that the retention time (3) A solution of Morphine Hydrochloride Hydrate (1 in of morphine is about 7 minutes. 50) responds to the Qualitative Tests <1.09> (2) for chloride. System suitability— Optical rotation <2.49> [a]20: －111 – －1169(0.5 g calcu- System performance: When the procedure is run with 20 D lated on the anhydrous basis, water, 25 mL, 100 mm). mL of the sample solution under the above operating conditions, morphine, the internal standard and atropine are pH <2.54> The pH of a solution obtained by dissolving eluted in this order, and the resolution between morphine 0.10 g of Morphine Hydrochloride Hydrate in 10 mL of and the internal standard is not less than 3. water is between 4.0 and 6.0. System repeatability: When the test is repeated 6 times Purity (1) Clarity and color of solution—Dissolve 0.40 g with 20 mL of the standard solution under the above operat- of Morphine Hydrochloride Hydrate in 10 mL of water: the ing conditions, the relative standard deviation of the ratios solution is clear. When perform the test with this solution as of the peak area of atropine to that of the internal standard directed under Ultraviolet-visible Spectrophotometry <2.24>, is not more than 1.0z. the absorbance at 420 nm is not more than 0.12. Containers and storage Containers—Hermetic containers, (2) Sulfate <1.14>—Dissolve 0.20 g of Morphine Hydro- and colored containers may be used. chloride Hydrate in 5 mL of water, and add 2 to 3 drops of Storage—Light-resistant. barium chloride TS: no turbidity is produced. (3) Meconic acid—Dissolve 0.20 g of Morphine Hydro- chloride Hydrate in 5 mL of water, and add 5 mL of dilute Morphine Hydrochloride Hydrate hydrochloric acid and 2 drops of iron (III) chloride TS: no red color develops. モルヒネ塩酸塩水和物 (4) Related substances—Dissolve 0.1 g of Morphine Hy- drochloride Hydrate in 10 mL of diluted ethanol (95) (1 in 2), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add diluted ethanol (95) (1 in 2) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mLeach of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog- C H NO .HCl.3H O: 375.84 17 19 3 2 raphy. Develop the plate with a mixture of ethanol (99.5), (5R,6S)-4,5-Epoxy-17-methyl-7,8-didehydromorphinan- toluene, acetone and ammonia solution (28) (14:14:7:1) to a 3,6-diol monohydrochloride trihydrate distance of about 15 cm, and air-dry the plate. Examine [6055-06-7] under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot from the sample solution are Morphine Hydrochloride Hydrate contains not less not more intense than the spot from the standard solution. than 98.0z and not more than 102.0z of morphine hydrochloride (C17H19NO3.HCl: 321.80), calculated on Water <2.48> 13–15z (0.1 g, direct titration). the anhydrous basis. Residue on ignition <2.44> Not more than 0.1z (0.5 g). Description Morphine Hydrochloride Hydrate occurs as Assay Weigh accurately about 0.5 g of Morphine Hydro- white, crystals or crystalline powder. chloride Hydrate, dissolve in 3.0 mL of formic acid, add 100 It is freely soluble in formic acid, soluble in water, spar- mL of a mixture of acetic anhydride and acetic acid (100) ingly soluble in methanol, and slightly soluble in ethanol (7:3), mix, and titrate <2.50> with 0.1 mol/L perchloric acid (95). VS (potentiometric titration). Perform a blank determina- It gradually becomes yellow-brown by light. tion, and make any necessary correction. Identification (1) Determine the absorption spectrum of a Each mL of 0.1 mol/L perchloric acid VS solution of Morphine Hydrochloride Hydrate (1 in 10,000) ＝ 32.18 mg of C H NO .HCl as directed under Ultraviolet-visible Spectrophotometry 17 19 3 <2.24>, and compare the spectrum with the Reference Spec- Containers and storage Containers—Tight containers. trum 1: both spectra exhibit similar intensities of absorption Storage—Light-resistant. at the same wavelengths. Separately, determine the absorption spectrum of a solution of Morphine Hydrochloride in dilute sodium hydroxide TS (1 in 10,000) as directed under Morphine Hydrochloride Injection Ultraviolet-visible Spectrophotometry, and compare the spectrum with the Reference Spectrum 2: both spectra ex- モルヒネ塩酸塩注射液 hibit similar intensities of absorption at the same wavelengths. Morphine Hydrochloride Injection is an aqueous so- (2) Determine the infrared absorption spectrum of Mor- lution for injection. phine Hydrochloride Hydrate as directed in the potassium It contains not less than 93.0z and not more than bromide disk method under Infrared Spectrophotometry 107.0z of the labeled amount of morphine hydrochlo- <2.25>, and compare the spectrum with the Reference Spec- 1132 Morphine Hydrochloride Tablets / Official Monographs JP XVI ride hydrate (C17H19NO3.HCl.3H2O: 375.84). the pH to 3.0 with sodium hydroxide TS. To 240 mL of this solution add 70 mL of tetrahydrofuran, and mix. Method of preparation Prepare as directed under Injec- Flow rate: Adjust the flow rate so that retention time of tions, with Morphine Hydrochloride Hydrate. morphine is about 10 minutes. Description Morphine Hydrochloride Injection is a clear, System suitability— colorless or pale yellow-brown liquid. System performance: When the procedure is run with 20 It gradually becomes yellow-brown by light. mL of the standard solution under the above operating con- pH:2.5–5.0 ditions, morphine and the internal standard are eluted in this order with the resolution between these peaks being not less Identification Take a volume of Morphine Hydrochloride than 3. Injection, equivalent to 0.04 g of Morphine Hydrochloride System repeatability: When the test is repeated 6 times Hydrate according to the labeled amount, add water to make with 20 mL of the standard solution under the above operat- 20 mL, and use this solution as the sample solution. To 5 mL ing conditions, the relative standard deviation of the ratios of the sample solution add water to make 100 mL, and deter- of the peak area of morphine to that of the internal standard mine the absorption spectrum as directed under Ultraviolet- is not more than 1.0z. visible Spectrophotometry <2.24>: it exhibits a maximum between 283 nm and 287 nm. And to 5 mL of the sample solu- Containers and storage Containers—Hermetic containers, tion add dilute sodium hydroxide TS to make 100 mL, and and colored containers may be used. determine the absorption spectrum: it exhibits a maximum Storage—Light-resistant. between 296 nm and 300 nm. Bacterial endotoxins <4.01> Less than 1.5 EU/mg. Morphine Hydrochloride Tablets Extractable volume <6.05> It meets the requirement. モルヒネ塩酸塩錠 Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Morphine Hydrochloride Tablets contain not less Insoluble particulate matter <6.07> It meets the require- than 93.0z and not more than 107.0z of the ment. labeled amount of morphine hydrochloride hydrate Sterility <4.06> Perform the test according to the Mem- (C17H19NO3.HCl.3H2O: 375.84). brane filtration method: it meets the requirement. Method of preparation Prepare as directed under Tablets, Assay Take exactly a volume of Morphine Hydrochloride with Morphine Hydrochloride Hydrate. Injection, equivalent to about 80 mg of morphine hydrochlo- Identification Weigh a quantity of powdered Morphine ride hydrate (C H NO .HCl.3H O), and add water to make 17 19 3 2 Hydrochloride Tablets equivalent to 0.01 g of Morphine Hy- exactly 20 mL. Pipet 5 mL of this solution, add exactly 10 drochloride Hydrate, add 100 mL of water, shake for 10 mL of the internal standard solution and water to make 50 minutes, and filter. Determine the absorption spectrum of mL, and use this solution as the sample solution. Separately, the filtrate as directed under Ultraviolet-visible Spectropho- weigh accurately about 25 mg of morphine hydrochloride for tometry <2.24>: it exhibits a maximum between 283 nm and assay, dissolve in exactly 10 mL of the internal standard so- 287 nm. And weigh a quantity of powdered Morphine Hy- lution, add water to make 50 mL, and use this solution as the drochloride Tablets equivalent to 0.01 g of Morphine Hydro- standard solution. Perform the test with 20 mLeachofthe chloride Hydrate, add 100 mL of dilute sodium hydroxide sample solution and standard solution as directed under Liq- TS, shake for 10 minutes, and filter. Determine the absorp- uid Chromatography <2.01> according to the following con- tion spectrum of the filtrate: it exhibits a maximum between ditions, and calculate the ratios, Q and Q , of the peak area T S 296 nm and 300 nm. of morphine to that of the internal standard. Uniformity of dosage units <6.02> Perform the test accord- Amount (mg) of morphine hydrochloride ing to the following method: it meets the requirement of the (C H NO .HCl.3H O) 17 19 3 2 Content uniformity test. ＝ M × Q /Q × 4 × 1.168 S T S To 1 tablet of Morphine Hydrochloride Tablets add ex-

MS: Amount (mg) of morphine hydrochloride for assay, actly 1 mL of the internal standard solution per 2 mg of mor- calculated on the anhydrous basis phine hydrochloride hydrate (C17H19NO3.HCl.3H2O), disperse the tablet into a small particles using ultrasonic waves, Internal standard solution—A solution of etilefrine hydro- then treat with ultrasonic waves for 15 minutes with occa- chloride (1 in 500). sional stirring, and add water to make V mL so that each mL Operating conditions— contains about 0.4 mg of morphine hydrochloride hydrate Detector: An ultraviolet absorption photometer (wave- (C H NO .HCl.3H O). Filter the solution, and use the fil- length: 285 nm). 17 19 3 2 trate as the sample solution. Then, proceed as directed in the Column: A stainless steel column 4.6 mm in inside diame- Assay. ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Amount (mg) of morphine hydrochloride hydrate

Column temperature: A constant temperature of about (C17H19NO3.HCl.3H2O) 409C. ＝ MS × QT/QS × V/50 × 1.168 Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in M : Amount (mg) of morphine hydrochloride for assay, 500 mL of diluted phosphoric acid (1 in 1000), and adjust S JP XVI Official Monographs / Mosapride Citrate Hydrate 1133

calculated on the anhydrous basis Amount (mg) of morphine hydrochloride (C H NO .HCl.3H O) Internal standard solution—A solution of etilefrine hydro- 17 19 3 2 ＝ M × Q /Q × 1.168 chloride (1 in 500). S T S M : Amount (mg) of morphine hydrochloride for assay, Dissolution <6.10> When the test is performed at 50 revolu- S calculated on the anhydrous basis tions per minute according to the Paddle method using 900 mL of water as the dissolution medium, the dissolution rate Internal standard solution—A solution of etilefrine hydro- in 15 minutes of Morphine Hydrochloride Tablets is not less chloride (1 in 500). than 85z. Operating conditions— Start the test with 1 tablet of Morphine Hydrochloride Detector: An ultraviolet absorption photometer (wave- Tablets, withdraw not less than 20 mL of the medium at the length: 285 nm). specified minute after starting the test, and filter through a Column: A stainless steel column 4.6 mm in inside diame- membrane filter with a pore size not exceeding 0.45 mm. Dis- ter and 15 cm in length, packed with octadecylsilanized silica card the first 10 mL of the filtrate, and use the subsequent gel for liquid chromatography (5 mminparticlediameter). filtrate as the sample solution. Separately, weigh accurately Column temperature: A constant temperature of about about 28 mg of morphine hydrochloride for assay (sepa- 409C. rately, determine the water <2.48> inthesamemanneras Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in Morphine Hydrochloride Hydrate), and dissolve in water to 500 mL of diluted phosphoric acid (1 in 1000), and adjust make exactly 100 mL. Pipet 2 mL of this solution, add water the pH to 3.0 with sodium hydroxide TS. To 240 mL of this to make exactly 50 mL, and use this solution as the standard solution add 70 mL of tetrahydrofuran, and mix. solution. Perform the test with exactly 25 mLeachofthe Flow rate: Adjust the flow rate so that the retention time sample solution and standard solution as directed under Liq- of morphine is about 10 minutes. uid Chromatography <2.01> according to the following con- System suitability— ditions, and determine the peak areas, AT and AS,ofmor- System performance: When the procedure is run with 20 phine in each solution. mL of the standard solution under the above operating conditions, morphine and the internal standard are eluted in this Dissolution rate (z) with respect to the labeled amount of order with the resolution between these peaks being not less morphine hydrochloride hydrate (C H NO .HCl.3H O) 17 19 3 2 than 3. ＝ M × A /A × 1/C × 36 × 1.168 S T S System repeatability: When the test is repeated 6 times

MS: Amount (mg) of morphine hydrochloride for assay, with 20 mL of the standard solution under the above operat- calculated on the anhydrous basis ing conditions, the relative standard deviation of the ratios C: Labeled amount (mg) of morphine hydrochloride hy- of the peak area of morphine to that of the internal standard

drate (C17H19NO3.HCl.3H2O) in 1 tablet is not more than 1.0z. Operating conditions— Containers and storage Containers—Tight containers. Proceed as directed in the operating conditions in the Storage—Light-resistant. Assay. System suitability— System performance: When the procedure is run with 25 Mosapride Citrate Hydrate mL of the standard solution under the above operating conditions, the number of theoretical plates and the symmetry モサプリドクエン酸塩水和物 factor of the peak of morphine are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of morphine is not more than 2.0z.

Assay Take not less than 20 Morphine Hydrochloride C21H25ClFN3O3.C6H8O7.2H2O: 650.05 Tablets, weigh accurately, and powder. Weigh accurately a 4-Amino-5-chloro-2-ethoxy-N-{[(2RS)- quantity of the powder, equivalent to about 20 mg of mor- 4-(4-fluorobenzyl)morpholin-2-yl]methyl}benzamide phine hydrochloride hydrate (C17H19NO3.HCl.3H2O), add monocitrate dihydrate exactly 10 mL of the internal standard solution, extract the [636582-62-2] mixture with ultrasonic waves for 10 minutes, and add water to make 50 mL. Filter this solution, and use the filtrate as Mosapride Citrate Hydrate contains not less than the sample solution. Separately, weigh accurately about 25 98.5z and not more than 101.0z of mosapride citrate mg of morphine hydrochloride for assay, dissolve in exactly (C21H25ClFN3O3.C6H8O7: 614.02), calculated on the 10 mL of the internal standard solution, add water to make anhydrous basis. 50 mL, and use this solution as the standard solution. Per- Description Mosapride Citrate Hydrate occurs as a white form the test with 20 mL each of the sample solution and to yellowish white crystalline powder. standard solution as directed under Liquid Chromatography It is freely soluble in N,N-dimethylformamide and in ace- <2.01> according to the following conditions, and calculate tic acid (100), sparingly soluble in methanol, slightly soluble the ratios, Q and Q , of the peak area of morphine to that T S in ethanol (99.5), and practically insoluble in water. of the internal standard. 1134 Mosapride Citrate Powder / Official Monographs JP XVI

A solution of Mosapride Citrate Hydrate in N,N- dimethylformamide (1 in 20) shows no optical rotation. Time after injection Mobile phase A Mobile phase B of sample (min) (volz) (volz) Identification (1) Determine the absorption spectrum of a solution of Mosapride Citrate Hydrate in methanol (1 in 0–35 80→ 45 20 → 55 50,000) as directed under Ultraviolet-visible Spectropho- tometry <2.24>, and compare the spectrum with the Refer- Flow rate: 1.0 mL per minute. ence Spectrum: both spectra exhibit similar intensities of ab- Time span of measurement: Beginning after the solvent sorption at the same wavelengths. peak to 35 minutes after injection. (2) Determine the infrared absorption spectrum of System suitability— Mosapride Citrate Hydrate as directed in the potassium bro- Test for required detectability: Pipet 4 mL of the standard mide disk method under Infrared Spectrophotometry <2.25>, solution, and add methanol to make exactly 20 mL. Confirm and compare the spectrum with the Reference Spectrum: that the peak area of mosapride obtained from 5 mLofthis both spectra exhibit similar intensities of absorption at the solution is equivalent to 15 to 25z of that of mosapride same wave numbers. from 5 mL of the standard solution. (3) A solution of Mosapride Citrate Hydrate in N,N- System performance: When the procedure is run with 5 mL dimethylformamide (1 in 10) responds to the Qualitative of the standard solution under the above operating condi- Tests <1.09> (1) for citrate. tions, the number of theoretical plates and the symmetry fac- Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of tor of the peak of mosapride are not less than 40,000 and not Mosapride Citrate Hydrate in a platinum crucible according more than 1.5, respectively. to Method 4, and perform the test. Prepare the control solu- System repeatability: When the test is repeated 6 times tion with 2.0 mL of Standard Lead Solution (not more than with 5 mL of the standard solution under the above operating 20 ppm). conditions, the relative standard deviation of the peak area (2) Related substances—Dissolve 0.10 g of Mosapride of mosapride is not more than 5.0z. Citrate Hydrate in 50 mL of methanol, and use this solution (3) Residual solvent—Being specified separately. as the sample solution. Pipet 1 mL of the sample solution, Water <2.48> 5.0–6.5z (0.5 g, volumetric titration, back and add methanol to make exactly 50 mL. Pipet 1 mL of this titration). solution, add methanol to make exactly 20 mL, and use this solution as the standard solution. Perform the test with ex- Residue on ignition <2.44> Not more than 0.1z (1 g, plati- actly 5 mL each of the sample solution and standard solution num crucible). as directed under Liquid Chromatography <2.01> according Assay Weigh accurately 0.5 g of Mosapride Citrate Hy- to the following conditions. Determine each peak area of drate, dissolve in 70 mL of acetic acid (100), and titrate both solutions by the automatic integration method: the area <2.50> with 0.1 mol/L perchloric acid VS (potentiometric of the peak having the relative retention time of about 0.47 titration). Perform a blank determination in the same man- with respect to mosapride from the sample solution is not ner, and make any necessary correction. larger than 3 times the peak area of mosapride from the standard solution, and the area of each peak other than the Each mL of 0.1 mol/L perchloric acid VS peak of mosapride and other than the peak mentioned above ＝ 61.40 mg of C21H25ClFN3O3.C6H8O7 from the sample solution is not larger than the peak area of Containers and storage Containers—Well-closed contain- mosapride from the standard solution. Furthermore, the ers. total area of the peaks other than the peak of mosapride from the sample solution is not larger than 5 times the peak area of mosapride from the standard solution. Mosapride Citrate Powder Operating conditions— Detector: An ultraviolet absorption photometer (wave- モサプリドクエン酸塩散 length: 274 nm). Column: A stainless steel column 4.6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica Mosapride Citrate Powder contains not less than gel for liquid chromatography (5 mm in particle diameter). 93.0z and not more than 107.0z of the labeled Column temperature: A constant temperature of about amount of mosapride citrate (C21H25ClFN3O3.C6H8O7: 409C. 614.02). Mobile phase A: Dissolve 8.82 g of trisodium citrate dihy- Method of preparation Prepare as directed under Granules drate in 800 mL of water, adjust the pH to 4.0 with dilute or Powders, with Mosapride Citrate Hydrate. hydrochloric acid, and add water to make 1000 mL. Mobile phase B: Acetonitrile. Identification (1) Powder Mosapride Citrate Powder. To Flowing of the mobile phase: Control the gradient by mix- a portion of the powder, equivalent to 10 mg of mosapride ing the mobile phases A and B as directed in the following citrate (C21H25ClFN3O3.C6H8O7) according to the labeled table. amount, add 10 mL of dilute acetic acid, shake for 10 minutes, and filter. To 5 mL of the filtrate add 0.3 mL of Dragendorff's TS: an orange precipitate is formed. (2) Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet- visible Spectrophotometry <2.24>: it exhibits maxima be- JP XVI Official Monographs / Mosapride Citrate Powder 1135 tween 271 nm and 275 nm and between 306 nm and 310 nm. that each mL contains about 20 mg of mosapride citrate (C H ClFN O .C H O ), and use this solution as the sam- Purity Related substances—Powder Mosapride Citrate 21 25 3 3 6 8 7 ple solution. Then, proceed as directed in the Assay. Powder. To a portion of the powder, equivalent to 10 mg of mosapride citrate (C21H25ClFN3O3.C6H8O7) according to the Amount (mg) of mosapride citrate (C21H25ClFN3O3.C6H8O7) labeled amount, moisten with 1 mL of water, then add 9 mL ＝ MS × AT/AS × V?/V × 1/50 of methanol, shake for 20 minutes, centrifuge, and use the M : Amount (mg) of mosapride citrate for assay, calcu- supernatant liquid as the sample solution. Pipet 1 mL of the S lated on the anhydrous basis sample solution, and add methanol to make exactly 20 mL. Pipet 2 mL of this solution, add methanol to make exactly Dissolution <6.10> When the test is performed at 50 revolu- 20 mL, and use this solution as the standard solution. Per- tions per minute according to the Paddle method, using 900 form the test with exactly 10 mL each of the sample solution mL of the 2nd fluid for dissolution test as the dissolution and standard solution as directed under Liquid Chromatog- medium, the dissolution rate in 45 minutes of Mosapride raphy <2.01> according to the following conditions, and de- Citrate Powder is not less than 70z. termine each peak area by the automatic integration method: Start the test with an amount of Mosapride Citrate the area of the two peaks, having the relative retention time Powder, equivalent to about 2.5 mg of mosapride citrate of about 0.60 and about 0.85 with respect to mosapride ob- (C21H25ClFN3O3.C6H8O7) according to the labeled amount, tained from the sample solution, is not larger than the peak withdraw not less than 20 mL of the medium at the specified area of mosapride from the standard solution, the area of minute after starting the test, and filter through a membrane other than the peak of mosapride and the peaks mentioned filter with a pore size not exceeding 0.45 mm. Discard the above is not larger than 2/5 times the peak area of first 10 mL of the filtrate, and use the subsequent filtrate as mosapride from the standard solution, and the total area of the sample solution. Separately, weigh accurately about 30 the peak other than mosapride is not larger than 2 times the mg of mosapride citrate for assay (separately determine the peak area of mosapride from the standard solution. water <2.48> in the same manner as Mosapride Citrate Hy- Operating conditions— drate), and dissolve in the mobile phase to make exactly 100 Detector, column, column temperature, mobile phases A mL. Pipet 2 mL of this solution, add the mobile phase to and B, and flow rate: Proceed as directed in the operating make exactly 200 mL, and use this solution as the standard conditions in the Purity (2) under Mosapride Citrate Hy- solution. Perform the test with exactly 50 mLeachofthe drate. sample solution and standard solution as directed under Liq- Flowing of mobile phase: Control the gradient by mixing uid Chromatography <2.01>, and determine the peak areas, the mobile phases A and B as directed in the following table. AT and AS, of mosapride of both solutions. Dissolution rate (z) with respect to the labeled amount Time after injection Mobile phase A Mobile phase B of mosapride citrate (C21H25ClFN3O3.C6H8O7) of sample (min) (vol ) (vol ) z z ＝ MS/MT × AT/AS × 1/C × 9

0–40 85–45 15–55 MS: Amount (mg) of mosapride citrate for assay, calculated on the anhydrous basis Time span of measurement: For 40 minutes after sample MT: Amount (g) of sample injection, beginning after the solvent peak. C: Labeled amount (mg) of mosapride citrate System suitability— (C21H25ClFN3O3.C6H8O7)in1g Test for required detectability: To exactly 1 mL of the Operating conditions— standard solution add methanol to make exactly 25 mL. Detector: An ultraviolet absorption photometer (wave- Confirm that the peak area of mosapride obtained with 10 length: 274 nm). mL of this solution is equivalent to 3.0 to 5.0z of that with Column: A stainless steel column 4.6 mm in inside diame- 10 mL of the standard solution. ter and 15 cm in length, packed with octadecylsilanized silica System performance: When the procedure is run with 10 gel for liquid chromatography (5 mminparticlediameter). mL of the standard solution under the above operating con- Column temperature: A constant temperature of about ditions, the number of theoretical plates and the symmetry 409C. factor of the peak of mosapride are not less than 40,000 and Mobile phase: Dissolve 8.82 g of trisodium citrate dihy- not more than 1.5, respectively. drate in 800 mL of water, adjust to pH 3.3 with dilute hydro- System repeatability: When the test is repeated 6 times chloric acid, and add water to make 1000 mL. To 240 mL of with 10 mL of the standard solution under the above operat- this solution add 90 mL of methanol and 70 mL of aceto- ing conditions, the relative standard deviation of the peak nitrile. area of mosapride is not more than 3.0z. Flow rate: Adjust the flow rate so that the retention time Uniformity of dosage units <6.02> Perform the test accord- of mosapride is about 9 minutes. ing to the following method: the powder in single-unit con- System suitability— tainer meets the requirement of the Content uniformity test. System performance: When the procedure is run with 50 To the total content of 1 container of Mosapride Citrate mL of the standard solution under the above operating con- Powder add 5 mL of water, and shake. Then, add 20 mL of ditions, the number of theoretical plates and the symmetry methanol, shake for 20 minutes, and add methanol to make factor of the peak of mosapride are not less than 4000 and exactly 50 mL. Centrifuge this solution, pipet V mL of the not more than 2.0, respectively. supernatant liquid, add methanol to make exactly V?mL so System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat- 1136 Mosapride Citrate Tablets / Official Monographs JP XVI ing conditions, the relative standard deviation of the peak peaks having the relative retention times of about 0.60 and area of mosapride is not more than 2.0z. about 0.85 with respect to mosapride from the sample solution is not larger than the peak area of mosapride from the Assay Powder Mosapride Citrate Powder. Weigh accu- standard solution, and the area of each peak other than the rately a portion of the powder, equivalent to about 10 mg of peak of mosapride and other than those mentioned above mosapride citrate (C H ClFN O .C H O ), moisten with 2 21 25 3 3 6 8 7 from the sample solution is not larger than 2/5 times the mL of water, add 70 mL of methanol, shake for 20 minutes, peak area of mosapride from the standard solution. Further- then add methanol to make exactly 100 mL, and centrifuge. more, the total area of the peaks other than mosapride from Pipet 10 mL of the supernatant liquid, add methanol to the sample solution is not larger than 2 times the peak area make exactly 50 mL, and use this solution as the sample of mosapride from the standard solution. solution. Separately, weigh accurately about 53 mg of Operating conditions— mosapride citrate for assay (separately determine the water Detector, column, column temperature, mobile phase A, <2.48> in the same manner as Mosapride Citrate Hydrate), mobile phase B, and flow rate: Proceed as directed in the op- and dissolve in methanol to make exactly 100 mL. To 2 mL erating conditions in the Purity (2) under Mosapride Citrate of this solution add methanol to make exactly 50 mL, and Hydrate. use this solution as the standard solution. Determine the ab- Flowing of the mobile phase: Control the gradient by mix- sorbances, A and A , of the sample solution and the stand- T S ing the mobile phases A and B as directed in the following ard solution at 273 nm as directed under Ultraviolet-visible table. Spectrophotometry <2.24>.

Amount (mg) of mosapride citrate (C21H25ClFN3O3.C6H8O7) Time after injection Mobile phase A Mobile phase B ＝ MS × AT/AS × 1/5 of sample (min) (volz) (volz) M : Amount (mg) of mosapride citrate for assay, calcu- S 0–40 85→ 45 15 → 55 lated on the anhydrous basis

Containers and storage Containers—Tight containers. Time span of measurement: Beginning after the solvent peak to 40 minutes after injection. System suitability— Mosapride Citrate Tablets Test for required detectability: Pipet 1 mL of the standard solution, and add methanol to make exactly 25 mL. Confirm モサプリドクエン酸塩錠 that the peak area of mosapride obtained from 10 mLofthis solution is equivalent to 3.0 to 5.0z of that of mosapride Mosapride Citrate Tablets contain not less than from 10 mL of the standard solution. 95.0z and not more than 105.0z of the labeled System performance: When the procedure is run with 10 mL of the standard solution under the above operating con- amount of mosapride citrate (C21H25ClFN3O3.C6H8O7: 614.02). ditions, the number of theoretical plates and the symmetry factor of the peak of mosapride are not less than 40,000 and Method of preparation Prepare as directed under Tablets, not more than 1.5, respectively. with Mosapride Citrate Hydrate. System repeatability: When the test is repeated 6 times Identification (1) To an amount of powdered Mosapride with 10 mL of the standard solution under the above operat- Citrate Tablets, equivalent to 10 mg of mosapride citrate ing conditions, the relative standard deviation of the peak area of mosapride is not more than 3.0z. (C21H25ClFN3O3.C6H8O7) according to the labeled amount, add 10 mL of dilute acetic acid, shake for 10 minutes, and Uniformity of dosage units <6.02> Perform the test accord- filter. To 5 mL of the filtrate add 0.3 mL of Dragendorff's ing to the following method: it meets the requirement of the TS: an orange precipitate is formed. Content uniformity test. (2) Determine the absorption spectrum of the sample so- To 1 tablet of Mosapride Citrate Tablets add 5 mL of lution obtained in the Assay as directed under Ultraviolet- water, and shake well to disintegrate. Add 20 mL of metha- visible Spectrophotometry <2.24>: it exhibits maxima be- nol, shake for 20 minutes, and add methanol to make exactly tween 271 nm and 275 nm, and between 306 nm and 310 nm. 50 mL. Centrifuge this solution, pipet V mL of the superna- Purity Related substances—Powder not less than 20 tablets tant liquid, add methanol to make exactly V?mL so that of Mosapride Citrate Tablets. Moisten a portion of the each mL contains about 20 mg of mosapride citrate powder, equivalent to 10 mg of mosapride citrate (C21H25ClFN3O3.C6H8O7), and use this solution as the sample solution. Proceed as directed in the Assay. (C21H25ClFN3O3.C6H8O7) according to the labeled amount, with 1 mL of water. Add 9 mL of methanol, shake for 20 Amount (mg) of mosapride citrate (C21H25ClFN3O3.C6H8O7) minutes, centrifuge, and use the supernatant liquid as the ＝ MS × AT/AS × V?/V × 1/50 sample solution. Pipet 1 mL of this solution, add methanol to make exactly 20 mL. Pipet 2 mL of the sample solution, MS: Amount (mg) of mosapride citrate for assay, calcu- add methanol to make exactly 20 mL, and use this solution lated on the anhydrous basis as the standard solution. Perform the test with exactly 10 mL Dissolution <6.10> When the test is performed at 50 revolu- each of the sample solution and standard solution as directed tions per minute according to the Paddle method using the under Liquid Chromatography <2.01> according to the fol- sinker, using 900 mL of 2nd fluid for dissolution test as the lowing conditions. Determine each peak area of both solu- dissolution medium, the dissolution rate in 45 minutes of tions by the automatic integration method: the area of the JP XVI Official Monographs / Mupirocin Calcium Hydrate 1137

Mosapride Citrate Tablets is not less than 80z. mosapride citrate for assay (separately, determine the water Start the test with 1 tablet of Mosapride Citrate Tablets, <2.48> in the manner as Mosapride Citrate Hydrate), and dis- withdraw not less than 20 mL of the medium at the specified solve in methanol to make exactly 100 mL. Pipet 2 mL of minute after starting the test, and filter through a membrane this solution, add methanol to make exactly 50 mL, and use filter with a pore size not exceeding 0.45 mm. Discard the this solution as the standard solution. Perform the test with first 10 mL of the filtrate, pipet V mL of the subsequent fil- the sample solution and standard solution as directed under trate, add the dissolution medium to make exactly V?mL so Ultraviolet-visible Spectrophotometry <2.24>, and determine that each mL contains about 2.8 mg of mosapride citrate the absorbances, AT and AS,at273nm. (C H ClFN O .C H O ) according to the labeled amount, 21 25 3 3 6 8 7 Amount (mg) of mosapride citrate (C H ClFN O .C H O ) and use this solution as the sample solution. Separately, 21 25 3 3 6 8 7 ＝ M × A /A × 1/5 weigh accurately about 30 mg of mosapride citrate for assay S T S

(separately, determine the water <2.48> in the same manner MS: Amount (mg) of mosapride citrate for assay, calcu- as Mosapride Citrate Hydrate), and dissolve in the mobile lated on the anhydrous basis phase to make exactly 100 mL. Pipet 2 mL of this solution, Containers and storage Containers—Tight containers. add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> ac- Freeze-dried Live Attenuated cording to the following conditions, and determine the peak Mumps Vaccine areas, AT and AS, of mosapride of both solutions. 乾燥弱毒生おたふくかぜワクチン Dissolution rate (z) with respect to the labeled amount

of mosapride citrate (C21H25ClFN3O3.C6H8O7) ＝ MS × AT/AS × V?/V × 1/C × 9 Freeze-dried Live Attenuated Mumps Vaccine is a dried preparation containing live attenuated mumps MS: Amount (mg) of mosapride citrate for assay, calculated on the anhydrous basis virus. It conforms to the requirements of Freeze-dried Live C: Labeled amount (mg) of mosapride citrate Attenuated Mumps Vaccine in the Minimum Require- (C H ClFN O .C H O )in1tablet 21 25 3 3 6 8 7 ments of Biologic Products. Operating conditions— Description Freeze-dried Live Attenuated Mumps Vaccine Detector: An ultraviolet absorption photometer (wave- becomes a clear, colorless, yellowish or reddish liquid on ad- length: 274 nm). dition of solvent. Column: A stainless steel column 4.6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about Mupirocin Calcium Hydrate 409C. ムピロシンカルシウム水和物 Mobile phase: Dissolve 8.82 g of trisodium citrate dihydrate in 800 mL of water, adjust the pH to 3.3 with dilute hydrochloric acid, and add water to make 1000 mL. To 240 mL of this solution add 90 mL of methanol and 70 mL of acetonitrile. Flow rate: Adjust the flow rate so that the retention time C H CaO .2H O: 1075.34 of mosapride is about 9 minutes. 52 86 18 2 Monocalcium bis[9-((2E)-4-{(2S,3R,4R,5S)-5- System suitability— [(2S,3S,4S,5S)-2,3-epoxy-5-hydroxy-4-methylhexyl]-3,4- System performance: When the procedure is run with 50 dihydroxy-3,4,5,6-tetrahydro-2H-pyran-2-yl}-3-methylbut- mL of the standard solution under the above operating con- 2-enoyloxy)nonanoate] dihydrate ditions, the number of theoretical plates and the symmetry [115074-43-6] factor of the peak of mosapride are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times Mupirocin Calcium Hydrate is the calcium salt of a substance having antibacterial activity produced by the with 50 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak growth of Pseudomonas fluorescens. It contains not less than 895 mg (potency) and not area of mosapride is not more than 2.0z. more than 970 mg (potency) per mg, calculated on the Assay Weigh accurately the mass of not less than 20 anhydrous basis. The potency of Mupirocin Calcium Mosapride Citrate Tablets, and powder. Weigh accurately a Hydrate is expressed as mass (potency) of mupirocin portion of the powder, equivalent to about 10 mg of (C26H44O9: 500.62). mosapride citrate (C H ClFN O .C H O ), and moisten 21 25 3 3 6 8 7 Description Mupirocin Calcium Hydrate occurs as a white with 2 mL of water. Add 70 mL of methanol, shake for 20 powder and has a bitter taste. minutes, add methanol to make exactly 100 mL, and centri- It is freely soluble in methanol and slightly soluble in fuge. Pipet 10 mL of the supernatant liquid, add methanol water and in ethanol (95). to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 53 mg of 1138 Mupirocin Calcium Hydrate / Official Monographs JP XVI

Identification (1) To 1 mL of a solution of Mupirocin the Assay. Calcium Hydrate in methanol (1 in 200) add 4 mL of Time span of measurement: About 3 times as long as the hydroxylamine perchlorate-ethanol TS and 1 mL of N,N?- retention time of mupirocin beginning after the solvent peak. dicyclohexylcarbodiimide-ethanol TS, shake well, and allow System suitability— to stand in lukewarm water for 20 minutes. After cooling, Test for required detection: Pipet 1 mL of the sample solu- add 1 mL of iron (III) perchorate hexahydrate-ethanol TS to tion (2), and add a mixture of 0.1 mol/L acetic acid-sodium the solution, and shake: a dark purple color develops. acetate buffer solution, pH 4.0, and a solution of tetra- (2) Determine the absorption spectrum of a solution of hydrofuran (3 in 4) (1:1) to make exactly 20 mL. Confirm Mupirocin Calcium Hydrate (1 in 50,000) as directed under that the peak area of mupirocin obtained from 20 mLofthis Ultraviolet-visible Spectrophotometry <2.24>:itexhibitsa solution is equivalent to 4 to 6z of that obtained from 20 mL maximum between 219 nm and 224 nm. of the sample solution (2). (3) Determine the infrared absorption spectrum of System performance: Proceed as directed in the system Mupirocin Calcium Hydrate as directed in the paste method suitability in the Assay. under Infrared Spectrophotometry <2.25>: it exhibits absorp- System repeatability: When the test is repeated 6 times tion at the wave numbers of about 1708 cm－1, 1648 cm－1, with 20 mL of the sample solution (2) under the above oper- 1558 cm－1,1231cm－1, 1151cm－1 and 894 cm－1. ating conditions, the relative standard deviation of the peak (4) A solution of Mupirocin Calcium Hydrate (3 in 1000) areas of mupirocin is not more than 2.0z. responds to the Qualitative Tests <1.09> (3) for calcium salt. (2) Inorganic salt from manufacturing process—Being

20 specified separately. Optical rotation <2.49> [a]D : －16 – －209(1 g calculated on the anhydrous basis, methanol, 20 mL, 100 mm). Water <2.48> Not less than 3.0z and not more than 4.5z (0.5 g, volumetric titration, direct titration). Purity (1) Related substances—Dissolve 50 mg of Mupirocin Calcium Hydrate in a mixture of 0.1 mol/L acetic Assay Weigh accurately an amount of Mupirocin Calcium acid-sodium acetate buffer solution, pH 4.0, and a solution Hydrate and Mupirocin Lithium RS, equivalent to about 20 of tetrahydrofuran (3 in 4) (1:1) to make 10 mL, and use this mg (potency), dissolve in a mixture of 0.1 mol/L acetic acid- solution as the sample solution (1). Pipet 2 mL of the sample sodium acetate buffer solution, pH 4.0 and a solution of solution (1), add a mixture of 0.1 mol/L acetic acid-sodium tetrahydrofuran (3 in 4) (1:1) to make exactly 200 mL, and acetate buffer solution, pH 4.0, and a solution of tetra- use these solutions as the sample solution and the standard hydrofuran (3 in 4) (1:1) to make exactly 100 mL, and use solution. Preserve these solutions at a temperature between this solution as the sample solution (2). Preserve these sam- 49Cand89C. Perform the test with exactly 20 mLofthe ple solutions at a temperature between 49Cand89C. Per- sample solution and standard solution as directed under form the test with exactly 20 mL of the sample solution (1) Liquid Chromatography <2.01> according to the following and the sample solution (2) as directed under Liquid Chro- conditions, and determine the peak areas, AT and AS,of matography <2.01> according to the following conditions, mupirocin of each solution. and determine the areas of each peak of the sample solution Amount [mg (potency)] of mupirocin (C H O ) (1) and the sample solution (2) by the automatic integration 26 44 9 ＝ M × A /A × 1000 method. Calculate the amount of the related substances by S T S the following formula: the amount of principal related sub- MS: Amount [mg (potency)] of Mupirocin Lithium RS stance (appeared at about 0.7 of the relative retention time to Operating conditions— mupirocin) is not more than 4.0z, and the total amount of Detector: An ultraviolet absorption photometer (wave- related substances (the total area of the peaks other than of length: 240 nm). the solvent and mupirocin) is not more than 6.0z. Column: A stainless steel column 4.6 mm in inside diame- Amount (z) of principal related substance ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticlediameter). A P × 100 ＝ i × 100 × Column temperature: A constant temperature of about A ＋ Am A × 100 100 － 409C. A ＋ A m Mobile phase: Dissolve 7.71 g of ammonium acetate in Total amount (z) of related substances 750 mL of water, adjust the pH to 5.7 with acetic acid (100), and add water to make 1000 mL. To 300 mL of this solution A P × 100 ＝ × 100 × add 100 mL of tetrahydrofuran. A ＋ Am A × 100 100 － Flow rate: Adjust the flow rate so that the retention time A ＋ A m of mupirocin is about 12.5 minutes. A: Total peak areas other than of the solvent and mupiro- System suitability— cin from the sample solution (1) System performance: Dissolve about 20 mg of Mupirocin Ai: Peak area of the relative retention time of about 0.7 to Lithium RS and about 5 mg of ethyl parahydroxybenzoate in mupirocin from the sample solution (1) a mixture of 0.1 mol/L acetic acid-sodium acetate buffer Am: A value of 50 times of peak area of mupirocin from solution, pH 4.0 and a solution of tetrahydrofuran (3 in 4) the sample solution (2) (1:1) to make 200 mL. When the procedure is run with 20 mL P: Potency per mg obtained from the assay of this solution under the above operating conditions, Operating conditions— mupirocin and ethyl parahydroxybenzoate are eluted in this Detector, column, column temperature, mobile phase, and order with the resolution between these peaks being not less flow rate: Proceed as directed in the operating conditions in than 12. JP XVI Official Monographs / Nabumetone 1139

System repeatability: When the test is repeated 6 times Operating conditions— with 20 mL of the standard solution under the above operat- Detector, column, column temperature, mobile phase, and ing conditions, the relative standard deviation of the peak flow rate: Proceed as directed in the operating conditions in areas of mupirocin is not more than 1.0z. the Assay under Mupirocin Calcium Hydrate. Time span of measurement: About 5 times as long as the Containers and storage Containers—Tight containers. retention time of mupirocin, beginning after the solvent peak. System suitability— Mupirocin Calcium Ointment Test for required detectability: To exactly 1 mL of the standard solution add a mixture of 0.1 mol/L acetic acid- ムピロシンカルシウム軟膏 sodium acetate buffer solution, pH 4.0 and diluted tetrahydrofuran (3 in 4) (1:1) to make exactly 20 mL. Confirm Mupirocin Calcium Ointment is an oily ointment that the peak area of mupirocin obtained with 20 mLofthis preparation. solution is equivalent to 4 to 6z of that with 20 mLofthe Mupirocin Calcium Ointment contains not less than standard solution. 95.0z and not more than 105.0z of the labeled po- System performance: Proceed as directed in the system tency of mupirocin (C26H44O9: 500.62). suitability in the Assay under Mupirocin Calcium Hydrate. System repeatability: When the test is repeated 6 times Method of preparation Prepare as directed under Oint- with 20 mL of the standard solution under the above operat- ments, with Mupirocin Calcium Hydrate. ing conditions, the relative standard deviation of the peak Identification To an amount of Mupirocin Calcium Oint- area of mupirocin is not more than 2.0z. ment, equivalent to 10 mg (potency) of Mupirocin Calcium Assay Weigh accurately an amount of Mupirocin Calcium Hydrate according to the labeled amount, add 5 mL of Ointment, equivalent to about 2 mg (potency) of Mupirocin water,andwarmonawaterbathat609C for 10 minutes Calcium Hydrate, add exactly 10 mL of diluted tetra- while occasional shaking. After cooling, filter, and to 1 mL hydrofuran (3 in 4), and shake vigorously. To this solution of the filtrate add water to make 100 mL. Determine the ab- add exactly 10 mL of 0.1 mol/L acetic acid-sodium acetate sorption spectrum of this solution as directed under Ultravi- buffer solution, pH 4.0, shake vigorously, filter through a olet-visible Spectrophotometry <2.24>:itexhibitsamaxi- glass wool filter, and use the filtrate as the sample solution. mum between 220 nm and 224 nm. Separately, weigh accurately an amount of Mupirocin Lithi- Purity Related substances—To an amount of Mupirocin um RS, equivalent to about 20 mg (potency), dissolve in a Calcium Ointment, equivalent to 50 mg (potency) of mixture of 0.1 mol/L acetic acid-sodium acetate buffer solu- Mupirocin Calcium Hydrate according to the labeled tion, pH 4.0 and diluted tetrahydrofuran (3 in 4) (1:1) to amount, add 5 mL of diluted tetrahydrofuran (3 in 4), and make exactly 200 mL, and use this solution as the standard shake vigorously. Then, add 5 mL of 0.1 mol/L acetic acid- solution. Then, proceed as directed in the Assay under sodium acetate buffer solution, pH 4.0, shake vigorously, Mupirocin Calcium Hydrate. filter through a glass wool filter, and use the filtrate as the Amount [mg (potency)] of mupirocin (C H O ) sample solution. Pipet 2 mL of the sample solution, add a 26 44 9 ＝ M × A /A × 1/10 mixture of 0.1 mol/L acetic acid-sodium acetate buffer solu- S T S tion, pH 4.0 and diluted tetrahydrofuran (3 in 4) (1:1) to MS: Amount [mg (potency)] of Mupirocin Lithium RS make exactly 100 mL, and use this solution as the standard Containers and storage Containers—Tight containers. solution. Perform the test with exactly 20 mLeachofthe sample solution and standard solution as directed under Liq- uid Chromatography <2.01> according to the following conditions, and determine the area of the peak other than Nabumetone mupirocin obtained from the sample solution and the peak ナブメトン area of mupirocin from the standard solution by the automatic integration method. Calculate the amount of each related substance using the following equation: the amount of the related substance having the relative retention time of about 0.7 to mupirocin is not more than 4.0z, the amount of the related substance other than that is not more than 1.5 C H O :228.29 z, and the total amount of the related substances is not 15 16 2 4-(6-Methoxynaphthalen-2-yl)butan-2-one more than 6.0z. [42924-53-8] Amount (z) of each related substance

＝ A/(SA ＋ Am) × 100 Nabumetone contains not less than 98.0z and not more than 101.0z of C H O ,calculatedonthe A: Peak area of each related substance obtained from the 15 16 2 anhydrous basis. sample solution. SA: Total area of the peaks other than mupirocin ob- Description Nabumetone occurs as white to yellowish tained from the sample solution. white crystals or a crystalline powder.

Am: Amount of 50 times the peak area of mupirocin ob- It is soluble in acetonitrile, sparingly soluble in methanol tained from the standard solution. and in ethanol (99.5), and practically insoluble in water. 1140 Nabumetone / Official Monographs JP XVI

Identification (1) Determine the absorption spectrum of a peak. solution of Nabumetone in methanol (1 in 30,000) as di- System suitability— rected under Ultraviolet-visible Spectrophotometry <2.24>, Test for required detectability: Pipet 2 mL of the standard and compare the spectrum with the Reference Spectrum or solution, and add acetonitrile to make exactly 10 mL. Con- the spectrum of a solution of Nabumetone RS prepared in firm that the peak area of nabumetone obtained from 10 mL the same manner as the sample solution: both spectra exhibit of this solution is equivalent to 14 to 26z of that from 10 mL similar intensities of absorption at the same wavelengths. of the standard solution. (2) Determine the infrared absorption spectrum of System performance: Proceed as directed in the system Nabumetone as directed in the potassium bromide disk suitability in the Assay. method under Infrared Spectrophotometry <2.25>,and System repeatability: When the test is repeated 6 times compare the spectrum with the Reference Spectrum or the with 10 mL of the standard solution under the above operat- spectrum of Nabumetone RS: both spectra exhibit similar ing conditions, the relative standard deviation of the peak intensities of absorption at the same wave numbers. area of nabumetone is not more than 5.0z. Melting point <2.60> 79–849C. Water <2.48> Not more than 0.2z (1 g, volumetric titration, direct titration). Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Nabumetone according to Method 2, and perform the test. Residue on ignition <2.44> Not more than 0.1z (1 g). Prepare the control solution with 1.0 mL of Standard Lead Assay Weigh accurately about 20 mg each of Nabumetone Solution (not more than 10 ppm). and Nabumetone RS (separately determine the water <2.48> (2) Related substances—Dissolve 20 mg of Nabumetone in the same manner as Nabumetone), dissolve them in aceto- in 20 mL of acetonitrile, and use this solution as the sample nitrile to make exactly 20 mL, and use these solutions as the solution. Pipet 5 mL of the sample solution, add acetonitrile sample solution and the standard solution, respectively. Per- to make exactly 50 mL. Pipet 1 mL of this solution, add form the test with exactly 10 mL each of the sample solution acetonitrile to make exactly 20 mL, and use this solution as and standard solution as directed under Liquid Chromatog- the standard solution. Perform the test with exactly 10 mL raphy <2.01> according to the following conditions, and de- each of the sample solution and standard solution as directed termine the peak area of nabumetone, A and A , from each under Liquid Chromatography <2.01> according to the fol- T S solution. lowing conditions. Determine each peak area of both solutions by the automatic integration method: the peak area of Amount (mg) of C15H16O2 ＝ MS × AT/AS the related substance G obtained from the sample solution is M : Amount (mg) of Nabumetone RS, calculated on the not larger than 3/5 times the peak area of nabumetone from S anhydrous basis the standard solution, and each peak area other than nabumetone and the related substance G is not larger than 1/5 Operating conditions— times the peak area of nabumetone from the standard solu- Detector: An ultraviolet absorption photometer (wave- tion. Furthermore, the total area of the peaks other than length: 254 nm). nabumetone is not larger than 1.6 times the peak area of Column: A stainless steel column 4.6 mm in inside diame- nabumetone from the standard solution. For these calcula- ter and 15 cm in length, packed with octadecylsilanized silica tions, use each peak area of the related substances A, B, C, gel for liquid chromatography (4 mminparticlediameter). D, E, F and G, which are having the relative retention time Column temperature: A constant temperature of about of about 0.73, 0.85, 0.93, 1.2, 1.9, 2.6 and 2.7 with respect 409C. to nabumetone, after multiplying by their relative response Mobile phase: To 600 mL of a mixture of water and acetic factors, 0.12, 0.94, 0.25, 0.42, 1.02, 0.91 and 0.1, respec- acid (100) (999:1) add 400 mL of a mixture of acetonitrile tively. and tetrahydrofuran (7:3). Operating conditions— Flow rate: Adjust the flow rate so that the retention time Detector, column, and column temperature: Proceed as of nabumetone is about 10 minutes. directed in the operating conditions in the Assay. System suitability— Mobile phase A: A mixture of water and acetic acid (100) System performance: When the procedure is run with 10 (999:1). mL of the standard solution under the above operating con- Mobile phase B: A mixture of acetonitrile and tetra- ditions, the number of theoretical plates and the symmetry hydrofuran (7:3). factor of the peak of nabumetone are not less than 6000 and Flowing of mobile phase: Control the gradient by mixing not more than 1.5, respectively. the mobile phases A and B as directed in the following table. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- Time after injection Mobile phase A Mobile phase B ing conditions, the relative standard deviation of the peak of sample (min) (volz) (volz) area of nabumetone is not more than 1.0z. Containers and storage Containers—Tight containers. 0–12 60 40 12–28 60→ 20 40 → 80

Flow rate: 1.3 mL per minute. Time span of measurement: About 3 times as long as the retention time of nabumetone, beginning after the solvent JP XVI Official Monographs / Nabumetone Tablets 1141

methanol, shake for 30 minutes, and then add methanol to Nabumetone Tablets make exactly 100 mL. Centrifuge this solution, pipet 5 mL of the supernatant liquid, add exactly 5 mL of the internal ナブメトン錠 standard solution, then add methanol to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of Nabumetone RS (separately deter- Nabumetone Tablets contain not less than 95.0z mine the water <2.48> in the same manner as Nabumetone), and not more than 105.0z of the labeled amount of dissolve by adding 50 mL of methanol and exactly 20 mL of nabumetone (C H O : 228.29). 15 16 2 the internal standard solution, then add methanol to make Method of preparation Prepare as directed under Tablets, 200 mL, and use this solution as the standard solution. Per- with Nabumetone. form the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography Identification To a quantity of powdered Nabumetone <2.01> according to the following conditions, and calculate Tablets, equivalent to 80 mg of Nabumetone according to the ratios, Q and Q , of the peak area of nabumetone to the labeled amount, add 50 mL of methanol, shake for 10 T S that of the internal standard. minutes and centrifuge the solution. To 1 mL of the supernatant liquid, add methanol to make 50 mL, and determine the Amount (mg) of nabumetone (C15H16O2) absorption spectrum of this solution as directed under Ultra- ＝ MS × QT/QS × 5 violet-visible Spectrophotometry <2.24>: it exhibits maxima M : Amount (mg) of Nabumetone RS, calculated on the between 259 nm and 263 nm, between 268 nm and 272 nm, S anhydrous basis between 316 nm and 320 nm, and between 330 nm and 334 nm. Internal standard solution—Dissolve 0.12 g of 2-ethylhexyl parahydroxybenzoate in methanol to make 100 mL. Uniformity of dosage units <6.02> It meets the requirement Operating conditions— of the Mass variation test. Detector: An ultraviolet absorption photometer (wave- Dissolution <6.10> When the test is performed at 75 revolu- length: 254 nm). tions per minute according to the Paddle method, using 900 Column: A stainless steel column 4 mm in inside diameter mL of a solution of polysorbate 80 (dissolving 3 g of poly- and 15 cm in length, packed with octadecylsilanized silica gel sorbate 80 in water to make 100 mL) as the dissolution me- for liquid chromatography (5 mm in particle diameter). dium, the dissolution rate in 60 minutes of Nabumetone Column temperature: A constant temperature of about Tablets is not less than 70z. 259C. Start the test with 1 tablet of Nabumetone Tablets, with- Mobile phase: A mixture of acetonitrile, water and acetic draw not less than 20 mL of the medium at the specified acid (100) (550:450:1). minute after starting the test, and filter through a membrane Flow rate: Adjust the flow rate so that the retention time filter with a pore size not exceeding 0.5 mm. Discard the first of nabumetone is about 6 minutes. 10 mL of the filtrate, pipet V mL of the subsequent filtrate, System suitability— add a solution, prepared by adding to 20 mL of ethanol System performance: When the procedure is run with 10 (99.5) the dissolution medium to make 50 mL, to make mL of the standard solution under the above operating con- exactly V?mL so that each mL contains about 89 mgof ditions, nabumetone and the internal standard are eluted in nabumetone (C15H16O2) according to the labeled amount, this order with the resolution between these peaks being not and use this solution as the sample solution. Separately, less than 13. weigh accurately about 22 mg of Nabumetone RS (separately System repeatability: When the test is repeated 6 times determine the water <2.48> in the same manner as Nabume- with 10 mL of the standard solution under the above operat- tone), and dissolve in ethanol (99.5) to make exactly 100 mL. ing conditions, the relative standard deviation of the ratio of Pipet 10 mL of this solution, add the dissolution medium to the peak area of nabumetone to that of the internal standard make exactly 25 mL, and use this solution as the standard is not more than 1.0z. solution. Determine the absorbances, A and A ,at331nm T S Containers and storage Containers—Well-closed contain- of the sample solution and standard solution as directed ers. under Ultraviolet-visible Spectrophotometry <2.24>,usinga solution prepared by adding to 20 mL of ethanol (99.5) the dissolution medium to make 50 mL as the blank. Dissolution rate (z) with respect to the labeled amount

of nabumetone (C15H16O2) ＝ MS × AT/AS × V?/V × 1/C × 360

MS: Amount (mg) of Nabumetone RS, calculated on the anhydrous basis

C: Labeled amount (mg) of nabumetone (C15H16O2)in1 tablet Assay Weigh accurately not less than 20 tablets of Nabumetone Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.2 g of nabumetone

(C15H16O2), add 10 mL of water and shake, add 40 mL of 1142 Nadolol / Official Monographs JP XVI

sitions of the principal spot and the spots other than the Nadolol principal spot from the sample solution. Scratch and collect the silica gel of the positions of the plate corresponding to ナドロール the principal spot and the spots other than the principal spot. To the silica gel collected from the principal spot add exactly 30 mL of ethanol (95), and to the silica gel from the spots other than the principal spot add exactly 10 mL of ethanol (95). After shaking them for 60 minutes, centrifuge, and determine the absorbances of these supernatant liquids at 278 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Separately, proceed in the same manner with each position of the silica gel from the control solution cor- C H NO : 309.40 17 27 4 responding to the principal spot and the spots other than the R1 ＝ OH, R2 ＝ H principal spot of the sample solution, and perform a blank (2RS,3SR)-5-{(2SR)-3-[(1,1-Dimethylethyl)amino]- determination to make correction. Amount of the related 2-hydroxypropyloxy}-1,2,3,4-tetrahydronaphthalene- substances calculated by the following equation is not more 2,3-diol than 2.0z. R1 ＝ H, R2 ＝ OH (2RS,3SR)-5-{(2RS)-3-[(1,1-Dimethylethyl)amino]- Amount (z) of related substances ＝ Ab/(Ab ＋ 3Aa) × 100

2-hydroxypropyloxy}-1,2,3,4-tetrahydronaphthalene- Aa: Corrected absorbance of the principle spot 2,3-diol Ab: Corrected absorbance of the spots other than the prin- [42200-33-9] ciple spot

Nadolol, when dried, contains not less than 98.0z Loss on drying <2.41> Not more than 1.0z (1 g, in vacuum, 609C, 3 hours). of C17H27NO4. Description Nadolol occurs as a white to yellow-brownish Residue on ignition <2.44> Not more than 0.1z (1 g). white crystalline powder. Isomer ratio Prepare a paste with 0.01 g of Nadolol as di- It is freely soluble in methanol and in acetic acid (100), rected in the paste method under Infrared Spectrophotome- soluble in ethanol (95), and slightly soluble in water and in try <2.25> so that its transmittance at an absorption band at a chloroform. wave number of about 1585 cm－1 is 25 to 30z,anddeter- A solution of Nadolol in methanol (1 in 100) shows no op- mine the infrared absorption spectrum between 1600 cm－1 －1 tical rotation. and 1100 cm . Determine the absorbances, A1265 and A1250, Melting point: about 1379C. from the transmittances, T1265 and T1250, at wave numbers of －1 －1 Identification (1) Determine the absorption spectrum of a about 1265 cm (racemic substance A) and 1250 cm solution of Nadolol in methanol (1 in 5000) as directed under (racemic substance B), respectively: the ratio A1265/A1250 is Ultraviolet-visible Spectrophotometry <2.24>,andcompare between 0.72 and 1.08. the spectrum with the Reference Spectrum: both spectra Assay Weigh accurately about 0.28 g of Nadolol, previ- exhibit similar intensities of absorption at the same wave- ously dried, dissolve in 50 mL of acetic acid (100), and titrate lengths. <2.50> with 0.1 mol/L perchloric acid VS until the color of (2) Determine the infrared absorption spectrum of the solution changes from purple through blue to green-blue Nadolol, previously dried, as directed in the potassium (indicator: 3 drops of crystal violet TS). Perform a blank de- bromide disk method under Infrared Spectrophotometry termination, and make any necessary correction. <2.25>: it exhibits absorption at the wave numbers of about 1585 cm－1,1460cm－1, 1092 cm－1, 935 cm－1 and 770 cm－1. Each mL of 0.1 mol/L perchloric acid VS ＝ 30.94 mg of C17H27NO4 Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Nadolol according to Method 2, and perform the test. Pre- Containers and storage Containers—Tight containers. pare the control solution with 2.0 mL of Standard Lead So- Storage—Light-resistant. lution (not more than 20 ppm). (2) Related substances—Dissolve 0.5 g of Nadolol in 10 mL of a mixture of methanol and chloroform (1:1), and use this solution as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatog- raphy <2.03>. Spot 100 mL each of the sample solution and a mixture of methanol and chloroform (1:1) as a control solution with 25 mm each of width at an interval of about 10 mm onthestartinglineofaplate0.25 mm in thickness of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of acetone, chloroform and diluted ammonia TS (1 in 3) (8:1:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm), and confirm the po- JP XVI Official Monographs / Nafamostat Mesilate 1143

tained from the sample solution is not larger than 1/5 times Nafamostat Mesilate the peak area of nafamostat from the standard solution. Furthermore, the total area of the peaks other than ナファモスタットメシル酸塩 nafamostat is not larger than the peak area of nafamostat from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 260 nm). Column: A stainless steel column 4.6 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticlediameter).

C19H17N5O2.2CH4O3S: 539.58 Column temperature: A constant temperature of about 6-Amidinonaphthalen-2-yl 4-guanidinobenzoate 409C. bis(methanesulfonate) Mobile phase: Dissolve 6.07 g of sodium 1-heptane sul- [82956-11-4] fonate in 1000 mL of diluted acetic acid (100) (3 in 500). To 700 mL of this solution add 300 mL of acetonitrile. Nafamostat Mesilate, when dried, contains not Flow rate: Adjust the flow rate so that the retention time less than 99.0z and not more than 101.0z of of nafamostat is about 7 minutes. C19H17N5O2.2CH4O3S. Time span of measurement: About 4 times as long as the retention time of nafamostat, beginning after the solvent Description Nafamostat Mesilate occurs as a white crystal- peak. line powder. System suitability— It is freely soluble in formic acid, soluble in water, and Test for required detectability: Pipet 5 mL of the standard practically insoluble in ethanol (99.5). solution, and add the mobile phase to make exactly 50 mL. It dissolves in 0.01 mol/L hydrochloric acid TS. Pipet 15 mL of this solution, and add the mobile phase to Melting point: about 2629C (with decomposition). make exactly 100 mL. Confirm that the peak area of Identification (1) Determine the absorption spectrum of a nafamostat obtained from 10 mL of this solution is equiva- solution of Nafamostat Mesilate in 0.01 mol/L hydrochloric lent to 1.1 to 1.9z of that from 10 mL of the standard solu- acid TS (1 in 200,000) as directed under Ultraviolet-visible tion. Spectrophotometry <2.24>, and compare the spectrum with System performance: Dissolve 0.1 g of nafamostat mesi- the Reference Spectrum: both spectra exhibit similar intensi- late in the mobile phase to make 100 mL. To 10 mL of this ties of absorption at the same wavelengths. solution add the mobile phase to make 100 mL. To 5 mL of (2) Determine the infrared absorption spectrum of this solution add 5 mL of a solution of 6-amidino-2- Nafamostat Mesilate as directed in the potassium bromide naphthol methanesulfonate in the mobile phase (1 in disk method under Infrared Spectrophotometry <2.25>,and 20,000). When the procedure is run with 10 mL of this solu- compare the spectrum with the Reference Spectrum: both tion under the above operating conditions, 6-amidino-2- spectra exhibit similar intensities of absorption at the same naphthol and nafamostat are eluted in this order with the wave numbers. resolution between these peaks being not less than 6. (3) A 0.1-g portion of Nafamostat Mesilate responds to System repeatability: When the test is repeated 6 times the Qualitative Tests <1.09> (1) for mesilate. with 10 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak pH <2.54> The pH of a solution prepared by dissolving area of nafamostat is not more than 2.0z. 1.0 g of Nafamostat Mesilate in 50 mL of water is between 4.7 and 5.7. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Purity (1) Clarity and color of solution—A solution pre- paredbydissolving1.0gofNafamostatMesilatein50mL Residue on ignition <2.44> Not more than 0.1z (1 g). of water is clear and colorless. Assay Weigh accurately about 0.25 g of Nafamostat Mesi- (2) Heavy metals <1.07>—Proceed with 2.0 g of late, previously dried, dissolve in 4 mL of formic acid, add Nafamostat Mesilate according to Method 4, and perform 50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L the test. Prepare the control solution with 2.0 mL of Stand- perchloric acid VS (potentiometric titration). Perform a ard Lead Solution (not more than 10 ppm). blank determination in the same manner, and make any nec- (3) Related substances—Conduct this procedure using essary correction. light-resistant vessels. Dissolve 0.10 g of Nafamostat Mesi- late in 100 mL of the mobile phase, and use this solution as Each mL of 0.1 mol/L perchloric acid VS the sample solution. Pipet 10 mL of the sample solution, add ＝ 26.98 mg of C19H17N5O2.2CH4O3S the mobile phase to make exactly 100 mL. Then pipet 5 mL Containers and storage Containers—Tight containers. of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of each solution by the automatic integration method: the area of each peak other than nafamostat ob- 1144 Nalidixic Acid / Official Monographs JP XVI

dixic acid is not larger than 2.5 times the peak area of nali- Nalidixic Acid dixic acid with the standard solution. Operating conditions— ナリジクス酸 Detector: An ultraviolet absorption photometer (wavelength: 260 nm). Column: A stainless steel column 4.6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticlediameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 6.24 g of sodium dihydrogen phos- C H N O : 232.24 12 12 2 3 phate dihydrate in 950 mL of water, adjust the pH to 2.8 1-Ethyl-7-methyl-4-oxo-1,4-dihydro-1,8-naphthyridine-3- with phosphoric acid, and add water to make 1000 mL. To carboxylic acid 300 mL of this solution add 200 mL of methanol. [389-08-2] Flow rate: Adjust the flow rate so that the retention time of nalidixic acid is about 19 minutes. Nalidixic Acid, when dried, contains not less than Time span of measurement: About 3 times as long as the 99.0z and not more than 101.0z of C H N O . 12 12 2 3 retention time of nalidixic acid beginning after the solvent Description Nalidixic Acid occurs as white to light yellow peak. crystals or crystalline powder. System suitability— It is sparingly soluble in N,N-dimethylformamide, very Test for required detectability: Pipet 5 mL of the standard slightly soluble in ethanol (99.5), and practically insoluble in solution, and add water to make exactly 10 mL. Confirm water. that the peak area of nalidixic acid obtained with 10 mLof It dissolves in sodium hydroxide TS. this solution is equivalent to 40 to 60z of that with 10 mLof the standard solution. Identification (1) Determine the absorption spectrum of a System performance: Dissolve 25 mg of methyl parahy- solution of Nalidixic Acid in 0.01 mol/L sodium hydroxide droxybenzoate in 100 mL of a mixture of water and metha- TS (1 in 200,000) as directed under Ultraviolet-visible Spec- nol (1:1). To 1 mL of this solution add water to make 10 trophotometry <2.24>, and compare the spectrum with the mL. To 5 mL of this solution add 5 mL of the standard solu- Reference Spectrum: both spectra exhibit similar intensities tion. When the procedure is run with 10 mL of this solution of absorption at the same wavelengths. under the above operating conditions, methyl parahydroxy- (2) Determine the infrared absorption spectrum of Nali- benzoate and nalidixic acid areelutedinthisorderwiththe dixic Acid, previously dried, as directed in the potassium resolution between these peaks being not less than 13. bromide disk method under Infrared Spectrophotometry System repeatability: When the test is repeated 6 times <2.25>, and compare the spectrum with the Reference Spec- with 10 mL of the standard solution under the above operat- trum: both spectra exhibit similar intensities of absorption at ing conditions, the relative standard deviation of the peak the same wave numbers. area of nalidixic acid is not more than 2.0z. Melting point <2.60> 225–2319C. Loss on drying <2.41> Not more than 0.20z (1 g, 1059C, Purity (1) Chloride <1.03>—To 2.0 g of Nalidixic Acid 3 hours). add 50 mL of water, warm at 709C for 5 minutes, cool Residue on ignition <2.44> Not more than 0.2z (1 g). quickly, and filter. To 25 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL, and perform the Assay Weigh accurately about 0.3 g of Nalidixic Acid, pre- test using this solution as the test solution. Prepare the con- viously dried, dissolve in 50 mL of N,N-dimethylformamide, trol solution with 0.35 mL of 0.01 mol/L hydrochloric acid and titrate <2.50> with 0.1 mol/L tetramethyl ammonium hy- VS (not more than 0.012z). droxide VS (potentiometric titration). Separately, to 50 mL (2) Heavy metals <1.07>—Proceed with 1.0 g of Nalidixic of N,N-dimethylformamide add 13 mL of a mixture of Acid according to Method 2, and perform the test. Prepare water and methanol (89:11), perform a blank determination the control solution with 2.0 mL of Standard Lead Solution with the solution, and make any necessary correction. (not more than 20 ppm). Each mL of 0.1 mol/L tetramethyl ammonium hydroxide VS (3) Related substances—Dissolve 20 mg of Nalidixic ＝ 23.22 mg of C H N O Acid in 20 mL of 0.01 mol/L sodium hydroxide TS. To 5 12 12 2 3 mL of this solution, add water to make 10 mL, and use this Containers and storage Containers—Tight containers. solution as the sample solution. Pipet 2 mL of the sample solution, add water to make exactly 1000 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than nalidixic acid with the sample solution is not larger than the peak area of nalidixic acid with the standard solution, and the total area of the peaks other than the peak of nali- JP XVI Official Monographs / Naphazoline Hydrochloride 1145

ferrate (III) TS on the plate: the number of the spot other Naloxone Hydrochloride than the principal spot from the sample solution is not more than 1 and it is not more intense than the spot from the ナロキソン塩酸塩 standard solution. Loss on drying <2.41> Not more than 2.0z [0.1 g, 1059C, 5 hours. Use a desiccator (phosphorus (V) oxide) for cooling]. Residue on ignition <2.44> Not more than 0.2z (0.1 g). Assay Weigh accurately about 0.3 g of Naloxone Hydro- chloride, dissolve in 80 mL of acetic acid (100) by warming. After cooling, add 80 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric C H NO .HCl: 363.84 19 21 4 titration). Perform a blank determination, and make any (5R,14S)-17-Allyl-4,5-epoxy-3,14-dihydroxymorphinan- necessary correction. 6-one monohydrochloride [357-08-4] Each mL of 0.1 mol/L perchloric acid VS

＝ 36.38 mg of C19H21NO4.HCl Naloxone Hydrochloride contains not less than Containers and storage Containers—Tight containers. 98.5z of C H NO .HCl,calculatedonthedried 19 21 4 Storage—Light-resistant. basis. Description Naloxone Hydrochloride occurs as white to yellowish white, crystals or crystalline powder. Naphazoline Hydrochloride It is freely soluble in water, soluble in methanol, slightly soluble in ethanol (99.5) and in acetic acid (100), and very ナファゾリン塩酸塩 slightly soluble in acetic anhydride. It is hygroscopic. It is gradually colored by light. Identification (1) Determine the absorption spectrum of a solution of Naloxone Hydrochloride (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>,and compare the spectrum with the Reference Spectrum: both C14H14N2.HCl: 246.74 spectra exhibit similar intensities of absorption at the same 2-(Naphthalen-1-ylmethyl)-4,5-dihydro-1H-imidazole wavelengths. monohydrochloride (2) Determine the infrared absorption spectrum of [550-99-2] Naloxone Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectropho- Naphazoline Hydrochloride, when dried, contains tometry <2.25>, and compare the spectrum with the Refer- not less than 98.5z of C14H14N2.HCl. ence Spectrum: both spectra exhibit similar intensities of Description Naphazoline Hydrochloride occurs as a white, absorption at the same wave numbers. crystalline powder. It is odorless, and has a bitter taste. (3) A solution of Naloxone Hydrochloride (1 in 50) It is freely soluble in water, soluble in ethanol (95) and in responds to the Qualitative Tests <1.09> (2) for chloride. acetic acid (100), very slightly soluble in acetic anhydride, 25 Optical rotation <2.49> [a]D : －170 – －1819(0.25 g calcu- and practically insoluble in diethyl ether. latedonthedriedbasis,water,10mL,100mm). Melting point: 255 – 2609C (with decomposition). pH <2.54> Dissolve 0.10 g of Naloxone Hydrochloride in Identification (1) To 10 mL of a solution of Naphazoline 10 mL of freshly boiled and cooled water: the pH of the so- Hydrochloride (1 in 100) add 5 mL of bromine TS, and boil: lution is between 4.5 and 5.5. a deep purple color develops. (2) To 30 mL of a solution of Naphazoline Hydrochlo- Purity Related substances—Conduct this procedure as ride (1 in 100) add 2 mL of sodium hydroxide TS, and ex- rapidly as possible without exposure to light, using light- tract with two 25-mL portions of diethyl ether. Evaporate resistant containers. Dissolve 0.08 g of Naloxone Hydrochlo- the combined diethyl ether extracts to dryness with the aid of ride in 10 mL of methanol, and use this solution as the a current of air. Dry the residue at 809C for 1 hour: the sample solution. Pipet 1 mL of the sample solution, add residue melts <2.60> between 1179Cand1209C. methanol to make exactly 200 mL, and use this solution as (3) Dissolve 0.02 g of the residue obtained in (2) in 2 to 3 the standard solution. Perform the test with these solutions drops of dilute hydrochloric acid and 5 mL of water, and as directed under Thin-layer Chromatography <2.03>. Spot add 2 mL of Reinecke salt TS: a red-purple, crystalline pre- 10 mL each of the sample solution and standard solution on a cipitate is formed. plate of silica gel for thin-layer chromatography. Develop (4) A solution of Naphazoline Hydrochloride (1 in 10) with a mixture of ammonia-saturated 1-butanol TS and responds to the Qualitative Tests <1.09> for chloride. methanol (20:1) to a distance of about 12 cm, and air-dry the plate. Spray evenly iron (III) chloride-potassium hexacyano- pH <2.54> Dissolve 0.10 g of Naphazoline Hydrochloride 1146 Naphazoline Nitrate / Official Monographs JP XVI in 10 mL of freshly boiled and cooled water: the pH of the between 5.0 and 7.0. solution is between 5.0 and 7.0. Melting point <2.60> 167 – 1709C. Purity (1) Clarity and color of solution—Dissolve 1.0 g Purity (1) Clarity and color of solution—Dissolve 0.5 g of Naphazoline Hydrochloride in 10 mL of water: the solu- of Naphazoline Nitrate in 50 mL of water: the solution is tion is clear and colorless. clear and colorless. (2) Heavy metals <1.07>—Proceed with 1.0 g of Napha- (2) Heavy metals <1.07>—Proceed with 1.0 g of Napha- zoline Hydrochloride according to Method 2, and perform zoline Nitrate according to Method 2, and perform the test. the test. Prepare the control solution with 2.0 mL of Stand- Prepare the control solution with 2.0 mL of Standard Lead ard Lead Solution (not more than 20 ppm). Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.4 g of Naphazoline Hy- Assay Weigh accurately about 0.4 g of Naphazoline drochloride, previously dried, dissolve in 50 mL of a mixture Nitrate, previously dried, dissolve in 10 mL of acetic acid of acetic anhydride and acetic acid (100) (7:3), and titrate (100) and 40 mL of acetic anhydride, and titrate <2.50> with <2.50> with 0.1 mol/L perchloric acid VS (potentiometric 0.1 mol/L perchloric acid VS (indicator: 3 drops of crystal titration). Perform a blank determination, and make any violet TS). Perform a blank determination, and make any necessary correction. necessary correction. Each mL of 0.1 mol/L perchloric acid VS Each mL of 0.1 mol/L perchloric acid VS ＝ 24.67 mg of C14H14N2.HCl ＝ 27.33 mg of C14H14N2.HNO3 Containers and storage Containers—Tight containers. Containers and storage Containers—Tight containers. Storage—Light-resistant. Storage—Light-resistant.

Naphazoline Nitrate Naphazoline and Chlorpheniramine ナファゾリン硝酸塩 Solution

ナファゾリン・クロルフェニラミン液

Naphazoline and Chlorpheniramine Solution contains not less than 0.045 w/vz and not more than 0.055 w/vz of naphazoline nitrate (C14H14N2.HNO3: C14H14N2.HNO3: 273.29 273.29), and not less than 0.09 w/vz and not 2-(Naphthalen-1-ylmethyl)-4,5-dihydro-1H-imidazole more than 0.11 w/vz of chlorpheniramine maleate mononitrate (C16H19ClN2.C4H4O4: 390.86). [5144-52-5] Method of preparation Naphazoline Nitrate, when dried, contains not less Naphazoline Nitrate 0.5 g than 98.5z of C14H14N2.HNO3. Chlorpheniramine Maleate 1 g Chlorobutanol 2 g Description Naphazoline Nitrate occurs as a white, crystal- Glycerin 50 mL line powder. It is odorless, and has a bitter taste. Purified Water or Purified It is freely soluble in acetic acid (100), soluble in ethanol Water in Containers a sufficient quantity (95), sparingly soluble in water, slightly soluble in acetic anhydride, and practically insoluble in diethyl ether. To make 1000 mL Identification (1) To 10 mL of a solution of Naphazoline Dissolve, and mix the above ingredients. Nitrate (1 in 100) add 5 mL of bromine TS, and boil: a deep Description Naphazoline and Chlorpheniramine Solution purple color develops. is a clear, colorless liquid. (2) To 20 mL of a solution of Naphazoline Nitrate (1 in 100) add 5 mL of sodium hydroxide TS, and extract with Identification (1) To 20 mL of Naphazoline and Chlor- two 25-mL portions of diethyl ether. Combine the diethyl pheniramine Solution add 2 mL of a solution of potassium ether extracts, evaporate to dryness with the aid of a current hydroxide (7 in 10) and 5 mL of pyridine, and heat at 1009C of air, and dry the residue at 809C for 1 hour: the residue so for 5 minutes: a red color is produced (chlorobutanol). obtained melts <2.60> between 1179Cand1209C. (2) Place 10 mL of Naphazoline and Chlorpheniramine (3) A solution of Naphazoline Nitrate (1 in 20) responds Solution in a glass-stoppered test tube, add 10 mL of ethanol to the Qualitative Tests <1.09> for nitrate. (95), 2 mL of sodium hydroxide TS and 1 mL of a solution of copper (II) chloride dihydrate in ethanol (95) (1 in 10), pH <2.54> Dissolve 0.1 g of Naphazoline Nitrate in 10 mL and shake: a blue color is produced (glycerin). of freshly boiled and cooled water: the pH of the solution is JP XVI Official Monographs / Naproxen 1147

(3) To 20 mL of Naphazoline and Chlorpheniramine Flow rate: Adjust the flow rate so that the retention time Solution add 5 mL of sodium hydroxide TS, extract with 10 of chlorpheniramine is about 10 minutes. mL of diethyl ether, and separate the diethyl ether layer. Selection of column: Proceed with 10 mL of the standard Take 5 mL of this solution, distil off the solvent, dissolve the solution under the above operating conditions. Use a column residue in 5 mL of methanol, and use this solution as the giving well-resolved peaks of the internal standard, naphazo- sample solution. Separately, dissolve 0.01 g each of napha- line and chlorpheniramine in this order. zoline nitrate and Chlorpheniramine Maleate RS in 10 mL Containers and storage Containers—Tight containers. and 5 mL of methanol, respectively, and use these solutions Storage—Light-resistant. as standard solutions (1) and (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solutions on a plate of silica gel with fluorescent indicator Naproxen for thin-layer chromatography. Develop the plate with a ナプロキセン mixture of chloroform, methanol, acetone and ammonia solution (28) (73:15:10:2) to a distance of about 10 cm, and air- dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): two spots from the sample solution exhibit the same Rf values as the spots from standard solutions (1) and (2). Spray evenly Dragendorff's TS on the plate: the C H O :230.26 spots from standard solutions (1) and (2) and the cor- 14 14 3 (2S)-2-(6-Methoxynaphthalen-2-yl)propanoic acid responding spot from the sample solutions reveal an orange [22204-53-1] color. Assay Pipet 4 mL of Naphazoline and Chlorpheniramine Naproxen, when dried, contains not less than 98.5z Solution, add exactly 4 mL of the internal standard solution, of C14H14O3. then add water to make 10 mL, and use this solution as the Description Naproxen occurs as white crystals or crystal- sample solution. Weigh accurately about 50 mg of naphazoline powder. It is odorless. line nitrate for assay, dried at 1059C for 2 hours, and about It is freely soluble in acetone, soluble in methanol, in 0.1 g of Chlorpheniramine Maleate RS, dried at 1059Cfor3 ethanol (99.5) and in chloroform, sparingly soluble in diethyl hours, dissolve in water to make exactly 100 mL. Pipet 4 mL ether, and practically insoluble in water. of this solution, add exactly 4 mL of the internal standard It dissolves in sodium hydroxide TS. solution, then add water to make 10 mL, and use this solution as the standard solution. Perform the test with 10 mL Identification (1) Dissolve 0.01 g of Naproxen in 5 mL of each of the sample solution and standard solutions as di- methanol, add 5 mL of water, then add 2 mL of potassium rected under Liquid Chromatography <2.01> according to iodide TS and 5 mL of a solution of potassium iodate (1 in the following conditions, and calculate the ratios, QTa and 100), and shake: a yellow to yellow-brown color develops. QTb, of the peak height of naphazoline and chlorphenira- To this solution add 5 mL of chloroform, and shake: a light mine to that of the internal standard of the sample solution, red-purple color develops in the chloroform layer. and the ratios, QSa and QSb, of the peak height of naphazo- (2) To 1 mL of a solution of Naproxen in ethanol (99.5) line and chlorpheniramine to that of the internal standard of (1 in 300) add 4 mL of hydroxylamine perchlorate-dehy- the standard solution. drated ethanol TS and 1 mL of N,N?-dicyclohexylcarbodiimide-dehydrated ethanol TS, shake well, and allow to stand Amount (mg) of naphazoline nitrate (C H N .HNO ) 14 14 2 3 in lukewarm water for 20 minutes. After cooling, add 1 mL ＝ M × Q /Q × 1/25 Sa Ta Sa of iron (III) perchlorate-dehydrated ethanol TS, and shake: Amount (mg) of chlorpheniramine maleate a red-purple color develops.

(C16H19ClN2.C4H4O4) (3) Determine the absorption spectrum of a solution of ＝ MSb × QTb/QSb × 1/25 Naproxen in ethanol (99.5) (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>,andcompare M : Amount (mg) of naphazoline nitrate for assay Sa the spectrum with the Reference Spectrum: both spectra M : Amount (mg) of Chlorpheniramine Maleate RS Sb exhibit similar intensities of absorption at the same wave- Internal standard solution—A solution of ethenzamide in lengths. methanol (1 in 1000). (4) Determine the infrared absorption spectrum of Operating conditions— Naproxen, previously dried, as directed in the potassium Detector: An ultraviolet absorption photometer (wave- bromide disk method under Infrared Spectrophotometry length: 254 nm). <2.25>, and compare the spectrum with the Reference Spec- Column: A stainless steel column, about 4 mm in inside trum: both spectra exhibit similar intensities of absorption at diameter and 25 to 30 cm in length, packed with octadecyl- the same wave numbers. silanized silica gel for liquid chromatography (5 mminparti- Optical rotation <2.49> [a]25: ＋63.0 – ＋68.59(after dry- cle diameter). D ing, 0.1 g, chloroform, 10 mL, 100 mm). Column temperature: Room temperature. Mobile phase: A mixture of acetonitrile and a solution of Melting point <2.60> 154 – 1589C. sodium laurylsulfate (1 in 500) in diluted phosphoric acid Purity (1) Clarity of solution—Dissolve 2.0 g of Napro- (1 in 1000) (1:1). xen in 20 mL of acetone: the solution is clear. Perform the 1148 Nateglinide / Official Monographs JP XVI test with this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: the absorbance at 400 nm is not Nateglinide more than 0.070. (2) Heavy metals <1.07>—Proceed with 2.0 g of Napro- ナテグリニド xen according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (3) Arsenic <1.11>—Prepare the test solution with 2.0 g of Naproxen according to Method 3, and perform the test (not more than 1 ppm). (4) Related substances—Conduct this procedure without exposure to daylight, using light-resistant vessels. Dissolve C H NO :317.42 0.10 g of Naproxen in 10 mL of a mixture of chloroform and 19 27 3 N-[trans-4-(1-Methylethyl)cyclohexanecarbonyl]-D-phenylalanine ethanol (99.5) (1:1), and use this solution as the sample solu- [105816-04-4] tion. Pipet 2 mL of the sample solution, and add a mixture of chloroform and ethanol (99.5) (1:1) to make exactly 100 Nateglinide, when dried, contains not less than mL. Pipet 5 mL of this solution, add a mixture of chlo- 98.0z and not more than 102.0z of C H NO . roform and ethanol (99.5) (1:1) to make exactly 50 mL, and 19 27 3 use this solution as the standard solution. Perform the test Description Nateglinide occurs as a white crystalline pow- with these solutions as directed under Thin-layer Chroma- der. tography <2.03>. Spot 10 mL each of the sample solution and It is freely soluble in methanol and in ethanol (99.5), standard solution on a plate of silica gel with fluorescent in- sparingly soluble in acetonitrile, and practically insoluble in dicator for thin-layer chromatography. Develop the plate water. with a mixture of hexane, dichloromethane, tetrahydrofuran It dissolves in dilute sodium hydroxide TS. and acetic acid (100) (50:30:17:3) to a distance of about 12 Identification (1) Determine the absorption spectrum of a cm, and air-dry the plate. Examine under ultraviolet light solution of Nateglinide in methanol (1 in 1000) as directed (main wavelength: 254 nm): the spots other than the princi- under Ultraviolet-visible Spectrophotometry <2.24>,and pal spot and the spot of the starting point from the sample compare the spectrum with the Reference Spectrum or the solution are not more intense than the spot from the stand- spectrum of a solution of Nateglinide RS prepared in the ard solution. same manner as the sample solution: both spectra exhibit Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, similar intensities of absorption at the same wavelengths. 3 hours). (2) Determine the infrared absorption spectrum of Nateglinide as directed in the potassium bromide disk Residue on ignition <2.44> Not more than 0.1z (1 g). method under Infrared Spectrophotometry <2.25>,and Assay Weigh accurately about 0.5g of Naproxen, previ- compare the spectrum with the Reference Spectrum or the ously dried, add 100 mL of diluted methanol (4 in 5), dis- spectrum of Nateglinide RS: both spectra exhibit similar solve by gentle warming if necessary, and titrate <2.50> with intensities of absorption at the same wave numbers. If any 0.1 mol/L sodium hydroxide VS (indicator: 3 drops of phe- difference appears between the spectra, recrystallize the nolphthalein TS). Perform a blank determination, and make sample and the reference standard according to the method any necessary correction. otherwise specified, filter and dry the crystals, and perform the test with the crystals. Each mL of 0.1 mol/L sodium hydroxide VS 20 ＝ 23.03 mg of C14H14O3 Optical rotation <2.49> [a]D : －36.5 – －40.09(after drying 0.2 g, dilute sodium hydroxide TS, 20 mL, 100 mm). Containers and storage Containers—Well-closed containers. Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Storage—Light-resistant. Nateglinide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 0.25 g of Nateglinide in 20 mL of acetonitrile. To 4 mL of this solution add the mobile phase to make 25 mL, and use this solution as the sample solution. Pipet 2.5 mL of the sample solution, and add the mobile phase to make exactly 50 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than nateglinide from the sample solution is not larger than the peak area of nateglinide from the standard solution. JP XVI Official Monographs / Nateglinide Tablets 1149

Operating conditions— System repeatability: When the test is repeated 6 times Detector, column, column temperature, mobile phase, and with 10 mL of the standard solution under the above operat- flow rate: Proceed as directed in the operating conditions in ing conditions, the relative standard deviation of the ratio of the Assay. the peak area of nateglinide to that of the internal standard Time span of measurement: About 4 times as long as the is not more than 1.0z. retention time of nateglinide, beginning after the solvent Containers and storage Containers—Well-closed contain- peak. ers. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating conditions, the number of theoretical plates and the symmetry Nateglinide Tablets factor of the peak of nateglinide are not less than 6000 and ナテグリニド錠 not more than 1.2, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- Nateglinide Tablets contain not less than 96.0z and ing conditions, the relative standard deviation of the peak not more than 104.0z of the labeled amount of area of nateglinide is not more than 2.0z. nateglinide (C19H27NO3: 317.42). (3) Residual solvent—Being specified separately. Method of preparation Prepare as directed under Tablets, Loss on drying <2.41> Not more than 0.2z (1 g, 1059C, with Nateglinide. 2 hours). Identification To an amount of powdered Nateglinide Residue on ignition <2.44> Not more than 0.1z (1 g). Tablets, equivalent to 20 mg of Nateglinide according to the labeled amount, add 20 mL of methanol, shake, and filter. Assay Weigh accurately about 0.1 g of Nateglinide, previ- Determine the absorption spectrum of the filtrate as directed ously dried, and dissolve in acetonitrile to make exactly 20 under Ultraviolet-visible Spectrophotometry <2.24>:itexhib- mL. Pipet 5 mL of this solution, add exactly 5 mL of the in- its maxima between 246 nm and 250 nm, between 251 nm ternal standard solution, add the mobile phase to make 50 and 255 nm, between 257 nm and 261 nm and between 262 mL, and use this solution as the sample solution. Separately, nm and 266 nm. weigh accurately about 50 mg of Nateglinide RS, previously dried, and dissolve in acetonitrile to make exactly 20 mL. Uniformity of dosage units <6.02> Perform the test accord- Pipet 10 mL of this solution, add exactly 5 mL of the inter- ing to the following method: it meets the requirement of the nal standard solution, add the mobile phase to make 50 mL, Content uniformity test. and use this solution as the standard solution. Perform the To 1 tablet of Nateglinide Tablets add 10 mL of 0.05 test with 10 mL each of the sample solution and standard mol/L sodium dihydrogen phosphate TS adjusted to pH 2.5 solution as directed under Liquid Chromatography <2.01> with phosphoric acid, shake to disintegrate the tablet, and according to the following conditions, and calculate the disperse to fine particles with the aid of ultrasonic waves. ratios, QT and QS, of the peak area of nateglinide to that of Add exactly 3V/50 mL of the internal standard solution, add the internal standard. 3V/5 mL of acetonitrile, shake for 10 minutes, and add acetonitrile to make V mL so that each mL contains about Amount (mg) of C19H27NO3 ＝ MS × QT/QS × 2 0.6 mg of nateglinide (C19H27NO3). Filter the solution MS: Amount (mg) of Nateglinide RS through a membrane filter with a pore size not exceeding 0.45 mm, and discard the first 5 mL of the filtrate. To 8 mL Internal standard solution—A solution of propyl parahy- of the subsequent filtrate add the mobile phase to make 10 droxybenzoate in the mobile phase (1 in 500). mL, and use this solution as the sample solution. Separately, Operating conditions— weigh accurately about 50 mg of Nateglinide RS, previously Detector: An ultraviolet absorption photometer (wave- dried at 1059C for 2 hours, and dissolve in acetonitrile to length: 210 nm). make exactly 10 mL. Pipet 6 mL of this solution, add exactly Column: A stainless steel column 4.6 mm in inside diame- 3 mL of the internal standard solution, and add the mobile ter and 15 cm in length, packed with octadecylsilanized silica phase to make 25 mL. To 8 mL of this solution add the gel for liquid chromatography (5 mm in particle diameter). mobile phase to make 20 mL, and use this solution as the Column temperature: A constant temperature of about standard solution. Perform the test with 10 mLeachofthe 409C. sample solution and standard solution as directed under Mobile phase: Adjust 0.05 mol/L sodium dihydrogen Liquid Chromatography <2.01>, and calculate the ratios, Q phosphate TS to pH 2.5 with phosphoric acid. To 550 mL of T and Q , of the peak area of nateglinide to that of the internal this solution add 450 mL of acetonitrile for liquid chroma- S standard. tography.

Flow rate: Adjust the flow rate so that the retention time Amount (mg) of nateglinide (C19H27NO3) of nateglinide is about 10 minutes. ＝ MS × QT/QS × 3V/250 System suitability— M : Amount (mg) of Nateglinide RS System performance: When the procedure is run with 10 S mL of the standard solution under the above operating con- Internal standard solution—A solution of propyl parahy- ditions, the internal standard and nateglinide are eluted in droxybenzoate in acetonitrile (1 in 250). this order with the resolution between these peaks being not Operating conditions— less than 19. Proceed as directed in the operating conditions in the 1150 Nateglinide Tablets / Official Monographs JP XVI

Assay. the internal standard solution, shake for 10 minutes, and System suitability— add acetonitrile to make V mL so that each mL contains

System performance: When the procedure is run with 10 about 6 mg of nateglinide (C19H27NO3). Filter this solution mL of the standard solution under the above operating con- through a membrane filter with a pore size not exceeding ditions, the internal standard and nateglinide are eluted in 0.45 mm, discard the first 5 mL of the filtrate, to 4 mL of the this order with the resolution between these peaks being not subsequent filtrate add the mobile phase to make 50 mL, and less than 19. use this solution as the sample solution. Separately, weigh System repeatability: When the test is repeated 6 times accurately about 60 mg of Nateglinide RS, previously dried with 10 mL of the standard solution under the above operat- at 1059C for 2 hours, add exactly 1 mL of the internal standing conditions, the relative standard deviation of the ratio of ard solution, and add acetonitrile to make 10 mL. To 4 mL the peak area of nateglinide to that of the internal standard of this solution add the mobile phase to make 50 mL, and is not more than 1.0z. use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution Dissolution <6.10> When the test is performed at 50 revolu- as directed under Liquid Chromatography <2.01> according tions per minute according to the Paddle method, using 900 to the following conditions, and calculate the ratios, Q and mL of the 2nd fluid for dissolution test as the dissolution T Q , of the peak area of nateglinide to that of the internal medium, the dissolution rate in 45 minutes of a 30-mg tablet S standard. and that in 30 minutes of a 90-mg tablet of Nateglinide

Tablets is not less than 75z, respectively. Amount (mg) of nateglinide (C19H27NO3)in1tablet Start the test with 1 tablet of Nateglinide Tablets, with- ＝ MS × QT/QS × V/200 draw not less than 20 mL of the medium at the specified M : Amount (mg) of Nateglinide RS minute after starting the test, and filter through a membrane S filter with a pore size not exceeding 0.45 mm. Discard the Internal standard solution—A solution of propyl parahy- first 5 mL of the filtrate, pipet V mL of the subsequent droxybenzoate in acetonitrile (3 in 125). filtrate, add the dissolution medium to make exactly V?mL Operating conditions— so that each mL contains about 33 mg of nateglinide Detector: An ultraviolet absorption photometer (wave-

(C19H27NO3) according to the labeled amount, and use this length: 210 nm). solution as the sample solution. Separately, weigh accurately Column: A stainless steel column 4.6 mm in inside diame- about 33 mg of Nateglinide RS, previously dried at 1059C ter and 15 cm in length, packed with octadecylsilanized silica for 2 hours, and dissolve in acetonitrile to make exactly 100 gel for liquid chromatography (5 mminparticlediameter). mL. Pipet 5 mL of this solution, add the mobile phase to Column temperature: A constant temperature of about make exactly 50 mL, and use this solution as the standard 409C. solution. Perform the test with exactly 10 mLeachofthe Mobile phase: Adjust to pH 2.5 of 0.05 mol/L sodium di- sample solution and standard solution as directed under hydrogen phosphate TS with phosphoric acid. To 550 mL of Liquid Chromatography <2.01> according to the following this solution add 450 mL of acetonitrile. conditions, and determine the peak areas, AT and AS,of Flow rate: Adjust the flow rate so that the retention time nateglinide of both solutions. of nateglinide is about 10 minutes. System suitability— Dissolution rate (z) with respect to the labeled amount System performance: When the procedure is run with 10 of nateglinide (C H NO ) 19 27 3 mL of the standard solution under the above operating con- ＝ M × A /A × V?/V × 1/C × 90 S T S ditions, the internal standard and nateglinide are eluted in

MS: Amount (mg) of Nateglinide RS this order with the resolution between these peaks being not C: Labeled amount (mg) of nateglinide (C19H27NO3)in1 less than 19. tablet System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- Operating conditions— ing conditions, the relative standard deviation of the ratio of Proceed as directed in the operating conditions in the the peak area of nateglinide to that of the internal standard Assay. is not more than 1.0z. System suitability— System performance: When the procedure is run with 10 Containers and storage Containers—Tight containers. mL of the standard solution under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of nateglinide are not less than 8000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of nateglinide is not more than 2.0z. Assay To 20 Nateglinide Tablets add V/5 mL of 0.05 mol/L sodium dihydrogen phosphate TS adjusted to pH 2.5 with phosphoric acid, shake to disintegrate the tablets, and disperse to fine particles with the aid of ultrasonic waves. Then, add V/2 mL of acetonitrile and exactly V/10 mL of JP XVI Official Monographs / Neostigmine Methylsulfate Injection 1151

dried, dissolve each in the mobile phase to make exactly 50 Neostigmine Methylsulfate mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly ネオスチグミンメチル硫酸塩 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to

the following conditions, and determine the peak areas, AT and AS, of neostigmine in each solution.

Amount (mg) of C13H22N2O6S ＝ MS × AT/AS

MS: Amount (mg) of Neostigmine Methylsulfate RS Operating conditions—

C13H22N2O6S: 334.39 Detector: An ultraviolet absorption photometer (wave- 3-(Dimethylcarbamoyloxy)-N,N,N- length: 259 nm). trimethylanilinium methyl sulfate Column: A stainless steel column 4.6 mm in inside diame- [51-60-5] ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticlediameter). Neostigmine Methylsulfate, when dried, contains Column temperature: A constant temperature of about not less than 98.0z and not more than 102.0z of 259C. C13H22N2O6S. Mobile phase: Dissolve 3.12 g of sodium dihydrogenphosphate dihydrate in 1000 mL of water, adjust to pH 3.0 with Description Neostigmine Methylsulfate occurs as a white, phosphoric acid, and add 0.871 g of sodium 1-pentanesul- crystalline powder. fonate to dissolve. To 890 mL of this solution add 110 mL of It is very soluble in water, and freely soluble in acetonitrile acetonitrile. and in ethanol (95). Flow rate: Adjust the flow rate so that the retention time Identification (1) Determine the absorption spectrum of a of neostigmine is about 9 minutes. solution of Neostigmine Methylsulfate (1 in 2000) as directed System suitability— under Ultraviolet-visible Spectrophotometry <2.24>,and System performance: Dissolve 25 mg of Neostigmine compare the spectrum with the Reference Spectrum or the Methylsulfate and 4 mg of dimethylaminophenol in 50 mL spectrum of a solution of Neostigmine Methylsulfate RS of the mobile phase. When the procedure is run with 10 mL prepared in the same manner as the sample solution: both of this solution under the above operating conditions, spectra exhibit similar intensities of absorption at the same dimethylaminophenol and neostigmine are eluted in this wavelengths. order with the resolution between these peaks being not less (2) Determine the infrared absorption spectrum of than 6. Neostigmine Methylsulfate, previously dried, as directed in System repeatability: When the test is repeated 6 times the potassium bromide disk method under Infrared Spectro- with 10 mL of the standard solution under the above operat- photometry <2.25>, and compare the spectrum with the ing conditions, the relative standard deviation of the peak Reference Spectrum or the spectrum of dried Neostigmine areas of neostigmine methylsulfate is not more than 1.0z. Methylsulfate RS: both spectra exhibit similar intensities of Containers and storage Containers—Tight containers. absorption at the same wave numbers. pH <2.54> Dissolve 1.0 g of Neostigmine Methylsulfate in 10 mL of freshly boiled and cooled water: the pH of the so- Neostigmine Methylsulfate Injection lution is between 3.0 and 5.0. ネオスチグミンメチル硫酸塩注射液 Melting point <2.60> 145–1499C. Purity (1) Clarity and color of solution—Dissolve 1.0 g Neostigmine Methylsulfate Injection is an aqueous of Neostigmine Methylsulfate in 10 mL of water: the solu- solution for injection. tion is clear and colorless. It contains not less than 93.0z and not more than (2) Sulfate—Dissolve 0.20 g of Neostigmine Methylsul- 107.0z of the labeled amount of neostigmine methyl- fate in 10 mL of water, add 1 mL of dilute hydrochloric acid sulfate (C H N O S: 334.39). and 1 mL of barium chloride TS: no turbidity is produced 13 22 2 6 immediately. Method of preparation Prepare as directed under Injec- (3) Dimethylaminophenol—Dissolve 0.10 g of Neostig- tions, with Neostigmine Methylsulfate. mine Methylsulfate in 5 mL of water, add 1 mL of sodium Description Neostigmine Methylsulfate Injection is a clear, hydroxide TS, and while cooling with ice, add 1 mL of colorless liquid. diazobenzenesulfonic acid TS: no color develops. It is slowly affected by light. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, pH: 5.0 – 6.5 3 hours). Identification Take a volume of Neostigmine Methylsul- Residue on ignition <2.44> Not more than 0.1z (1 g). fate Injection equivalent to 5 mg of neostigmine methylsulfate according to the labeled amount, add water to make 10 Assay Weigh accurately about 25 mg each of Neostigmine mL if necessary, and determine the absorption spectrum of Methylsulfate and Neostigmine Methylsulfate RS, previously this solution as directed under Ultraviolet-visible Spectro- 1152 Nicardipine Hydrochloride / Official Monographs JP XVI photometry <2.24>: it exhibits a maximum between 257 nm Nicardipine Hydrochloride, previously dried, as directed in and 261 nm. the potassium bromide disk method under Infrared Spectro- photometry <2.25>, and compare the spectrum with the Ref- Bacterial endotoxins <4.01> Less than 5 EU/mg. erence Spectrum: both spectra exhibit similar intensities of Extractable volume <6.05> It meets the requirement. absorption at the same wave numbers. (3) Dissolve 0.02 g of Nicardipine Hydrochloride in 10 Foreign insoluble matter <6.06> Perform the test according mL of water and 3 mL of nitric acid: the solution responds to Method 1: it meets the requirement. to the Qualitative Tests <1.09> for chloride. Insoluble particulate matter <6.07> It meets the require- Melting point <2.60> 167 – 1719C. ment. Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Sterility <4.06> Perform the test according to the Mem- Nicardipine Hydrochloride according to Method 4, and per- brane filtration method: it meets the requirement. form the test. Prepare the control solution with 2.0 mL of Assay Use Neostigmine Methylsulfate Injection as the Standard Lead Solution (not more than 20 ppm). sample solution. Separately, weigh accurately about 25 mg (2) Related substances—Conduct this procedure without of Neostigmine Methylsulfate RS, previously dried at 1059C exposure to daylight, using light-resistant vessels. Dissolve for 3 hours, dissolve in the mobile phase to make exactly 50 0.10 g of Nicardipine Hydrochloride in 50 mL of the mobile mL, and use this solution as the standard solution. Proceed phase, and use this solution as the sample solution. Pipet 1 as directed in the Assay under Neostigmine Methylsulfate. mL of the sample solution, add the mobile phase to make exactly 50 mL, then take exactly 1 mL of this solution, add the Amount (mg) of neostigmine methylsulfate (C H N O S) 13 22 2 6 mobile phase to make exactly 10 mL, and use this solution as ＝ M × A /A S T S the standard solution. Perform the test with exactly 10 mL

MS: Amount (mg) of Neostigmine Methylsulfate RS each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol- Containers and storage Containers—Hermetic containers. lowing conditions. Determine each peak area of both solu- Storage—Light-resistant. tions by the automatic integration method: the area of each peak other than the peak of nicardipine from the sample solution is not larger than the peak area of nicardipine from Nicardipine Hydrochloride the standard solution, and the total area of each peak other than the peak of nicardipine is not larger than 2 times the ニカルジピン塩酸塩 peak area of nicardipine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 254 nm). Column: A stainless steel column 4.6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticlediameter). Column temperature: A constant temperature of about 309C. C H N O .HCl: 515.99 26 29 3 6 Mobile phase: A mixture of a solution of perchloric acid 2-[Benzyl(methyl)amino]ethyl methyl (4RS)- (43 in 50,000) and acetonitrile (3:2). 2,6-dimethyl-4-(3-nitrophenyl)-1,4-dihydropyridine- Flow rate: Adjust the flow rate so that the retention time 3,5-dicarboxylate monohydrochloride of nicardipine is about 6 minutes. [54527-84-3] Time span of measurement: About 4 times as long as the retention time of nicardipine beginning after the solvent Nicardipine hydrochloride, when dried, contains not peak. less than 98.5z of C H N O .HCl. 26 29 3 6 System suitability— Description Nicardipine Hydrochloride occurs as a pale Test for required detection: To exactly 2 mL of the stand- greenish yellow crystalline powder. ard solution add the mobile phase to make exactly 20 mL. It is freely soluble in methanol and in acetic acid (100), Confirm that the peak area of nicardipine obtained from 10 sparingly soluble in ethanol (99.5), and slightly soluble in mL of this solution is equivalent to 8 to 12z of that from 10 water, in acetonitrile and in acetic anhydride. mL of the standard solution. A solution of Nicardipine Hydrochloride in methanol (1 in System performance: Dissolve 2 mg each of Nicardipine 20) shows no optical rotation. Hydrochloride and nifedipine in 50 mL of the mobile phase. It is gradually affected by light. When the procedure is run with 10 mL of this solution under the above operating conditions, nicardipine and nifedipine Identification (1) Determine the absorption spectrum of a are eluted in this order with the resolution between these solution of Nicardipine Hydrochloride in ethanol (99.5) (1 in peaks being not less than 3. 100,000) as directed under Ultraviolet-visible Spectropho- System repeatability: When the test is repeated 6 times tometry <2.24>, and compare the spectrum with the Refer- with 10 mL of the standard solution under the above operat- ence Spectrum: both spectra exhibit similar intensities of ab- ing conditions, the relative standard deviation of the peak sorption at the same wavelengths. areas of nicardipine is not more than 3z. (2) Determine the infrared absorption spectrum of JP XVI Official Monographs / Nicardipine Hydrochloride Injection 1153

Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, ard solution. 2 hours). Operating conditions— Detector, column, column temperature, mobile phase, and Residue on ignition <2.44> Not more than 0.1z (1 g). flow rate: Proceed as directed in the operating conditions in Assay Conduct this procedure without exposure to day- the Assay. light, using light-resistant vessels. Weigh accurately about Time span of measurement: About 3 times as long as the 0.9 g of Nicardipine Hydrochloride, previously dried, dis- retention time of nicardipine beginning after the solvent solve in 100 mL of a mixture of acetic anhydride and acetic peak. acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric System suitability— acid VS (potentiometric titration). Perform a blank determi- Test for required detectability: To exactly 2 mL of the nation, and make any necessary correction. standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of nicardipine obtained Each mL of 0.1 mol/L perchloric acid VS from 10 mL of this solution is equivalent to 8 to 12z of that ＝ 51.60 mg of C H N O .HCl 26 29 3 6 from 10 mL of the standard solution. Containers and storage Containers—Well-closed contain- System performance: Proceed as directed in the system ers. suitability in the Assay. Storage—Light-resistant. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak Nicardipine Hydrochloride areas of nicardipine is not more than 1.0z. Injection Bacterial endotoxins <4.01> Less than 8.33 EU/mg. Extractable volume <6.05> It meets the requirement. ニカルジピン塩酸塩注射液 Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Nicardipine Hydrochloride Injection is an aqueous solution for injection. Insoluble particulate matter <6.07> It meets the require- It contains not less than 93.0z and not more than ment. 107.0z of the labeled amount of nicardipine hydro- Sterility <4.06> Perform the test according to the Mem- chloride (C H N O .HCl: 515.99). 26 29 3 6 brane filtration method: it meets the requirement. Method of preparation Prepare as directed under Injec- Assay Conduct the procedure without exposure to day- tions, with Nicardipine Hydrochloride. light using light-resistant vessels. To an exact volume of Description Nicardipine Hydrochloride Injection occurs as Nicardipine Hydrochloride Injection, equivalent to about 2 a clear pale yellow liquid. mg of nicardipine hydrochloride (C26H29N3O6.HCl), add ex- It is gradually changed by light. actly 5 mL of the internal standard solution and methanol to make 50 mL, and use this solution as the sample solution. Identification To a volume of Nicardipine Hydrochloride Separately, weigh accurately about 50 mg of nicardipine hy- Injection, equivalent to 1 mg of Nicardipine Hydrochloride drochloride for assay, previously dried at 1059C for 2 hours, according to the labeled amount, add ethanol (99.5) to make dissolve in methanol to make exactly 50 mL. Pipet 2 mL of 100 mL. Determine the absorption spectrum of this solution this solution, add exactly 5 mL of the internal standard solu- as directed under Ultraviolet-visible Spectrophotometry tion and methanol to make 50 mL, and use this solution as <2.24>: it exhibits maxima between 235 nm and 239 nm, and the standard solution. Perform the test with 10 mLeachof between 351 nm and 355 nm. the sample solution and standard solution as directed under pH <2.54> 3.0–4.5 Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, Q and Q , of the peak Purity Related substances—Conduct the procedure with- T S area of nicardipine to that of the internal standard. out exposure to day-light using light-resistant vessels. To a volume of Nicardipine Hydrochloride Injection, equivalent Amount (mg) of nicardipine hydrochloride to 5 mg of Nicardipine Hydrochloride according to the (C26H29N3O6.HCl) labeled amount, add the mobile phase to make 10 mL, and ＝ MS × QT/QS × 1/25 use this solution as the sample solution. To exactly 2 mL of M : Amount (mg) of nicardipine hydrochloride for assay the sample solution add the mobile phase to make exactly S 100 mL, and use this solution as the standard solution. Per- Internal standard solution—A solution of di-n-butyl phtha- form the test with exactly 10 mL each of the sample solution late in methanol (1 in 625). and standard solution as directed under Liquid Chromatog- Operating conditions— raphy <2.01> according to the following conditions, and de- Detector: An ultraviolet absorption photometer (wave- termine the peak areas of these solutions by the automatic length: 254 nm). integration method: the areas of the peaks other than nicar- Column: A stainless steel column 4.6 mm in inside diame- dipine from the sample solution are not larger than the peak ter and 15 cm in length, packed with octadecylsilanized silica area of nicardipine from the standard solution, and the total gel for liquid chromatography (5 mminparticlediameter). of the areas of the peaks other than nicardipine is not larger Column temperature: A constant temperature of about than 2 times of the peak area of nicardipine from the stand- 409C. 1154 Nicergoline / Official Monographs JP XVI

Mobile phase: Dissolve 1.36 g of potassium dihydrogen Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of phosphate in water to make 1000 mL. To 320 mL of this so- Nicergoline according to Method 2, and perform the test. lution add 680 mL of methanol. Prepare the control solution with 2.0 mL of Standard Lead Flow rate: Adjust the flow rate so that the retention time Solution (not more than 10 ppm). of nicardipine is about 8 minutes. (2) Related substances—Dissolve 25 mg of Nicergoline in System suitability— 25 mL of acetonitrile, and use this solution as the sample so- System performance: When the procedure is run with 10 lution. Pipet 1 mL of the sample solution, and add aceto- mL of the standard solution under the above operating con- nitrile to make exactly 100 mL. Pipet 10 mL of this solution, ditions, nicardipine and the internal standard are eluted in add acetonitrile to make exactly 50 mL, and use this solution this order with the resolution between these peaks being not as the standard solution. Perform the test with exactly 20 mL less than 6. each of the sample solution and standard solution as directed System repeatability: When the test is repeated 5 times under Liquid Chromatography <2.01> according to the fol- with 10 mL of the standard solution under the above operat- lowing conditions, and determine each peak area by the au- ing conditions, the relative standard deviation of the peak tomatic integration method: the area of the peak, having the areas of nicardipine is not more than 1.0z. relative retention time of about 0.5 with respect to nicergoline, is not larger than 4 times the peak area of nicergoline Containers and storage Containers—Hermetic containers. from the standard solution, and the area of the peak other Colored containers may be used. than nicergoline and other than the peak mentioned above is Storage—Light-resistant. not larger than 2.5 times the peak area of nicergoline from the standard solution. The peak which area is larger than the peak area of nicergoline from the standard solution is not Nicergoline more than two peaks, and the total area of the peaks other than the peak of nicergoline is not larger than 7.5 times the ニセルゴリン peak area of nicergoline from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 288 nm). Column: A stainless steel column 4.6 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticlediameter). Column temperature: A constant temperature of about 259C. Mobile phase: Adjust the pH of 0.05 mol/L potassium dihydrogen phosphate TS to 7.0 with triethylamine. To 350 mL of this solution add 350 mL of methanol and 300 mL of C H BrN O : 484.39 24 26 3 3 acetonitrile. [(8R,10S)-10-Methoxy-1,6-dimethylergolin-8-yl]methyl Flow rate: Adjust the flow rate so that the retention time 5-bromopyridine-3-carboxylate of nicergoline is about 25 minutes. [27848-84-6] Time span of measurement: About 2 times as long as the retention time of nicergoline beginning after the solvent Nicergoline, when dried, contains not less than peak. 98.5z and not more than 101.0z of C H BrN O . 24 26 3 3 System suitability— Description Nicergoline occurs as white to light yellow, Test for required detectability: To 1 mL of the sample so- crystals or crystalline powder. lution add acetonitrile to make exactly 50 mL, and use this It is soluble in acetonitrile, in ethanol (99.5) and in acetic solution as the solution for system suitability test. Pipet 5 anhydride, and practically insoluble in water. mL of the solution for system suitability test, and add aceto- It is gradually colored to light brown by light. nitrile to make exactly 100 mL. Confirm that the peak area Melting point: about 1369C (with decomposition). of nicergoline obtained with 20 mL of this solution is equivalent to 3 to 7z of that with 20 mL of the solution for system Identification (1) Determine the absorption spectrum of a suitability test. solution of Nicergoline in ethanol (99.5) (1 in 100,000) as System performance: When the procedure is run with 20 directed under Ultraviolet-visible Spectrophotometry <2.24>, mL of the sample solution under the above operating condi- and compare the spectrum with the Reference Spectrum: tions, the number of theoretical plates and the symmetry fac- both spectra exhibit similar intensities of absorption at the tor of the peak of nicergoline are not less than 8000 and not same wavelengths. more than 2.0, respectively. (2) Determine the infrared absorption spectrum of Nicer- System repeatability: When the test is repeated 6 times goline as directed in the potassium bromide disk method with 20 mL of the standard solution under the above operat- under Infrared Spectrophotometry <2.25>, and compare the ing conditions, the relative standard deviation of the peak spectrum with the Reference Spectrum: both spectra exhibit area of nicergoline is not more than 4.0z. similar intensities of absorption at the same wave numbers. Loss on drying <2.41> Not more than 0.5z (2 g, in vacu- Optical rotation <2.49> [a]20: ＋5.2 – ＋6.29(after drying, D um, 609C, 2 hours). 0.5 g, ethanol (95), 10 mL, 100 mm). Residue on ignition <2.44> Not more than 0.1z (1 g). JP XVI Official Monographs / Nicergoline Powder 1155

Assay Weigh accurately about 0.4 g of Nicergoline, previ- the above operating conditions, the relative standard devia- ously dried, add 10 mL of acetic anhydride, and warm to tion of the peak area of nicergoline is not more than 1.5z. dissolve. After cooling, add 40 mL of nitrobenzene, and Uniformity of dosage unit <6.02> The Nicergoline Powder titrate <2.50> with 0.1 mol/L perchloric acid VS until the in single-unit container meets the requirement of the Mass color of the solution changes to blue-green from red through variation test. a blue-purple (indicator: 10 drops of neutral red TS). Per- form a blank determination in the same manner, and make Dissolution <6.10> When the test is performed at 50 revolu- any necessary correction. tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution Each mL of 0.1 mol/L perchloric acid VS medium, the dissolution rate in 15 minutes of Nicergoline ＝ 24.22 mg of C H BrN O 24 26 3 3 Powder is not less than 80z. Containers and storage Containers—Well-closed contain- Start the test with an accurately weighed amount of ers. Nicergoline Powder, equivalent to about 5 mg of nicergoline

Storage—Light-resistant. (C24H26BrN3O3) according to the labeled amount, withdraw not less than about 20 mL of the medium at the specified minute after starting the test, and filter through a laminated Nicergoline Powder polyester fiber. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa- ニセルゴリン散 rately, weigh accurately about 50 mg of nicergoline for assay, previously dried in vacuum at 609C for 2 hours, and dissolve in 0.1 mol/L hydrochloric acid TS to make exactly Nicergoline Powder contains not less than 95.0z 50 mL. Pipet 5 mL of this solution, and add the dissolution and not more than 105.0z of the labeled amount of medium to make exactly 100 mL. Pipet 10 mL of this solu- nicergoline (C H BrN O : 484.39). 24 26 3 3 tion, add the dissolution medium to make exactly 100 mL, Method of preparation Prepare as directed under Granules and use this solution as the standard solution. Determine the or Powders, with Nicergoline. absorbances at 225 nm, AT1 and AS1, and at 250 nm, AT2 and A , of the sample solution and standard solution as directed Identification Vigorously shake for 10 minutes a quantity S2 under Ultraviolet-visible Spectrophotometry <2.24>,using of Nicergoline Powder, equivalent to 10 mg of Nicergoline the dissolution medium as the blank. according to the labeled amount, with 20 mL of diluted ethanol (4 in 5), and centrifuge for 10 minutes. To 2 mL of Dissolution rate (z) with respect to the labeled amount the supernatant liquid add ethanol (99.5) to make 100 mL. of nicergoline (C24H26BrN3O3) Determine the absorption spectrum of this solution as di- ＝ MS/MT × (AT1 － AT2)/(AS1 － AS2) × 1/C × 9 rected under Ultraviolet-visible Spectrophotometry <2.24>:it M : Amount (mg) of nicergoline for assay exhibits maxima between 226 nm and 230 nm, and between S M : Amount (g) of sample 286 nm and 290 nm. T C: Labeled amount (mg) of nicergoline (C24H26BrN3O3)in Purity Related substances—Perform the test with 20 mLof 1g the sample solution obtained in the Assay as directed under Assay Weigh accurately a quantity of Nicergoline Powder, Liquid Chromatography <2.01> according to the following equivalent to about 20 mg of nicergoline (C H BrN O ), conditions. Determine each peak area by the automatic in- 24 26 3 3 add exactly 20 mL of a mixture of acetonitrile and water tegration method, and calculate the amount of substances (17:3), vigorously shake for 10 minutes, centrifuge for 10 other than nicergoline by the area percentage method: the minutes, and use the supernatant liquid as the sample solu- total amount of them is not more than 2.0z. tion. Separately, weigh accurately about 20 mg of nicer- Operating conditions— goline for assay, previously dried in vacuum at 609Cfor2 Detector, column, column temperature, mobile phase, and hours, dissolve in exactly 20 mL of the mixture of aceto- flow rate: Proceed as directed in the operating conditions in nitrile and water (17:3), and use this solution as the standard the Assay. solution. Perform the test with exactly 20 mLeachofthe Time span of measurement: About 2 times as long as the sample solution and standard solution as directed under retention time of nicergoline after the solvent peak. Liquid Chromatography <2.01> according to the following System suitability— conditions, and determine the peak areas, A and A ,of Test for required detectability: To 1 mL of the standard T S nicergoline. solution obtained in the Assay add a mixture of acetonitrile and water (17:3) to make 50 mL, and use this solution as the Amount (mg) of nicergoline (C24H26BrN3O3) solution for system suitability test. Pipet 5 mL of the solu- ＝ MS × AT/AS tion for system suitability test, add the mixture of aceto- M : Amount (mg) of nicergoline for assay nitrile and water (17:3) to make exactly 100 mL. Confirm S that the peak area of nicergoline obtained with 20 mLofthis Operating conditions— solution is equivalent to 3 to 7z of that with 20 mLoftheso- Detector: An ultraviolet absorption photometer (wave- lution for system suitability test. length: 288 nm). System performance: Proceed as directed in the system Column: A stainless steel column 4.6 mm in inside diame- suitability in the Assay. ter and 25 cm in length, packed with octadecylsilanized silica System repeatability: When the test is repeated 6 times gel for liquid chromatography (5 mminparticlediameter). with 20 mL of the solution for system suitability test under Column temperature: A constant temperature of about 1156 Nicergoline Tablets / Official Monographs JP XVI

409C. that the peak area of nicergoline obtained with 20 mLofthis Mobile phase: Adjust the pH of 0.05 mol/L potassium di- solution is equivalent to 3 to 7z of that with 20 mLoftheso- hydrogen phosphate TS to 7.0 with triethylamine. To 350 lution for system suitability test. mL of this solution add 350 mL of methanol and 300 mL of System performance: Proceed as directed in the system acetonitrile. suitability in the Assay. Flow rate: Adjust the flow rate so that the retention time System repeatability: When the test is repeated 6 times of nicergoline is about 25 minutes. with 20 mL of the solution for system suitability test under System suitability— the above operating conditions, the relative standard devia- System performance: When the procedure is run with 20 tion of the peak area of nicergoline is not more than 1.5z. mL of the standard solution under the above operating con- Uniformity of dosage units <6.02> Perform the test accord- ditions, the number of theoretical plates and the symmetry ing to the following method: it meets the requirement of the factor of the peak of nicergoline are not less than 8000 and Content uniformity test. not more than 2.0, respectively. To 1 tablet of Nicergoline Tablets add exactly 25 mL of System repeatability: When the test is repeated 6 times diluted ethanol (4 in 5), disperse to fine particles with the aid with 20 mL of the standard solution under the above operat- of ultrasonic wave, and shake for 5 minutes. Centrifuge this ing conditions, the relative standard deviation of the peak solution for 10 minutes, pipet exactly 4 mL of the superna- area of nicergoline is not more than 1.0z. tant liquid, add diluted ethanol (4 in 5) to make exactly 25 Containers and storage Containers—Tight containers. mL, and use this solution as the sample solution. Separately, Storage—Light-resistant. weigh accurately about 10 mg of nicergoline for assay, previously dried in vacuum at 609C for 2 hours, and dissolve in exactly 25 mL of diluted ethanol (4 in 5). Pipet 4 mL of this Nicergoline Tablets solution, add diluted ethanol (4 in 5) to make exactly 50 mL, and use this solution as the standard solution. Determine the

ニセルゴリン錠 absorbances at 288 nm, AT1 and AS1, and at 340 nm, AT2 and AS2, of the sample solution and the standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Nicergoline Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of nicer- Amount (mg) of nicergoline (C24H26BrN3O3) goline (C24H26BrN3O3: 484.39). ＝ MS × (AT1 － AT2)/(AS1 － AS2) × 1/2

Method of preparation Prepare as directed under Tablets, MS: Amount (mg) of nicergoline for assay with Nicergoline. Dissolution Being specified separately. Identification Take a quantity of powdered Nicergoline Assay Weigh accurately the mass of not less than 20 Nicer- Tablets, equivalent to 10 mg of Nicergoline according to the goline Tablets, and powder. Weigh accurately a portion of labeled amount, add 20 mL of ethanol (99.5), shake vigor- the powder, equivalent to about 20 mg of nicergoline ously for 10 minutes, and filter through a 0.45-mm pore-size (C H BrN O ), add exactly 20 mL of a mixture of aceto- membrane filter. To 2 mL of the filtrate add ethanol (99.5) 24 26 3 3 nitrile and water (17:3), vigorously shake for 10 minutes, to make 100 mL. Determine the absorption spectrum of this centrifuge for 10 minutes, and use the supernatant liquid as solution as directed under Ultraviolet-visible Spectropho- the sample solution. Separately, weigh accurately about 20 tometry <2.24>: it exhibits maxima between 226 nm and 230 mg of nicergoline for assay, previously dried in vacuum at nm, and between 286 nm and 290 nm. 609C for 2 hours, dissolve in exactly 20 mL of the mixture of Purity Related substances—Perform the test with 20 mLof acetonitrile and water (17:3), and use this solution as the the sample solution obtained in the Assay as directed under standard solution. Perform the test with exactly 20 mLeach Liquid Chromatography <2.01> according to the following of the sample solution and standard solution as directed conditions. Determine each peak area by the automatic in- under Liquid Chromatography <2.01> according to the fol- tegration method, and calculate the amount of substances lowing conditions, and determine the peak areas, AT and AS, other than nicergoline by the area percentage method: the of nicergoline. total amount of them is not more than 2.0z. Amount (mg) of nicergoline (C H BrN O ) ＝ M × A /A Operating conditions— 24 26 3 3 S T S

Detector, column, column temperature, mobile phase, and MS: Amount (mg) of nicergoline for assay flow rate: Proceed as directed in the operating conditions in Operating conditions— the Assay. Detector: An ultraviolet absorption photometer (wave- Time span of measurement: About 2 times as long as the length: 288 nm). retention time of nicergoline beginning after the solvent Column: A stainless steel column 4.6 mm in inside diame- peak. ter and 25 cm in length, packed with octadecylsilanized silica System suitability— gel for liquid chromatography (5 mminparticlediameter). Test for required detectability: To 1 mL of the standard Column temperature: A constant temperature of about solution obtained in the Assay add a mixture of acetonitrile 409C. and water (17:3) to make 50 mL, and use this solution as the Mobile phase: Adjust the pH of 0.05 mol/L potassium di- solution for system suitability test. Pipet 5 mL of the solu- hydrogen phosphate TS to 7.0 with triethylamine. To 350 tion for system suitability test, add the mixture of aceto- mL of this solution add 350 mL of methanol and 300 mL of nitrile and water (17:3) to make exactly 100 mL. Confirm acetonitrile. JP XVI Official Monographs / Niceritrol 1157

Flow rate: Adjust the flow rate so that the retention time pare the control solution with 2.0 mL of Standard Lead So- of nicergoline is about 25 minutes. lution (not more than 20 ppm). System suitability— (3) Arsenic <1.11>—Prepare the test solution with 1.0 g System performance: When the procedure is run with 20 of Niceritrol according to Method 3, and perform the test. mL of the standard solution under the above operating con- Use 10 mL of a solution of magnesium nitrate hexahydrate ditions, the number of theoretical plates and the symmetry in ethanol (95) (1 in 10) (not more than 2 ppm). factor of the peak of nicergoline are not less than 8000 and (4) Pyridine—Dissolve 0.5 g of Niceritrol in N,N- not more than 2.0, respectively. dimethylformamide to make exactly 10 mL, and use this System repeatability: When the test is repeated 6 times solution as the sample solution. Separately, weigh accurately with 20 mL of the standard solution under the above operat- about 0.1 g of pyridine, and add N,N-dimethylformamide to ing conditions, the relative standard deviation of the peak make exactly 100 mL. Pipet 1 mL of this solution, add N,N- area of nicergoline is not more than 1.0z. dimethylformamide to make exactly 100 mL, then pipet 0.5 mL of this solution, add N,N-dimethylformamide to make Containers and storage Containers—Tight containers. exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 2 mL each of the sample solution and standard solution as directed under Gas Chroma- Niceritrol tography <2.02> according to the following conditions. Determine each peak area of pyridine in both solutions: the ニセリトロール peak area of pyridine from the sample solution is not larger than the peak area of pyridine from the standard solution. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A column 3 mm in inside diameter and 3 m in length, packed with polyethylene glycol 20M for gas chromatography coated at the ratio of 10z on acid-treated and silanized siliceous earth for gas chromatography (150 to 180 mm in particle diameter). Column temperature: A constant temperature of about C H N O : 556.52 29 24 4 8 1609C. Pentaerythritol tetranicotinate Carrier gas: Nitrogen. [5868-05-3] Flow rate: Adjust the flow rate so that the retention time of pyridine is about 2 minutes. Niceritrol, when dried, contains not less than 99.0z System suitability— of C H N O . 29 24 4 8 System performance: When the procedure is run with 2 mL Description Niceritrol occurs as a white to pale yellowish of the standard solution under the above operating condi- white powder. It is odorless, and has a slightly bitter taste. tions, the number of theoretical steps of the peak of pyridine It is freely soluble in chloroform, soluble in N,N- is not less than 1500 steps. dimethylformamide, very slightly soluble in ethanol (95), System repeatability: When the test is repeated 6 times and practically insoluble in water and in diethyl ether. with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak areas Identification (1) Determine the absorption spectrum of a of pyridine is not more than 3.0z. solution of Niceritrol in 0.1 mol/L hydrochloric acid TS (1 (5) Free acids—Transfer about 1 g of Niceritrol, weighed in 100,000) as directed under Ultraviolet-visible Spectropho- accurately, to a separator, dissolve in 20 mL of chloroform, tometry <2.24>, and compare the spectrum with the Refer- and extract with 20 mL and then 10 mL of water while shak- ence Spectrum: both spectra exhibit similar intensities of ing well. Combine the whole extracts, and titrate <2.50> with absorption at the same wavelengths. 0.01 mol/L sodium hydroxide VS (indicator: 3 drops of phe- (2) Determine the infrared absorption spectrum of nolphthalein TS). Perform a blank determination, make any Niceritrol, previously dried, as directed in the potassium necessary correction, and calculate the amount of free acid bromide disk method under Infrared Spectrophotometry by the following equation: it is not more than 0.1z. <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at Each mL of 0.01 mol/L sodium hydroxide VS the same wave numbers. ＝ 1.231 mg of C6H5NO2 Melting point <2.60> 162–1659C. (6) Related substances—Dissolve 0.10 g of Niceritrol in 10 mL of chloroform, and use this solution as the sample so- Purity (1) Chloride <1.03>—To 2.0 g of Niceritrol add 50 lution. Pipet 1 mL of the sample solution, and add chlo- mL of water, and warm at 709C for 20 minutes, while shak- roform to make exactly 20 mL. Pipet exactly 2 mL of this so- ing occasionally. After cooling, filter, and to 25 mL of the lution, add chloroform to make exactly 20 mL, and use this filtrate add 6 mL of dilute nitric acid and water to make 50 solution as the standard solution. Perform the test with these mL. Perform the test using this solution as the test solution. solutions as directed under Thin-layer Chromatography Prepare the control solution with 1.0 mL of 0.01 mol/L hy- <2.03>. Spot 10 mL each of the sample solution and standard drochloric acid VS (not more than 0.036z). solution on a plate of silica gel with fluorescent indicator for (2) Heavy metals <1.07>—Proceed with 1.0 g of thin-layer chromatography. Develop the plate with a mixture Niceritrol according to Method 2, and perform the test. Pre- of chloroform and ethanol (95) (4:1) to a distance of about 1158 Nicomol / Official Monographs JP XVI

10 cm, and air-dry the plate. Examine under ultraviolet light mide disk method under Infrared Spectrophotometry <2.25>, (main wavelength: 254 nm): the spots other than the princi- and compare the spectrum with the Reference Spectrum: pal spot from the sample solution are not more intense than both spectra exhibit similar intensities of absorption at the the spot from the standard solution. same wave numbers. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, Melting point <2.60> 181 – 1859C. 3 hours). Purity (1) Clarity and color of solution—Dissolve 1.0 g Residue on ignition <2.44> Not more than 0.1z (1 g). of Nicomol in 10 mL of 1 mol/L hydrochloric acid TS: the solution is clear and colorless. Assay Weigh accurately about 1 g of Niceritrol, previously (2) Acidity—To 1.0 g of Nicomol add 50 mL of freshly dried, add exactly 25 mL of 0.5 mol/L sodium hydroxide boiled and cooled water, shake for 5 minutes, filter, and to VS, boil gently for 20 minutes under a reflux condenser with 25 mL of the filtrate add 0.60 mL of 0.01 mol/L sodium hy- a carbon dioxide absorber (soda lime). After cooling, titrate droxide VS and 2 drops of phenolphthalein TS: a red color <2.50> immediately the excess sodium hydroxide with 0.5 develops. mol/L hydrochloric acid VS (indicator: 3 drops of phenol- (3) Chloride <1.03>—Dissolve 0.6 g of Nicomol in 15 mL phthalein TS). Perform a blank determination. of dilute nitric acid, and add water to make 50 mL. Perform Each mL of 0.5 mol/L sodium hydroxide VS the test using this solution as the test solution. Prepare the

＝ 69.57 mg of C29H24N4O8 control solution as follows: to 0.40 mL of 0.01 mol/L hydrochloric acid VS add 15 mL of dilute nitric acid and water to Containers and storage Containers—Well-closed contain- make 50 mL (not more than 0.024z). ers. (4) Heavy metals <1.07>—Proceed with 1.0 g of Nicomol according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not Nicomol more than 20 ppm). (5) Arsenic <1.11>—Prepare the test solution with 1.0 g ニコモール of Nicomol according to Method 3, and perform the test (not more than 2 ppm). (6) Related substances—Dissolve 0.20 g of Nicomol in 20 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add chloroform to make exactly 20 mL. Pipet 2 mL of this solution, add chloroform to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot

C34H32N4O9: 640.64 10 mL each of the sample solution and standard solution on a (2-Hydroxycyclohexane-1,1,3,3-tetrayl)tetramethyl plate of silica gel with fluorescent indicator for thin-layer tetranicotinate chromatography. Develop the plate with a mixture of [27959-26-8] dichloromethane, ethanol (95), acetonitrile and ethyl acetate (5:3:1:1) to a distance of about 10 cm, and air-dry the plate. Nicomol, when dried, contains not less than 98.0z Examine under ultraviolet light (main wavelength: 254 nm): of C34H32N4O9. the spots other than the principal spot from the sample solution are not more intense than the spot from the standard so- Description Nicomol occurs as a white, crystalline powder. lution. It is odorless and tasteless. It is soluble in chloroform, and practically insoluble in Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, water, in ethanol (95) and in diethyl ether. 4 hours). It dissolves in dilute hydrochloric acid and in dilute nitric Residue on ignition <2.44> Not more than 0.1z (1 g). acid. Assay Weigh accurately about 1.5 g of Nicomol, previ- Identification (1) Mix 0.01 g of Nicomol with 0.02 g of 1- ously dried, add exactly 40 mL of 0.5 mol/L sodium hydrox- chloro-2,4-dinitrobenzene, add 2 mL of dilute ethanol, heat ide VS, and boil gently under a reflux condenser connected in a water bath for 5 minutes, cool, and add 4 mL of potas- to a carbon dioxide absorption tube (soda lime) for 10 sium hydroxide-ethanol TS: a dark red color develops. minutes. After cooling, titrate <2.50> immediately the excess (2) Dissolve 0.1 g of Nicomol in 5 mL of dilute hydro- sodium hydroxide with 0.25 mol/L sulfuric acid VS (indica- chloric acid, and add 5 drops of Reinecke salt TS: a light red tor: 3 drops of phenolphthalein TS). Perform a blank deter- precipitate is formed. mination. (3) Determine the absorption spectrum of a solution of Nicomolin1mol/LhydrochloricacidTS(1in100,000)as Each mL of 0.5 mol/L sodium hydroxide VS directed under Ultraviolet-visible Spectrophotometry <2.24>, ＝ 80.08 mg of C34H32N4O9 and compare the spectrum with the Reference Spectrum: Containers and storage Containers—Tight containers. both spectra exhibit similar intensities of absorption at the same wavelengths. (4) Determine the infrared absorption spectrum of Nicomol, previously dried, as directed in the potassium bro- JP XVI Official Monographs / Nicorandil 1159

mine the absorbances, AT and AS, of the sample solution Nicomol Tablets and standard solution at 262 nm as directed under Ultravio- let-visible Spectrophotometry <2.24>. ニコモール錠 Amount (mg) of nicomol (C34H32N4O9) ＝ MS × AT/AS × 25/2 Nicomol Tablets contain not less than 95.0z and M : Amount (mg) of nicomol for assay not more than 105.0z of the labeled amount of S nicomol (C34H32N4O9: 640.64). Containers and storage Containers—Tight containers. Method of preparation Prepare as directed under Tablets, with Nicomol. Nicorandil Identification To a portion of powdered Nicomol Tablets, equivalent to 0.5 g of Nicomol according to the labeled ニコランジル amount, add 20 mL of chloroform, shake, and filter. Evaporate the filtrate on a water bath to dryness. Proceed with the residue as directed in the Identification (1) and (2) under Nicomol. Uniformity of dosage units <6.02> It meets the requirement C H N O : 211.17 of the Mass variation test. 8 9 3 4 N-[2-(Nitrooxy)ethyl]pyridine-3-carboxamide Dissolution <6.10> When the test is performed at 75 revolu- [65141-46-0] tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution Nicorandil contains not less than 98.5z and not medium, the dissolution rate in 60 minutes of Nicomol more than 101.0z of C8H9N3O4,calculatedonthe Tablets is not less than 75z. anhydrous basis. Start the test with 1 tablet of Nicomol Tablets, withdraw Description Nicorandil occurs as white crystals. not less than 20 mL of the medium at the specified minute It is freely soluble in methanol, in ethanol (99.5) and in after starting the test, and filter through a membrane filter acetic acid (100), soluble in acetic anhydride, and sparingly with a pore size not exceeding 0.8 mm. Discard the first 10 soluble in water. mL of the filtrate, pipet V mL of the subsequent filtrate, add Melting point: about 929C (with decomposition). the dissolution medium to make exactly V?mL so that each mL contains about 18 mg of nicomol (C34H32N4O9) according Identification (1) Determine the absorption spectrum of a to the labeled amount, and use this solution as the sample solution of Nicorandil (1 in 50,000) as directed under Ultra- solution. Separately, weigh accurately about 0.1 g of violet-visible Spectrophotometry <2.24>, and compare the nicomol for assay, previously dried at 1059C for 4 hours, spectrum with the Reference Spectrum: both spectra exhibit dissolve in the dissolution medium to make exactly 100 mL, similar intensities of absorption at the same wavelengths. then pipet 2 mL of this solution, add the dissolution medium (2) Determine the infrared absorption spectrum of to make exactly 100 mL, and use this solution as the stand- Nicorandil as directed in the potassium bromide disk method ard solution. Determine the absorbances, AT and AS,ofthe under Infrared Spectrophotometry <2.25>, and compare the sample solution and standard solution at 262 nm as directed spectrum with the Reference Spectrum: both spectra exhibit under Ultraviolet-visible Spectrophotometry <2.24>. similar intensities of absorption at the same wave numbers. Dissolution rate (z) with respect to the labeled amount Purity (1) Sulfate <1.14>—Dissolve 2.0 g of Nicorandil in

of nicomol (C34H32N4O9) 20 mL of dilute ethanol, add 1 mL of dilute hydrochloric ＝ MS × AT/AS × V?/V × 1/C × 18 acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution M : Amount (mg) of nicomol for assay S with 0.40 mL of 0.005 mol/L sulfuric acid VS, 20 mL of C: Labeled amount (mg) of nicomol (C H N O )in1 34 32 4 9 dilute ethanol and 1 mL of dilute hydrochloric acid, and tablet dilute with water to make 50 mL (not more than 0.010z). Assay Weigh accurately not less than 20 Nicomol Tablets (2) Heavy metals <1.07>—Proceed with 2.0 g of Nicoran- and powder. Weigh accurately a portion of the powder, dil according to Method 2, and perform the test. Prepare the equivalent to about 1 g of nicomol (C34H32N4O9), add 100 control solution with 2.0 mL of Standard Lead Solution (not mL of 1 mol/L hydrochloric acid TS, shake well, add water more than 10 ppm). to make exactly 500 mL, and filter. Discard the first 50 mL (3) Related substances—Dissolve 20 mg of Nicorandil in of the filtrate, pipet 2 mL of the subsequent filtrate, add 50 10 mL of the mobile phase, and use this solution as the sam- mL of 1 mol/L hydrochloric acid TS and water to make ex- ple solution. Perform the test with 10 mL of the sample solu- actly 250 mL, and use this solution as the sample solution. tion as directed under Liquid Chromatography <2.01> ac- Separately, weigh accurately about 80 mg of nicomol for cording to the following conditions, and determine each assay, previously dried at 1059C for 4 hours, dissolve in 50 peak area by the automatic integration method: the peak mL of 1 mol/L hydrochloric acid TS, and add water to make area of N-(2-hydroxyethyl)isonicotinamide nitric ester, hav- exactly 100 mL. Pipet 2 mL of this solution, add 20 mL of 1 ing the relative retention time of about 0.86 with respect to mol/L hydrochloric acid TS and water to make exactly 100 nicorandil, is not more than 0.5z of the peak area of mL, and use this solution as the standard solution. Deter- nicorandil, the area of all other peaks is less than 0.1z,and 1160 Nicotinamide / Official Monographs JP XVI the sum area of the peaks other than nicorandil and N-(2- hydroxyethyl)isonicotinamide nitric ester is not more than Nicotinamide 0.25z of the total peak area. Operating conditions— ニコチン酸アミド Detector: An ultraviolet absorption photometer (wavelength: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticlediameter). Column temperature: A constant temperature of about C H N O: 122.12 259C. 6 6 2 Pyridine-3-carboxamide Mobile phase: A mixture of water, tetrahydrofuran, [98-92-0] triethylamine and trifluoroacetic acid (982:10:5:3). Flow rate: Adjust the flow rate so that the retention time Nicotinamide, when dried, contains not less than of nicorandil is about 18 minutes. 98.5z and not more than 102.0z of C H N O. Time span of measurement: About 3 times as long as the 6 6 2 retention time of nicorandil beginning after the solvent peak. Description Nicotinamide occurs as white crystals or crys- System suitability— talline powder. It is odorless, and has a bitter taste. Test for required detectability: Measure exactly 1 mL of It is freely soluble in water and in ethanol (95), and the sample solution, add the mobile phase to make exactly slightly soluble in diethyl ether. 500 mL, and use this solution as the solution for system Identification (1) Mix 5 mg of Nicotinamide with 0.01 g suitability test. Pipet 1 mL of the solution for system suita- of 1-chloro-2,4-dinitrobenzene, heat gently for 5 to 6 bility test, and add the mobile phase to make exactly 20 mL. seconds, and fuse the mixture. Cool, and add 4 mL of potas- Confirm that the peak area of nicorandil obtained with 10 sium hydroxide-ethanol TS: a red color is produced. mL of this solution is equivalent to 2 to 8z of that with 10 (2) To 0.02 g of Nicotinamide add 5 mL of sodium mL of the solution for system suitability test. hydroxide TS, and boil carefully: the gas evolved turns System performance: Dissolve 10 mg of N-(2-hydrox- moistened red litmus paper blue. yethyl)isonicotinamide nitric ester in the mobile phase to (3) Dissolve 0.02 g of Nicotinamide in water to make make 100 mL. To 1 mL of this solution add 10 mL of the 1000 mL. Determine the absorption spectrum of the solul- sample solution. When the procedure is run with this solution as directed under Ultraviolet-visible Spectrophotometry tion under the above operating conditions, N-(2-hydrox- <2.24>, and compare the spectrum with the Reference Spec- yethyl)isonicotinamide nitric ester and nicorandil are eluted trum or the spectrum of a solution of Nicotinamide RS pre- in this order with the resolution between these peaks being pared in the same manner as the sample solution: both spec- not less than 3.0. tra exhibit similar intensities of absorption at the same wave- System repeatability: When the test is repeated 6 times lengths. with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia- pH <2.54> Dissolve 1.0 g of Nicotinamide in 20 mL of tion of the peak area of nicorandil is not more than 1.5z. water: the pH of this solution is between 6.0 and 7.5. Water <2.48> Not more than 0.1z (2 g, volumetric titra- Melting point <2.60> 128 – 1319C tion, direct titration). Purity (1) Clarity and color of solution—Dissolve 1.0 g Residue on ignition <2.44> Not more than 0.1z (1 g). of Nicotinamide in 20 mL of water: the solution is clear and colorless. Assay Weigh accurately about 0.3 g of Nicorandil, dissolve (2) Chloride <1.03>—Take 0.5 g of Nicotinamide, and in 30 mL of a mixture of acetic anhydride and acetic acid perform the test. Prepare the control solution with 0.30 mL (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid of 0.01 mol/L hydrochloric acid VS (not more than VS (potentiometric titration). Perform a blank determina- 0.021z). tion in the same manner, and make any necessary correction. (3) Sulfate <1.14>—Take 1.0 g of Nicotinamide, and per- Each mL of 0.1 mol/L perchloric acid VS form the test. Prepare the control solution with 0.40 mL of

＝ 21.12 mg of C8H9N3O4 0.005 mol/L sulfuric acid VS (not more than 0.019z). (4) Heavy metals <1.07>—Proceed with 1.0 g of Nicotin- Containers and storage Containers—Tight containers. amide according to Method 1, and perform the test. Prepare Storage—At a temperature between 29Cand89C. the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). (5) Readily carbonizable substances <1.15>—Take 0.20 g of Nicotinamide, and perform the test. The solution has no more color than Matching Fluid A. Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 25 mg each of Nicotinamide JP XVI Official Monographs / Nicotinic Acid 1161 and Nicotinamide RS, both previously dried, dissolve sepa- It dissolves in sodium hydroxide TS and in sodium car- rately in 3 mL of water, and add the mobile phase to make bonate TS. exactly 100 mL. Pipet 8 mL each of these solutions, and add Identification (1) Triturate 5 mg of Nicotinic Acid with the mobile phase to make exactly 50 mL. Pipet 5 mL each of 0.01 g of 1-chloro-2,4-dinitrobenzene, and fuse the mixture these solutions, add exactly 5 mL of the internal standard so- by gentle heating for 5 to 6 seconds. Cool, and add 4 mL of lution, and use these solutions as the sample solution and the potassium hydroxide-ethanol TS: a dark red color is pro- standard solution, respectively. Perform the test with 20 mL duced. each of the sample solution and standard solution as directed (2) Dissolve 0.02 g of Nicotinic Acid in water to make under Liquid Chromatography <2.01> according to the fol- 1000 mL. Determine the absorption spectrum of the solution lowing conditions, and calculate the ratios, Q and Q ,of T S as directed under Ultraviolet-visible Spectrophotometry the peak area of nicotinamide to that of the internal stand- <2.24>, and compare the spectrum with the Reference Spec- ard. trum or the spectrum of a solution of Nicotinic Acid RS

Amount (g) of nicotinamide (C6H6N2O) ＝ MS × QT/QS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same M : Amount (mg) of dried Nicotinamide RS S wavelengths. Internal standard solution—A solution of nicotinic acid (1 in pH <2.54> Dissolve 0.20 g of Nicotinic Acid in 20 mL of 25,000). water: the pH of this solution is between 3.0 and 4.0. Operating conditions— Detector: An ultraviolet absorption photometer (wave- Melting point <2.60> 234 – 2389C length: 254 nm). Purity (1) Clarity and color of solution—Dissolve 0.20 g Column: A stainless steel column 4.6 mm in inside diame- of Nicotinic Acid in 20 mL of water: the solution is clear and ter and 25 cm in length, packed with octadecylsilanized silica colorless. gel for liquid chromatography (5 mm in particle diameter). (2) Chloride <1.03>—Perform the test with 0.5 g of Nico- Column temperature: A constant temperature of about tinic Acid. Prepare the control solution with 0.30 mL of 0.01 259C. mol/L hydrochloric acid VS (not more than 0.021z). Mobile phase: Dissolve 1 g of sodium 1-heptane sulfonate (3) Sulfate <1.14>—Dissolve 1.0 g of Nicotinic Acid in 3 in water to make 1000 mL. To 700 mL of this solution add mL of dilute hydrochloric acid and water to make 50 mL. 300 mL of methanol. Perform the test using this solution as the test solution. Pre- Flow rate: Adjust the flow rate so that the retention time pare the control solution with 0.40 mL of 0.005 mol/L sulfu- of nicotinamide is about 7 minutes. ric acid VS and 3 mL of dilute hydrochloric acid, and dilute System suitability— with water to make 50 mL (not more than 0.019z). System performance: When the procedure is run with 20 (4) Nitro compounds—Dissolve 1.0 g of Nicotinic Acid mL of the standard solution under the above operating con- in 8 mL of sodium hydroxide TS, and add water to make 20 ditions, nicotinic acid and nicotinamide are eluted in this mL: the solution has no more color than Matching Fluid A. order with the resolution between these peaks being not less (5) Heavy metals <1.07>—Proceed with 1.0 g of Nicotinic than 5. Acid according to Method 2, and perform the test. Prepare System repeatability: When the test is repeated 6 times the control solution with 2.0 mL of Standard Lead Solution with 20 mL of the standard solution under the above operat- (not more than 20 ppm). ing conditions, the relative standard deviation of the ratio of the peak area of nicotinamide to that of the internal stand- Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, ard is not more than 1.0z. 1 hour). Containers and storage Containers—Tight containers. Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.3 g of Nicotinic Acid, previously dried, dissolve in 50 mL of water, and titrate <2.50> Nicotinic Acid with 0.1 mol/L sodium hydroxide VS (indicator: 5 drops of phenolphthalein TS). ニコチン酸 Each mL of 0.1 mol/L sodium hydroxide VS

＝ 12.31 mg of C6H5NO2 Containers and storage Containers—Well-closed containers.

C6H5NO2: 123.11 Pyridine-3-carboxylic acid [59-67-6]

Nicotinic Acid, when dried, contains not less than 99.5z of C6H5NO2. Description Nicotinic Acid occurs as white crystals or crystalline powder. It is odorless, and has a slightly acid taste. It is sparingly soluble in water, slightly soluble in ethanol (95), and very slightly soluble in diethyl ether. 1162 Nicotinic Acid Injection / Official Monographs JP XVI

Amount (mg) of nicotinic acid (C6H5NO2) Nicotinic Acid Injection ＝ MS × QT/QS M : Amount (mg) of Nicotinic Acid RS ニコチン酸注射液 S Internal standard solution—A solution of caffeine in the mobile phase (1 in 1000). Nicotinic Acid Injection is an aqueous solution for Operating conditions— injection. Detector: An ultraviolet absorption photometer (wave- It contains not less than 95.0z and not more length: 260 nm). than 110.0z of the labeled amount of nicotinic acid Column: A stainless steel column 4.6 mm in inside diame- (C H NO : 123.11). 6 5 2 ter and 15 cm in length, packed with octadecylsilanized silica Method of preparation Prepare as directed under Injec- gel for liquid chromatography (5 mminparticlediameter). tions, with Nicotinic Acid. It may contain Sodium Car- Column temperature: A constant temperature of about bonate or Sodium Hydroxide as a solubilizer. 359C. Mobile phase: Dissolve 1.1 g of sodium 1-octane sulfonate Description Nicotinic Acid Injection is a clear, colorless in a mixture of 0.05 mol/L sodium dihydrogenphosphate liquid. TS, pH 3.0 and methanol (4:1) to make 1000 mL. pH:5.0–7.0 Flow rate: Adjust the flow rate so that the retention time Identification (1) To a volume of Nicotinic Acid Injec- of caffeine is about 9 minutes. tion, equivalent to 0.1 g of Nicotinic Acid according to the System suitability— labeled amount, add 0.3 mL of dilute hydrochloric acid, and System performance: When the procedure is run with 10 evaporate on a water bath to 2 mL. After cooling, collect the mL of the standard solution under the above operating con- crystals formed, wash with small portions of ice-cold water ditions, nicotinic acid and the internal standard are eluted in until the last washing shows no turbidity on the addition of this order with the resolution between these peaks being not silver nitrate TS, and dry at 1059C for 1 hour: the crystals less than 10. melt <2.60> between 2349Cand2389C. With the crystals, System repeatability: When the test is repeated 6 times proceed as directed in the Identification (1) under Nicotinic with 10 mL of the standard solution under the above operat- Acid. ing conditions, the relative standard deviation of the ratios (2) Dissolve 0.02 g of the dried crystals obtained in (1) in of the peak area of nicotinic acid to that of the internal water to make 1000 mL, and determine the absorption spec- standard is not more than 1.0z. trum as directed under Ultraviolet-visible Spectrophotome- Containers and storage Containers—Hermetic containers. try <2.24>: it exhibits a maximum between 261 nm and 263 nm, and a minimum between 235 nm and 239 nm. Sepa- rately, determine the absorbances of this solution, A1 and Nifedipine A2, at each wavelength of maximum and minimum absorption, respectively: the ratio A /A is between 0.35 and 0.39. 2 1 ニフェジピン Bacterial endotoxins <4.01> Less than 3.0 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the requirement.

Sterility <4.06> Perform the test according to the Mem- C17H18N2O6: 346.33 brane filtration method: it meets the requirement. Dimethyl 2,6-dimethyl-4-(2-nitrophenyl)-1,4- dihydropyridine-3,5-dicarboxylate Assay Measure exactly a volume of Nicotinic Acid Injec- [21829-25-4] tion, equivalent to about 0.1 g of nicotinic acid (C6H5NO2), and add the mobile phase to make exactly 100 mL. Pipet 10 Nifedipine contains not less than 98.0z and not mL of this solution, add exactly 10 mL of the internal stand- more than 102.0z of C H N O ,calculatedonthe ard solution, then add the mobile phase to make 100 mL, 17 18 2 6 dried basis. and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of Nicotinic Acid RS, previ- Description Nifedipine occurs as a yellow, crystalline pow- ously dried at 1059C for 1 hour, and dissolve in the mobile der. It is odorless and tasteless. phase to make exactly 100 mL. Pipet 10 mL of this solution, It is freely soluble in acetone and in dichloromethane, add exactly 10 mL of the internal standard solution, then sparingly soluble in methanol, in ethanol (95) and in acetic add the mobile phase to make 100 mL, and use this solution acid (100), slightly soluble in diethyl ether, and practically as the standard solution. Perform the test with 10 mLeachof insoluble in water. the sample solution and standard solution as directed under It is affected by light. Liquid Chromatography <2.01> according to the following Identification (1) Dissolve 0.05 g of Nifedipine in 5 mL conditions, and calculate the ratios, Q and Q , of the peak T S of ethanol (95), and add 5 mL of hydrochloric acid and 2 g area of nicotinic acid to that of the internal standard. JP XVI Official Monographs / Nilvadipine 1163 of zinc powder. Allow to stand for 5 minutes, and filter. ethyl acetate (3:2) to a distance of about 10 cm, and air-dry Perform the test with the filtrate as directed under Qualita- the plate. Examine under ultraviolet light (main wavelength: tive Tests <1.09> for primary aromatic amines: a red-purple 254 nm): the spot from the sample solution, corresponding color develops. to that from the standard solution, is not more intense than (2) Determine the absorption spectrum of a solution of the spot from the standard solution. Nifedipine in methanol (1 in 100,000) as directed under Ul- Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, traviolet-visible Spectrophotometry <2.24>, and compare the 2 hours). spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Residue on ignition <2.44> Not more than 0.1z (1 g). (3) Determine the infrared absorption spectrum of Assay The procedure should be performed under protec- Nifedipine, previously dried, as directed in the potassium tion from direct sunlight in light-resistant vessels. Weigh ac- bromide disk method under Infrared Spectrophotometry curately about 0.12 g of Nifedipine, and dissolve in metha- <2.25>, and compare the spectrum with the Reference Spec- nol to make exactly 200 mL. Measure exactly 5 mL of this trum: both spectra exhibit similar intensities of absorption at solution, and add methanol to make exactly 100 mL. Deter- the same wave numbers. mine the absorbance A of this solution at the wavelength of Melting point <2.60> 172–1759C. maximum absorption at about 350 nm as directed under Ul- traviolet-visible Spectrophotometry <2.24>. Purity (1) Clarity and color of solution—Dissolve 0.5 g of Nifedipine in 5 mL of acetone: the solution is clear and Amount (mg) of C17H18N2O6 ＝ A/142.3 × 40,000 yellow. Containers and storage Containers—Tight containers. (2) Chloride <1.03>—To 2.5 g of Nifedipine add 12 mL Storage—Light-resistant. of dilute acetic acid and 13 mL of water, and heat to boil. After cooling, filter, and discard the first 10 mL of the filtrate. To 5 mL of the subsequent filtrate add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test Nilvadipine using this solution as the test solution. Prepare the control ニルバジピン solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). (3) Sulfate <1.14>—To 4 mL of the filtrate obtained in (2) add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.45 mL of 0.005 mol/L sulfuric acid VS (not more than 0.054z). (4) Heavy metals <1.07>—Proceed with 2.0 g of Nifedi- pine according to Method 2, and perform the test. Prepare C19H19N3O6: 385.37 the control solution with 2.0 mL of Standard Lead Solution 3-Methyl 5-(1-methylethyl) (4RS)-2-cyano-6-methyl- (not more than 10 ppm). 4-(3-nitrophenyl)-1,4-dihydropyridine-3,5-dicarboxylate (5) Arsenic <1.11>—Prepare the test solution with 1.0 g [75530-68-6] of Nifedipine according to Method 3, and perform the test (not more than 2 ppm). Nilvadipine contains not less than 98.0z and not (6) Basic substances—The procedure should be per- more than 102.0z of C19H19N3O6. formed under protection from direct sunlight in light- Description Nilvadipine occurs as a yellow crystalline pow- resistant vessels. Dissolve 5.0 g of Nifedipine in 80 mL of a der. mixture of acetone and acetic acid (100) (5:3), and titrate It is freely soluble in acetonitrile, soluble in methanol, <2.50> with 0.02 mol/L perchloric acid VS (potentiometric sparingly soluble in ethanol (99.5), and practically insoluble titration). Perform a blank determination, and make any in water. necessary correction. Not more than 1.9 mL of 0.02 mol/L A solution of Nilvadipine in acetonitrile (1 in 20) shows no perchloric acid VS is consumed. optical rotation. (7) Dimethyl-2,6-dimethyl-4-(2-nitrosophenyl)-3,5- pyridinedicarboxylate—The procedure should be performed Identification (1) Determine the absorption spectrum of a under protection from direct sunlight in light-resistant ves- solution of Nilvadipine in ethanol (99.5) (1 in 100,000) as di- sels. Dissolve 0.15 g of Nifedipine in dichloromethane to rected under Ultraviolet-visible Spectrophotometry <2.24>, make exactly 10 mL, and use this solution as the sample so- and compare the spectrum with the Reference Spectrum or lution. Separately, dissolve 10 mg of dimethyl 2,6-dimethyl- the spectrum of a solution of Nilvadipine RS prepared in the 4-(2-nitrosophenyl)-3,5-pyridine-dicarboxylate for thin-layer same manner as the sample solution: both spectra exhibit chromatography in exactly 10 mL of dichloromethane. similar intensities of absorption at the same wavelengths. Measure exactly 1 mL of this solution, add dichloromethane (2) Determine the infrared absorption spectrum of Nil- to make exactly 20 mL, and use this solution as the standard vadipine as directed in the potassium bromide disk method solution. Perform the test with these solutions as directed under Infrared Spectrophotometry <2.25>, and compare the under Thin-layer Chromatography <2.03>. Spot 10 mLeach spectrum with the Reference Spectrum or the spectrum of of the sample solution and standard solution on a plate of Nilvadipine RS: both spectra exhibit similar intensities of silica gel with fluorescent indicator for thin-layer chromatog- absorption at the same wave numbers. raphy. Develop the plate with a mixture of cyclohexane and 1164 Nilvadipine Tablets / Official Monographs JP XVI

Melting point <2.60> 167–1719C ratios, QT and QS, of the peak area of nilvadipine to that of the internal standard. Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of

Nilvadipine according to Method 2, and perform the test. Amount (mg) of C19H19N3O6 ＝ MS × QT/QS Prepare the control solution with 2.0 mL of Standard Lead M : Amount (mg) of Nilvadipine RS Solution (not more than 10 ppm). S (2) Related substances—Dissolve 20 mg of Nilvadipine in Internal standard solution—A solution of acenaphthene in 20 mL of acetonitrile, and use this solution as the sample so- methanol (1 in 200). lution. Perform the test with 5 mL of the sample solution as Operating conditions— directed under Liquid Chromatography <2.01> according to Detector: An ultraviolet absorption photometer (wave- the following conditions. Determine each peak area by the length: 254 nm). automatic integration method, and calculate the amount of Column: A stainless steel column 4 mm in inside diameter them by the area percentage method: the amount of each and 15 cm in length, packed with octadecylsilanized silica gel related substance is not more than 0.3z, and the total of for liquid chromatography (5 mm in particle diameter). them is not more than 0.5z. Column temperature: A constant temperature of about Operating conditions— 259C. Detector: An ultraviolet absorption photometer (wave- Mobile phase: Dissolve 2.5 g of diammonium hydrogen length: 240 nm). phosphate in 1000 mL of water, add 10 mL of tetrabutylam- Column: A stainless steel column 4.6 mm in inside diame- monium hydoxide TS, adjust the pH to 7.0 with diluted ter and 15 cm in length, packed with octadecylsilanized silica phosphoric acid (1 in 10), and add 900 mL of acetonitrile. gel for liquid chromatography (5 mm in particle diameter). Flow rate: Adjust the flow rate so that the retention time Column temperature: A constant temperature of about of nilvadipine is about 12 minutes. 259C. System suitability— Mobile phase: A mixture of phosphate buffer solution, System performance: When the procedure is run with 5 mL pH 7.4, methanol and acetonitrile (32:27:18). of the standard solution under the above operating condi- Flow rate: Adjust the flow rate so that the retention time tions, nilvadipine and the internal standard are eluted in this of nilvadipine is about 12 minutes. order with the resolution between these peaks being not less Time span of measurement: About 2.5 times as long as the than 8. retention time of nilvadipine beginning after the solvent System repeatability: When the test is repeated 6 times peak. with 5 mL of the standard solution under the above operating System suitability— conditions, the relative standard deviation of the ratio of the Test for required detectability: Pipet 1 mL of the sample peak area of nilvadipine to that of the internal standard is solution, add acetonitrile to make exactly 100 mL, and use not more than 1.0z. this solution as the solution for system suitability test. Pipet Containers and storage Containers—Well-closed contain- 1 mL of the solution for system suitability test, and add ers. acetonitrile to make exactly 10 mL. Confirm that the peak area of nilvadipine obtained from 5 mL of this solution is equivalent to 7 to 13z of that from 5 mL of the solution for system suitability test. Nilvadipine Tablets System performance: When the procedure is run with 5 mL ニルバジピン錠 of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of nilvadipine is not less than Nilvadipine Tablets contain not less than 93.0z and 3300 and not more than 1.3, respectively. not more than 107.0z of the labeled amount of nil- System repeatability: Pipet 1 mL of the solution for sys- vadipine (C19H19N3O6: 385.37). tem suitability test, and add acetonitrile to make exactly 10 Method of preparation Prepare as directed under Tablets, mL. When the test is repeated 6 times with 5 mL of this solu- with Nilvadipine. tion under the above operating conditions, the relative standard deviation of the peak area of nilvadipine is not more Identification To a quantity of powdered Nilvadipine than 1.5z. Tablets, equivalent to 1 mg of Nilvadipine according to the labeled amount, add 100 mL of ethanol (99.5), shake for 10 Loss on drying <2.41> Not more than 0.1z (1 g, 1059C, minutes, centrifuge, and use the supernatant liquid as the 2 hours). sample solution. Determine the absorption spectrum of the Residue on ignition <2.44> Not more than 0.1z (1 g). sample solution as directed under Ultraviolet-visible Spectro- photometry <2.24>: it exhibits a maximum between 239 nm Assay Weigh accurately about 25 mg each of Nilvadipine and 243 nm and a maximum having a broad-ranging absorp- and Nilvadipine RS, dissolve in methanol to make exactly 25 tion between 371 nm and 381 nm. mL. Pipet 10 mL each of these solutions, add exactly 20 mL of the internal standard solution, 20 mL of water and metha- Uniformity of dosage units <6.02> Perform the test accord- nol to make 100 mL, and use these solutions as the sample ing to the following method: it meets the requirement of the solution and the standard solution, respectively. Perform the Content uniformity test. test with 5 mL each of the sample solution and standard solu- To 1 tablet of Nilvadipine Tablets add V mL of a mixture tion as directed under the Liquid Chromatography <2.01> of acetonitrile and water (7:3) so that each mL of the solu- according to the following conditions, and calculate the tion contains about 0.2 mg of nilvadipine (C19H19N3O6), add JP XVI Official Monographs / Nilvadipine Tablets 1165 exactly V mL of the internal standard solution, and disperse not more than 1.5, respectively. the particles with the aid of ultrasonic waves. Centrifuge for System repeatability: When the test is repeated 6 times 10 minutes, and use the supernatant liquid as the sample so- with 20 mL of the standard solution under the above operat- lution. Separately, weigh accurately about 20 mg of Nilvadi- ing conditions, the relative standard deviation of the peak pine RS, dissolve in the mixture of acetonitrile and water area of nilvadipine is not more than 1.5z. (7:3) to make exactly 20 mL. Pipet 5 mL of this solution, Assay Weigh accurately not less than 20 Nilvadipine add exactly 25 mL of the internal standard solution and the Tablets, and powder. Weigh accurately an amount of mixture of acetonitrile and water (7:3) to make 50 mL, and the powder, equivalent to about 5 mg of nilvadipine use this solution as the standard solution. Proceed as di- (C H N O ), add 10 mL of a mixture of acetonitrile and rected in the Assay. 19 19 3 6 water (7:3) and exactly 25 mL of the internal standard solu-

Amount (mg) of nilvadipine (C19H19N3O6) tion, shake for 15 minutes, and add the mixture of aceto- ＝ MS × QT/QS × V/100 nitrile and water (7:3) to make 50 mL. Centrifuge, and use the supernatant liquid as the sample solution. Separately, M : Amount (mg) of Nilvadipine RS S weigh accurately about 20 mg of Nilvadipine RS, dissolve in Internal standard solution—A solution of acenaphthene in the mixture of acetonitrile and water (7:3) to make exactly 20 acetonitrile (1 in 500). mL. Pipet 5 mL of this solution, add exactly 25 mL of the internal standard solution and the mixture of acetonitrile Dissolution <6.10> When the test is performed at 50 revolu- and water (7:3) to make 50 mL, and use this solution as the tions per minute according to the Paddle method, using 900 standard solution. Perform the test with 5 mLeachofthe mL of water as the dissolution medium, the dissolution rate sample solution and standard solution as directed under in 30 minutes of Nilvadipine Tablets is not less than 85z. Liquid Chromatography <2.01> according to the following Start the test with 1 tablet of Nilvadipine Tablets, conditions, and calculate the ratios, Q and Q , of the peak withdraw not less than 20 mL of the medium at the specified T S area of nilvadipine to that of the internal standard. minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard the first Amount (mg) of nilvadipine (C19H19N3O6) 10 mL of the filtrate, pipet 10 mL of the subsequent filtrate, ＝ MS × QT/QS × 1/4 add exactly 1 mL of methanol, and use this solution as the M : Amount (mg) of Nilvadipine RS sample solution. Separately, weigh accurately an amount of S Nilvadipine RS, equivalent to 10 times the labeled amount of Internal standard solution—A solution of acenaphthene in Nilvadipine Tablets, and dissolve in methanol to make ex- acetonitrile (1 in 500). actly 50 mL. Pipet 5 mL of this solution, and add methanol Operating conditions— to make exactly 100 mL. Pipet 1 mL of this solution, add Detector: An ultraviolet absorption photometer (wave- exactly 10 mL of water, and use this solution as the standard length: 254 nm). solution. Perform the test with exactly 20 mLeachofthe Column: A stainless steel column 4 mm in inside diameter sample solution and standard solution as directed under and 15 cm in length, packed with octadecylsilanized silica gel Liquid Chromatography <2.01> according to the following for liquid chromatography (5 mm in particle diameter). conditions, and determine the peak areas, AT and AS,of Column temperature: A constant temperature of about nilvadipine. 259C. Mobile phase: Dissolve 2.5 g of diammonium hydrogen Dissolution rate (z) with respect to the labeled amount phosphate in 1000 mL of water, add 10 mL of tetrabutylam- of nilvadipine (C H N O ) 19 19 3 6 monium hydoxide TS, adjust the pH to 7.0 with diluted ＝ M × A /A × 1/C × 9 S T S phosphoric acid (1 in 10), and add 900 mL of acetonitrile.

MS: Amount (mg) of Nilvadipine RS Flow rate: Adjust the flow rate so that the retention time C: Labeled amount (mg) of nilvadipine (C19H19N3O6)in1 of nilvadipine is about 12 minutes. tablet System suitability— System performance: When the procedure is run with 5 mL Operating conditions— of the standard solution under the above operating condi- Detector: An ultraviolet absorption photometer (wave- tions, nilvadipine and the internal standard are eluted in this length: 242 nm). order with the resolution between these peaks being not less Column: A stainless steel column 4 mm in inside diameter than 8. and 15 cm in length, packed with octadecylsilanized silica gel System repeatability: When the test is repeated 6 times for liquid chromatography (5 mminparticlediameter). with 5 mL of the standard solution under the above operating Column temperature: A constant temperature of about conditions, the relative standard deviation of the ratio of the 259C. peak area of nilvadipine to that of the internal standard is Mobile phase: A mixture of phosphate buffer solution, not more than 1.0z. pH 7.4, methanol and acetonitrile (7:7:6). Flow rate: Adjust the flow rate so that the retention time Containers and storage Containers—Well-closed contain- of nilvadipine is about 5 minutes. ers. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of nilvadipine are not less than 2000 and 1166 Nitrazepam / Official Monographs JP XVI

raphy. Develop the plate with a mixture of nitromethane and Nitrazepam ethyl acetate (17:3) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: ニトラゼパム 254 nm): the spots other than the principal spot from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.4 g of Nitrazepam, previ-

C15H11N3O3: 281.27 ously dried, and dissolve in 40 mL of acetic acid (100). 7-Nitro-5-phenyl-1,3-dihydro-2H-1,4-benzodiazepin-2-one Titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio- [146-22-5] metric titration). Perform a blank determination, and make any necessary correction. Nitrazepam, when dried, contains not less than Each mL of 0.1 mol/L perchloric acid VS 99.0z of C15H11N3O3. ＝ 28.13 mg of C15H11N3O3 Description Nitrazepam occurs as white to yellow crystals Containers and storage Containers—Tight containers. or crystalline powder. It is odorless. Storage—Light-resistant. It is freely soluble in acetic acid (100), soluble in acetone and in chloroform, slightly soluble in methanol, in ethanol (95) and in ethanol (99.5), very slightly soluble in diethyl ether, and practically insoluble in water. Nitrendipine Melting point: about 2279C (with decomposition). ニトレンジピン Identification (1) To 3 mL of a solution of Nitrazepam in methanol (1 in 500) add 0.1 mL of sodium hydroxide TS: a yellow color is produced. (2) To 0.02 g of Nitrazepam add 15 mL of dilute hydrochloric acid, boil for 5 minutes, cool, and filter: the filtrate responds to the Qualitative Tests <1.09> for primary aromatic amines. (3) Neutralize 0.5 mL of the filtrate obtained in (2) with C H N O : 360.36 sodium hydroxide TS, add 2 mL of ninhydrin TS, and heat 18 20 2 6 3-Ethyl 5-methyl (4RS)-2,6-dimethyl-4-(3-nitrophenyl)- on a water bath: a purple color is produced. 1,4-dihydropyridine-3,5-dicarboxylate (4) Determine the absorption spectrum of a solution of [39562-70-4] Nitrazepam in ethanol (99.5) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>,andcompare Nitrendipine, when dried, contains not less than the spectrum with the Reference Spectrum: both spectra 98.5z and not more than 101.0z of C H N O . exhibit similar intensities of absorption at the same wave- 18 20 2 6 lengths. Description Nitrendipine occurs as a yellow crystalline powder. Purity (1) Clarity and color of solution—Dissolve 0.10 g It is soluble in acetonitrile, sparingly soluble in methanol of Nitrazepam in 20 mL of acetone: the solution is clear and and in ethanol (99.5), and practically insoluble in water. pale yellow to light yellow in color. It is gradually colored to brownish yellow by light. (2) Heavy metals <1.07>—Proceedwith1.0gofNitra- A solution of Nitrendipine in acetonitrile (1 in 50) shows zepam according to Method 2, and perform the test. Prepare no optical rotation. the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Identification (1) Determine the absorption spectrum of a (3) Arsenic <1.11>—Prepare the test solution with 1.0 g solution of Nitrendipine in methanol (1 in 80,000) as directed of Nitrazepam according to Method 3, and perform the test under Ultraviolet-visible Spectrophotometry <2.24>,and (not more than 2 ppm). compare the spectrum with the Reference Spectrum: both (4) Related substances—Dissolve 0.25 g of Nitrazepam spectra exhibit similar intensities of absorption at the same in a 10 mL of mixture of methanol and chloroform (1:1), wavelengths. and use this solution as the sample solution. Pipet 1 mL of (2) Determine the infrared absorption spectrum of the sample solution, add a mixture of methanol and chlo- Nitrendipine as directed in the potassium bromide disk roform (1:1) to make exactly 20 mL, pipet 2 mL of this solu- method under Infrared Spectrophotometry <2.25>,andcom- tion, add a mixture of methanol and chloroform (1:1) to pare the spectrum with the Reference Spectrum: both spectra make exactly 50 mL, and use this solution as the standard exhibit similar intensities of absorption at the same wave solution. Perform the test with these solutions as directed numbers. under Thin-layer Chromatography <2.03>. Spot 10 mLeach Melting point <2.60> 157 – 1619C. of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog- Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of JP XVI Official Monographs / Nitrendipine Tablets 1167

Nitrendipine according to Method 4, and perform the test. Residue on ignition <2.44> Not more than 0.1z (1 g). Prepare the control solution with 2.0 mL of Standard Lead Assay Weigh accurately about 0.3 g of Nitrendipine, previ- Solution (not more than 10 ppm). ously dried, dissolve in 60 mL of a solution of sulfuric acid (2) Related substances—Conduct this procedure rapidly in ethanol (99.5) (3 in 100), add 50 mL of water, and titrate using light-resistant vessels. Dissolve 40 mg of Nitrendipine <2.50> with 0.1 mol/L serium (IV) tetraammonium sulfate in 5 mL of acetonitrile, add the mobile phase to make 25 VS until the red-orange color of the solution vanishes (indi- mL, and use this solution as the sample solution. Pipet 1 mL cator: 3 drops of 1,10-phenanthroline TS). Perform a blank of the sample solution, add the mobile phase to make exactly determination in the same manner, and make any necessary 100 mL, and use this solution as the standard solution. Per- correction. form the test immediately with exactly 10 mLeachofthe sample solution and standard solution as directed under Liq- Each mL of 0.1 mol/L serium (IV) tetraammonium uid Chromatography <2.01> according to the following con- sulfate VS ditions. Determine each peak area by the automatic integra- ＝ 18.02 mg of C18H20N2O6 tion method, and calculate the amount of related substances Containers and storage Containers—Tight containers. by the following equation: the amount of a related sub- Storage—Light-resistant. stance, having the relative retention time of about 0.8 with respect to nitrendipine, is not more than 1.0z, a related substance, having the relative retention time of about 1.3, is not more than 0.25z, and other related substances are not more Nitrendipine Tablets than 0.2z, respectively. The total amount of the substances ニトレンジピン錠 other than nitrendipine is not more than 2.0z. Amount (z) of related substance ＝ A /A T S Nitrendipine Tablets contain not less than 93.0z AT: Each peak area other than nitrendipine obtained from and not more than 107.0z of the labeled amount of the sample solution nitrendipine (C18H20N2O6: 360.36). A : Peak area of nitrendipine obtained from the standard S Method of preparation Prepare as directed under Tablets, solution with Nitrendipine. Operating conditions— Identification Shake a quantity of powdered Nitrendipine Detector: An ultraviolet absorption photometer (wave- Tablets, equivalent to 5 mg of Nitrendipine according to the length: 254 nm). labeled amount, with 70 mL of methanol, then add metha- Column: A stainless steel column 6 mm in inside diameter nol to make 100 mL, and centrifuge. To 5 mL of the super- and 15 cm in length, packed with octadecylsilanized silica gel natant liquid add methanol to make 20 mL, and determine for liquid chromatography (5 mminparticlediameter). the absorption spectrum of this solution as directed under Column temperature: A constant temperature of about Ultraviolet-visible Spectrophotometry <2.24>:itexhibits 259C. maxima between 234 nm and 238 nm, and between 350 nm Mobile phase: A mixture of water, tetrahydrofuran and and 354 nm. acetonitrile (14:6:5). Flow rate: Adjust the flow rate so that the retention time Uniformity of dosage units <6.02> Perform the test accord- of nitrendipine is about 12 minutes. ing to the following method: it meets the requirement of the Time span of measurement: About 2.5 times as long as the Content uniformity test. retention time of nitrendipine beginning after the solvent Conduct this procedure using light-resistant vessels. To 1 peak. tablet of Nitrendipine Tablets add 15 mL of diluted aceto- System suitability— nitrile (4 in 5), stir until the tablet is completely disintegrat- Test for required detectability: To exactly 2 mL of the ed, and further stir for 10 minutes. Add diluted acetonitrile standard solution add the mobile phase to make exactly 10 (4 in 5) to make exactly 20 mL, and centrifuge. Pipet V mL mL. Confirm that the peak area of nitrendipine obtained of the supernatant liquid, equivalent to about 1 mg of nitren- with 10 mL of this solution is equivalent to 14 to 26z of that dipine (C18H20N2O6), add exactly 5 mL of the internal stand- with 10 mL of the standard solution. ard solution, then add diluted acetonitrile (4 in 5) to make 25 System performance: Dissolve 10 mg of Nitrendipine and mL, and use this solution as the sample solution. Proceed as 3 mg of propyl parahydroxybenzoate in 5 mL of acetonitrile, directed in the Assay. and add the mobile phase to make 100 mL. When the proce- Amount (mg) of nitrendipine (C H N O ) dure is run with 5 mL of this solution under the above operat- 18 20 2 6 ＝ M × Q /Q × 1/V × 1/5 ing conditions, propyl parahydroxybenzoate and nitrendi- S T S pine are eluted in this order with the resolution between these MS: Amount (mg) of nitrendipine for assay peaks being not less than 6. Internal standard solution—A solution of propyl parahy- System repeatability: When the test is repeated 6 times droxybenzoate in diluted acetonitrile (4 in 5) (1 in 10,000). with 10 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak Dissolution <6.10> When the test is performed at 100 revo- area of nitrendipine is not more than 2.0z. lutions per minute according to the Paddle method, using 900 mL of the dissolution medium containing 3 g of polysor- Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, bate 80 in 5 L of water for 5-mg tablet and the dissolution 2 hours). medium containing 3 g of polysorbate 80 in 2000 mL of 1168 Nitrogen / Official Monographs JP XVI water for 10-mg tablet, the dissolution rate in 45 minutes of diluted acetonitrile (4:5) to make exactly 200 mL. Pipet 4 mL Nitrendipine Tablets is not less than 70z. of this solution, add exactly 10 mL of the internal standard Conduct this procedure using light-resistant vessels. Start solution and diluted acetonitrile (4:5) to make 50 mL, and the test with 1 tablet of Nitrendipine Tablets, withdraw not use this solution as the standard solution. Perform the test less than 20 mL of the medium at the specified minute after with 10 mL each of the sample solution and standard solution starting the test, and filter through a membrane filter with a as directed under Liquid Chromatography <2.01> according pore size not exceeding 0.45 mm. Discard the first 10 mL of to the following conditions, and calculate the ratios, QT and the filtrate, pipet the subsequent V mL, add the dissolution QS, of the peak area of nitrendipine to that of the internal medium to make exactly V?mL so that each mL contains standard. about 5.6 mg of nitrendipine (C H N O ) according to the 18 20 2 6 Amount (mg) of nitrendipine (C H N O ) labeled amount, and use this solution as the sample solution. 18 20 2 6 ＝ M × Q /Q × 1/50 Separately, weigh accurately about 28 mg of nitrendipine for S T S assay, previously dried at 1059C for 2 hours, dissolve in MS: Amount (mg) of nitrendipine for assay methanol to make exactly 100 mL, then pipet 5 mL of this Internal standard solution—A solution of propyl parahy- solution, and add the dissolution medium to make exactly 50 droxybenzoate in diluted acetonitrile (4 in 5) (1 in 10,000). mL. Pipet 5 mL of this solution, add the dissolution medium Operating conditions— to make exactly 25 mL, and use this solution as the standard Detector: An ultraviolet absorption photometer (wave- solution. Perform the test with exactly 20 mLeachofthe length: 254 nm). sample solution and standard solution as directed under Column: A stainless steel column 6 mm in inside diameter Liquid Chromatography <2.01> according to the following and 15 cm in length, packed with octadecylsilanized silica gel conditions, and determine the peak areas, A and A ,of T S for liquid chromatography (5 mm in particle diameter). nitrendipine. Column temperature: A constant temperature of about Dissolution rate (z) with respect to the labeled amount 259C.

of nitrendipine (C18H20N2O6) Mobile phase: A mixture of water, tetrahydrofuran and ＝ MS × AT/AS × V?/V × 1/C × 18 acetonitrile (14:6:5). Flow rate: Adjust the flow rate so that the retention time M : Amount (mg) of nitrendipine for assay S of nitrendipine is about 12 minutes. C: Labeled amount (mg) of nitrendipine (C H N O )in1 18 20 2 6 System suitability— tablet System performance: When the procedure is run with 10 Operating conditions— mL of the standard solution under the above operating con- Detector: An ultraviolet absorption photometer (wave- ditions, the internal standard and nitrendipine are eluted in length: 356 nm). this order with the resolution between these peaks being not Column: A stainless steel column 4.6 mm in inside diame- less than 6. ter and 15 cm in length, packed with octadecylsilanized silica System repeatability: When the test is repeated 6 times gel for liquid chromatography (5 mm in particle diameter). with 10 mL of the standard solution under the above operat- Column temperature: A constant temperature of about ing conditions, the relative standard deviation of the peak 259C. area of nitrendipine is not more than 1.0z. Mobile phase: A mixture of water, tetrahydrofuran and Containers and storage Containers—Tight containers. acetonitrile (14:6:5). Storage—Light-resistant. Flow rate: Adjust the flow rate so that the retention time of nitrendipine is about 9 minutes. System suitability— System performance: When the procedure is run with 20 Nitrogen mL of the standard solution under the above operating con- 窒素 ditions, the number of theoretical plates and the symmetry factor of the peak of nitrendipine are not less than 5000 and not more than 2.0, respectively. N2: 28.01 System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat- Nitrogen is the nitrogen produced by the air lique- ing conditions, the relative standard deviation of the peak faction separation method. area of nitrendipine is not more than 2.0z. It contains not less than 99.5 volz of N2. Assay Conduct this procedure using light-resistant vessels. Description Nitrogen is a colorless gas at room tempera- To 20 tablets of Nitrendipine Tablets add 150 mL of diluted ture and under atmospheric pressure, and is odorless. acetonitrile (4 in 5), stir until the tablets completely disinte- 1 mL of Nitrogen dissolves in 65 mL of water and in 9 mL grate, and stir for further 10 minutes. Add diluted aceto- of ethanol (95) at 209C and at a pressure of 101.3 kPa. nitrile (4 in 5) to make exactly 200 mL, and centrifuge. Pipet 1000 mL of Nitrogen at 09C and at a pressure of 101.3 a volume of the supernatant liquid, equivalent to about 2 mg kPa weighs 1.251 g. of nitrendipine (C H N O ), add exactly 10 mL of the inter- 18 20 2 6 Identification Introduce 1 mL each of Nitrogen and nitro- nal standard solution and diluted acetonitrile (4 in 5) to gen into a gas-measuring tube or syringe for gas chromatog- make 50 mL, and use this solution as the sample solution. raphy from a cylinder with a pressure-reducing valve, Separately, weigh accurately about 0.1 g of nitrendipine for through a directly connected polyvinyl chloride or stainless assay, previously dried at 1059C for 2 hours, and dissolve in JP XVI Official Monographs / Nitroglycerin Tablets 1169 steel tube. Perform the test with these gases as directed under (C3H5N3O9) according to the labeled amount, shake thor- Gas Chromatography <2.02> according to the following con- oughly with 12 mL of diethyl ether, filter, and use the filtrate ditions: the principal peak obtained form Nitrogen has the as the sample solution. Evaporate 5 mL of the sample solu- same retention time with the peak from nitrogen. tion, dissolve the residue in 1 to 2 drops of sulfuric acid, and Operating conditions— add 1 drop of diphenylamine TS: a deep blue color develops. Proceed as directed in the operating conditions in the (2) Evaporate 5 mL of the sample solution obtained in Assay. (1), add 5 drops of sodium hydroxide TS, heat over a low flame, and concentrate to about 0.1 mL. Cool, heat the Purity Oxygen—The peak area of oxygen obtained from residue with 0.02 g of potassium hydrogen sulfate: the odor Nitrogen in the Assay is not larger than 1/2 times that ob- of acrolein is perceptible. tained from the standard gas mixture. Purity Free nitrate ion—Transfer an accurately measured Assay Introduce 1.0 mL of Nitrogen into a gas-measuring quantity of powdered Nitroglycerin Tablets, equivalent to 20 tube or syringe for gas chromatography from a cylinder with mg of nitroglycerin (C H N O ) according to the labeled a pressure-reducing valve, through a directly connected poly- 3 5 3 9 amount, to a separator, add 40 mL of isopropylether and 40 vinyl chloride or stainless steel tube. Perform the test with mL of water, shake for 10 minutes, and allow the layers to this gas as directed under Gas Chromatography <2.02> ac- separate. Collect the aqueous layer, add 40 mL of isopropyl- cording to the following conditions. Measure the peak area ether, shake for 10 minutes, collect the aqueous layer, filter, A of oxygen. Separately, introduce 1.0 mL of oxygen into T and use the filtrate as the sample solution. Separately, trans- the gas mixer, add carrier gas to make exactly 100 mL, mix fer 10 mL of Standard Nitric Acid Solution to a separator, thoroughly, and use this as the standard gas mixture. Pro- add 30 mL of water and 40 mL of the isopropyl ether layer ceed with 1.0 mL of this mixture in the same manner under of the first extraction of the sample solution, shake for 10 Nitrogen, and measure the peak area A of oxygen. S minutes, continue the procedure in the same manner as the

Amount (volz)ofN2 ＝ 100 － AT/AS sample solution, and use the solution so obtained as the standard solution. Transfer 20 mL each of the sample solu- Operating conditions— tion and the standard solution to Nessler tubes, respectively, Detector: A thermal-conductivity detector. shake well with 30 mL of water and 0.06 g of Griess- Column: A column 3 mm in inside diameter and 3 m in Romijin's nitric acid reagent, allow to stand for 30 minutes, length, packed with zeolite for gas chromatography (250 to and observe the tubes horizontally: the sample solution has 355 mm in particle diameter; 0.5 nm in pore size). no more color than the standard solution. Column temperature: A constant temperature of about 509C. Uniformity fo dosage units <6.02> Perform the test accord- Carrier gas: Hydrogen or helium. ing to the following method: it meets the requirement of the Flow rate: Adjust the flow rate so that the retention time Content uniformity test. of oxygen is about 3 minutes. Transfer 1 tablet of Nitroglycerin Tablets to a glass- System suitability— stoppered centrifuge tube, and add exactly V mL of acetic System performance: Introduce 1.0 mL of oxygen into the acid (100) to provide a solution containing about 30 mgof gas mixer, add Nitrogen to make 100 mL, and mix thor- nitroglycerin (C3H5N3O9) per ml. Shake vigorously for 1 oughly. When the procedure is run with 1.0 mL of this mix- hour, and after disintegrating the tablet, centrifuge, and use ture under the above operating conditions, oxygen and nitro- the supernatant liquid as the sample solution. When the gen are eluted in this order with the resolution between these tablet does not disintegrate during this procedure, transfer 1 peaks being not less than 1.5. tablet of Nitroglycerin Tablets to a glass-stoppered centri- System repeatability: When the test is repeated 5 times fuge tube, wet the tablet with 0.05 mL of acetic acid (100), with 1.0 mL of the standard gas mixture under the above and grind down it with a glass rod. While rinsing the glass conditions, the relative standard deviation of the peak area rod, add acetic acid (100) to make exactly V mL of a solution of oxygen is not more than 2.0z. containing about 30 mgofnitroglycerin(C3H5N3O9)perml. Shake for 1 hour, centrifuge, and use the supernatant liquid Containers and storage Containers—Pressure-resistant as the sample solution. Separately, weigh accurately about cylinders. 90 mg of potassium nitrate, previously dried at 1059Cfor4 Storage—Not exceeding 409C. hours, dissolve in 5 mL of water, and add acetic acid (100) to make exactly 100 mL. Pipet 5 mL of the solution, add acetic acid (100) to make exactly 100 mL, and use this solution as Nitroglycerin Tablets the standard solution. Measure exactly 2 mL each of the sample solution and the standard solution, add 2 mL each of ニトログリセリン錠 salicylic acid TS shake, allow to stand for 15 minutes, and add 10 mL each of water. Render the solution alkaline with Nitroglycerin Tablets contain not less than 80.0z about 12 mL of a solution of sodium hydroxide (2 in 5) while and not more than 120.0z of the labeled amount of cooling in ice, and add water to make exactly 50 mL. Per- nitroglycerin (C3H5N3O9: 227.09). form the test with these solutions as directed under Ultravio- let-visible Spectrophotometry <2.24>, using a solution, pre- Method of preparation Prepare as directed under Tablets, pared with 2 mL of acetic acid (100) in the same manner, as with nitroglycerin. the blank. Determine the absorbances, AT and AS,ofthe Identification (1) Weigh a quantity of powdered subsequent solutions of the sample solution and the standard Nitroglycerin Tablets, equivalent to 6 mg of nitroglycerin solution at 410 nm, respectively. 1170 Nitrous Oxide / Official Monographs JP XVI

Amount (mg) of nitroglycerin (C3H5N3O9) 1000 mL of Nitrous Oxide at 09C and at a pressure of ＝ MS × AT/AS × V/2000 × 0.749 101.3 kPa weighs about 1.96 g.

MS: Amount (mg) of potassium nitrate Identification (1) A glowing splinter of wood held in Nitrous Oxide: it bursts into flame immediately. Calculate the average content from the contents of 10 (2) Transfer 1 mL each of Nitrous Oxide and nitrous tablets: it meets the requirements of the test when each con- oxide directly from metal cylinders with a pressure-reducing tent deviates from the average content by not more than valve to gas measuring tubes or syringes for gas chromatog- 25z. When there is 1 tablet showing a deviation exceeding raphy, using a polyvinyl chloride induction tube. Perform 25z and not exceeding 30z, determine the content of an the test with these gases as directed under Gas Chromatogra- additional 20 tablets in the same manner. Calculate the 30 phy <2.02> according to the conditions of the Assay: the deviations from the new average of all 30 tablets: it meets the retention time of the main peak from Nitrous Oxide coin- requirements of the test when 1 tablet may deviate from the cides with that of nitrous oxide. average content by between 25z and 30z, but no tablet deviates by more than 30z. Purity Maintain the containers of Nitrous Oxide between 189Cand229C for more than 6 hours before the test, and Disintegration <6.09> It meets the requirement, provided correct the volume at 209C and at a pressure of 101.3 kPa. that the time limit of the test is 2 minutes, and the use of the (1) Acidity or alkalinity—To 400 mL of freshly boiled disksisomitted. and cooled water add 0.3 mL of methyl red TS and 0.3 mL Assay Weigh accurately and disintegrate, by soft pressing, of bromothymol blue TS, and boil for 5 minutes. Transfer not less than 20 Nitroglycerin Tablets. Weigh accurately a 50 mL of this solution to each of three Nessler tubes marked portion of the powder, equivalent to about 3.5 mg of A, B and C. Add 0.10 mL of 0.01 mol/L hydrochloric acid nitroglycerin (C3H5N3O9), add exactly 50 mL of acetic acid VS to tube A, 0.20 mL of 0.01 mol/L hydrochloric acid VS (100), shake for 1 hour, filter, and use this filtrate as the to tube B, stopper each of the tubes, and cool. Pass 100 mL sample solution. Separately, weigh accurately about 90 mg of Nitrous Oxide through the solution in tube A for 15 of potassium nitrate, previously dried at 1059C for 4 hours, minutes, employing delivery tube with an orifice approxi- dissolve in 5 mL of water, and add acetic acid (100) to make mately 1 mm in diameter and extending to within 2 mm of exactly 100 mL. Pipet 10 mL of the solution, add acetic acid the bottom of the Nessler tube: the color of the solution in (100) to make exactly 100 mL, and use this solution as the tube A is not deeper orange-red than that of the solution in standard solution. Measure exactly 2 mL each of the sample tube B and not deeper yellow-green than that of the solution solution and the standard solution, to each solution add 2 in tube C. mL of salicylic acid TS, shake, allow to stand for 15 (2) Carbon dioxide—Pass 1000 mL of Nitrous Oxide minutes, and add 10 mL of water. Render the solution alka- through 50 mL of barium hydroxide TS in a Nessler tube, in line with about 12 mL of a solution of sodium hydroxide (2 the same manner as directed in (1): any turbidity produced in 5) while cooling in ice, and add water to make exactly 50 does not exceed that produced in the following control solu- mL. Perform the test with these solutions as directed under tion. Ultraviolet-visible Spectrophotometry <2.24>, using a solu- Control solution: To 50 mL of barium hydroxide TS in a tion, prepared with 2 mL of acetic acid (100) in the same Nessler tube add 1 mL of a solution of 0.1 g of sodium manner, as the blank. Determine the absorbances, AT and hydrogen carbonate in 100 mL of freshly boiled and cooled AS, of the subsequent solutions of the sample solution and water. the standard solution at 410 nm, respectively. (3) Oxidizing substances—Transfer 15 mL of potassium iodide-starchTStoeachoftwo Nessler tubes marked A and Amount (mg) of nitroglycerin (C H N O ) 3 5 3 9 B, add 1 drop of acetic acid (100) to each of the tubes, ＝ M × A /A × 1/20 × 0.749 S T S shake, and use these as solution A and solution B, respec-

MS: Amount (mg) of potassium nitrate tively. Pass 2000 mL of Nitrous Oxide through solution A for 30 minutes in the same manner as directed in (1): the Containers and storage Containers—Tight containers. color of solution A is the same as that of the stoppered, un- Storage—Light-resistant, and not exceeding 209C. treated solution B. (4) Potassium permanganate-reducing substance—Pour 50 mL of water into each of two Nessler tubes marked A and Nitrous Oxide B, add 0.10 mL of 0.02 mol/L potassium permanganate VS to each of the tubes, and use these as solution A and solution 亜酸化窒素 B, respectively. Pass 1000 mL of Nitrous Oxide through solution A in the manner as directed in (1): the color of solu-

N2O: 44.01 tion A is the same as that of solution B. (5) Chloride <1.03>—Pour 50 mL of water into each of Nitrous Oxide contains not less than 97.0 volz of two Nessler tubes marked A and B, add 0.5 mL of silver ni- N2O. trate TS to each of the tubes, shake, and use these as solution A and solution B, respectively. Pass 1000 mL of Nitrous Description Nitrous Oxide is a colorless gas at room tem- Oxide through solution A in the same manner as directed in perature and at atmospheric pressure, and is odorless. (1): the turbidity of solution A is the same as that of solution 1 mL of Nitrous Oxide dissolves in 1.5 mL of water and in B. 0.4 mL of ethanol (95) at 209C and at a pressure of 101.3 (6) Carbon monoxide—Introduce 5.0 mL of Nitrous kPa. It is soluble in diethyl ether and in fatty oils. Oxide into a gas-cylinder or a syringe for gas chromatogra- JP XVI Official Monographs / Nizatidine 1171 phy from a metal cylinder holding gas under pressure and fitted with a pressure-reducing valve, through a directly con- Nizatidine nected polyvinyl tube. Perform the test with this according to the Gas Chromatography <2.02> under the following con- ニザチジン ditions: no peak is observed at the same retention time as that of carbon monoxide. Operating conditions— Detector: A thermal-conductivity detector. Column: A column about 3 mm in inside diameter and about 3 m in length, packed with 300 to 500 mmzeolitefor gas chromatography (0.5 nm in pore size). C12H21N5O2S2:331.46 Column temperature: A constant temperature of about (1EZ)-N-{2-[({2-[(Dimethylamino)methyl]thiazol- 509C. 4-yl}methyl)sulfanyl]ethyl}-N?-methyl-2-nitroethene- Carrier gas: Hydrogen or helium. 1,1-diamine Flow rate: Adjust the flow rate so that the retention time [76963-41-2] of carbon monoxide is about 20 minutes. Selection of column: To 0.1 mL each of carbon monoxide Nizatidine, when dried, contains not less than and air in a gas mixer add carrier gas to make 100 mL, and 98.0z and not more than 101.0z of C12H21N5O2S2. mix well. Proceed with 5.0 mL of the mixed gas under the Description Nizatidine occurs as a white to pale yellowish above operating conditions. Use a column giving well- white crystalline powder, and has a characteristic odor. resolved peaks of oxygen, nitrogen and carbon monoxide in It is soluble in methanol, sparingly soluble in water, and this order. slightly soluble in ethanol (99.5). Detection sensitivity: Adjust the sensitivity so that the peak height of carbon monoxide obtained from 5.0 mL of Identification (1) Determine the absorption spectrum of a the mixed gas used in the selection of column is about 10 cm. solution of Nizatidine in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>,and Assay Withdraw Nitrous Oxide as directed in the Purity. compare the spectrum with the Reference Spectrum or the Introduce 1.0 mL of Nitrous Oxide into a gas-measuring spectrum of a solution of Nizatidine RS prepared in the same tube or syringe for gas chromatography from a metal cylin- manner as the sample solution: both spectra exhibit similar der under pressure through a pressure-reducing valve and a intensities of absorption at the same wavelengths. directly connected polyvinyl tube. Perform the test with this (2) Determine the infrared absorption spectrum of solution as directed under Gas Chromatography <2.02> ac- Nizatidine, previously dried, as directed in the potassium cording to the following conditions, and determine the peak bromide disk method under Infrared Spectrophotometry area A of air. Separately, introduce 3.0 mL of nitrogen into T <2.25>, and compare the spectrum with the Reference Spec- a gas mixer, add carrier gas to make exactly 100 mL, mix trum or the spectrum of dried Nizatidine RS: both spectra thoroughly, and use this as the standard mixed gas. Proceed exhibit similar intensities of absorption at the same wave with 1.0 mL of this mixture as directed in the case of Nitrous numbers. Oxide, and determine the peak area AS of nitrogen in the same manner. Melting point <2.60> 130 – 1359C(afterdrying).

Amount (volz)ofN2O ＝ 100 － 3 × AT/AS Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Nizatidine according to Method 4, and perform the test Operating conditions— using 3 mL of sulfuric acid. Prepare the control solution Detector: A thermal-conductivity detector. with 2.0 mL of Standard Lead Solution (not more than 10 Column: A column about 3 mm in inside diameter and ppm). about 3 m in length, packed with silica gel for gas chroma- (2) Related substances—Dissolve 50 mg of Nizatidine in tography (300 to 500 mminparticlediameter). 10 mL of a mixture of the mobile phase A and mobile phase Column temperature: A constant temperature of about B (19:6), and use this solution as the sample solution. Pipet 3 509C. mL of the sample solution, add the mixture of the mobile Carrier gas: Hydrogen or helium. phase A and mobile phase B (19:6) to make exactly 200 mL, Flow rate: Adjust the flow rate so that the retention time and use this solution as the standard solution. Perform the of nitrogen is about 2 minutes. test with exactly 50 mL each of the sample solution and Selection of column: To 3.0 mL of nitrogen in a gas mixer standard solution as directed under Liquid Chromatography add Nitrous Oxide to make 100 mL, and mix well. Proceed <2.01> according to the following conditions. Determine each with 1.0 mL of the mixed gas under the above operating con- peak area from both solutions by the automatic integration ditions. Use a column giving well-resolved peaks of nitrogen method: the area of the peaks other than nizatidine peak ob- and nitrous oxide in this order. tained from the sample solution is not larger than 1/5 times System repeatability: Repeat thetestfivetimeswiththe the nizatidine peak area from the standard solution. Further- standard mixed gas under the above operating conditions: more,thetotaloftheareasofpeaks other than the nizati- the relative standard deviation of the peak area of nitrogen is dine peak is not larger than the peak area of nizatidine from not more than 2.0z. the standard solution. Containers and storage Containers—Metal cylinders. Operating conditions— Storage—Not exceeding 409C. Detector: An ultraviolet absorption photometer (wavelength: 254 nm). 1172 Nizatidine Capsules / Official Monographs JP XVI

Column: A stainless steel column 4.6 mm in inside diame- Column temperature: A constant temperature of about ter and 25 cm in length, packed with octadecylsilanized silica 409C. gel for liquid chromatography (5 mm in particle diameter). Mobile phase: Dissolve 5.9 g of ammonium acetate in 760 Column temperature: A constant temperature of about mL of water, add 1 mL of diethylamine, and adjust to pH 259C. 7.5 with acetic acid (100). To this solution add 240 mL of Mobile phase A: Dissolve 5.9 g of ammonium acetate in methanol. 760 mL of water, add 1 mL of diethylamine, and adjust to Flow rate: Adjust the flow rate so that the retention time pH 7.5 with acetic acid (100). of nizatidine is about 10 minutes. Mobile phase B: Methanol. System suitability— Flowing of mobile phase: Control the gradient by mixing System performance: When the procedure is run with 10 the mobile phases A and B as directed in the following table. mL of the standard solution under the above operating conditions, the number of theoretical plates and the symmetry Time after injection Mobile phase A Mobile phase B factor of the peak of nizatidine are not less than 5000 and of sample (min) (volz) (volz) not more than 1.5, respectively. System repeatability: When the test is repeated 6 times 0–3 76 24 with 10 mL of the standard solution under the above operat- 3–20 76→ 50 24 → 50 ing conditions, the relative standard deviation of the peak 20–45 50 50 area of nizatidine is not more than 1.0z. Containers and storage Containers—Tight containers. Flow rate: 1.0 mL per minute. Time span of measurement: About 3 times as long as the retention time of nizatidine, beginning after the solvent Nizatidine Capsules peak. System suitability— ニザチジンカプセル Test for required detectability: Pipet 5 mL of the standard solution, and add a mixture of the mobile phase A and mobile phase B (19:6) to make exactly 25 mL. Confirm that the Nizatidine Capsules contain not less than 95.0z and peak area of nizatidine obtained from 50 mL of this solution not more than 105.0z of the labeled amount of nizati- is equivalent to 15 to 25z of that from 50 mL of the stand- dine (C12H21N5O2S2: 331.46). ard solution. Method of preparation Prepare as directed under Cap- System performance: When the procedure is run with 50 sules, with Nizatidine. mL of the standard solution under the above operating conditions, the number of theoretical plates and the symmetry Identification Take out the contents of Nizatidine Cap- factor of the peak of nizatidine are not less than 20,000 and sules, and powder. To a portion of the powder, equivalent to not more than 2.0, respectively. 50 mg of Nizatidine according to the labeled amount, add 50 System repeatability: When the test is repeated 6 times mL of methanol, shake well, and filter. Pipet 1 mL of the with 50 mL of the standard solution under the above operat- filtrate, and add methanol to make 100 mL. Determine the ing conditions, the relative standard deviation of the peak absorption spectrum of the solution as directed under Ultra- area of nizatidine is not more than 2.0z. violet-visible Spectrophotometry <2.24>: it exhibits maxima between 239 nm and 244 nm, and between 323 nm and 327 Loss on drying <2.41> Not more than 0.5z (2 g, 1009C, nm. 1 hour). Uniformity of dosage units <6.02> Perform the test accord- Residue on ignition <2.44> Not more than 0.1z (1 g). ing to the following method: it meets the requirement of the Assay Weigh accurately about 15 mg each of Nizatidine Content uniformity test. and Nizatidine RS, both previously dried, dissolve each in Take out the contents from 1 capsule of Nizatidine Cap- the mobile phase to make exactly 50 mL, and use these sules, add the mobile phase to make V mL so that each mL solutions as the sample solution and the standard solution, contains about 1.5 mg of nizatidine (C12H21N5O2S2). After respectively. Perform the test with exactly 10 mLeachofthe shaking vigorously for 10 minutes, centrifuge. Pipet 10 mL sample solution and standard solution as directed under of the supernatant liquid, add exactly 5 mL of the internal Liquid Chromatography <2.01> according to the following standard solution and add the mobile phase to make 50 mL, and use this solution as the sample solution. Then, proceed conditions. Determine the peak area of nizatidine, AT and as directed in the Assay. AS, from each solution.

Amount (mg) of nizatidine (C12H21N5O2S2) Amount (mg) of C12H21N5O2S2 ＝ MS × AT/AS ＝ MS × QT/QS × V/10 MS: Amount (mg) of Nizatidine RS MS: Amount (mg) of Nizatidine RS Operating conditions— Detector: An ultraviolet absorption photometer (wave- Internal standard solution—A solution of phenol in the mo- length: 254 nm). bile phase (1 in 100). Column: A stainless steel column 4.6 mm in inside diame- Dissolution <6.10> When the test is performed at 50 revolu- ter and 15 cm in length, packed with octadecylsilanized silica tions per minute according to the Paddle method, using a gel for liquid chromatography (5 mm in particle diameter). sinker, using 900 mL of water as the dissolution medium, the JP XVI Official Monographs / Noradrenaline 1173 dissolution rate in 15 minutes of Nizatidine Capsules is not of nizatidine is about 10 minutes. less than 80z. System suitability— Start the test with 1 capsule of Nizatidine Capsules, with- System performance: When the procedure is run with 10 draw not less than 10 mL of the medium at the specified mL of the standard solution under the above operating con- minute after starting the test, and filter through a membrane ditions, the internal standard and nizatidine are eluted in this filter with a pore size not exceeding 0.45 mm. Discard the order with the resolution between these peaks being not less first 2 mL of the filtrate, pipet V mL of the subsequent fil- than 3. trate, and add water to make exactly V?mL so that each mL System repeatability: When the test is repeated 6 times contains about 10 mg of nizatidine (C12H21N5O2S2) according with 10 mL of the standard solution under the above operat- to the labeled amount. Use this solution as the sample solu- ing conditions, the relative standard deviation of the ratio of tion. Separately, weigh accurately about 25 mg of Nizatidine the peak area of nizatidine to that of the internal standard is RS, previously dried at 1009C for 1 hour, and dissolve in not more than 1.0z. water to make exactly 100 mL. Pipet 2 mL of this solution, Containers and storage Containers—Tight containers. add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet- visible Spectrophotometry <2.24>, and determine the absor- Noradrenaline bances, AT and AS,at314nm. Norepinephrine Dissolution rate (z) with respect to the labeled amount

of nizatidine (C12H21N5O2S2) ノルアドレナリン＝ MS × AT/AS × V?/V × 1/C × 36

MS: Amount (mg) of Nizatidine RS C: Labeled amount (mg) of nizatidine (C12H21N5O2S2)in1 capsule Assay Take out the contents of not less than 10 Nizatidine

Capsules, weigh accurately the mass of the contents, and C8H11NO3: 169.18 powder. Weigh accurately a portion of the powder, equiva- 4-[(1RS)-2-Amino-1-hydroxyethyl]benzene-1,2-diol lent to about 0.15 g of nizatidine (C12H21N5O2S2), add ex- [51-41-2] actly 50 mL of the mobile phase, shake vigorously for 10 minutes, and centrifuge. Pipet 5 mL of the supernatant liq- Noradrenaline, when dried, contains not less than uid, add exactly 5 mL of the internal standard solution, add 98.0z of dl-norepinephrine (C8H11NO3). the mobile phase to make 50 mL, and use this solution as the Description Noradrenaline occurs as a white to light brown sample solution. Separately, weigh accurately about 15 mg or slightly reddish brown, crystalline powder. of Nizatidine RS, previously dried at 1009C for 1 hour, dis- It is freely soluble in acetic acid (100), very slightly soluble solve in 30 mL of the mobile phase, add exactly 5 mL of the in water, and practically insoluble in ethanol (95). internal standard solution, add the mobile phase to make 50 It dissolves in dilute hydrochloric acid. mL, and use this solution as the standard solution. Perform It gradually changes to brown by air and by light. the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> Identification (1) Determine the absorption spectrum of a according to the following conditions, and calculate the solution of Noradrenaline in 0.1 mol/L hydrochloric acid TS ratios, QT and QS, of the peak area of nizatidine to that of (3 in 100,000) as directed under Ultraviolet-visible Spectro- the internal standard. photometry <2.24>, and compare the spectrum with the Ref- erence Spectrum: both spectra exhibit similar intensities of Amount (mg) of nizatidine (C H N O S ) 12 21 5 2 2 absorption at the same wavelengths. ＝ M × Q /Q × 10 S T S (2) Determine the infrared absorption spectrum of

MS: Amount (mg) of Nizatidine RS Noradrenaline, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry Internal standard solution—A solution of phenol in the mo- <2.25>, and compare the spectrum with the Reference Spec- bile phase (1 in 100). trum: both spectra exhibit similar intensities of absorption at Operating conditions— the same wave numbers. Detector: An ultraviolet absorption photometer (wavelength: 254 nm). Purity (1) Clarity and color of solution—Dissolve 0.10 g Column: A stainless steel column 4.6 mm in inside diame- of Noradrenaline in 10 mL of 0.1 mol/L hydrochloric acid ter and 15 cm in length, packed with octadecylsilanized silica TS, and add water to make 100 mL: the solution is clear and gel for liquid chromatography (5 mm in particle diameter). colorless. Column temperature: A constant temperature of about (2) Arterenone—Dissolve 50 mg of Noradrenaline in 409C. 0.01 mol/L hydrochloric acid TS to make exactly 100 mL. Mobile phase: Dissolve 5.9 g of ammonium acetate in 760 Determine the absorbance of the solution at 310 nm as di- mL of water, add 1 mL of diethylamine, and adjust to pH rected under Ultraviolet-visible Spectrophotometry <2.24>:it 7.5 with acetic acid (100). To this solution add 240 mL of is not more than 0.1. methanol. (3) Adrenaline—Dissolve 10.0 mg of Noradrenaline in Flow rate: Adjust the flow rate so that the retention time 2.0 mL of diluted acetic acid (100) (1 in 2). Pipet 1 mL of 1174 Noradrenaline Injection / Official Monographs JP XVI this solution, add water to make 10 mL, then mix with 0.3 line Injection, equivalent to 10 mg of Noradrenaline accord- mL of a solution of sodium nitrite (1 in 100), and allow to ing to the labeled amount, add water to make exactly 20 mL, stand for 1 minute: the solution has no more color than the and determine the absorbance of this solution at 310 nm as following control solution. directed under Ultraviolet-visible Spectrophotometry <2.24>: Control solution: Dissolve 2.0 mg of Adrenaline Bitartrate the absorbance is not more than 0.10. RS and 90 mg of Noradrenaline Bitartrate RS in water to (2) Adrenaline—Measure a volume of Noradrenaline In- make exactly 10 mL. Measure exactly 1 mL of this solution, jection, equivalent to 5 mg of Noradrenaline according to add1.0mLofdilutedaceticacid(100)(1in2)andwaterto the labeled amount, add 1 mL of diluted acetic acid (100) make 10 mL, and proceed in the same manner. (1 in 2) and water to make exactly 10 mL, and proceed as directed in the Purity (3) under Noradrenaline. Loss on drying <2.41> Not more than 1.0z (1 g, in vacuum, silica gel, 18 hours). Bacterial endotoxins <4.01> Less than 300 EU/mg. Residue on ignition <2.44> Not more than 0.1z (1 g). Extractable volume <6.05> It meets the requirement. Assay Weigh accurately about 0.3 g of Noradrenaline, pre- Foreign insoluble matter <6.06> Perform the test according viously dried, dissolve in 50 mL of acetic acid for nona- to Method 1: it meets the requirement. queous titration by warming, if necessary, and titrate <2.50> Insoluble particulate matter <6.07> It meets the require- with 0.1 mol/L perchloric acid VS until the color of the solu- ment. tion changes from blue-purple through blue to blue-green (indicator: 2 drops of crystal violet TS). Perform a blank de- Sterility <4.06> Perform the test according to the Mem- termination, and make any necessary correction. brane filtration method: it meets the requirement. Each mL of 0.1 mol/L perchloric acid VS Assay Pipet a volume of Noradrenaline Injection, equiva-

＝ 16.92 mg of C8H11NO3 lent to about 5 mg of dl-noradrenaline (C8H11NO3), add water to make exactly 25 mL, and use this solution as the Containers and storage Containers—Tight containers. sample solution. Separately, weigh accurately about 10 mg Storage—Light-resistant, under nitrogen atmosphere, and of Noradrenaline Bitartrate RS, previously dried in a desic- in a cold place. cator (in vacuum, silica gel) for 24 hours, dissolve in water to make exactly 25 mL, and use this solution as the standard solution. Piper 5 mL each of the sample solution and the Noradrenaline Injection standard solution, add 0.2 mL each of starch TS, then add iodine TS dropwise with swirling until a persistent blue color Noradrenaline Hydrochloride Injection is produced. Add 2 mL of iodine TS, and shake. Adjust the Norepinephrine Hydrochloride Injection pH of the solution to 6.5 with 0.05 mol/L disodium Norepinephrine Injection hydrogenphosphate TS, add 10 mL of phosphate buffer solution, pH 6.5, and shake. Immediately after allowing to ノルアドレナリン注射液 stand for 3 minutes, add sodium thiosulfate TS dropwise until a red-purple color develops, then add water to make ex-

actly 50 mL. Determine the absorbances, AT and AS,ofthe Noradrenaline Injection is an aqueous solution for subsequent solutions of the sample solution and the standard injection. solution at 515 nm within 5 minutes as directed under Ultra- It contains not less than 90.0z and not more than violet-visible Spectrophotometry <2.24>. 110.0z of the labeled amount of dl-noradrenaline Amount (mg) of dl-noradrenaline (C H NO ) (C8H11NO3: 169.18). 8 11 3 ＝ MS × AT/AS × 0.502 Method of preparation Dissolve Noradrenaline in 0.01 mol/L hydrochloric acid TS, and prepare as directed under MS: Amount (mg) of Noradrenaline Bitartrate RS Injections. Containers and storage Containers—Hermetic containers, Description Norepinephrine Injection is a clear, colorless and colored containers may be used. liquid. Storage—Light-resistant. It gradually becomes a pale red color by light and by air. pH:2.3–5.0 Identification Transfer a volume of Noradrenaline Injec- tion, equivalent to 1 mg of Noradrenaline according to the labeled amount, to each of two test tubes A and B, and add 1 mL of water to each tube. Add 10 mL of potassium hydrogen phthalate buffer solution, pH 3.5, to A, and 10 mL of phosphate buffer solution, pH 6.5, to B. To each of these solutions add 1.0 mL of iodine TS, allow to stand for 5 minutes, and add 2.0 mL of sodium thiosulfate TS: no color or a pale red color develops in test tube A, and a deep red- purple color develops in test tube B. Purity (1) Arterenone—Measure a volume of Noradrena- JP XVI Official Monographs / Norfloxacin 1175

Norethisterone Norfloxacin

ノルエチステロンノルフロキサシン

C20H26O2:298.42 C16H18FN3O3: 319.33 17-Hydroxy-19-nor-17a-pregn-4-en-20-yn-3-one 1-Ethyl-6-fluoro-4-oxo-7-(piperazin-1-yl)- [68-22-4] 1,4-dihydroquinoline-3-carboxylic acid [70458-96-7] Norethisterone, when dried, contains not less than 97.0z and not more than 103.0z of C20H26O2. Norfloxacin, when dried, contains not less than 99.0z of C H FN O . Description Norethisterone occurs as a white to pale yel- 16 18 3 3 lowish white crystalline powder. It has no odor. Description Norfloxacin occurs as a white to pale yellow It is sparingly soluble in ethanol (95), in acetone, and in crystalline powder. tetrahydrofuran, slightly soluble in diethyl ether, and very It is freely soluble in acetic acid (100), slightly soluble in slightly soluble in water. ethanol (99.5) and in acetone, very slightly soluble in metha- It is affected by light. nol, and practically insoluble in water. It dissolves in dilute hydrochloric acid TS and in sodium Identification (1) To 2 mg of Norethisterone add 2 mL of hydroxide TS. sulfuric acid: the solution shows a red-brown color and a yel- It is hygroscopic. low-green fluorescence. Add 10 mL of water to this solution It is gradually colored by light. cautiously: a yellow color develops and a yellow-brown precipitate is formed. Identification (1) Dissolve 0.01 g of Norfloxacin in a so- (2) To 25 mg of Norethisterone add 3.5 mL of a solution lution of sodium hydroxide (1 in 250) to make 100 mL. To 5 of 0.05 g of hydroxylammonium chloride and 0.05 g of an- mL of this solution add a solution of sodium hydroxide (1 in hydrous sodium acetate trihydrate in 25 mL of methanol. 250) to make 100 mL. Determine the absorption spectrum of Heat under a reflux condenser on a water bath for 5 hours, this solution as directed under Ultraviolet-visible Spectro- cool, and add 15 mL of water. Collect the precipitate photometry <2.44>, and compare the spectrum with the Ref- formed, wash with 1 to 2 mL of water, recrystallize from erence Spectrum: both spectra exhibit similar intensities of methanol, and dry in a desiccator (in vacuum, silica gel) for absorption at the same wavelengths. 5 hours: the crystals melt <2.60> between 1129C and 1189C. (2) Dissolve a suitable amount of Norfloxacin in a suitable amount of acetone, evaporate the acetone under reduced Optical rotation <2.49> [a]20: －32 – －379(after drying, D pressure, and dry the residue. Determine the infrared ab- 0.25 g, acetone, 25 mL, 100 mm). sorption spectrum of the residue so obtained as directed in Melting point <2.60> 203–2099C. the potassium bromide disk method under Infrared Spectro- photometry <2.25>, and compare the spectrum with the Ref- Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu- erence Spectrum: both spectra exhibit similar intensities of um, silica gel, 4 hours). absorption at the same wave numbers. Residue on ignition <2.44> Not more than 0.1z (0.5 g). Purity (1) Sulfate <1.14>—Dissolve 1.0 g of Norfloxacin Assay Weigh accurately about 0.2 g of Norethisterone, in 7 mL of 0.5 mol/L sodium hydroxide TS and 23 mL of previously dried, dissolve in 40 mL of tetrahydrofuran, add water, and add 1 drop of phenolphthalein TS. Add gradually 10 mL of a solution of silver nitrate (1 in 20), and titrate diluted hydrochloric acid (1 in 3) to this solution until the red <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric color disappears, then add 0.5 mL of dilute hydrochloric titration). Perform a blank determination, and make any acid, and cool in ice for 30 minutes. Filter through a glass necessary correction. filter (G4), and wash the residue with 10 mL of water. Com- bine the filtrate and the washing, and add 1 mL of dilute hy- Each mL of 0.1 mol/L sodium hydroxide VS drochloric acid and water to make 50 mL. Perform the test ＝ 29.84 mg of C H O 20 26 2 using this solution as the test solution. Prepare the control Containers and storage Containers—Tight containers. solution as follows. To 0.50 mL of 0.005 mol/L sulfuric acid Storage—Light-resistant. VS add 7 mL of 0.5 mol/L sodium hydroxide TS and 1 drop of phenolphthalein TS, add diluted hydrochloric acid (1 in 3) until the red color disappears, then add 1.5 mL of dilute hydrochloric acid, 1 or 2 drops of bromophenol blue TS and water to make 50 mL (not more than 0.024z). (2) Heavy metals <1.07>—Proceed with 2.0 g of Norflox- acin according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution 1176 Norgestrel / Official Monographs JP XVI

(not more than 15 ppm). and practically insoluble in water. (3) Arsenic <1.11>—Prepare the test solution with 1.0 g Identification (1) Dissolve 1 mg of Norgestrel in 2 mL of of Norfloxacin according to Method 3, and perform the test ethanol (95), and add 1 mL of sulfuric acid: a red-purple (not more than 2 ppm). color develops. With this solution, examine under ultraviolet (4) Related substances—Conduct this procedure without light (main wavelength: 365 nm): the solution shows a red- exposure to light, using light-resistant vessels. Dissolve orange fluorescence. 0.10 g of Norfloxacin in 50 mL of a mixture of methanol and (2) Determine the infrared absorption spectrum of Nor- acetone (1:1), and use this solution as the sample solution. gestrel, previously dried, as directed in the potassium bro- Pipet 1 mL of the sample solution, add a mixture of metha- mide disk method under Infrared Spectrophotometry <2.25>, nol and acetone (1:1) to make exactly 100 mL. Pipet 2 mL of and compare the spectrum with the Reference Spectrum: this solution, add a mixture of methanol and acetone (1:1) to both spectra exhibit similar intensities of absorption at the make exactly 10 mL, and use this solution as the standard same wave numbers. solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mLeach Melting point <2.60> 206 – 2129C. of the sample solution and standard solution on a plate of Purity (1) Heavy metals <1.07>—Take 1.0 g of Norges- silica gel with fluorescent indicator for thin-layer chromatog- trel, heat gently to carbonize, cool, add 10 mL of a solution raphy (5 – 7 mm in particle diameter). Develop with a mix- of magnesium nitrate hexahydrate in ethanol (95) (1 in 10), ture of methanol, chloroform, toluene, diethylamine and and ignite the ethanol to burn. After cooling, add 1 mL of water (20:20:10:7:4) to a distance of about 9 cm, and air-dry sulfuric acid, proceed with this solution according to Method the plate. Examine under ultraviolet light (main wavelength: 4, and perform the test. Prepare the control solution with 2.0 254 nm and 366 nm): the number of the spot other than the mL of Standard Lead Solution (not more than 20 ppm). principal spot from the sample solution is not more than 2 (2) Related substances—Dissolve 30 mg of Norgestrel in and they are not more intense than the spot from the stand- 5 mL of chloroform, and use this solution as the sample so- ard solution. lution. Pipet 1 mL of the sample solution, add chloroform to Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, make exactly 100 mL, and use this solution as the standard 2 hours). solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mLeach Residue on ignition <2.44> Not more than 0.1z (1 g). of the sample solution and standard solution on a plate of Assay Weigh accurately about 0.5 g of Norfloxacin, previ- silica gel with fluorescent indicator for thin-layer chromatog- ously dried, dissolve in 50 mL of acetic acid (100), and titrate raphy. Develop the plate with a mixture of dichloromethane <2.50> with 0.1 mol/L perchloric acid VS (potentiometric and ethyl acetate (2:1) to a distance of about 10 cm, and air- titration). Perform a blank determination, and make any dry the plate. Examine under ultraviolet light (main wave- necessary correction. length: 254 nm): the spots other than the principal spot from the sample solution are not more intense than the spot from Each mL of 0.1 mol/L perchloric acid VS the standard solution. ＝ 31.93 mg of C16H18FN3O3 Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, Containers and storage Containers—Tight containers. 3 hours). Storage—Light-resistant. Residue on ignition <2.44> Not more than 0.2z (0.5 g). Assay Weigh accurately about 0.2 g of Norgestrel, previ- Norgestrel ously dried, dissolve in 40 mL of tetrahydrofuran, add 10 mL of a solution of silver nitrate (1 in 20), and titrate <2.50> ノルゲストレル with 0.1 mol/L sodium hydroxide VS (potentiometric titration). Perform a blank determination, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS

＝ 31.25 mg of C21H28O2 Containers and storage Containers—Well-closed contain-

C21H28O2:312.45 ers. 13-Ethyl-17-hydroxy-18,19-dinor-17a-pregn-4-en-20-yn- 3-one [6533-00-2]

Norgestrel, when dried, contains not less than 98.0z of C21H28O2. Description Norgestrel occurs as white crystals or crystalline powder. It is soluble in tetrahydrofuran and in chloroform, sparingly soluble in ethanol (95), slightly soluble in diethyl ether, JP XVI Official Monographs / Norgestrel and Ethinylestradiol Tablets 1177

solution as directed under Liquid Chromatography <2.01> Norgestrel and Ethinylestradiol according to the following conditions. Calculate the ratios, QTa and QTb, of the peak areas of norgestrel and ethinyles- Tablets tradiol to the peak area of the internal standard of the sample solution and also the ratios, Q and Q , of the peak ノルゲストレル・エチニルエストラジオール錠 Sa Sb areas of norgestrel and ethinylestradiol to the peak area of the internal standard of the standard solution. Norgestrel and Ethinylestradiol Tablets contain not Amount (mg) of norgestrel (C H O ) less than 90.0z and not more than 110.0z of the 21 28 2 ＝ MSa × QTa/QSa × 1/100 labeled amount of norgestrel (C21H28O2: 312.45) and ethinylestradiol (C20H24O2: 296.40). Amount (mg) of ethinylestradiol (C20H24O2) ＝ M × Q /Q × 1/100 Method of preparation Prepare as directed under Tablets, Sb Tb Sb with Norgestrel and Ethinylestradiol. MSa: Amount (mg) of Norgestrel RS M : Amount (mg) of Ethinylestradiol RS Identification (1) Weigh a quantity of Norgestrel and Sb Ethinylestradiol Tablets, equivalent to 10 mg of Norgestrel Internal standard solution—A solution of diphenyl in according to the labeled amount, previously powdered, add diluted methanol (7 in 10) (1 in 50,000). 10 mL of chloroform, shake for 10 minutes, and filter. To 2 Operating conditions— mL of the filtrate add 6 mL of sodium hydroxide TS, shake Proceed as directed in the operating conditions in the vigorously, and centrifuge. Take 1 mL of the chloroform Assay. layer, evaporate on a water bath to dryness, dissolve the System suitability— residue in 2 mL of ethanol (95), and add 1 mL of sulfuric Proceed as directed in the system suitability in the Assay. acid: a red-purple color develops. Examine under ultraviolet Dissolution <6.10> When the test is performed at 50 revolu- light (main wavelength: 365 nm): this solution shows a red- tions per minute according to the Paddle method, using 900 orange fluorescence (norgestrel). mL of water as the dissolution medium, the dissolution rate (2) Take 1 mL of the filtrate obtained in (1), evaporate in 45 minutes of Norgestrel and Ethinylestradiol Tablets is on a water bath to dryness, add 1 mL of boric acid-methanol not less than 70z. buffer solution to the residue, shake, and cool in ice. Add 1 Start the test with 1 tablet of Norgestrel and Ethinyles- mL of ice-cold diazo TS, shake, add 1 mL of sodium hy- tradiol Tablets, withdraw not less than 50 mL of the medium droxide TS, and shake: a red-orange color develops at the specified minute after starting the test, and filter (ethinylestradiol). through a membrane filter with a pore size not exceeding (3) Use the filtrate obtained in (1) as the sample solution. 0.8 mm. Discard the first 10 mL of the filtrate, pipet exactly Separately, dissolve 10 mg of Norgestrel RS and 1 mg of V mL of the subsequent filtrate, equivalent to about 17 mgof Ethinylestradiol RS, respectively, in 10 mL of chloroform, norgestrel (C H O ) and about 1.7 mg of ethinylestradiol and use these solutions as the standard solution (1) and the 21 28 2 (C H O ), transfer into a chromatography column [pre- standard solution (2). Perform the test with these solutions 20 24 2 pared by packing 0.36 g of octadecylsilanized silica gel for as directed under Thin-layer Chromatography <2.03>. Spot pretreatment (55 to 105 mm in particle diameter) in a tube 20 mL each of the sample solution and standard solutions (1) about 1 cm in inside diameter]. After washing the column and (2) on a plate of silica gel for thin-layer chromatogra- with 15 mL of water, elute with 3 mL of methanol, and phy. Develop the plate with a mixture of 1,2-dichloroethane, evaporate the effluent in a water bath to dryness at about methanol and water (368:32:1) to a distance of about 10 cm, 409C with the aid of a current air. Dissolve the residue in and air-dry the plate. Spray evenly a solution of p-toluene- exactly 2 mL of diluted methanol (7 in 10), and use this solu- sulfonate in ethanol (95) (1 in 5) on the plate, and heat at tion as the sample solution. Separately, weigh accurately 1059C for 5 minutes. Examine under ultraviolet light (main about 25 mg of Norgestrel RS and about 2.5 mg of wavelength: 365nm): two spots from the sample solution Ethinylestradiol RS, dissolve in diluted methanol (7 in 10) to show the similar color tone and Rf value to each spot from make exactly 100 mL, then pipet 3 mL of this solution, add the standard solutions (1) and (2). diluted methanol (7 in 10) to make exactly 100 mL, and use Uniformity of dosage units <6.02> Perform the test accord- this solution as the standard solution. Perform the test with ing to the following method: it meets the requirement of the exactly 50 mL each of the sample solution and standard Content uniformity test. solution as directed under Liquid Chromatography <2.01> Add 2 mL of diluted methanol (7 in 10) to 1 tablet of Nor- according to the following conditions. Determine the peak gestrel and Ethinylestradiol Tablets, add exactly 2 mL of the areas, ATa and ATb, of norgestrel and ethinylestradiol from internal standard solution, shake for 20 minutes, and centri- the sample solution, and the peak areas, ASa and ASb,of fuge. Filter the supernatant liquid through a membrane filter norgestrel and ethinylestradiol from the standard solution. with pore size of not more than 0.2 mm, and use this filtrate Dissolution rate (z) with respect to the labeled amount as the sample solution. Separately, weigh accurately quanti- of norgestrel (C H O ) ties of Norgestrel RS and of Ethinylestradiol RS, equivalent 21 28 2 ＝ M × A /A × 1/V × 1/C × 54 to 100 times each of the labeled amounts, dissolve in diluted Sa Ta Sa a methanol (7 in 10) to make exactly 200 mL. Pipet 2 mL of Dissolution rate (z) with respect to the labeled amount this solution, add exactly 2 mL of the internal standard solu- of ethinylestradiol (C20H24O2) tion, and use this solution as the standard solution. Perform ＝ MSb × ATb/ASb × 1/V × 1/Cb × 54 the test with 20 mL each of the sample solution and standard MSa: Amount (mg) of Norgestrel RS 1178 Nortriptyline Hydrochloride / Official Monographs JP XVI

MSb: Amount (mg) of Ethinylestradiol RS ing conditions, the relative standard deviation of the ratios Ca: Labeled amount (mg) of norgestrel (C21H28O2)in1 of the peak area of ethinylestradiol and norgestrel to that of tablet the internal standard are not more than 1.0z, respectively. C : Labeled amount (mg) of ethinylestradiol (C H O )in b 20 24 2 Containers and storage Containers—Tight containers. 1tablet Operating conditions— Proceed as directed in the operating conditions in the Nortriptyline Hydrochloride Assay. System suitability— ノルトリプチリン塩酸塩 Proceed as directed in the system suitability in the Assay. Assay Weigh accurately not less than 20 Norgestrel and Ethinylestradiol Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 1 mg of norgestrel (C21H28O2), add 4 mL of diluted methanol (7 in 10), add exactly 4 mL of the internal standard solution, shake for 20 C H N.HCl: 299.84 minutes, and centrifuge. Filter the supernatant liquid 19 21 3-(10,11-Dihydro-5H-dibenzo[a,d]cyclohepten-5- through a membrane filter with pore size of not more than ylidene)-N-methylpropylamine monohydrochloride 0.2 mm, and use this filtrate as the sample solution. Sepa- [894-71-3] rately, weigh accurately about 50 mg of Norgestrel RS and about 5 mg of Ethinylestradiol RS, and dissolve in diluted Nortriptyline Hydrochloride, when dried, contains methanol (7 in 10) to make exactly 200 mL. Pipet 4 mL of not less than 98.5z of C H N.HCl. this solution, add exactly 4 mL of the internal standard solu- 19 21 tion, and use this solution as the standard solution. Perform Description Nortriptyline Hydrochloride occurs as a white the test with 20 mL each of the sample solution and standard to yellowish white, crystalline powder. It is odorless, or has a solution as directed under Liquid Chromatography <2.01> faint, characteristic odor. according to the following conditions. Calculate the ratios, It is freely soluble in acetic acid (100) and in chloroform,

QTa and QTb, of the peak areas of norgestrel and ethinyles- soluble in ethanol (95), sparingly soluble in water, and prac- tradiol to the peak area of the internal standard of the sam- tically insoluble indiethylether. ple solution and also the ratios, QSa and QSb, of the peak The pH of a solution of Nortriptyline Hydrochloride (1 in areas of norgestrel and ethinylestradiol to the peak area of 100) is about 5.5. the internal standard of the standard solution. Melting point: 215 – 2209C.

Amount (mg) of norgestrel (C21H28O2) Identification (1) To 5 mL of a solution of Nortriptyline ＝ MSa × QTa/QSa × 1/50 Hydrochloride (1 in 100) add 1 mL of bromine TS: the color of the test solution disappears. Amount (mg) of ethinylestradiol (C H O ) 20 24 2 (2) To 5 mL of a solution of Nortriptyline Hydrochlo- ＝ M × Q /Q × 1/50 Sb Tb Sb ride (1 in 100) add 1 to 2 drops of a solution of quinhydrone

MSa: Amount (mg) of Norgestrel RS in methanol (1 in 40): a red color gradually develops. MSb: Amount (mg) of Ethinylestradiol RS (3) Determine the absorption spectrum of a solution of Nortriptyline Hydrochloride (1 in 100,000) as directed under Internal standard solution—A solution of diphenyl in Ultraviolet-visible Spectrophotometry <2.24>,andcompare diluted methanol (7 in 10) (1 in 50,000). the spectrum with the Reference Spectrum: both spectra Operating conditions— exhibit similar intensities of absorption at the same wave- Detector: Norgestrel—An ultraviolet absorption photome- lengths. ter (wavelength: 241 nm). (4) Determine the infrared absorption spectrum of Nor- Ethinylestradiol—A fluorophotometer (excitation wave- triptyline Hydrochloride, previously dried, as directed in the length: 281 nm, fluorescence wavelength: 305 nm). potassium chloride disk method under Infrared Spectropho- Column: A stainless steel column 4.6 mm in inside diame- tometry <2.25>, and compare the spectrum with the Refer- ter and 25 cm in length, packed with octadecylsilanized silica ence Spectrum: both spectra exhibit similar intensities of ab- gel for liquid chromatography (10 mminparticlediameter). sorption at the same wave numbers. Column temperature: A constant temperature of about (5) A solution of Nortriptyline Hydrochloride (1 in 100) 259C. responds to the Qualitative Tests <1.09> for chloride. Mobile phase: A mixture of acetonitrile and water (11:9). Flow rate: Adjust the flow rate so that the retention time Purity (1) Clarity and color of solution—Dissolve 0.10 g of norgestrel is about 10 minutes. of Nortriptyline Hydrochloride in 10 mL of water: the solu- System suitability— tion is clear and colorless to very light yellow. System performance: When the procedure is run with 20 (2) Heavy metals <1.07>—Proceedwith1.0gofNortrip- mL of the standard solution under the above operating con- tyline Hydrochloride according to Method 2, and perform ditions, ethinylestradiol, norgestrel and the internal standard the test. Prepare the control solution with 2.0 mL of Stand- are eluted in this order, and the resolution between the peaks ard Lead Solution (not more than 20 ppm). of norgestrel and the internal standard is not less than 8. (3) Arsenic <1.11>—Prepare the test solution with 1.0 g System repeatability: When the test is repeated 6 times of Nortriptyline Hydrochloride according to Method 3, and with 20 mL of the standard solution under the above operat- perform the test (not more than 2 ppm). JP XVI Official Monographs / Noscapine 1179

(4) Related substances—Dissolve 0.50 g of Nortriptyline solution of Noscapine in methanol (1 in 20,000) as directed Hydrochloride in 20 mL of chloroform, and use this solution under Ultraviolet-visible Spectrophotometry <2.24>,and as the sample solution. Pipet 2 mL of the sample solution, compare the spectrum with the Reference Spectrum: both and add chloroform to make exactly 100 mL. Pipet 5 mL of spectra exhibit similar intensities of absorption at the same this solution, add chloroform to make exactly 50 mL, and wavelengths. use this solution as the standard solution. Perform the test (2) Determine the infrared absorption spectrum of with these solutions as directed under Thin-layer Chroma- Noscapine, previously dried, as directed in the potassium tography <2.03>. Spot 4 mL each of the sample solution and bromide disk method under the Infrared Spectrophotometry standard solution on a plate of silica gel with fluorescent in- <2.25>, and compare the spectrum with the Reference Spec- dicator for thin-layer chromatography. Develop the plate trum: both spectra exhibit similar intensities of absorption at with a mixture of cyclohexane, methanol and diethylamine the same wave numbers. (8:1:1) to a distance of about 15 cm, and air-dry the plate. Optical rotation <2.49> [a]20: ＋42 – ＋489(after drying, Examine under ultraviolet light (main wavelength: 254 nm): D 0.5 g, 0.1 mol/L hydrochloric acid TS, 25 mL, 100 nm). the spots other than the principal spot from the sample solution are not more intense than the spot from the standard so- Melting point <2.60> 174 – 1779C. lution. Purity (1) Chloride <1.03>—Dissolve 0.7 g of Noscapine Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, in 20 mL of acetone, add 6 mL of dilute nitric acid and water 2 hours). to make 50 mL, and perform the test with this solution. Prepare the control solution as follows: To 0.4 mL of 0.01 Residue on ignition <2.44> Not more than 0.1z (1 g). mol/L hydrochloric acid add 20 mL of acetone, 6 mL of Assay Weigh accurately about 0.5 g of Nortriptyline Hy- dilute nitric acid and water to make 50 mL (not more than drochloride, previously dried, dissolve in 5 mL of acetic acid 0.02z). (100), add 50 mL of acetic anhydride, and titrate <2.50> with (2) Heavy metals <1.07>—Proceed with 2.0 g of Nosca- 0.1 mol/L perchloric acid VS (potentiometric titration). Per- pine according to Method 2, and perform the test. Prepare form a blank determination, and make any necessary correc- the control solution with 2.0 mL of Standard Lead Solution tion. (not more than 10 ppm). (3) Morphine—Dissolve 10 mg of Noscapine in 1 mL of Each mL of 0.1 mol/L perchloric acid VS water and 5 mL of 1-nitroso-2-naphthol TS with shaking, ＝ 29.98 mg of C H N.HCl 19 21 add 2 mL of a solution of potassium nitrate (1 in 10), and Containers and storage Containers—Well-closed contain- warm at 409C for 2 minutes. Add 1 mL of a solution of so- ers. dium nitrite (1 in 5000), and warm at 409C for 5 minutes. Storage—Light-resistant. After cooling, shake the solution with 10 mL of chloroform, centrifuge, and collect the aqueous layer: the solution so obtained has no more color than a pale red. Noscapine (4) Related substances—Dissolve 0.7 g of Noscapine in 50 mL of acetone, and use this solution as the sample solu- Narcotine tion. Pipet 5 mL of the sample solution, add acetone to make exactly 50 mL. Pipet 5 mL of this solution, add ace- ノスカピン tone to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetone, toluene, ethanol (99.5) and ammonia solution (28) (60:60:9:2) to a distance of about 10 cm, and air-dry the plate. Spray evenly dilute bismuth subnitrate-potassium iodide TS for spray on the plate: the spots other than the principal spot from the sample solution C H NO : 413.42 22 23 7 are not more intense than the spot from the standard solu- (3S)-6,7-Dimethoxy-3-[(5R)-4-methoxy-6-methyl- tion. 5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]isoquinolin- 5-yl]isobenzofuran-1(3H)-one Loss on drying <2.41> Not more than 0.5z (2 g, 1059C, [128-62-1] 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Noscapine, when dried, contains not less than 98.5z of C22H23NO7. Assay Weigh accurately about 0.8 g of Noscapine, previously dried, dissolve in 30 mL of acetic acid (100) and titrate Description Noscapine occurs as white crystals or crystal- <2.50> with 0.1 mol/L perchloric acid VS (indicator: 3 drops line powder. It is odorless and tasteless. of crystal violet TS). Perform a blank determination, and It is very soluble in acetic acid (100), slightly soluble in make any necessary correction. ethanol (95) and in diethyl ether, and practically insoluble in water. Each mL of 0.1 mol/L perchloric acid VS ＝ 41.34 mg of C H NO Identification (1) Determine the absorption spectrum of a 22 23 7 1180 Noscapine Hydrochloride Hydrate / Official Monographs JP XVI

Containers and storage Containers—Well-closed contain- chloride Hydrate in 1 mL of water, add 5 mL of 1-nitroso-2- ers. naphthol TS and 2 mL of a solution of potassium nitrate (1 Storage—Light-resistant. in 10), and warm at 409C for 2 minutes. Add 1 mL of a solution of sodium nitrite (1 in 5000), and warm at 409Cfor5 minutes. After cooling, shake the mixture with 10 mL of Noscapine Hydrochloride Hydrate chloroform, centrifuge, and separate the aqueous layer: the solution so obtained has no more color than a pale red color. Narcotine Hydrochloride Loss on drying <2.41> Not more than 9.0z (0.5 g, 1209C, 4 hours). ノスカピン塩酸塩水和物 Residue on ignition <2.44> Not more than 0.5z (1 g). Assay Weigh accurately about 0.5 g of Noscapine Hydro- chloride Hydrate, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS C H NO .HCl.xH O 22 23 7 2 ＝ 44.99 mg of C H NO .HCl (3S)-6,7-Dimethoxy-3-[(5R)-4-methoxy-6-methyl- 22 23 7 5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]isoquinolin- Containers and storage Containers—Well-closed contain- 5-yl]isobenzofuran-1(3H)-one monohydrochloride ers. hydrate Storage—Light-resistant. [912-60-7, anhydride]

Noscapine Hydrochloride Hydrate, when dried, Nystatin contains not less than 98.0z of noscapine hydrochloride C22H23NO7.HCl: 449.88. ナイスタチン Description Noscapine Hydrochloride Hydrate occurs as colorless or white crystals or crystalline powder. It is odor- Nystatin is a mixture of polyene macrolide sub- less, and has a bitter taste. stances having antifungal activity produced by the It is freely soluble in water, in acetic acid (100), and in ace- growth of Streptomyces noursei. tic anhydride, soluble in ethanol (95), and practically insolu- It contains not less than 4600 units (potency) per ble in diethyl ether. mg, calculated on the dried basis. The potency of Nys- tatin is expressed as the unit of nystatin (C H NO : Identification (1) To 1 mg of Noscapine Hydrochloride 47 75 17 926.09), and one unit corresponds to 0.27 mg of nysta- Hydrate add 1 drop of formaldehyde-sulfuric acid TS: a pur- tin (C H NO ). ple color, changing to yellow-brown, is produced. 47 75 17 (2) To 1 mg of Noscapine Hydrochloride Hydrate add 1 Description Nystatin occurs as a white to light yellow- drop of a solution of ammonium vanadate (V) in sulfuric brown powder. acid (1 in 200): an orange color is produced. It is soluble in formamide, sparingly soluble in methanol, (3) Dissolve 0.02 g of Noscapine Hydrochloride Hydrate slightly soluble in ethanol (95), and very slightly soluble in in 1 mL of water, and add 3 drops of sodium acetate TS: a water. white, flocculent precipitate is produced. It dissolves in sodium hydroxide TS. (4) Dissolve 1 mg of Noscapine Hydrochloride Hydrate Identification (1) Dissolve 1 mg of Nystatin in 5 mL of in 1 mL of diluted sulfuric acid (1 in 35), shake with 5 drops water and 1 mL of sodium hydroxide TS, heat for 2 minutes, of a solution of disodium chlomotropate dihydrate (1 in 50), and cool. To this solution add 3 mL of a solution of 4- and add 2 mL of sulfuric acid dropwise: a purple color is aminoacetophenone in methanol (1 in 200) and 1 mL of hy- produced. drochloric acid: a red-purple color develops. (5) Dissolve 0.1 g of Noscapine Hydrochloride Hydrate (2) To 10 mg of Nystatin add 50.25 mL of a mixture of in 10 mL of water, make the solution alkaline with ammonia diluted methanol (4 in 5) and sodium hydroxide TS (200:1), TS, and shake with 10 mL of chloroform. Separate the chlo- heat at not exceeding 509C to dissolve, then add diluted roform layer, wash with 5 mL of water, and filter. Distil methanol (4 in 5) to make 500 mL. Determine the absorption most of the filtrate on a water bath, add 1 mL of ethanol spectrum of this solution as directed under Ultraviolet- (99.5), and evaporate to dryness. Dry the residue at 1059C visible Spectrophotometry <2.24>, and compare the spectrum for 4 hours: the residue so obtained melts <2.60> between with the Reference Spectrum or the spectrum of a solution of 1749C and 1779C. Nystatin RS prepared in the same manner as the sample solu- (6) Make a solution of Noscapine Hydrochloride Hy- tion: both spectra exhibit similar intensities of absorption at drate (1 in 50) alkaline with ammonia TS, and filter the pre- thesamewavelengths. cipitate. Acidify the filtrate with dilute nitric acid: the solution responds to the Qualitative Tests <1.09> (2) for chloride. Purity Heavy metals <1.07>—Proceed with 1.0 g of Nysta- tin according to Method 4, and perform the test. Prepare the Purity Morphine—Dissolve 10 mg of Noscapine Hydro- JP XVI Official Monographs / Ofloxacin 1181 control solution with 2.0 mL of Standard Lead Solution (not A soluton of Ofloxacin in sodium hydroxide TS (1 in 20) more than 20 ppm). does not show optical rotation. It is changed in color by light. Loss on drying <2.41> Not more than 5.0z (0.3 g, in vacu- Melting point: about 2659C (with decomposition). um, 609C, 3 hours). Identification (1) Determine the absorption spectrum of a Assay Perform the test according to the Cylinder-plate solution of Ofloxacin in 0.1 mol/L hydrochloric acid TS (1 method as directed under Microbial Assay for Antibiotics in 150,000) as directed under Ultraviolet-visible Spectropho- <4.02> according to the following conditions. tometry <2.24>, and compare the spectrum with the Refer- (i) Test organism—Saccharomyces cerevisiae ATCC ence Spectrum: both spectra exhibit similar intensities of ab- 9763 sorption at the same wavelengths. (ii) Culture medium—Use the medium 2) Medium for (2) Determine the infrared absorption spectrum of test organism [12] under (1) Agar media for seed and base Ofloxacin as directed in the potassium bromide disk method layer. under Infrared Spectrophotometry <2.25>, and compare the (iii) Standard solutions—Use a light-resistant container. spectrum with the Reference Spectrum: both spectra exhibit Weigh accurately an amount of Nystatin RS equivalent to similar intensities of absorption at the same wave numbers. about 60,000 units, previously dried at 409C for 2 hours in vacuum (not more than 0.67 kPa), dissolve in formamide to Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of make a solution of 3000 units per mL, and use this solution Ofloxacin according to Method 4, and perform the test. Pre- as the standard stock solution. Keep the standard stock solu- pare the control solution with 2.0 mL of Standard Lead So- tion at 59C or below and use within 3 days. Take exactly a lution (not more than 10 ppm). suitable amount of the standard stock solution before use, (2) Related substances—Conduct this procedure without add phosphate buffer solution, pH 6.0 to make solutions so exposure to light. Dissolve 10 mg of Ofloxacin in 50 mL of a that each mL contains 300 units and 150 units, and use these mixture of water and acetonitrile (6:1), and use this solution solutions as the high concentration standard solution and the as the sample solution. Pipet 1 mL of the sample solution, low concentration standard solution, respectively. and add a mixture of water and acetonitrile (6:1) to make (iv) Sample solutions—Use a light-resistant container. exactly 20 mL. Pipet 1 mL of this solution, add a mixture of Weigh accurately an amount of Nystatin equivalent to about water and acetonitrile (6:1) to make exactly 10 mL, and use 60,000 units, dissolve in formamide to make a solution of this solution as the standard solution. Perform the test with 3000 units per mL, and use this solution as the sample stock exactly 10 mL each of the sample solution and standard solution. Take exactly a suitable amount of the sample stock solution as directed under Liquid Chromatography <2.01> solution, add phosphate buffer solution, pH 6.0 to make so- according to the following conditions, and determine each lutions so that each mL contains 300 units and 150 units, and peak area by the automatic integration method: the area of use these solutions as the high concentration sample solution the peak other than ofloxacin obtained from the sample so- and the low concentration sample solution, respectively. lution is not larger than 0.4 times the peak area of ofloxacin from the standard solution, and the total area of the peaks Containers and storage Containers—Tight containers. other than ofloxacin is not larger than the peak area from Storage—Light-resistant, and in a cold place. the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave- Ofloxacin length: 294 nm). Column: A stainless steel column 4.6 mm in inside diame- オフロキサシン ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticlediameter). Column temperature: A constant temperature of about 459C. Mobile phase: Dissolve 7.0 g of sodium perchlorate monohydrate and 4.0 g of ammonium acetate in 1300 mL of water, adjust the pH to 2.2 with phosphoric acid, and add 240 mL of acetonitrile. C H FN O :361.37 18 20 3 4 Flow rate: Adjust the flow rate so that the retention time (3RS)-9-Fluoro-3-methyl-10-(4-methylpiperazin-1-yl)- of ofloxacin is about 20 minutes. 7-oxo-2,3-dihydro-7H-pyrido[1,2,3-de]-[1,4]benzoxazine- Time span of measurement: About 1.8 times as long as the 6-carboxylic acid retention time of ofloxacin beginning after the solvent peak. [82419-36-1] System suitability— Test for required detectability: Measure 1 mL of the Ofloxacin, when dried, contains not less than standard solution, and add a mixture of water and aceto- 99.0z and not more than 101.0z of ofloxacin nitrile (6:1) to make exactly 20 mL. Confirm that the peak (C H FN O ). 18 20 3 4 area of ofloxacin obtained from 10 mL of this solution is Description Ofloxacin occurs as pale yellowish white to equivalent to 4 to 6z of that from 10 mL of the standard so- light yellowish white, crystals or crystalline powder. lution. It is freely soluble in acetic acid (100), slightly soluble in System performance: To 0.5 mL of the sample solution water, and very slightly soluble in acetonitrile and in ethanol add 1 mL of a solution of ofloxacin demethyl substance in a (99.5). mixture of water and acetonitrile (6:1) (1 in 20,000) and a 1182 Olive Oil / Official Monographs JP XVI mixture of water and acetonitrile (6:1) to make 100 mL. the former petroleum ether solution. Wash the petroleum When the procedure is run with 10 mL of this solution under ether solution repeatedly with 20-mL portions of water until the above operating conditions, ofloxacin demethyl sub- the washings show no more acidity to methyl orange TS. stance and ofloxacin are eluted in this order with the resolu- Then add 5 g of anhydrous sodium sulfate, shake, filter, tion between these peaks being not less than 2.5. wash anhydrous sodium sulfate with two 10-mL portions of System repeatability: When the test is repeated 6 times petroleum ether, filter the washings using the former separa- with 10 mL of the standard solution under the above operat- tor, combine the filtrates, distil the petroleum ether on a ing conditions, the relative standard deviation of the peak water bath, passing nitrogen. Dissolve the residue in acetone area of ofloxacin is not more than 2.0z. to make exactly 20 mL, and use this solution as the sample solution. Separately, dissolve 0.067 g of methyl behenate in Loss on drying <2.41> Not less than 0.2z (1 g, 1059C, acetone to make exactly 50 mL. Pipet 2 mL of this solution, 4 hours). add acetone to make exactly 20 mL, and use this solution as Residue on ignition <2.44> Not more than 0.1z (1 g). the standard solution. Perform the test with exactly 2 mL each of the sample solution and standard solution as directed Assay Weigh accurately about 0.3 g of Ofloxacin, previ- under Gas Chromatography <2.02> according to the follow- ously dried, dissolve in 100 mL of acetic acid (100), and ing conditions. Measure the peak heights, H and H ,of titrate <2.50> with 0.1 mol/L perchloric acid VS (potenti- T S methyl behenate of respective solutions: H is not higher ometric titration). Perform a blank determination, and make T than H . any necessary correction. S Operating conditions— Each mL of 0.1 mol/L perchloric acid VS Detector: A hydrogen flame-ionization detector.

＝ 36.14 mg of C18H20FN3O4 Column: A glass column about 3 mm in inside diameter and about 2 m in length, packed with silanized siliceous Containers and storage Containers—Tight containers. earth for gas chromatography (150 to 180 mminparticledi- Storage—Light-resistant. ameter), coated with polyethylene glycol 20 mol/L in a ratio of 5z. Column temperature: A constant temperature of about Olive Oil 2209C. Oleum Olivae Carrier gas: Nitrogen. Flow rate: Adjust the flow rate so that the retention time of methyl behenate is about 18 minutes. オリブ油 Detection sensitivity: Adjust the detection sensitivity so that the peak height of methyl behenate obtained from 2 mL Olive Oil is the fixed oil obtained by expression of the standard solution is 5 to 10 mm. from the ripe fruit of Olea europaea Linne(á Oleaceae). Containers and storage Containers—Tight containers. Description Olive Oil is a light yellow oil. It has a faint odor, which is not rancid, and has a bland taste. It is miscible with diethyl ether, with petroleum diethyl Omeprazole ether and with carbon disulfide. It is slightly soluble in ethanol (95). オメプラゾール The whole or a part of it congeals between 09Cand69C. Congealing point of the fatty acids: 17 – 269C

25 Specific gravity <1.13> d 25: 0.908 – 0.914 Acid value <1.13> Not more than 1.0. Saponification value <1.13> 186–194 C17H19N3O3S: 345.42 Unsaponifiable matters <1.13> Not more than 1.5z. (RS)-5-Methoxy-2-{[(4-methoxy-3,5-dimethylpyridin- 2-yl)methyl]sulfinyl}-1H-benzoimidazole Iodine value <1.13> 79–88 [73590-58-6] Purity (1) Drying oil—Mix 2 mL of Olive Oil with 10 mL of diluted nitric acid (1 in 4), add 1 g of powdered sodium Omeprazole, when dried, contains not less than nitrite little by little with thorough shaking, and allow to 99.0z and not more than 101.0z of C17H19N3O3S. stand in a cold place for 4 to 10 hours: the mixture congeals Description Omeprazole occurs as a white to yellowish to a white solid. white crystalline powder. (2) Peanut oil—Weigh exactly 1.0 g of Olive Oil, dissolve It is freely soluble in N,N-dimethylformamide, sparingly in 60 mL of sulfuric acid-hexane-methanol TS, boil for 2.5 soluble in ethanol (99.5), and practically insoluble in water. hours on a water bath under a reflux condenser, cool, trans- A solution of Omeprazole in N,N-dimethylformamide (1 fer to a separator, and add 100 mL of water. Wash the flask in 25) shows no optical rotation. with 50 mL of petroleum ether, add the washing to the sepa- It gradually turns yellowish white on exposure to light. rator, shake, allow to stand, and separate the petroleum Melting point: about 1509C (with decomposition). ether layer. Extract the water layer with another 50 mL of petroleum ether, and combine the petroleum ether layer with Identification (1) Add phosphate buffer solution, pH JP XVI Official Monographs / Powdered Opium 1183

7.4, to 1 mL of a solution of Omeprazole in ethanol (99.5) (1 from 10 mL of the solution for system suitability test. in 1000) to make 50 mL. Determine the absorption spectrum System performance: Dissolve 10 mg of Omeprazole and of this solution as directed under Ultraviolet-visible Spectro- 25 mg of 1,2-dinitrobenzene in 5 mL of sodium borate solu- photometry <2.24>, and compare the spectrum with the Ref- tion (19 in 5000) and 95 mL of ethanol (99.5). When the erence Spectrum: both spectra exhibit similar intensities of procedure is run with 10 mL of this solution under the above absorption at the same wavelengths. conditions, omeprazole and 1,2-dinitrobenzene are eluted in (2) Determine the infrared absorption spectrum of this order with the resolution between these peaks being not Omeprazole as directed in the potassium bromide disk less than 10. method under Infrared Spectrophotometry <2.25>,andcom- System repeatability: When the test is repeated 6 times pare the spectrum with the Reference Spectrum: both spectra with 10 mL of the solution for system suitability test under exhibit similar intensities of absorption at the same wave the above operating conditions, the relative standard devia- numbers. tion of the peak area of omeprazole is not more than 2.0z. Purity (1) Clarity and color of solution—Dissolve 0.5 g Loss on drying <2.41> Not more than 0.2z (1 g, in vacu- of Omeprazole in 25 mL of N,N-dimethylformamide: the um, phosphorus (V) oxide, 509C, 2 hours). solution is clear and colorless or light yellow. Perform the Residue on ignition <2.44> Not more than 0.1z (1 g). test with this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: the absorbance at 420 nm is not Assay Weigh accurately about 0.4 g of Omeprazole, previ- more than 0.3. ously dried, dissolve in 70 mL of N,N-dimethylformamide, (2) Heavy metals <1.07>—Proceed with 2.0 g of Omepra- and titrate <2.50> with 0.1 mol/L tetramethylammonium hy- zole according to Method 2, and perform the test. Prepare droxide VS (potentiometric titration). Separately, perform a the control solution with 2.0 mL of Standard Lead Solution blank determination using the same method on a solution (not more than 10 ppm). consisting of 70 mL of N,N-dimethylformamide and 12 mL (3) Related substances—Conduct the procedure soon of water, and make any necessary correction. after preparation of the sample solution. Dissolve 50 mg of Each mL of 0.1 mol/L tetramethylammonium hydroxide VS Omeprazole in 50 mL of the mobile phase, and use this solu- ＝ 34.54 mg of C H N O S tion as the sample solution. Perform the test with 10 mLof 17 19 3 3 the sample solution as directed under Liquid Chromatogra- Containers and storage Containers—Tight containers. phy <2.01> according to the following conditions. Determine Storage—Light-resistant, in a cold place. each of the peak areas of the sample solution by the automatic integration method, and calculate the amounts of them by the area percentage method: each of the amount of Powdered Opium the peaks other than omeprazole is not more than 0.1z,and the total amount of the peaks other than omeprazole is not Opium Pulveratum more than 0.5z. Operating conditions— アヘン末 Detector: An ultraviolet absorption photometer (wavelength: 280 nm). Powdered Opium is a homogeneous powder of Column: A stainless steel column 4.6 mm in inside diame- opium obtained from Papaver somniferum Linneá ter and 15 cm in length, packed with octylsilanized silica gel (Papaveraceae). Starch or Lactose Hydrate may be for liquid chromatography (5 mminparticlediameter). added. Column temperature: A constant temperature of about Powdered Opium contains not less than 9.5z and 259C. not more than 10.5z of morphine (C H NO : Mobile phase: Dissolve 2.83 g of disodium hydrogen phos- 17 19 3 285.34). phate dodecahydrate and 0.21 g of sodium dihydrogen phosphate dihydrate in water to make 1000 mL. If necessary, Description Powdered Opium occurs as a yellow-brown to adjust the pH to 7.6 with diluted phosphoric acid (1 in 100). dark brown powder. Add 11 volumes of acetonitrile to 29 volumes of this solu- Identification (1) To 0.1 g of Powdered Opium add 5 mL tion. of diluted ethanol (7 in 10), dissolve by treating with ultra- Flow rate: Adjust the flow rate so that the retention time sonic waves for 10 minutes, and add diluted ethanol (7 in 10) of omeprazole is about 8 minutes. to make 10 mL. Filter this solution, and use the filtrate as Time span of measurement: About 10 times as long as the the sample solution. Separately, dissolve 25 mg of Morphine retention time of omeprazole, beginning after the solvent Hydrochloride Hydrate, 12 mg of Codeine Phosphate Hy- peak. drate, 2 mg of Papaverine Hydrochloride, and 12 mg of System suitability— Noscapine Hydrochloride Hydrate separately in 25 mL of di- Test for required detectability: Pipet 5 mL of the sample luted ethanol (7 in 10), and use these solutions as the stand- solution, and add the mobile phase to make exactly 100 mL. ard solution (1), the standard solution (2), the standard solu- Pipet 5 mL of this solution, add the mobile phase to make tion (3) and the standard solution (4), respectively. Perform exactly 50 mL, and use this solution as the solution for sys- the test with these solutions as directed under Thin-layer tem suitability test. Pipet 5 mL of the solution for system Chromatography <2.03>. Spot 10 mL each of the sample solu- suitability test, and add the mobile phase to make exactly 25 tion and standard solutions on a plate of silica gel for thin- mL. Confirm that the peak area of omeprazole obtained layer chromatography. Develop the plate with a mixture of from 10 mL of this solution is equivalent to 15 to 25z of that acetone, toluene, ethanol (99.5) and ammonia water (28) 1184 Diluted Opium Powder / Official Monographs JP XVI

(20:20:3:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the Diluted Opium Powder plate: each spot from the sample solution shows the same color tone and Rf value of each spot obtained from the アヘン散 standard solution (1), the standard solution (2), the standard solution (3), and the standard solution (4) (morphine, Diluted Opium Powder contains not less than codeine, papaverine and noscapine), respectively. 0.90z and not more than 1.10z of morphine (2) To 0.1 g of Powdered Opium add 5 mL of water, and (C H NO : 285.34). shake the mixture for 5 minutes. Filter, to the filtrate add 1 17 19 3 mL of a solution of hydroxylammonium chloride (3 in 10) Method of preparation and 1 drop of iron (III) chloride TS, and shake: a red-brown Powdered Opium 100 g color is produced. To this solution add immediately 5 mL of Starch or a suitable diluent a sufficient quantity diethyl ether, and shake: the diethyl ether layer has no red- purple color (meconic acid). To make 1000 g Loss on drying <2.41> Not more than 8.0z (1 g, 1059C, Prepare as directed under Powders, with the above ingre- 5 hours). dients. Lactose Hydrate should not be used. Assay Place about 5 g of Powdered Opium, accurately Description Diluted Opium Powder occurs as a light brown weighed, in a mortar, and triturate it with exactly 10 mL of powder. water. Add 2 g of calcium hydroxide and exactly 40 mL of Identification (1) Proceed with 1 g of Diluted Opium water, and stir the mixture for 20 minutes. Filter, and shake Powder as directed in the Identification (1) under Powdered 30 mL of the filtrate with 0.1 g of magnesium sulfate hepta- Opium. hydrate for 1 minute. To the mixture add 0.3 g of calcium (2) Proceed with 1 g of Diluted Opium Powder as di- hydroxide, shake for 1 minute, and allow to stand for 1 rected in the Identification (2) under Powdered Opium. hour. Filter, place 20 mL of the filtrate, exactly measured, in a glass-stoppered flask, and add 10 mL of diethyl ether and Assay Place about 50 g of Diluted Opium Powder, accu- 0.3 g of ammonium chloride. Shake vigorously with caution. rately weighed, in a glass-stoppered flask, and stir with 250 When crystals begin to separate out, shake for 30 minutes mL of dilute ethanol in a water bath at 409C for 1 hour. with a mechanical shaker, and set aside overnight at a tem- Filter the mixture through a glass filter (G3). Transfer the perature of 59Cto109C. Decant the diethyl ether layer and residue on the filter to the first glass-stoppered flask, and filter first, and then the water layer through filter paper 7 cm add 50 mL of dilute ethanol. Stir the mixture in a water bath in diameter. Wash the adhering crystals in the flask with at 409C for 10 minutes, and filter through the same glass three 5-mL portions of water saturated with diethyl ether, filter. Repeat the extraction with three 50-mL portions of di- and wash the crystals on the filter paper with each of these lute ethanol. Evaporate the combined filtrate in a mortar to washings. Wash the top of the glass-stoppered flask and the dryness on a water bath. Add 10 mL of ethanol (99.5) to the upper part of the filter paper with final 5 mL of water satu- residue, evaporate to dryness again, and, after cooling, rated with diethyl ether. Transfer the crystals and the filter triturate it with exactly 10 mL of water. Proceed with this so- paper to a beaker. Dissolve the crystals remaining in the lution as directed in Assay under Powdered Opium. glass-stoppered flask with the aid of 15 mL of 0.05 mol/L Each mL of 0.05 mol/L sulfuric acid VS sulfuric acid VS, accurately measured, and pour the solution ＝ 28.53 mg of C H NO into the beaker. Wash the glass-stoppered flask with four 17 19 3 5-mL portions of water, and add the washings to the solu- Containers and storage Containers—Tight containers. tion in the beaker. Titrate <2.50> the excess sulfuric acid with 0.1 mol/L sodium hydroxide VS (indicator: 4 drops of methyl red-methylene blue TS). Opium Tincture

Each mL of 0.05 mol/L sulfuric acid VS アヘンチンキ＝ 28.53 mg of C17H19NO3

Containers and storage Containers—Tight containers. Opium Tincture contains not less than 0.93 w/vz and not more than 1.07 w/vz of morphine (C17H19NO3: 285.34). Method of preparation Powdered Opium 100 g 35 volz Ethanol a sufficient quantity To make 1000 mL Prepare as directed under Tinctures, with the above ingredients. May be prepared with an appropriate quantity of Ethanol and Purified Water or Purified Water in Containers in place of 35 volz Ethanol. Description Opium Tincture is a dark red-brown liquid. JP XVI Official Monographs / Opium Alkaloids Hydrochlorides 1185

It is affected by light. 3.0 and 4.0. Identification (1) To 1 mL of Opium Tincure add diluted Purity (1) Clarity and color of solution—Dissolve 0.5 g ethanol (7 in 10) to make 10 mL, filter, and use the filtrate as of Opium Alkaloids Hydrochlorides in 10 mL of water: the the sample solution. Proceed as directed in the Identification solution is clear, and its absorbance <2.24> at 420 nm is not (1) under Powdered Opium. more than 0.20. (2) Evaporate 1 mL of Opium Tincture to dryness on a (2) Meconic acid—Dissolve 0.1 g of Opium Alkaloids water bath, and proceed with the residue as directed in the Hydrochlorides in 2 mL of water, and pour into a polyethy- Identification (2) under Powdered Opium. lene column 1 cm in inside diameter, packed with about 0.36 g of aminopropylsilanized silica gel for pretreatment Alcohol number <1.01> Not less than 3.5 (Method 1). (55 – 105 mm in particle diameter) and previously washed Assay Evaporate 50 mL of Opium Tincture, accurately through with 5 mL of water. Then, wash the column with 5 measured, on a water bath to dryness. Add 10 mL of ethanol mL of water, 5 mL of methanol and 10 mL of 0.1 mol/L hy- (99.5) to the residue, evaporate to dryness again, cool, and drochloric acid in this order, then elute with 2 mL of 1 triturate with exactly 10 mL of water. Proceed with this solu- mol/L hydrochloric acid, and use the eluate as the test solution as directed in the Assay under Powdered Opium. tion. To the test solution add 2 mL of dilute sodium hydroxide TS and 1 drop of iron (III) chloride TS: no red color de- Each mL of 0.05 mol/L sulfuric acid VS velops. ＝ 28.53 mg of C17H19NO3 Loss on drying <2.41> Not more than 6.0z (0.5 g, 1209C, Containers and storage Containers—Tight containers. 8 hours). Storage—Light-resistant. Residue on ignition <2.44> Not more than 0.5z (0.5 g). Assay Weigh accurately about 0.1 g of Opium Alkaloids Opium Alkaloids Hydrochlorides Hydrochlorides, and dissolve in water to make exactly 50 mL, and use this solution as the sample solution. Separately, アヘンアルカロイド塩酸塩 weigh accurately about 60 mg of morphine hydrochloride for assay, dissolve in water to make exactly 50 mL, and use this Opium Alkaloids Hydrochlorides consist of the hy- solution as the standard solution. Perform the test with ex- drochlorides of some of the main alkaloids obtained actly 20 mL each of the sample solution and standard solu- from opium. tion as directed under Liquid Chromatography <2.01> ac- It contains not less than 47.0z and not more than cording to the following conditions, and determine the peak 52.0z of morphine (C17H19NO3: 285.34), and not less areas of morphine, codeine, papaverine, thebaine, narceine than 35.0z and not more than 41.0z of other opium and noscapine, AT1, AT2, AT3, AT4, AT5 and AT6, from the alkaloids. sample solution, and the peak area of morphine, AS,from the standard solution. Description Opium Alkaloids Hydrochlorides occur as a white to light brown powder. Amount (mg) of morphine (C17H19NO3) It is soluble in water, and slightly soluble in ethanol (99.5). ＝ MS × AT1/AS × 0.887 It is colored by light. Amount (mg) of other opium alkaloids

Identification (1) Dissolve 0.1 g of Opium Alkaloids Hy- ＝ MS × {(AT2 ＋ 0.29AT3 ＋ 0.20AT4 drochlorides in 10 mL of diluted ethanol (1 in 2), and use ＋ 0.19AT5 ＋ AT6)/AS} × 0.887 this solution as the sample solution. Separately, dissolve 60 M : Amount (mg) of morphine hydrochloride for assay, mg of Morphine Hydrochloride Hydrate, 40 mg of Nosca- S calculated on the anhydrous basis pine Hydrochloride Hydrate, 10 mg of Codein Phosphate Hydrate and 10 mg of Papaverine Hydrochloride in 10 mL The relative retention time of codine, papaverine, the- each of diluted ethanol (1 in 2), and use these solutions as the baine, narceine and noscapine with respect to morphine ob- standard solutions (1), (2), (3) and (4), respectively. Perform tained under the following operating conditions are as the test with these solutions as directed under Thin-layer follows. Chromatography <2.03>. Spot 20 mLeachofthesamplesolu- tion and standard solutions on a plate of silica gel with fluo- Component Relative retention time rescent indicator for thin-layer chromatography. Develop codeine 1.1 the plate with a mixture of acetone, toluene, ethanol (99.5) papaverine 1.9 and ammonia solution (28) (20:20:3:1) to a distance of about thebaine 2.5 10 cm, and air-dry the plate. Examine under ultraviolet light narceine 2.8 (main wavelength: 254 nm): each spot from the sample solu- noscapine 3.6 tion is the same in color tone and Rf value with the corresponding spot from the standard solutions (1), (2), (3) and Operating conditions— (4) (morphine, noscapine, codeine and papaverine). Detector: An ultraviolet absorption photometer (wave- (2) A solution of Opium Alkaloids Hydrochlorides (1 in length: 285 nm). 50) responds to the Qualitative Tests <1.09> (2) for chloride. Column: A stainless steel column 4.6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica pH <2.54> Dissolve 1.0 g of Opium Alkaloids Hydrochlo- gel for liquid chromatography (5 mminparticlediameter). rides in 50 mL of water: the pH of the solution is between Column temperature: A constant temperature of about 1186 Opium Alkaloids Hydrochlorides Injection / Official Monographs JP XVI

409C. mL, and use this solution as the standard solution. Perform Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in the test with 20 mL each of the sample solution and standard 500 mL of diluted phosphoric acid (1 in 1000), and adjust solution as directed under Liquid Chromatography <2.01> the pH to 3.0 with sodium hydroxide TS. To 240 mL of this according to the following conditions, and calculate the solution add 70 mL of tetrahydrofuran, and mix. ratios, QT and QS, of the peak area of morphine to that of Flow rate: Adjust the flow rate so that the retention time the internal standard. of morphine is about 10 minutes. Amount (mg) of morphine (C H NO ) System suitability— 17 19 3 ＝ M × Q /Q × 0.887 System performance: Dissolve 60 mg of Morphine Hydro- S T S chloride Hydrate, 10 mg of Codeine Phosphate Hydrate, 10 MS: Amount (mg) of morphine hydrochloride for assay, mg of Papaverine Hydrochloride and 40 mg of Noscapine calculated on the anhydrous basis Hydrochloride Hydrate in water to make 50 mL. When the Internal standard solution—A solution of etilefrine hydro- procedure is run with 20 mL of this solution under the above chloride (1 in 500). operating conditions, morphine, codeine, papaverine and Operating conditions— noscapine are eluted in this order with the complete separa- Detector: An ultraviolet absorption photometer (wave- tion between these peaks and with the resolution between the length: 285 nm). peaks of morphine and codeine being not less than 1.5. Column: A stainless steel column 4.6 mm in inside diame- System repeatability: When the test is repeated 6 times ter and 15 cm in length, packed with octadecylsilanized silica with 20 mL of the standard solution under the above operat- gel for liquid chromatography (5 mminparticlediameter). ing conditions, the relative standard deviation of the peak Column temperature: A constant temperature of about area of morphine is not more than 1.0z. 409C. Containers and storage Containers—Tight containers. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in Storage—Light-resistant. 500 mL of diluted phosphoric acid (1 in 1000), and adjust the pH to 3.0 with sodium hydroxide TS. To 240 mL of this solution add 70 mL of tetrahydrofuran, and mix. Opium Alkaloids Hydrochlorides Flow rate: Adjust the flow rate so that the retention time of morphine is about 10 minutes. Injection System suitability— System performance: When the procedure is run with 20 アヘンアルカロイド塩酸塩注射液 mL of the standard solution under the above operating conditions, morphine and the internal standard are eluted in this Opium Alkaloids Hydrochlorides Injection is an order with the resolution between these peaks being not less aqueous solution for injection. than 3. It contains not less than 0.90 w/vz and not more System repeatability: When the test is repeated 6 times than 1.10 w/vz of morphine (C17H19NO3: 285.34). with 20 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios Method of preparation of the peak area of morphine to that of the internal standard Opium Alkaloids Hydrochlorides 20 g is not more than 1.0z. Water for Injection or Sterile Water Containers and storage Containers—Hermetic containers, for Injection in Containers a sufficient quantity and colored containers may be used. To make 1000 mL Storage—Light-resistant. Prepare as directed under Injections, with the above ingredients. Description Opium Alkaloids Hydrochlorides Injection is a clear, colorless or light brown liquid. It is affected by light. pH:2.5–3.5 Identification To 1 mL of Opium Alkaloids Hydrochlo- rides Injection add 1 mL of ethanol (99.5), mix, and use this solution as the sample solution, and proceed as directed in the Identification (1) under Opium Alkaloids Hydrochlo- rides. Extractable volume <6.05> It meets the requirement. Assay Pipet 2 mL of Opium Alkaloids Hydrochlorides In- jection, add exactly 10 mL of the internal standard solution and water to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of morphine hydrochloride for assay, and dissolve in exactly 10 mL of the internal standard solution, add water to make 50 JP XVI Official Monographs / Opium Alkaloids and Atropine Injection 1187

add water to make 50 mL, and use this solution as the stand- Opium Alkaloids and Atropine ard solution. Perform the test with 20 mLeachofthesample solution and standard solution as directed under Liquid Injection Chromatography <2.01> according to the following conditions, and calculate the ratios, Q and Q , of the peak area アヘンアルカロイド・アトロピン注射液 T S of morphine to that of the internal standard. Amount (mg) of morphine (C H NO ) Opium Alkaloids and Atropine Injection is an 17 19 3 ＝ M × Q /Q × 0.887 aqueous solution for injection. S T S It contains not less than 0.90 w/vz and not more MS: Amount (mg) of morphine hydrochloride for assay, than 1.10 w/vz of morphine (C17H19NO3: 285.34), calculated on the anhydrous basis and not less than 0.027 w/v and not more than 0.033 z Internal standard solution—A solution of ethylefrine hydro- w/v of atropine sulfate Hydrate [(C H NO ) . z 17 23 3 2 chloride (1 in 500). H2SO4.H2O: 694.84]. Operating conditions— Method of preparation Detector: An ultraviolet absorption photometer (wavelength: 285 nm). Opium Alkaloids Hydrochlorides 20 g Column: A stainless steel column 4.6 mm in inside diame- Atropine Sulfate Hydrate 0.3 g ter and 15 cm in length, packed with octadecylsilanized silica Water for Injection or Sterile Water gel for liquid chromatography (5 mminparticlediameter). for Injection in Containers a sufficient quantity Column temperature: A constant temperature of about To make 1000 mL 409C. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in Prepare as directed under Injections, with the above ingre- 500 mL of diluted phosphoric acid (1 in 1000), and adjust dients. the pH to 3.0 with sodium hydroxide TS. To 240 mL of this Description Opium Alkaloids and Atropine Injection is a solution add 70 mL of tetrahydrofuran, and mix. colorless or light brown, clear liquid. Flow rate: Adjust the flow rate so that the retention time It is affected by light. of morphine is about 10 minutes. pH:2.5–3.5 System suitability— System performance: When the procedure is run with 20 Identification (1) To 1 mL of Opium Alkaloids and Atro- mL of the standard solution under the above operating con- pine Injection add 1 mL of ethanol (99.5), mix, and use this solution as the sample solution. Proceed with the sample so- ditions, morphine and the internal standard are eluted in this order with the resolution between these peaks being not less lution as directed in the Identification (1) under Opium than 3. Alkaloids Hydrochlorides. (2) To 2 mL of Opium Alkaloids and Atropine Injection System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat- add 2 mL of ammonia TS, extract with 10 mL of diethyl ether, and filter the diethyl ether layer. Evaporate the filtrate ing conditions, the relative standard deviation of the ratios of the peak area of morphine to that of the internal standard on a water bath to dryness, add 1 mL of ethanol (99.5) to the is not more than 2.0 . residue, and heat to dissolve. Allow to stand this solution in z an ice water for 30 minutes with occasional shaking. After (2) Atropine sulfate hydrate—Pipet 2 mL of Opium Alkaloids and Atropine Injection, add exactly 2 mL of the crystals are formed, use the supernatant liquid as the sample solution. Separately, dissolve 0.03 g of Atropine Sulfate RS internal standard solution, and add 10 mL of diluted dilute hydrochloric acid (1 in 10). Shake this solution with two in 100 mL of water, proceed with 2 mL of this solution in the 10-mL portions of dichloromethane. Remove the dichloro- same manner as for the sample solution, and use a solution so obtained as the standard solution. Perform the test with methane layer, to the water layer add 2 mL of ammonia TS, immediately add 20 mL of dichloromethane, shake vigor- these solutions as directed under Thin-layer Chromatogra- ously, filter the dichloromethane extract through filter paper phy <2.03>. Spot 10 mL each of the sample solution and on which 5 g of anhydrous sodium sulfate is placed, and standard solution on a plate of silica gel for thin-layer chro- evaporate the filtrate to dryness under reduced pressure. To matography. Develop the plate with a mixture of methanol and ammonia water (28) (200:3) to a distance of about 10 the residue add 0.5 mL of 1,2-dichloromethane and 0.5 mL of bis-trimethylsilylacetamide, stopper tightly, warm in a cm, and air-dry the plate. Spray evenly Dragendorff's TS for water bath at 60 C for 15 minutes, and use this solution as spraying on the plate: a spot of about 0.2 Rf value among 9 the sample solution. Separately, weigh accurately about 30 the several spots from the sample solution and an orange mg of Atropine Sulfate RS (determine separately the loss on colored spot from the standard solution show the same color <2.41> tone, and have the same Rf value (atropine). drying under the same conditions as Atropine Sulfate Hydrate), and dissolve in water to make exactly 100 mL. Extractable volume <6.05> It meets the requirements. Pipet 2 mL of this solution, and add exactly 2 mL of the internal standard solution. Proceed with this solution in the Assay (1) Morphine—Pipet 2 mL of Opium Alkaloids same manner as directed for the sample solution, and use and Atropine Injection, add exactly 10 mL of the internal standard solution, then add water to make 50 mL, and use this solution as the standard solution. Perform the test with 2 mL each of the sample solution and standard solution as this solution as the sample solution. Separately, weigh accu- directed under Gas Chromatography <2.02> according to the rately about 25 mg of morphine hydrochloride for assay, dis- following conditions, and calculate the ratios, Q and Q ,of solve in exactly 10 mL of the internal standard solution, then T S 1188 Opium Alkaloids and Scopolamine Injection / Official Monographs JP XVI the peak area of atropine to that of the internal standard. It is affected by light. pH: 2.5 – 3.5 Amount (mg) of atropine sulfate hydrate

[(C17H23NO3)2.H2SO4.H2O] Identification (1) To 1 mL of Opium Alkaloids and ＝ MS × QT/QS × 1/50 × 1.027 Scopolamine Injection add 1 mL of water and 2 mL of ethanol (99.5), mix, and use this solution as the sample solu- M : Amount (mg) of Atropine Sulfate RS, calculated on S tion. Proceed with the sample solution as directed in the the dried basis Identification (1) under Opium Alkaloids Hydrochlorides. Internal standard solution—A solution of homatropine (2) To 1 mL of Opium Alkaloids and Scopolamine Injec- hydrobromide (1 in 4000). tion add 1 mL of water and 2 mL of ammonia TS, extract Operating conditions— with 10 mL of diethyl ether, and filter the diethyl ether layer. Detector: A hydrogen flame-ionization detector. Evaporate the filtrate on a water bath to dryness, add 1 mL Column: A glass column 3 mm in inside diameter and of ethanol (99.5) to the residue, and heat to dissolve. Allow 1.5 m in length, packed with 180 to 250 mm siliceous earth to stand this solution in an ice water for 30 minutes with oc- for gas chromatography coated in 1 to 3z with 50z phenyl- casional shaking. After crystals are formed, use the superna- methyl silicone polymer for gas chromatography. tant liquid as the sample solution. Separately, dissolve 0.03 g Column temperature: A constant temperature of about of Scopolamine Hydrobromide RS in 100 mL of water. To 2 2109C. mL of this solution add 2 mL of ammonia TS, proceed with Carrier gas: Nitrogen or helium. this solution in the same manner as for the sample solution, Flow rate: Adjust the flow rate so that the retention time and use a solution so obtained as the standard solution. Per- of atropine is about 5 minutes. form the test with these solutions as directed under Thin- System suitability— layer Chromatography <2.03>. Spot 10 mLeachofthesample System performance: When the procedure is run with 2 mL solution and standard solution on a plate of silica gel for of the standard solution under the above operating condi- thin-layer chromatography. Develop the plate with a mixture tions, the internal standard and atropine are eluted in this of methanol and ammonia water (28) (200:3) to a distance of order with the resolution between these peaks being not less about 10 cm, and air-dry the plate. Spray evenly Dragendor- than 3. ff's TS for spraying on the plate: a spot of about 0.7 Rf System repeatability: When the test is repeated 5 times value among the several spots from the sample solution and with 2 mL of the standard solution under the above operating an orange colored spot from the standard solution show the conditions, the relative standard deviation of the ratios of same color tone, and have the same Rf value (scopolamine). the peak area of atropine to that of the internal standard is Extractable volume <6.05> It meets the requirements. not more than 2.0z. Assay (1) Morphine—Pipet 1 mL of Opium Alkaloids Containers and storage Containers—Hermetic containers, and Scopolamine Injection, add 10 mL of the internal stand- and colored containers may be used. ard solution and water to make 50 mL, and use this solution Storage—Light-resistant. as the sample solution. Separately, weigh accurately about 25 mg of morphine hydrochloride for assay, dissolve in exactly 10 mL of the internal standard solution, add water to Opium Alkaloids and Scopolamine make 50 mL, and use this solution as the standard solution. Injection Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography アヘンアルカロイド・スコポラミン注射液 <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of morphine to that of the internal standard. Opium Alkaloids and Scopolamine Injection is an aqueous solution for injection. Amount (mg) of morphine (C17H19NO3) It contains not less than 1.80 w/vz and not more ＝ MS × QT/QS × 0.887 than 2.20 w/vz of morphine (C17H19NO3: 285.34) M : Amount (mg) of morphine hydrochloride for assay, and not less than 0.054 w/v and not more than S z calculated on the anhydrous basis 0.066 w/vz of scopolamine hydrobromide hydrate (C17H21NO4.HBr.3H2O: 438.31). Internal standard solution—A solution of etilefrin hydrochloride (1 in 500). Method of preparation Operating conditions— Opium Alkaloids Hydrochlorides 40 g Detector: An ultraviolet absorption photometer (wave- Scopolamine Hydrobromide Hydrate 0.6 g length: 285 nm). Water for Injection or Sterile Water Column: A stainless steel column 4.6 mm in inside diame- for Injection in Containers a sufficient quantity ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticlediameter). To make 1000 mL Column temperature: A constant temperature of about Prepare as directed under Injections, with the above ingre- 409C. dients. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in Description Opium Alkaloids and Scopolamine Injection is 500 mL of diluted phosphoric acid (1 in 1000), and adjust the pH to 3.0 with sodium hydroxide TS. To 240 mL of this a clear, colorless to light brown liquid. solution add 70 mL of tetrahydrofuran, and mix. JP XVI Official Monographs / Weak Opium Alkaloids and Scopolamine Injection 1189

Flow rate: Adjust the flow rate so that the retention time this order with the resolution between these peaks being not of morphine is about 10 minutes. less than 6. System suitability— System repeatability: When the test is repeated 5 times System performance: When the procedure is run with 20 with 2 mL of the standard solution under the above operating mL of the standard solution under the above operating con- conditions, the relative standard deviation of the ratios of ditions, morphine and the internal standard are eluted in this the peak area of scopolamine to that of the internal standard order with the resolution between these peaks being not less is not more than 2.0z. than 3. Containers and storage Containers—Hermetic containers, System repeatability: When the test is repeated 6 times and colored containers may be used. with 20 mL of the standard solution under the above operat- Storage—Light-resistant. ing conditions, the relative standard deviation of the ratios of the peak area of morphine to that of the internal standard is not more than 2.0z. (2) Scopolamine hydrobromide hydrate—Pipet 2 mL of Weak Opium Alkaloids and Opium Alkaloids and Scopolamine Injection, and add ex- Scopolamine Injection actly 2 mL of the internal standard solution. To this solution add 10 mL of diluted dilute hydrochloric acid (1 in 10), and 弱アヘンアルカロイド・スコポラミン注射液 shake with two 10-mL portions of dichloromethane. Remove the dichloromethane layer, to the water layer add 2 mL of Weak Opium Alkaloids and Scopolamine Injection ammonia TS, add immediately 20 mL of dichloromethane, shake vigorously, filter the dichloromethane extract through is an aqueous solution for injection. It contains not less than 0.90 w/v and not more a filter paper on which 5 g of anhydrous sodium sulfate is z than 1.10 w/v of morphine (C H NO : 285.34) placed, and evaporate the filtrate to dryness under reduced z 17 19 3 and not less than 0.027 w/v and not more than pressure. To the residue add 0.5 mL of 1,2-dichloroethane z 0.033 w/v of scopolamine hydrobromide hydrate and 0.5 mL of bis-trimethyl silyl acetamide, stopper tightly, z (C H NO .HBr.3H O: 438.31). warm in a water bath at 609C for 15 minutes, and use this 17 21 4 2 solution as the sample solution. Separately, weigh accurately Method of preparation about 60 mg of Scoporamine Hydrobromide RS (determine Opium Alkaloids Hydrochlorides 20 g separately the loss on drying <2.41> under the same condi- Scopolamine Hydrobromide Hy- tions as Scopolamine Hydrobromide Hydrate), and dissolve drate 0.3 g in water to make exactly 100 mL. Pipet 2 mL of this solu- Water for Injection or Sterile Water tion, add exactly 2 mL of the internal standard solution. for Injection in Containers a sufficient quantity Proceed with this solution in the same manner as for the sample solution, and use thus obtained solution as the stand- To make 1000 mL ard solution. Perform the test with 2 mLeachofthesample Prepare as directed under Injections, with the above ingre- solution and standard solution as directed under Gas Chro- dients. matography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of Description Weak Opium Alkaloids and Scopolamine In- scopolamine to that of the internal standard. jection is a clear, colorless or light brown liquid. It is affected by light. Amount (mg) of scopolamine hydrobromide hydrate pH: 2.5 – 3.5 (C17H21NO4.HBr.3H2O) ＝ MS × QT/QS × 1/50 × 1.141 Identification (1) To 1 mL of Opium Alkaloids and Scopolamine Injection add 1 mL of ethanol (99.5), mix, and M : Amount (mg) of Scopolamine Hydrobromide RS, cal- S use this solution as the sample solution. Proceed with the culatedonthedriedbasis sample solution as directed in the Identification (1) under Internal standard solution—A solution of homatropine Opium Alkaloids Hydrochlorides. hydrobromide (1 in 4000). (2) To 2 mL of Weak Opium Alkaloids and Scopolamine Operating conditions— Injection add 2 mL of ammonia TS, extract with 10 mL of Detector: A hydrogen flame-ionization detector. diethyl ether, and filter the diethyl ether layer. Evaporate the Column: A glass column 3 mm in inside diameter and filtrate on a water bath to dryness, add 1 mL of ethanol 1.5 m in length, packed with 180 to 250 mm siliceous earth (99.5) to the residue, and heat to dissolve. Allow to stand for gas chromatography coated in 1 to 3z with 50z phenyl- this solution in an ice water for 30 minutes with occasional methyl silicone polymer for gas chromatography. shaking. After crystals are formed, use the supernatant liq- Column temperature: A constant temperature of about uid as the sample solution. Separately, dissolve 0.03 g of 2109C. Scopolamine Hydrobromide RS in 100 mL of water, proceed Carrier gas: Nitrogen or helium. with 2 mL of this solution in the same manner as for the Flow rate: Adjust the flow rate so that the retention time sample solution, and use a solution so obtained as the stand- of scopolamine is about 8 minutes. ard solution. Perform the test with these solutions as di- System suitability— rected under Thin-layer Chromatography <2.03>. Spot 10 mL System performance: When the procedure is run with 2 mL each of the sample solution and standard solution on a plate of the standard solution under the above operating condi- of silica gel for thin-layer chromatography. Develop the tions, the internal standard and scopolamine are eluted in plate with a mixture of methanol and ammonia water (28) 1190 Orange Oil / Official Monographs JP XVI

(200:3) to a distance of about 10 cm, and air-dry the plate. under reduced pressure. To the residue add 0.5 mL of 1,2- Spray evenly Dragendorff's TS for spraying on the plate: a dichloroethane and 0.5 mL of bis-trimethyl silyl acetamide, spot of about 0.7 Rf value among the several spots from the stopper tightly, warm in a water bath at 609Cfor15 sample solution and an orange colored spot from the stand- minutes, and use this solution as the sample solution. ard solution show the same color tone, and have the same Rf Separately, weigh accurately about 60 mg of Scoporamine value (scopolamine). Hydrobromide RS (separately determine the loss on drying <2.41> under the same conditions as Scopolamine Hydro- Extractable volume <6.05> It meets the requirements. bromide Hydrate), and dissolve in water to make exactly 100 Assay (1) Morphine—Pipet 2 mL of Weak Opium mL. Pipet 2 mL of this solution, add exactly 2 mL of the Alkaloids and Scopolamine Injection, add exactly 10 mL of internal standard solution. Proceed with this solution in the the internal standard solution and water to make 50 mL, and same manner as for the sample solution, and use so obtained use this solution as the sample solution. Separately, weigh solution as the standard solution. Perform the test with 2 mL accurately about 25 mg of morphine hydrochloride for each of the sample solution and standard solution as directed assay, dissolve in exactly 10 mL of the internal standard so- under Gas Chromatography <2.02> according to the follow- lution, add water to make 50 mL, and use this solution as the ing conditions, and calculate the ratios, QT and QS,ofthe standard solution. Perform the test with 20 mLeachofthe peak area of scopolamine to that of the internal standard. sample solution and standard solution as directed under Liq- Amount (mg) of scopolamine hydrobromide hydrate uid Chromatography <2.01> according to the following con- (C H NO .HBr.3H O) ditions, and calculate the ratios, Q and Q , of the peak area 17 21 4 2 T S ＝ M × Q /Q × 1/50 × 1.141 of morphine to that of the internal standard. S T S M : Amount (mg) of Scopolamine Hydrobromide RS, Amount (mg) of morphine (C H NO ) S 17 19 3 calculated on the dried basis ＝ MS × QT × QS × 0.887 Internal standard solution—A solution of homatropine M : Amount (mg) of morphine hydrochloride for assay, S hydrobromide (1 in 4000). calculated on the anhydrous basis Operating conditions— Internal standard solution—A solution of etilefrin hydro- Detector: A hydrogen flame-ionization detector. chloride (1 in 500). Column: A glass column 3 mm in inside diameter and Operating conditions— 1.5 m in length, packed with 180 to 250 mm siliceous earth Detector: An ultraviolet absorption photometer (wave- for gas chromatography coated in 1 to 3z with 50z phenyl- length: 285 nm). methyl silicone polymer for gas chromatography. Column: A stainless steel column 4.6 mm in inside diame- Column temperature: A constant temperature of about ter and 15 cm in length, packed with octadecylsilanized silica 2109C. gel for liquid chromatography (5 mm in particle diameter). Carrier gas: Nitrogen or helium. Column temperature: A constant temperature of about Flow rate: Adjust the flow rate so that the retention time 409C. of scopolamine is about 8 minutes. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in System suitability— 500 mL of diluted phosphoric acid (1 in 1000), and adjust System performance: When the procedure is run with 2 mL the pH to 3.0 with sodium hydroxide TS. To 240 mL of this of the standard solution under the above operating condi- solution add 70 mL of tetrahydrofuran, and mix. tions, the internal standard and scopolamine are eluted in Flow rate: Adjust the flow rate so that the retention time this order with the resolution between these peaks being not of morphine is about 10 minutes. less than 6. System suitability— System repeatability: When the test is repeated 5 times System performance: When the procedure is run with 20 with 2 mL of the standard solution under the above operating mL of the standard solution under the above operating con- conditions, the relative standard deviation of the ratios of ditions, morphine and the internal standard are eluted in this the peak area of scopolamine to that of the internal standard order with the resolution between these peaks being not less is not more than 2.0z. than 3. Containers and storage Containers—Hermetic containers, System repeatability: When the test is repeated 6 times and colored containers may be used. with 20 mL of the standard solution under the above operat- Storage—Light-resistant. ing conditions, the relative standard deviation of the ratios of the peak area of morphine to that of the internal standard is not more than 2.0z. (2) Scopolamine hydrobromide hydrate—Pipet 4 mL of Orange Oil Weak Opium Alkaloids and Scopolamine Injection, and add exactly 2 mL of the internal standard solution. To this solu- Oleum Aurantii tion add 10 mL of diluted dilute hydrochloric acid (1 in 10), オレンジ油 and shake with two 10-mL portions of dichloromethane. Remove the dichloromethane layer, to the water layer add 2 mL of ammonia TS, add immediately 20 mL of dichloro- Orange Oil is the essential oil obtained by expression methane, shake vigorously, filter the dichloromethane ex- from the peel of the edible fruit of Citrus species tract through a filter paper on which 5 g of anhydrous sodi- (Rutaceae). um sulfate is placed, and evaporate the filtrate to dryness Description Orange Oil is a yellow to yellow-brown liquid. JP XVI Official Monographs / Oxapium Iodide 1191

It has a characteristic, aromatic odor, and a slightly bitter clear, and has no more color than the following control solu- taste. tion. It is miscible with an equal volume of ethanol (95) with Control solution: To 3 mL of Matching Fluid T add 1 mL turbidity. of diluted hydrochloric acid (1 in 40). (2) Heavy metals <1.07>—Proceed with 2.0 g of Orci- Refractive index <2.45> n20: 1.472 – 1.474 D prenaline Sulfate according to Method 2, and perform the 20 Optical rotation <2.49> a D : ＋85 – ＋999(100 mm). test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Specific gravity <1.13> d 20: 0.842 – 0.848 20 (3) Orciprenalone—Dissolve 0.200 g of Orciprenaline Purity Heavy metals <1.07>—Proceed with 1.0 mL of Sulfate in 0.01 mol/L hydrochloric acid TS to make exactly Orange Oil according to Method 2, and perform the test. 20 mL. Perform the test with this solution as directed under Prepare the control solution with 4.0 mL of Standard Lead Ultraviolet-visible Spectrophotometry <2.24>: the absor- Solution (not more than 40 ppm). bance at 328 nm is not more than 0.075. Containers and storage Containers—Tight containers. Loss on drying <2.41> Not more than 1.5z (1 g, in vacu- Storage—Light-resistant. um, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Orciprenaline Sulfate Assay Weigh accurately about 0.7 g of Orciprenaline Sul- fate, dissolve in 100 mL of acetic acid (100) by warming on a オルシプレナリン硫酸塩 water bath, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS

＝ 52.06 mg of (C11H17NO3)2.H2SO4 Containers and storage Containers—Tight containers. Storage—Light-resistant. (C11H17NO3)2.H2SO4: 520.59 5-{(1RS)-1-Hydroxy- 2-[(1-methylethyl)amino]ethyl}benzene-1,3-diol hemisulfate Oxapium Iodide [5874-97-5] オキサピウムヨウ化物 Orciprenaline Sulfate contains not less than 98.5z of (C11H17NO3)2.H2SO4,calculatedonthedriedbasis. Description Orciprenaline Sulfate occurs as white crystals or crystalline powder. It is freely soluble in water, slightly soluble in ethanol (95) and in acetic acid (100), and practically insoluble in diethyl ether. C H INO : 471.42 A solution of Orciprenaline Sulfate (1 in 20) shows no op- 22 34 2 1-(2-Cyclohexyl-2-phenyl-1,3-dioxolan-4-ylmethyl)-1- tical rotation. methylpiperidinium iodide Melting point: about 2209C (with decomposition). [6577-41-9] Identification (1) Determine the absorption spectrum of a solution of Orciprenaline Sulfate in 0.01 mol/L hydrochloric Oxapium Iodide, when dried, contains not less than acid TS (1 in 10,000) as directed under Ultraviolet-visible 98.5z of C22H34INO2. Spectrophotometry <2.24>, and compare the spectrum with Description Oxapium Iodide occurs as a white, crystalline the Reference Spectrum: both spectra exhibit similar inten- powder. sities of absorption at the same wavelengths. It is soluble in acetonitrile, in methanol and in ethanol (2) Determine the infrared absorption spectrum of Orci- (95), slightly soluble in water, in acetic anhydride and in ace- prenaline Sulfate, previously dried, as directed in the potas- tic acid (100), and practically insoluble in diethyl ether. sium bromide disk method under Infrared Spectrophotome- A solution of Oxapium Iodide in methanol (1 in 100) does try <2.25>, and compare the spectrum with the Reference not show optical rotation. Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Identification (1) Determine the infrared absorption spec- (3) A solution of Orciprenaline Sulfate (1 in 100) re- trum of Oxapium Iodide, previously dried, as directed in the sponds to the Qualitative Tests <1.09> for sulfate. paste method under Infrared Spectrophotometry <2.25>,and compare the spectrum with the Reference Spectrum: both pH <2.54> Dissolve 1.0 g of Orciprenaline Sulfate in 10 mL spectra exhibit similar intensities of absorption at the same of water: the pH of this solution is between 4.0 and 5.5. wave numbers. Purity (1) Clarity and color of solution—Dissolve 1.0 g (2) Dissolve 0.1 g of Oxapium Iodide in 10 mL of metha- of Orciprenaline Sulfate in 10 mL of water: the solution is nol, and add 2 mL of dilute nitric acid and 2 mL of silver 1192 Oxaprozin / Official Monographs JP XVI nitrate TS: a greenish yellow precipitate is formed. Melting point <2.60> 198–2039C. Oxaprozin Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of オキサプロジン Oxapium Iodide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.05 g of Oxapium Iodide in 100 mL of a mixture of water and acetonitrile (1:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of water and C H NO :293.32 acetonitrile (1:1) to make exactly 50 mL, and use this solu- 18 15 3 3-(4,5-Diphenyloxazol-2-yl)propanoic acid tion as the standard solution. Perform the test with exactly [21256-18-8] 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to Oxaprozin, when dried, contains not less than the following conditions. Determine each peak area of each 98.5z of C H NO . solution by the automatic integration method: the total area 18 15 3 of the peaks other than the peak of oxapium from the Description Oxaprozin occurs as a white to yellowish white sample solution is not larger than the area of the peak of crystalline powder. oxapium from the standard solution. It is sparingly soluble in methanol and in ethanol (95), Operating conditions— slightly soluble in diethyl ether, and practically insoluble in Detector: An ultraviolet absorption photometer (wave- water. length: 254 nm). It is gradually affected by light. Column: A stainless steel column about 4 mm in inside di- Identification Determine the infrared absorption spectrum ameter and about 15 cm in length, packed with octadecyl- of Oxaprozin, previously dried, as directed in the potassium silanized silica gel for liquid chromatography (5 mminparti- bromide disk method under Infrared Spectrophotometry cle diameter). <2.25>, and compare the spectrum with the Reference Spec- Column temperature: A constant temperature of 209Cto trum: both spectra exhibit similar intensities of absorption at 309C. the same wave numbers. Mobile phase: To 57 mL of acetic acid (100) and 139 mL 1z of triethylamine add water to make 1000 mL. To 50 mL of Absorbance <2.24> E1cm (285 nm): 455 – 495 (after drying, this solution add 500 mL of acetonitril, 10 mL of dilute ace- 10 mg, methanol, 1000 mL). tic acid and 440 mL of water. Melting point <2.60> 161 – 1659C. Flow rate: Adjust the flow rate so that the retention time of oxapium is about 4 minutes. Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Selection of column: Dissolve 0.05 g of Oxapium Iodide Oxaprozin according to Method 4, and perform the test. and 3 mg of benzophenone in 100 mL of the mobile phase. Prepare the control solution with 2.0 mL of Standard Lead Proceed with 20 mL of this solution under the above operat- Solution (not more than 10 ppm). ing conditions, and calculate the resolution. Use a column (2) Arsenic <1.11>—Prepare the test solution with 2.0 g giving elution of oxapium and benzophenone in this order of Oxaprozin according to Method 3, and perform the test with the resolution between these peaks being not less than 5. (not more than 1 ppm). Detection sensitivity: Adjust the detection sensitivity so (3) Related substances—Dissolve 0.10 g of Oxaprozin in that the peak height of oxapium obtained from 50 mLofthe 10 mL of methanol, and use this solution as the sample solu- standard solution composes 5 to 15z of the full scale. tion. Pipet 1 mL of the sample solution, add methanol to Time span of measurement: About 6 times as long as the make exactly 100 mL, and use this solution as the standard retention time of oxapium beginning after the peak of iodide solution (1). Pipet 5 mL, 3 mL and 1 mL of this solution, ion. add methanol to each to make exactly 10 mL, and use these solutions as the standard solutions (2), (3) and (4), respec- Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, tively. Perform the test with these solutions as directed under 4 hours). Thin-layer Chromatography <2.03>. Spot 10 mLeachofthe Residue on ignition <2.44> Not more than 0.1z (1 g). sample solution and standard solutions (1), (2), (3) and (4) on a plate of silica gel with fluorescent indicator for thin- Assay Weigh accurately about 0.7 g of Oxapium Iodide, layer chromatography. Develop the plate with a mixture of previously dried, dissolve in 50 mL of a mixture of acetic an- ethyl acetate and acetic acid (100) (99:1) to a distance of hydride and acetic acid (100) (9:1), and titrate <2.50> with 0.1 about 15 cm, and air-dry the plate. Examine under ultravio- mol/L perchloric acid VS (potentiometric titration, platinum let light (main wavelength: 254 nm): the total intensity of the electrode). Perform a blank determination, and make any spots other than the principal spot from the sample solution necessary correction. is not more than 1.0z calculated on the basis of intensities Each mL of 0.1 mol/L perchloric acid VS of the spots from the standard solutions (1), (2), (3) and (4). ＝ 47.14 mg of C H INO 22 34 2 Loss on drying <2.41> Not more than 0.3z (1 g, 1059C, Containers and storage Containers—Tight containers. 2 hours). Storage—Light-resistant. Residue on ignition <2.44> Not more than 0.3z (1 g). JP XVI Official Monographs / Oxazolam 1193

Assay Weigh accurately about 0.5 g of Oxaprozin, previ- Oxazolam in ethanol (95) (1 in 100,000) as directed under ously dried, dissolve in 50 mL of ethanol (95), and titrate Ultraviolet-visible Spectrophotometry <2.24>,andcompare <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric the spectrum with the Reference Spectrum: both spectra titration). Perform a blank determination, and make any exhibit similar intensities of absorption at the same wave- necessary correction. lengths. (5) Proceed with Oxazolam as directed under Flame Each mL of 0.1 mol/L sodium hydroxide VS Coloration Test <1.04> (2), and perform the test: a green ＝ 29.33 mg of C H NO 18 15 3 color appears. Containers and storage Containers—Tight containers. Absorbance <2.24> E1z (246 nm): 410 – 430 (after drying, Storage—Light-resistant. 1cm 1 mg, ethanol (95), 100 mL). Purity (1) Chloride <1.03>—To 1.0 g of Oxazolam add 50 Oxazolam mL of water, allow to stand for 1 hour with occasional shaking, and filter. To 25 mL of this filtrate add 6 mL of dilute オキサゾラム nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.20 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.014z). (2) Heavy metals <1.07>—Proceed with 1.0 g of Oxazol- am according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm).

C18H17ClN2O2:328.79 (3) Arsenic <1.11>—Place 1.0 g of Oxazolam in a Kjel- 10-Chloro-2-methyl-11b-phenyl-2,3,7,11b- dahl flask, add 5 mL of sulfuric acid and 5 mL of nitric acid, tetrahydro[1,3]oxazolo[3,2-d][1,4]benzodiazepin- and heat gently. Repeat the addition of 2 to 3 mL of nitric 6(5H)-one acid at times, and continue to heat until a colorless to light [24143-17-7] yellow solution is obtained. After cooling, add 15 mL of saturated ammonium oxalate monohydrate solution, heat Oxazolam, when dried, contains not less than the solution until dense white fumes are evolved, and evapo- 99.0z of C18H17ClN2O2. rate to a volume of 2 to 3 mL. After cooling, dilute with waterto10mL,andperformthetestwiththissolutionas Description Oxazolam occurs as white crystals or crystal- the test solution (not more than 2 ppm). line powder. (4) Related substances—Dissolve 0.05 g of Oxazolam in It is odorless and tasteless. 10 mL of dichloromethane, and use this solution as the It is freely soluble in acetic acid (100), soluble in 1,4- sample solution. Pipet 1 mL of the sample solution, add dioxane and in dichloromethane, slightly soluble in ethanol dichloromethane to make exactly 200 mL, and use this solu- (95) and in diethyl ether, and practically insoluble in water. tion as the standard solution. Perform the test with these so- It dissolves in dilute hydrochloric acid. lutions as directed under Thin-layer Chromatography <2.03>. It gradually changes in color by light. Spot 10 mL each of the sample solution and standard solu- Melting point: about 1879C (with decomposition). tion on a plate of silica gel with fluorescent indicator for Identification (1) Dissolve 0.01 g of Oxazolam in 10 mL thin-layer chromatography. Immediately air-dry, develop of ethanol (95) by heating, and add 1 drop of hydrochloric the plate with a mixture of toluene and acetone (8:1) to a dis- acid: a light yellow color develops, and the solution shows a tance of about 10 cm, and air-dry the plate. Examine under yellow-green fluorescence under ultraviolet light (main wave- ultraviolet light (main wavelength: 254 nm): the spots other length: 365 nm). Add 1 mL of sodium hydroxide TS to this than the principal spot from the sample solution are not solution: the color and fluorescence of this solution disap- more intense than the spot from the standard solution. pear immediately. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, (2) Dissolve 0.01 g of Oxazolam in 5 mL of dilute hydro- 3 hours). chloric acid by heating in a water bath for 10 minutes. After cooling, 1 mL of this solution responds to the Qualitative Residue on ignition <2.44> Not more than 0.1z (1 g). Tests <1.09> for primary aromatic amines. Assay Weigh accurately about 0.65 g of Oxazolam, previ- (3) Place 2 g of Oxazolam in a 200-mL flask, add 50 mL ously dried, dissolve in 100 mL of a mixture of acetic acid of ethanol (95) and 25 mL of 6 mol/L hydrochloric acid TS, (100) and 1,4-dioxane (1:1). Titrate <2.50> with 0.1 mol/L and boil under a reflux condenser for 5 hours. After cooling, perchloric acid VS until the color of the solution changes neutralize with a solution of sodium hydroxide (1 in 4), and from purple through blue to blue-green (indicator: 2 drops extract with 30 mL of dichloromethane. Dehydrate with 3 g of crystal violet TS). Perform a blank determination, and of anhydrous sodium sulfate, filter, and evaporate the make any necessary correction. dichloromethane of the filtrate. Dissolve the residue in 20 mL of methanol by heating on a water bath, and cool imme- Each mL of 0.1 mol/L perchloric acid VS diately in an ice bath. Collect the crystals, and dry in vacuum ＝ 32.88 mg of C18H17ClN2O2 at 609C for 1 hour: the crystals melt <2.60> between 969C Containers and storage Containers—Tight containers. and 1009C. Storage—Light-resistant. (4) Determine the absorption spectrum of a solution of 1194 Oxethazaine / Official Monographs JP XVI

methanol to make exactly 10 mL, then add 0.1 mL of a solu- Oxethazaine tion of 1-fluoro-2,4-dinitrobenzene in methanol (1 in 25), shake well, and heat at 609C for 20 minutes: the solution has Oxetacaine no more color than the following control solution. Control solution: To 0.10 g of 2-aminoethanol add metha- オキセサゼイン nol to make exactly 200 mL, pipet 1 mL of this solution, and add methanol to make exactly 10 mL. Proceed as directed above. Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). C H N O : 467.64 28 41 3 3 Assay Weigh accurately about 0.9 g of Oxethazaine, previ- 2,2?-(2-Hydroxyethylimino)bis[N-(1,1-dimethyl-2- ously dried, dissolve in 50 mL of acetic acid (100), and titrate phenylethyl)-N-methylacetamide] <2.50> with 0.1 mol/L perchloric acid VS (indicator: 2 drops [126-27-2] of crystal violet TS). Perform a blank determination, and make any necessary correction. Oxethazaine, when dried, contains not less than 98.5z of C28H41N3O3. Each mL of 0.1 mol/L perchloric acid VS ＝ 46.76 mg of C H N O Description Oxethazaine occurs as a white to pale yellow- 28 41 3 3 ish white, crystalline powder. Containers and storage Containers—Tight containers. Identification (1) Determine the absorption spectrum of a solution of Oxethazaine in ethanol (95) (1 in 2500) as directed under Ultraviolet-visible Spectrophotometry <2.24>, Oxprenolol Hydrochloride and compare the spectrum with the Reference Spectrum: オクスプレノロール塩酸塩 both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Oxethazaine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>,andcompare the spectrum with the Reference Spectrum: both spectra C H NO .HCl: 301.81 exhibits similar intensities of absorption at the same wave 15 23 3 (2RS)-1-[2-(Allyloxy)phenoxy]- numbers. 3-(1-methylethyl)aminopropan-2-ol monohydrochloride Melting point <2.60> 101–1049C. [6452-73-9] Purity (1) Chloride <1.03>—Dissolve 1.0 g of Oxetha- Oxprenolol Hydrochloride, when dried, contains zaine in 20 mL of ethanol (95), add 6 mL of dilute nitric acid not less than 98.5z of C H NO .HCl. and water to make 50 mL. Perform the test using this solu- 15 23 3 tion as the test solution. Prepare the control solution with Description Oxprenolol Hydrochloride occurs as a white, 0.30 mL of 0.01 mol/L hydrochloric acid VS, 20 mL of crystalline powder. ethanol (95), 6 mL of dilute nitric acid and water to make 50 It is very soluble in water, freely soluble in ethanol (95) mL (not more than 0.011z). and in acetic acid (100), slightly soluble in acetic anhydride, (2) Heavy metals <1.07>—Proceed with 2.0 g of Oxetha- and practically insoluble in diethyl ether. zaine according to Method 2, and perform the test. Prepare Identification (1) To 2 mL of a solution of Oxprenolol the control solution with 2.0 mL of Standard Lead Solution Hydrochloride (1 in 100) add 1 drop of copper (II) sulfate TS (not more than 10 ppm). and 2 mL of sodium hydroxide TS: a blue-purple color de- (3) Related substances—Dissolve 0.40 g of Oxethazaine velops. To this solution add 1 mL of diethyl ether, shake in 10 mL of ethanol (95), and use this solution as the sample well, and allow to stand: a red-purple color develops in the solution. Pipet 1 mL of the sample solution, add ethanol (95) diethyl ether layer, and a blue-purple color develops in the to make exactly 100 mL, and use this solution as the stand- water layer. ard solution. Perform the test with these solutions as di- (2) To 3 mL of a solution of Oxprenolol Hydrochloride rected under Thin-layer Chromatography <2.03>. Spot 10 mL (1 in 150) add 3 drops of Reinecke salt TS: a light red pre- each of the sample solution and standard solution on a plate cipitate is formed. of silica gel with fluorescent indicator for thin-layer chroma- (3) Determine the infrared absorption spectrum of Ox- tography. Develop the plate with a mixture of isopropyl- prenolol Hydrochloride, previously dried, as directed in the ether, tetrahydrofuran, methanol and ammonia solution (28) potassium chloride disk method under Infrared Spectropho- (24:10:5:1) to a distance of about 10 cm, and air-dry the tometry <2.25>, and compare the spectrum with the Refer- plate. Examine under ultraviolet light (main wavelength: 254 ence Spectrum: both spectra exhibit similar intensities of ab- nm): the spots other than the principal spot from the sample sorption at the same wave numbers. solution are not more intense than the spot from the stand- (4) A solution of Oxprenolol Hydrochloride (1 in 50) re- ard solution. sponds to the Qualitative Tests <1.09> for chloride. (4) 2-Aminoethanol—To 1.0 g of Oxethazaine add JP XVI Official Monographs / Oxprenolol Hydrochloride 1195 pH <2.54> Dissolve 1.0 g of Oxprenolol Hydrochloride in 10 mL of water: the pH of this solution is between 4.5 and Oxybuprocaine Hydrochloride 6.0. Benoxinate Hydrochloride Melting point <2.60> 107–1109C. Purity (1) Clarity and color of solution—Dissolve 1.0 g オキシブプロカイン塩酸塩 of Oxprenolol Hydrochloride in 10 mL of water: the solution is clear and colorless. (2) Heavy metals <1.07>—Proceed with 2.0 g of Ox- prenolol Hydrochloride according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Stand- ard Lead Solution (not more than 10 ppm). (3) Arsenic <1.11>—Prepare the test solution with 1.0 g of Oxprenolol Hydrochloride according to Method 3, and C17H28N2O3.HCl: 344.88 perform the test (not more than 2 ppm). 2-(Diethylamino)ethyl 4-amino-3-butyloxybenzoate (4) Related substances—Dissolve 0.25 g of Oxprenolol monohydrochloride Hydrochloride in 10 mL of water, and use this solution as [5987-82-6] the sample solution. Pipet 4 mL of the sample solution, and add water to make exactly 100 mL. Pipet 5 mL of this solu- Oxybuprocaine Hydrochloride, when dried, con- tion, add water to make exactly 100 mL, and use this solu- tains not less than 99.0z of C17H28N2O3.HCl. tion as the standard solution. Perform the test with these so- Description Oxybuprocaine Hydrochloride occurs as white lutions as directed under Thin-layer Chromatography <2.03>. crystals or crystalline powder. It is odorless, and has a saline Spot 10 mL each of the sample solution and standard solu- taste. It exhibits anesthetic properties when placed on the tion on a plate of silica gel with fluorescent indicator for tongue. thin-layer chromatography. Develop the plate in a devel- It is very soluble in water, freely soluble in ethanol (95) oping chamber saturated with ammonia vapor with a mix- and in chloroform, and practically insoluble in diethyl ether. ture of chloroform and methanol (9:1) to a distance of about The pH of a solution of Oxybuprocaine Hydrochloride 10 cm, and air-dry the plate. Examine under ultraviolet light (1 in 10) is between 5.0 and 6.0. (main wavelength: 254 nm): the spots other than the princi- It is gradually colored by light. pal spot from the sample solution are not more intense than the spot from the standard solution. Identification (1) Dissolve 0.01 g of Oxybuprocaine Hy- drochloride in 1 mL of dilute hydrochloric acid and 4 mL of Loss on drying <2.41> Not more than 0.5z (1 g, 809C, water. This solution responds to the Qualitative Tests <1.09> 3 hours). for primary aromatic amines. Residue on ignition <2.44> Not more than 0.1z (1 g). (2) Dissolve 0.1 g of Oxybuprocaine Hydrochloride in 8 mL of water, and add 3 mL of ammonium thiocyanate TS: Assay Weigh accurately about 0.6 g of Oxprenolol Hydro- an oily substance is produced. Rub the inner surface of the chloride, previously dried, dissolvein50mLofamixtureof container with a glass rod: white crystals are formed. Collect acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> the crystals so obtained, recrystallize from water, and dry in with 0.1 mol/L perchloric acid VS (potentiometric titration). a desiccator (in vacuum, phosphorus (V) oxide) for 5 hours: Perform a blank determination, and make any necessary the crystals melt <2.60> between 1039Cand1069C. correction. (3) Determine the absorption spectrum of a solution of Each mL of 0.1 mol/L perchloric acid VS Oxybuprocaine Hydrochloride (1 in 100,000) as directed

＝ 30.18 mg of C15H23NO3.HCl under Ultraviolet-visible Spectrophotometry <2.24>,and compare the spectrum with the Reference Spectrum: both Containers and storage Containers—Tight containers. spectra exhibit similar intensities of absorption at the same wavelengths. (4) A solution of Oxybuprocaine Hydrochloride (1 in 10) responds to the Qualitative Tests <1.09> for chloride. Melting point <2.60> 158 – 1629C. Purity (1) Clarity and color of solution—Dissolve 1.0 g of Oxybuprocaine Hydrochloride in 10 mL of water: the solution is clear and colorless. (2) Heavy metals <1.07>—Proceed with 1.0 g of Oxybu- procaine Hydrochloride according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (3) Related substances—Dissolve 0.25 g of Oxybu- procaine Hydrochloride in 10 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add chloroform to make exactly 20 mL. Pipet 1 mL of this solution, add chloroform to make exactly 1196 Oxycodone Hydrochloride Hydrate / Official Monographs JP XVI

50 mL, and use this solution as the standard solution. Per- trum: both spectra exhibit similar intensities of absorption at form the test with these solutions as directed under Thin- thesamewavelengths. layer Chromatography <2.03>. Spot 10 mLeachofthesample (2) Determine the infrared absorption spectrum of Oxyc- solution and standard solution on a plate of silica gel for odone Hydrochloride Hydrate as directed in the potassium thin-layer chromatography. Develop the plate with a mixture bromide disk method under Infrared Spectrophotometry of chloroform, ethanol (95) and formic acid (7:2:1) to a dis- <2.25>, and compare the spectrum with the Reference Spec- tance of about 10 cm, and air-dry the plate. Spray evenly 4- trum: both spectra exhibit similar intensities of absorption at dimethylaminobenzaldehyde TS for spraying on the plate: the same wave numbers. the spots other than the principal spot from the sample solu- (3) A solution of Oxycodone Hydrochloride Hydrate tion are not more intense than the spot from the standard so- (1 in 50) responds to the Qualitative Tests <1.09> (2) for lution. chloride.

20 Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, Optical rotation <2.49> [a]D : －140 – －1499(0.5 g, calcu- 2 hours). lated on the anhydrous basis, water, 25 mL, 100 mm). Residue on ignition <2.44> Not more than 0.1z (1 g). Purity (1) Clarity and color of solution—Dissolve 0.5 g of Oxycodone Hydrochloride Hydrate in 10 mL of water: Assay Weigh accurately about 0.6 g of Oxybuprocaine Hy- the solution is clear and colorless. drochloride, previously dried, dissolve in 50 mL of a mixture (2) Morphine—Dissolve 10 mg of Oxycodone Hydro- of acetic anhydride and acetic acid (100) (7:3), and titrate chloride Hydrate in 1 mL of water, add 5 mL of 1-nitroso-2- <2.50> with 0.1 mol/L perchloric acid VS (potentiometric naphthole TS and 2 mL of a solution of potassium nitrate titration). Perform a blank determination, and make any (1 in 10), and warm at 409C for 2 minutes. To this solution necessary correction. add 1 mL of a solution of sodium nitrite (1 in 5000), and Each mL of 0.1 mol/L perchloric acid VS warm at 409C for 5 minutes. After cooling, add 10 mL of

＝ 34.49 mg of C17H28N2O3.HCl chloroform, shake, centrifuge, and separate the water layer: the color of the solution is not more intense than a pale red. Containers and storage Containers—Well-closed contain- (3) Codeine—Dissolve 10 mg of Oxycodone Hydrochlo- ers. ride Hydrate in 5 mL of sulfuric acid, add 1 drop of iron Storage—Light-resistant. (III) chloride TS, and warm: no blue color is produced. Add 1 drop of nitric acid: no red color develops. (4) Thebaine—Dissolve 0.10 g of Oxycodone Hydrochlo- Oxycodone Hydrochloride Hydrate ride Hydrate in 2 mL of diluted hydrochloric acid (1 in 10), and heat the solution in a water bath for 25 minutes. After オキシコドン塩酸塩水和物 cooling, add 0.5 mL of 4-aminoantipyrine hydrochloride TS and 0.5 mL of a solution of potassium hexacyanoferrate (III) (1 in 100), and shake. Then shake the solution with 2 mL of ammonia TS and 3 mL of chloroform: no red color develops in the chloroform layer. Water <2.48> 12–15z (0.2 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (0.5 g). C18H21NO4.HCl.3H2O: 405.87 (5R)-4,5-Epoxy-14-hydroxy-3-methoxy-17- Assay Weigh accurately about 0.5 g of Oxycodone Hydro- methylmorphinan-6-one monohydrochloride trihydrate chloride Hydrate, dissolve in 50 mL of a mixture of acetic [124-90-3, anhydride] anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per- Oxycodone Hydrochloride Hydrate contains not less form a blank determination, and make any necessary correc- than 98.0z of C18H21NO4.HCl (mol. wt.: 351.83), cal- tion. culated on the anhydrous basis. Each mL of 0.1 mol/L perchloric acid VS

Description Oxycodone Hydrochloride Hydrate occurs as a ＝ 35.18 mg of C18H21NO4.HCl white, crystalline powder. Containers and storage Containers—Tight containers. It is freely soluble in water, in methanol and in acetic acid Storage—Light-resistant. (100), sparingly soluble in ethanol (95), slightly soluble in acetic anhydride, and practically insoluble in diethyl ether. The pH of a solution dissolved 1.0 g of Oxycodone Hy- drochloride Hydrate in 10 mL of water is between 3.8 and 5.8. It is affected by light. Identification (1) Determine the absorption spectrum of a solution of Oxycodone Hydrochloride Hydrate (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- JP XVI Official Monographs / Compound Oxycodone and Atropine Injection 1197

Amount (mg) of oxycodone hydrochloride hydrate Compound Oxycodone Injection (C18H21NO4.HCl.3H2O) ＝ MSa × QTa/QSa × 1/25 × 1.154 Compound Hycodenone Injection Amount (mg) of hydrocotarnine hydrochloride hydrate (C H NO .HCl.H O) 複方オキシコドン注射液 12 15 3 2 ＝ MSb × QTb/QSb × 1/25 × 1.070 M : Amount (mg) of oxycodone hydrochloride for assay, Compound Oxycodone Injection is an aqueous solu- Sa calculated on the anhydrous basis tion for injection. M : Amount (mg) of hydrocotarnine hydrochloride for It contains not less than 0.74 w/vz and not more Sb assay than 0.86 w/vz of oxycodone hydrochloride hydrate (C18H21NO4.HCl.3H2O: 405.87), and not less than Internal standard solution—Dissolve 0.02 g of phenacetin in 0.18 w/vz and not more than 0.22 w/vz of hydroco- 10 mL of ethanol (95), and add water to make 100 mL. tarnine hydrochloride hydrate (C12H15NO3.HCl.H2O: Operating conditions— 275.73). Detector: An ultraviolet absorption photometer (wavelength: 285 nm). Method of preparation Column: A stainless steel column about 4 mm in inside di- Oxycodone Hydrochloride Hydrate 8 g ameter and about 15 cm in length, packed with octadecyl- Hydrocotarnine Hydrochloride Hydrate 2 g silanized polyvinyl alcohol gel polymer for liquid chroma- Water for Injection or Sterile Water tography (5 mm in particle diameter). for Injection in Containers a sufficient quantity Column temperature: A constant temperature of about To make 1000 mL 259C. Mobile phase: To 500 mL of 0.05 mol/L disodium hydro- Prepare as directed under Injections, with the above ingre- gen phosphate TS add 0.05 mol/L sodium dihydrogen phos- dients. phate TS, and adjust the pH to 8.0. To 300 mL of this solu- Description Compound Oxycodone Injection is a clear, tion add 200 mL of acetonitrile, and mix. colorless to pale yellow liquid. Flow rate: Adjust the flow rate so that the retention time It is affected by light. of oxycodone is about 8 minutes. pH:2.5–4.0 Selection of column: Proceed with 10 mL of the standard solution under the above operating conditions, and use a Identification (1) To 1 mL of Compound Oxycodone In- column giving elution of the internal standard, oxycodone jection add 1 mL of 2,4-dinitrophenylhydrazine-ethanol TS: and hydrocotarnine in this order, with complete separation a yellow precipitate is formed (oxycodone). of these peaks. (2) Evaporate 1 mL of Compound Oxycodone Injection on a water bath. Dissolve the residue in 2 mL of sulfuric Containers and storage Containers—Hermetic containers, acid: a yellow color is produced. Heat the solution: it and colored containers may be used. changes to red, and then to deep orange-red (hydrocotar- Storage—Light-resistant. nine). (3) Evaporate 1 mL of Compound Oxycodone Injection on a water bath. Dissolve the residue in 3 mL of sulfuric Compound Oxycodone and acid, add 2 drops of a solution of tannic acid in ethanol (95) Atropine Injection (1 in 20), and allow to stand: a deep green color is produced (hydrocotarnine). Hycoato Injection Extractable volume <6.05> It meets the requirement. 複方オキシコドン・アトロピン注射液 Assay Pipet 2 mL of Compound Oxycodone Injection, add exactly 10 mL of the internal standard solution, and use this solution as the sample solution. Separately, weigh accurately Compound Oxycodone and Atropine Injection is an about 0.4 g of oxycodone hydrochloride for assay and about aqueous solution for injection. 0.1 g of hydrocotarnine hydrochloride for assay previously It contains not less than 0.74 w/vz and not more dried at 1059C for 3 hours, and dissolve in water to make ex- than 0.86 w/vz of oxycodone hydrochloride hydrate actly 50 mL. Pipet 2 mL of this solution, add exactly 10 mL (C18H21NO4.HCl.3H2O: 405.87), not less than 0.18 of the internal standard solution, and use this solution as the w/vz and not more than 0.22 w/vz of hydrocotar- standard solution. Perform the test with 10 mLeachofthe nine hydrochloride hydrate (C12H15NO3.HCl.H2O: sample solution and standard solution as directed under Liq- 275.73), and not less than 0.027 w/vz and not uid Chromatography <2.01> according to the following con- more than 0.033 w/vz of atropine sulfate hydrate [(C17H23NO3)2.H2SO4.H2O: 694.83]. ditions. Calculate the ratios, QTa and QTb, of the peak area of oxycodone and hydrocotarnine to that of the internal standard from the sample solution, and the ratios, QSa and QSb, of the peak area of oxycodone and hydrocotarnine to that of the internal standard from the standard solution. 1198 Compound Oxycodone and Atropine Injection / Official Monographs JP XVI

Method of preparation Amount (mg) of oxycodone hydrochloride hydrate (C H NO .HCl.3H O) Oxycodone Hydrochloride Hydrate 8 g 18 21 4 2 ＝ M × Q /Q × 1/25 × 1.154 Hydrocotarnine Hydrochloride Hydrate 2 g Sa Ta Sa Atropine Sulfate Hydrate 0.3 g Amount (mg) of hydrocotarnine hydrochloride

Water for Injection or Sterile Water hydrate (C12H15NO3.HCl.H2O) for Injection in Containers a sufficient quantity ＝ MSb × QTb/QSb × 1/25 × 1.070

To make 1000 mL MSa: Amount (mg) of oxycodone hydrochloride for assay, Prepare as directed under Injections, with the above ingre- calculated on the anhydrous basis dients. MSb: Amount (mg) of hydrocotarnine hydrochloride for assay Description Compound Oxycodone and Atropine Injection is a colorless or pale yellow, clear liquid. Internal standard solution—Dissolve 0.02 g of phenacetin in It is affected by light. 10 mL of ethanol (95), and add water to make 100 mL. pH:2.5–4.0 Operating conditions— Detector: An ultraviolet absorption photometer (wave- Identification (1) To 1 mL of Compound Oxycodone and length: 285 nm). Atropin Injection add 1 mL of 2,4-dinitrophenylhydrazine- Column: A stainless steel column about 4 mm in inside di- ethanol TS: a yellow precipitate is formed (oxycodone). ameter and about 15 cm in length, packed with octadecyl- (2) Evaporate 1 mL of Compound Oxycodone and silanized polyvinyl alcohol gel polymer for liquid chroma- Atropin Injection on a water bath, and dissolve the residue tography (5 mm in particle diameter). in 2 mL of sulfuric acid: a yellow color is produced. Heat the Column temperature: A constant temperature of about solution: it changes to red, and then to deep orange-red 259C. (hydrocotarnine). Mobile phase: To 500 mL of 0.05 mol/L disodium (3) Evaporate 1 mL of Compound Oxycodone and hydrogenphosphate TS add 0.05 mol/L sodium dihydrogen- Atropin Injection on a water bath. Dissolve the residue in 3 phosphate TS, and adjust the pH to 8.0. To 300 mL of this mL of sulfuric acid, add 2 drops of a solution of tannic acid solution add 200 mL of acetonitrile, and mix. in ethanol (95) (1 in 20), and allow to stand: a deep green Flow rate: Adjust the flow rate so that the retention time color is produced (hydrocotarnine). of oxycodone hydrochloride is about 8 minutes. (4) To 1 mL of Compound Oxycodone and Atropine In- Selection of column: Proceed with 10 mL of the standard jection add 0.5 mL of 2,4-dinitrophenylhydrazine-ethanol solution under the above operating conditions, and use a TS, and allow to stand for 1 hour. Centrifuge, and add ace- column giving elution of the internal standard, oxycodone tone to the supernatant liquid until no more precipitate is and hydrocotarine in this order with complete separation of produced. Allow to stand for 20 minutes, and centrifuge. To these peaks. the supernatant liquid add potassium hydroxide TS until the (2) Atropine sulfate hydrate—Pipet 2 mL of Compound liquid is light purple. Shake the liquid with 5 mL of dichloro- Oxycodone and Atropine Injection, and add exactly 2 mL of methane, and separate the dichloromethane layer. Take 0.5 the internal standard solution. To this solution add 10 mL of mL of the dichloromethane layer, and evaporate to dryness diluteddilutehydrochloricacid(1in10)and2mLofammo- on a water bath. Add 5 drops of fuming nitric acid to the nia TS, immediately add 20 mL of dichloromethane, shake residue, and evaporate to dryness on a water bath. Cool, dis- vigorously, filter the dichloromethane layer through filter solve the residue in 1 mL of N,N-dimethylformamide, and paper on which 5 g of anhydrous sodium sulfate is placed, add 6 drops of tetraethylammonium hydroxide TS: a red- and evaporate the filtrate to dryness under reduced pressure. purple color is produced (atropine). To the residue add 0.5 mL of 1,2-dichloromethane and Extractable volume <6.05> It meets the requirement. 0.5 mL of bis-trimethylsilylacetamide, stopper tightly, warm in a water bath at 609C for 15 minutes, and use this solution Assay (1) Oxycodone hydrochloride hydrate and as the sample solution. Separately, weigh accurately about hydrocotarnine hydrochloridehydrate—Pipet2mLofCom- 30 mg of Atropine Sulfate RS (separately determine the loss pound Oxycodone and Atropine Injection, add exactly 10 on drying <2.41> under the same conditions as Atropine Sul- mL of the internal standard solution, and use this solution as fate Hydrate), and dissolve in water to make exactly 100 mL. the sample solution. Separately, weigh accurately about 0.4 g Pipet 2 mL of this solution, and add exactly 2 mL of the in- of oxycodone hydrochloride for assay and about 0.1 g of ternal standard solution. Proceed with this solution in the hydrocotarnine hydrochloride for assay previously dried at same manner as directed for the sample solution, and use so 1059C for 3 hours, and dissolve in water to make exactly 50 obtained solution as the standard solution. Perform the test mL. Pipet 2 mL of this solution, add exactly 10 mL of the with 2 mL each of the sample solution and standard solution internal standard solution, and use this solution as the stand- as directed under Gas Chromatography <2.02> according to ard solution. Perform the test with 10 mLeachofthesample the following conditions, and calculate the ratios, QT and solution and standard solution as directed under Liquid QS, of the peak area of atropine to that of the internal stand- Chromatography <2.01> according to the following condi- ards. tions. Calculate the ratios, QTa and QSb, of the peak area of oxycodone and hydrocotarnine to that of the internal stand- Amount (mg) of atropine sulfate hydrate [(C17H23NO3)2.H2SO4.H2O] ard from the sample solution, and the ratios, QSa and QSb,of the peak area of oxycodone and hydrocotarnine to that of ＝ MS × QT/QS × 1/50 × 1.027 the internal standard from the standard solution. MS: Amount (mg) of Atropine Sulfate RS, calculated on JP XVI Official Monographs / Oxygen 1199

the dried basis roform and diethyl ether (3:2) successively, combine the extracts in a tared vessel, and evaporate the combined extract Internal standard solution—A solution of homatropine on a water bath. Dry the residue over silica gel to constant hydrobromide (1 in 4000). mass: the mass of the residue is not more than 50 mg. Operating conditions— (5) Nonvolatile residue—Evaporate 20.0 mL of Oxydol Detector: A hydrogen flame-ionization detector. on a water bath to dryness, and dry the residue at 1059Cfor Column: A glass column about 3 mm in inside diameter 1 hour: the mass of the residue is not more than 20 mg. and about 1.5 m in length, packed with 180- to 250-mm siliceous earth for gas chromatography coated with 1 to 3z Assay Pipet 1.0 mL of Oxydol, transfer it to a flask con- of 50z phenyl-methylsilicone polymer. taining 10 mL of water and 10 mL of dilute sulfuric acid, Column temperature: A constant temperature of about and titrate <2.50> with 0.02 mol/L potassium permanganate 2109C. VS. Carrier gas: Nitrogen or helium. Each mL of 0.02 mol/L potassium permanganate VS Flow rate: Adjust the flow rate so that the retention time ＝ 1.701 mg of H O of atropine is about 5 minutes. 2 2 Selection of column: Proceed with 2 mL of the standard Containers and storage Containers—Tight containers. solution under the above operating conditions, and calculate Storage—Light-resistant, and not exceeding 309C. the resolution. Use a column giving elution of the internal standard and atropine in this order with the resolution between these peaks being not less than 3. Oxygen Containers and storage Containers—Hermetic containers, 酸素 and colored containers may be used. Storage—Light-resistant.

O2: 32.00 Oxydol Oxygen is oxygen produced by the air liquification separation method. オキシドール It contains not less than 99.5 v/vz of O2. Description Oxygen is a colorless gas under atmospheric Oxydol contains not less than 2.5 w/vz and not pressure, and is odorless. more than 3.5 w/vz of hydrogen peroxide (H2O2: 1 mL of Oxygen dissolves in 32 mL of water, and in 7 mL 34.01). It contains suitable stabilizers. of ethanol (95) at 209C and at a pressure of 101.3 kPa. 1000 mL of Oxygen at 09C and at a pressure of 101.3 kPa Description Oxydol occurs as a clear, colorless liquid. It is weighs 1.429 g. odorless or has an odor resembling that of ozone. It gradually decomposes upon standing or upon vigorous Identification Transfer 1 mL each of Oxygen and oxygen agitation. directly from cylinders with a pressure-reducing valve to gas- It rapidly decomposes when in contact with oxidizing sub- measuring tubes or syringes for gas chromatography, using a stances as well as reducing substances. polyvinyl chloride induction tube. Perform the test with It, when alkalized, decomposes with effervescence. these gases as directed under Gas Chromatography <2.02> It is affected by light. according to the following conditions: the retention time of pH:3.0–5.0 principal peak obtained from Oxygen is the same as that of 20 Specific gravity d 20: about 1.01 the peak obtained from oxygen. Operating conditions— Identification 1 mL of Oxydol responds to the Qualitative Proceed as directed in the operating conditions in the Tests <1.09> for peroxide. Purity. Purity (1) Acidity—To 25.0 mL of Oxydol add 2 drops of Purity Nitrogen—Transfer 1.0 mL of Oxygen directly from phenolphthalein TS and 2.5 mL of 0.1 mol/L sodium hy- cylinder with a pressure-reducing valve to gas-measuring droxide VS: a red color develops. tube or syringe for gas chromatography, using a polyvinyl (2) Heavy metals <1.07>—To 5.0 mL of Oxydol add 20 chloride induction tube. Perform the test with this gas as mL of water and 2 mL of ammonia TS, evaporate on a water directed under Gas Chromatography <2.02> according to the bath to dryness, dissolve the residue in 2 mL of dilute acetic following conditions, and determine the peak area A of acid by heating, add water to make 50 mL, and perform the T nitrogen. Introduce 0.50 mL of nitrogen into the gas mixer, test using this solution as the test solution. Prepare the draw carrier gas into the mixer to make exactly 100 mL, control solution with 2 mL of dilute acetic acid, 2.5 mL of allow to mix thoroughly and use this gas as the standard Standard Lead Solution and water to make 50 mL (not more mixed gas. Perform the test in the same manner with 1.0 mL than 5 ppm). of this mixture as directed above, and determine the peak (3) Arsenic <1.11>—To 1.0 mL of Oxydol add 1 mL of area A of nitrogen: A is not larger than A . ammonia TS, evaporate on a water bath to dryness, take the S T S Operating conditions— residue, prepare the test solution according to Method 1, and Detector: A thermal-conductivity detector. perform the test (not more than 2 ppm). Column: A column 3 mm in inside diameter and 3 m in (4) Organic stabilizer—Extract 100 mL of Oxydol with length, packed with zeolite for gas chromatography 250- to 50-mL, 25-mL and 25-mL portions of a mixture of chlo- 1200 Oxymetholone / Official Monographs JP XVI

355-mm in particle diameter (a porosity of 0.5 nm). constant, and designate this as V (mL). With fresh ammo- Column temperature: A constant temperature of about nium chloride-ammonia TS in C, repeat the procedure at 509C. least four times, and measure the volume of residual gas. Carrier gas: Hydrogen or helium. Calculate the volume of Oxygen and V in the following Flow rate: Adjust the flow rate so that the retention time formula on the basis of the gas volume at 209Candat of nitrogen is about 5 minutes. 101.3 kPa. System suitability— Volume (mL) of oxygen (O ) System performance: Introduce 0.5 mL of nitrogen into a 2 ＝ volume of Oxygen (mL) － V (mL) gas mixer, add Oxygen to make 100 mL, and mix thoroughly. When the test is run with 1.0 mL of the mixture Containers and storage Containers—Cylinders. under the above operating conditions, oxygen and nitrogen Storage—Not exceeding 409C. are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 5 times Oxymetholone with 1.0 mL of the standard mixed gas under the above operating conditions, the relative standard deviation of the peak オキシメトロン area of nitrogen is not more than 2.0z. Assay (i) Apparatus—The apparatus is shown diagram- matically in the accompanying figure. A is a 100-mL gas buret having a two-way stopcock a, b – c, d – e and e – f are graduated in 0.1 mL, and c – d is graduated in 2 mL. A is properly connected with a leveling tube B by a thick rubber tube. Fill ammonium chloride-ammonia TS up to the middle C H O :332.48 of A and B. Place in the absorption ball g of the gas pipette 21 32 3 17b-Hydroxy-2-hydroxymethylene-17a-methyl-5a- C a coil of copper wire, not more than 2 mm in diameter, androstan-3-one which extends to the uppermost portion of the bulb, add 125 [434-07-1] mL of ammonium chloride-ammonia TS, and stopper with a rubber stopper i. Connect C with A using the thick rubber Oxymetholone, when dried, contains not less than tube. 97.0z and not more than 103.0z of C21H32O3. Description Oxymetholone occurs as a white to pale yellowish white, crystalline powder. It is odorless. It is freely soluble in chloroform, soluble in 1,4-dioxane, sparingly soluble in methanol, in ethanol (95) and in acetone, slightly soluble in diethyl ether, and practically insoluble in water. It is gradually colored and decomposed by light. Identification (1) Dissolve 2 mg of Oxymetholone in 1 mL of ethanol (95), and add 1 drop of iron (III) chloride TS: a purple color develops. (2) Dissolve 0.01 g of Oxymetholone in methanol to make 50 mL. To 5 mL of the solution add 5 mL of sodium hydroxide-methanol TS and methanol to make 50 mL. De- termine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>,and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (3) Determine the infrared absorption spectrum of Oxymetholone as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>,andcompare the spectrum with the Reference Spectrum: both spectra (ii) Procedure—Open a, set B downward and draw the exhibit similar intensities of absorption at the same wave liquid in g to the stopcock opening a. Then close a. Open a numbers. to the intake tube h, and fill A and h with ammonium chlo- 20 ride-ammonia TS by lifting B. Close a, connect h with a con- Optical rotation <2.49> [a]D : ＋34 – ＋389(after drying, tainer of Oxygen, open a, set B downward and measure 0.2g,1,4-dioxane,10mL,100mm). accurately 100 mL of Oxygen. Open a toward C, and trans- Melting point <2.60> 175 – 1829C. fer the Oxygen to g by lifting B. Close a, and rock C gently for 5 minutes. Open a, draw the residual gas back into A by Purity (1) Clarity and color of solution—Dissolve 0.5 g setting B downward, and measure the volume of the residual of Oxymetholone in 25 mL of 1,4-dioxane: the solution is gas. Repeat the procedure until the volume of residual gas is clear, and shows a colorless to pale yellow color. JP XVI Official Monographs / Oxytetracycline Hydrochloride 1201

(2) Related subslances—Dissolve 50 mg of Oxymetho- Description Oxytetracycline Hydrochloride occurs as yel- lone in 5 mL of chloroform, and use this solution as the sam- low, crystals or crystalline powder. ple solution. Pipet 1 mL of the sample solution, add chlo- It is freely soluble in water, and slightly soluble in ethanol roform to make exactly 200 mL, and use this solution as the (99.5). standard solution. Perform the test with these solutions as Identification (1) Determine the absorption spectrum of a directed under Thin-layer Chromatography <2.03>. Spot 10 solution of Oxytetracycline Hydrochloride in 0.1 mol/L hy- mL each of the sample solution and standard solution on a drochloric acid TS (1 in 50,000) as directed under Ultra- plate of silica gel for thin-layer chromatography, and air-dry violet-visible Spectrophotometry <2.24>, and compare the the spot. Develop immediately the plate with a mixture of spectrum with the Reference Spectrum or the spectrum of a toluene and ethanol (99.5) (49:1) to a distance of about 12 solution of Oxytetracycline Hydrochloride RS prepared in cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid the same manner as the sample solution: both spectra exhibit TS on the plate, and heat at 1009C for 3 to 5 minutes: any similar intensities of absorption at the same wavelengths. spot other than the principal spot and starting point ob- (2) Dissolve 20 mg of Oxytetracycline Hydrochloride in 3 tained from the sample solution is not more intense than the mL of water, and add 1 drop of silver nitrate TS: a white spot from the standard solution. turbidity is produced. Loss on drying <2.41> Not more than 1.0z (0.5 g, in vacu- Optical rotation <2.49> [a]20: －188 – －2009(0.25 g calcu- um, phosphorus (V) oxide, 4 hours). D lated on the dried basis, 0.1 mol/L hydrochloric acid, 25 Residue on ignition <2.44> Not more than 0.1z (0.5 g). mL, 100 mm). Assay Weigh accurately about 40 mg of Oxymetholone, Purity (1) Heavy metals <1.07>—Proceed with 0.5 g of previously dried, and dissolve in methanol to make exactly Oxytetracycline Hydrochloride according to Method 2, and 50 mL. Pipet 5 mL of this solution, and add methanol to perform the test. Prepare the control solution with 2.5 mL of make exactly 50 mL. To exactly measured 5 mL of this solu- Standard Lead Solution (not more than 50 ppm). tion add 5 mL of sodium hydroxide-methanol TS and (2) Related substances—Dissolve 20 mg of Oxytetracy- methanol to make exactly 50 mL. Determine the absorbance cline Hydrochloride in 0.01 mol/L hydrochloric acid TS to A of this solution at the wavelength of maximum absorption make exactly 25 mL, and use this solution as the sample so- at about 315 nm as directed under Ultraviolet-visible Spec- lution. Separately, dissolve 20 mg of 4-epioxytetracycline in trophotometry <2.24>, using a solution, prepared by adding 0.01 mol/L hydrochloric acid TS to make exactly 25 mL, methanol to 5 mL of sodium hydroxide-methanol TS to and use this solution as 4-epioxytetracycline stock solution. make 50 mL, as the blank. Separately, dissolve 20 mg of tetracycline hydrochloride in 0.01 mol/L hydrochloric acid TS to make exactly 25 mL, Amount (mg) of C H O ＝ A/541 × 50,000 21 32 3 and use this solution as tetracycline hydrochloride stock so- Containers and storage Containers—Tight containers. lution. Separately, dissolve 8 mg of b-apooxytetracycline in 5 Storage—Light-resistant. mL of 0.01 mol/L sodium hydroxide TS, add 0.01 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as b-apooxytetracycline stock solution. Pipet 1 mL Oxytetracycline Hydrochloride of 4-epioxytetracycline stock solution, 4 mL of tetracycline hydrochloride stock solution and 40 mL of b-apooxytetracy- オキシテトラサイクリン塩酸塩 cline stock solution, add 0.01 mol/L hydrochloric acid TS to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mLeachofthe sample solution and standard solution as directed under Liq- uid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the peak areas of 4-epioxytetracycline and tetracycline obtained from the sample solution are not larger than each of the peak area from the standard solution, and C H N O .HCl: 496.89 22 24 2 9 the total area of the peaks, a-apooxytetracycline having the (4S,4aR,5S,5aR,6S,12aS)-4-Dimethylamino- relative retention time of about 2.1 with respect to oxytetra- 3,5,6,10,12,12a-hexahydroxy-6-methyl-1,11- cycline, b-apooxytetracycline and the peaks, which appear dioxo-1,4,4a,5,5a,6,11,12a-octahydrotetracene-2- between a-apooxytetracycline and b-apooxytetracycline, is carboxamide monohydrochloride not larger than the peak area of b-apooxytetracycline from [2058-46-0] the standard solution. The peak area of 2-acetyl-2-decarbox- amide oxytetracycline, which appears after the principal Oxytetracycline Hydrochloride is the hydrochloride peak, obtained from the sample solution is not larger than 4 of a tetracycline substance having antibacterial activity times the peak area of 4-epioxytetracycline from the stand- produced by the growth of Streptomyces rimosus. ard solution. It contains not less than 880 mg (potency) and not Operating conditions— more than 945 mg (potency) per mg, calculated on the Detector: An ultraviolet absorption photometer (wave- dried basis. The potency of Oxytetracycline Hydro- length: 254 nm). chloride is expressed as mass (potency) of oxytetracy- Column: A stainless steel column 4.6 mm in inside diame- cline (C H N O : 460.43). 22 24 2 9 ter and 25 cm in length, packed with styrene-divinylbenzene 1202 Oxytetracycline Hydrochloride / Official Monographs JP XVI copolymer for liquid chromatography (8 mminparticledi- 20 mL of this solution under the above operating conditions, ameter). the relative standard deviation of the peak area of 4-epiox- Column temperature: A constant temperature of about ytetracycline is not more than 2.0z. 609C. Loss on drying <2.41> Not more than 2.0z (1 g, in vacu- Mobile phase A: Mix 60 mL of 0.33 mol/L potassium um, 609C, 3 hours). dihydrogen phosphate TS, 100 mL of a solution of tetrabutylammonium hydrogensulfate (1 in 100), 10 mL of a solution Residue on ignition <2.44> Not more than 0.5z (1 g). of disodium dihydrogen ethylenediamine tetraacetate dihy- Assay Weigh accurately an amount of Oxytetracycline Hy- drate (1 in 2500) and 200 mL of water, and adjust the pH to drochloride and Oxytetracycline Hydrochloride RS, equiva- 7.5 with 2 mol/L sodium hydroxide TS. To this solution add lent to about 50 mg (potency), and dissolve each in diluted 30 g of t-butanol and water to make 1000 mL. hydrochloric acid (1 in 100) to make exactly 50 mL. Pipet 5 Mobile phase B: Mix 60 mL of 0.33 mol/L potassium mL each of these solutions, add diluted methanol (3 in 20) to dihydrogen phosphate TS, 50 mL of a solution of tetrabut- make exactly 50 mL, and use these solutions as the sample ylammonium hydrogensulfate (1 in 100), 10 mL of a solution solution and the standard solution. Perform the test with ex- of disodium dihydrogen ethylenediamine tetraacetate dihy- actly 20 mL each of the sample solution and standard solu- drate (1 in 2500) and 200 mL of water, and adjust the pH to tion as directed under Liquid Chromatography <2.01> ac- 7.5 with 2 mol/L sodium hydroxide TS. To this solution add cording to the following conditions, and determine the peak 100 g of t-butanol and water to make 1000 mL. areas, A and A , of oxytetracycline. Flowing of the mobile phase: Control the gradient by mix- T S ing the mobile phases A and B as directed in the following Amount [mg (potency)] of oxytetracycline (C22H24N2O9) table. ＝ MS × AT/AS × 1000

MS: Amount [mg (potency)] of Oxytetracycline Hydro- Time after injection Mobile phase A Mobile phase B chloride RS of sample (min) (volz) (volz) Operating conditions— 0–20 70→ 10 30 → 90 Detector: An ultraviolet absorption photometer (wave- 20–35 10→ 20 90 → 80 length: 263 nm). Column: A stainless steel column 4.6 mm in inside diame- Flow rate: 1.0 mL/min. ter and 25 cm in length, packed with strongly acidic ion ex- Time span of measurement: About 3.5 times as long as the change resin for liquid chromatography (5 mminparticledi- retention time of oxytetracycline beginning after the solvent ameter). peak. Column temperature: A constant temperature of about System suitability— 309C. Test for required detectability: Pipet 1 mL of 4-epiox- Mobile phase: Dissolve 3.402 g of potassium dihydrogen ytetracycline stock solution, and add 0.01 mol/L hydrochlo- phosphate and 9.306 g of disodium dihydrogen ethylene- ricacidTStomakeexactly200mL.Pipet4mLofthissolu- diamine tetraacetate dihydrate in 700 mL of water, add 300 tion, and add 0.01 mol/L hydrochloric acid TS to make mL of methanol, and adjust the pH to 4.5 with dilute hydro- exactly 20 mL. Confirm that the peak area of 4-epiox- chloric acid. ytetracycline obtained from 20 mL of this solution is equiva- Flow rate: Adjust the flow rate so that the retention time lent to 14 to 26z of that from 20 mL of the standard solu- of oxytetracycline is about 7 minutes. tion. System suitability— System performance: Dissolve 8 mg of a-apooxytetracy- System performance: When the procedure is run with 20 cline in 5 mL of 0.01 mol/L sodium hydroxide TS, add 0.01 mL of the standard solution under the above operating con- mol/LhydrochloricacidTStomake100mL,andusethis ditions, the theoretical plates and the symmetrical coefficient solution as a-apooxytetracycline stock solution. Mix 3 mL of of the peak of oxytetracycline are not less than 1000 and not the sample solution, 2 mL of 4-epioxytetracycline stock solu- more than 2.0, respectively. tion, 6 mL of tetracycline hydrochloride stock solution, 6 System repeatability: When the test is repeated 6 times mL of b-apooxytetracycline stock solution and 6 mL of a- with 20 mL of the standard solution under the above operat- apooxytetracycline stock solution, and add 0.01 mol/L hy- ing conditions, the relative standard deviation of the peak drochloric acid TS to make 50 mL. When the procedure is area of oxytetracycline is not more than 1.0z. run with 20 mL of this solution under the above operating Containers and storage Containers—Tight containers. conditions, 4-epioxytetracycline, oxytetracycline, tetracy- Storage—Light-resistant. cline, a-apooxytetracycline and b-apooxytetracycline are eluted in this order with the resolutions between the peaks, 4- epioxytetracycline and oxytetracycline, oxytetracycline and tetracycline, and a-apooxytetracycline and b-apooxytetracycline being not less than 4, not less than 5 and not less than 4, respectively, and the symmetry factor of the peak of oxytetracycline is not more than 1.3. System repeatability: Pipet 1 mL of 4-epioxytetracycline stock solution, and add 0.01 mol/L hydrochloric acid TS to make exactly 200 mL. When the test is repeated 6 times with JP XVI Official Monographs / Oxytocin 1203

ter and 8 cm in length, packed with strongly acidic ion- Oxytocin exchange resin for liquid chromatography (sodium type) composed with a sulfonated polystyrene copolymer (3 mmin オキシトシン particle diameter). Column temperature: A constant temperature of about 579C. Chemical reaction bath temperature: A constant tempera-

C43H66N12O12S2: 1007.19 ture of about 1309C. [50-56-6] Color developing time: About 1 minute. Mobile phase: Prepare mobile phases A, B and C accord- Oxytocin is a synthetic peptide having the property ing to the following table. of causing the contraction of uterine smooth muscle. It contains not less than 540 oxytocin Units and not Mobile phase A B C more than 600 oxytocin Units per mg, calculated on Citric acid mono- the dehydrated and de-acetic acid basis. 19.80 g 22.00 g 6.10 g hydrate Description Oxytocin occurs as a white powder. Trisodium citrate 6.19 g 7.74 g 26.67 g It is very soluble in water, and freely soluble in ethanol dihydrate (99.5). Sodium chloride 5.66 g 7.07 g 54.35 g It dissolves in hydrochloric acid TS. Ethanol (99.5) 260.0 mL 20.0 mL — The pH of a solution prepared by dissolving 0.10 g of Benzyl alcohol — — 5.0 mL Oxytocin in 10 mL of freshly boiled and cooled water is be- Thiodiglycol 5.0 mL 5.0 mL — tween 4.0 and 6.0. Lauromacrogol 4.0 mL 4.0 mL 4.0 mL It is hygroscopic. solution (1 in 4) Capryric acid 0.1 mL 0.1 mL 0.1 mL Identification Determine the absorption spectrum of a so- Water a sufficient a sufficient a sufficient lution of Oxytocin (1 in 2000) as directed under Ultraviolet- amount amount amount visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in- Total amount 2000 mL 1000 mL 1000 mL tensities of absorption at the same wavelengths. pH 3.3 3.2 4.9 Constituent amino acids Put about 1 mg of Oxytocin in a test tube for hydrolysis, add 6 mol/L hydrochloric acid TS Flowing of the mobile phase: Control the gradient by mix- to dissolve, replace the air in the tube with Nitrogen, seal the ing the mobile phases A, B and C as directed in the following tube under reduced pressure, and heat at 110 to 1159Cfor table. 16 hours. After cooling, open the tube, evaporate the hydrolyzate to dryness under reduced pressure, add 2 mL of 0.02 mol/L hydrochloric acid TS to dissolve the residue, and Time after Mobile Mobile Mobile use this solution as the sample solution. Separately, weigh injection of phase phase phase sample (min) A(volz) B(volz) C(volz) accurately about 27 mg of L-aspartic acid, about 24 mg of L-threonine, about 21 mg of L-serine, about 29 mg of L- 0–9 100 0 0 glutamic acid, about 23 mg of L-proline, about 15 mg of 9–25 0 100 0 glycine, about 18 mg of L-alanine, about 23 mg of L-valine, 25–61 0 100→ 00→ 100 about 48 mg of L-cystine, about 30 mg of methionine, about 61–80 0 0 100 26 mg of L-isoleucine, about 26 mg of L-leucine, about 36 mg of L-tyrosine, about 33 mg of phenylalanine, about 37 Reaction reagent: Mix 407 g of lithium acetate dihydrate, mg of L-lysine hydrochloride, about 42 mg of L-histidine hy- 245 mL of acetic acid (100) and 801 mL of 1-methoxy-2- drochloride monohydrate and about 42 mg of L-arginine hydrochloride, dissolve them in 10 mL of 1 mol/L hydrochlo- propanol, add water to make 2000 mL, stir for more than 10 minutes while passing Nitrogen, and use this solution as So- ric acid TS, and add water to make exactly 100 mL. Pipet 5 lution A. Separately, to 1957 mL of 1-methoxy-2-propanol mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test add 77 g of ninhydrin and 0.134 g of sodium borohydride, stir for more than 30 minutes while passing Nitrogen, and with exactly 20 mL each of the sample solution and standard use this solution as Solution B. Mix Solution A and Solution solution as directed under Liquid Chromatography <2.01> Bbeforeuse. according to the following conditions, and calculate the re- Flow rate of mobile phase: About 0.26 mL per minute. spective molar ratios with respect to leucine: 0.95 – 1.05 for aspartic acid, 0.95 – 1.05 for glutamic acid, 0.95 – 1.05 for Flow rate of reaction reagent: About 0.3 mL per minute. System suitability— proline, 0.95 – 1.05 for glycine, 0.80 – 1.10 for isoleucine, 0.80 – 1.05 for tyrosine and 0.80 – 1.05 for cystine, and not System performance: When the procedure is run with 20 mL of the standard solution under the above operating con- more than 0.01 each for others. ditions, aspartic acid, threonine, serine, glutamic acid, pro- Operating conditions— Detector: A visible spectrophotometer (wavelength: 440 line, glycine, alanine, valine, cystine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine and argi- nm and 570 nm). Column: A stainless steel column 4.6 mm in inside diame- nine are eluted in this order with the resolutions between the 1204 Oxytocin / Official Monographs JP XVI peaks of threonine and serine, glycine and alanine, and Operating conditions— isoleucine and leucine being not less than 1.5, 1.4 and 1.2, Detector, column, column temperature, mobile phase, respectively. flowing of mobile phase, and flow rate: Proceed as directed System repeatability: When the test is repeated 3 times in the operating conditions in the Assay. with 20 mL of the standard solution under the above operat- Time span of measurement: About 2.5 times as long as the ing conditions, the relative standard deviations of the peak retention time of oxytocin. area of aspartic acid, proline, valine and arginine are not System suitability— more than 2.0z, respectively. Test for required detectability: Measure exactly 1 mL of the sample solution, add the mobile phase A to make exactly Purity (1) Acetic acid—Weigh accurately about 15 mg of 100 mL, and use this solution as the solution for system Oxytocin, dissolve in the internal standard solution to make suitability test. Pipet 1 mL of the solution for system suita- exactly 10 mL, and use this solution as the sample solution. bility test, and add the mobile phase A to make exactly 10 Separately, weigh accurately about 1 g of acetic acid (100), mL. Confirm that the peak area of oxytocin obtained from add the internal standard solution to make exactly 100 mL. 50 mL of this solution is equivalent to 5 to 15z of that from Pipet 2 mL of this solution, add the internal standard solu- 50 mL of the solution for system suitability test. tion to make exactly 200 mL, and use this solution as the System performance: Dissolve an adequate amount of standard solution. Perform the test with 10 mLeachofthe oxytocin and vasopressin in the mobile phase A, so that each sample solution and standard solution as directed under Liq- mL contains about 0.1 mg each of them. When the proce- uid Chromatography <2.01> according to the following con- dure is run with 50 mL of this solution under the above oper- ditions, and calculate the ratios, Q and Q , of the peak area T S ating conditions, vasopressin and oxytocin are eluted in this of acetic acid to that of the internal standard: the amount of order with the resolution between these peaks being not less acetic acid is not less than 6.0z and not more than 10.0z. than 14, and the symmetry factor of the peak of oxytocin is

Amount (z) of acetic acid (C2H4O2) not more than 1.5. ＝ MS/MT × QT/QS × 1/10 System repeatability: When the test is repeated 6 times with 50 mL of the solution for system suitability test under M : Amount (mg) of acetic acid (100) S the above operating conditions, the relative standard devia- M : Amount (mg) of the sample T tion of the peak area of oxytocin is not more than 2.0z. Internal standard solution—A solution of propionic acid in Water <2.48> Not more than 5.0z (50 mg, coulometric the mobile phase (1 in 10,000). titration). Operating conditions— Detector: An ultraviolet absorption photometer (wave- Assay Weigh accurately an amount of Oxytocin, equiva- length: 210 nm). lent to about 13,000 Units, dissolve in the mobile phase A to Column: A stainless steel column 4.6 mm in inside diame- make exactly 100 mL, and use this solution as the sample ter and 15 cm in length, packed with octadecylsilanized silica solution. Separately, dissolve 1 bottle of the Oxytocin RS in gel for liquid chromatography (5 mm in particle diameter). the mobile phase A to make a known concentration solution Column temperature: A constant temperature of about containing each mL contains about 130 Units, and use this 409C. solution as the standard solution. Perform the test with Mobile phase: To 0.7 mL of phosphoric acid add 900 mL exactly 25 mL each of the sample solution and standard of water, adjust the pH to 3.0 with 8 mol/L sodium hydrox- solution as directed under Liquid Chromatography <2.01> ide TS, and add water to make 1000 mL. To 950 mL of this according to the following conditions, and determine the solution add 50 mL of methanol. peak areas, AT and AS, of oxytocin. Flow rate: Adjust the flow rate so that the retention time Units per mg of Oxytocin, calculated on the dehydrated of acetic acid is about 3 minutes. and de-acetic acid basis System suitability— ＝ M /M × A /A × 100 System performance: When the procedure is run with 10 S T T S mL of the standard solution under the above operating con- MS: Units per mL of the standard solution ditions, acetic acid and propionic acid are eluted in this order MT: Amount (mg) of sample, calculated on the dehy- with the resolution between these peaks being not less than drated and de-acetic acid basis 14. Operating conditions— System repeatability: When the test is repeated 6 times Detector: An ultraviolet absorption photometer (wave- with 10 mL of the standard solution under the above operat- length: 220 nm). ing conditions, the relative standard deviation of the ratio of Column: A stainless steel column 4.6 mm in inside diame- the peak area of acetic acid to that of the internal standard is ter and 15 cm in length, packed with octadecylsilanized silica not more than 2.0z. gel for liquid chromatography (5 mminparticlediameter). (2) Related substances—Dissolve 25 mg of Oxytocin in Column temperature: A constant temperature of about 100 mL of the mobile phase A, and use this solution as the 259C. sample solution. Perform the test with 50 mLofthesample Mobile phase A: Dissolve 15.6 g of sodium dihydrogen solution as directed under Liquid Chromatography <2.01> phosphate dihydrate in 1000 mL of water. according to the following conditions, determine each peak Mobile phase B: A mixture of water and acetonitrile (1:1). area by the automatic integration method, and calculate the Flowing of the mobile phase: Control the gradient by mix- amount of them by the area percentage method: the amount ing the mobile phases A and B as directed in the following of each peak other than Oxytocin is not more than 1.5z, table. and the total of them is not more than 5.0z. JP XVI Official Monographs / Oxytocin Injection 1205

determine the peak areas, AT and AS, of oxytocin. Time after injection Mobile phase A Mobile phase B of sample (min) (volz) (volz) Units per mL of Oxytocin Injection ＝ MS × AT/AS × b/a 0–30 70→ 40 30 → 60 M : Units per mL of the standard solution 30–30.1 40→ 70 60 → 30 S a: Volume (mL) of sample 30.1–45 70 30 b: Total volume of the sample solution prepared by dilut- ing with the diluent Flow rate: About 1.0 mL per minute. Diluent: Dissolve 5 g of chlorobutanol, 1.1 g of sodium System suitability— acetate trihydrate, 5 g of acetic acid (100) and 6 System performance: Dissolve 2 mg each of oxytocin and mL of ethanol (99.5) in water to make 1000 mL. vasopressin in 20 mL of the mobile phase A. When the procedure is run with 25 mL of this solution under the above Operating conditions— operating conditions, vasopressin and oxytocin are eluted in Detector: An ultraviolet absorption photometer (wave- this order with the resolution between these peaks being not length: 220 nm). less than 14, and the symmetry factor of the peak of oxyto- Column: A stainless steel column 4.6 mm in inside diame- cin is not more than 1.5. ter and 15 cm in length, packed with octadecylsilanized silica System repeatability: When the test is repeated 6 times gel for liquid chromatography (5 mminparticlediameter). with 25 mL of the standard solution under the above operat- Column temperature: A constant temperature of about ing conditions, the relative standard deviation of the peak 259C. area of oxytocin is not more than 1.0z. Mobile phase A: Dissolve 15.6 g of sodium dihydrogen phosphate dihydrate in 1000 mL of water. Containers and storage Containers—Tight containers. Mobile phase B: A mixture of water and acetonitrile (1:1). Storage—At 2 to 89C. Flowing of the mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Oxytocin Injection Time after injection Mobile phase A Mobile phase B オキシトシン注射液 of sample (min) (volz) (volz)

Oxytocin Injection is an aqueous solution for injec- 0–30 70→ 40 30 → 60 tion. 30–30.1 40→ 70 60 → 30 It contains not less than 90.0z and not more than 30.1 – 45 70 30 110.0z of the labeled oxytocin Units. Flow rate: About 1.0 mL per minute. Method of preparation Prepare as directed under Injec- System suitability— tions, with Oxytocin. System performance: Dissolve 2 mg each of oxytocin and Description Oxytocin Injection is a colorless, clear liquid. vasopressin in 100 mL of the mobile phase A. When the procedure is run with 100 mL of this solution under the pH <2.54> 2.5–4.5 above operating conditions, vasopressin and oxytocin are Bacterial endotoxins <4.01> Less than 10 EU/oxytocin eluted in this order with the resolution between these peaks Unit. being not less than 14, and the symmetry factor of the peak of oxytocin is not more than 1.5. Extractable volume <6.05> It meets the requirement. System repeatability: When the test is repeated 6 times Foreign insoluble matter <6.06> Perform the test according with 100 mL of the standard solution under the above operat- to the Method 1: it meets the requirement. ing conditions, the relative standard deviation of the peak area of oxytocin is not more than 2.0z. Insoluble particulate matter <6.07> It meets the requirement. Containers and storage Containers—Hermetic containers. Storage—In a cold place, and avoid freezing. Sterility <4.06> Perform the test according to the Mem- brane filtration method: it meets the requirement. Assay Measure exactly a portion of Oxytocin Injection according to the labeled Units, dilute with the diluent so that each mL contains about 1 Unit, and use this solution as the sample solution. Separately, dissolve 1 bottle of Oxytocin RS in the mobile phase A to make exactly 20 mL. Pipet a suitable volume of this solution, dilute with the diluent to make a known concentration solution so that each mL contains about 1 Unit, and use this solution as the standard solution. Perform the test with exactly 100 mL each of the sample solution and standard solution as directed under Liquid Chroma- tography <2.01> according to the following conditions, and 1206 Ozagrel Sodium / Official Monographs JP XVI

of the amount other than ozagrel is not more than 0.2z, Ozagrel Sodium and the total amount other than ozagrel is not more than 0.5z. オザグレルナトリウム Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wavelength: 220 nm).

C13H11N2NaO2: 250.23 Time span of measurement: About 2 times as long as the Monosodium (2E)-3-[4-(1H-imidazol- retention time of ozagrel, beginning after the solvent peak. 1-ylmethyl)phenyl]prop-2-enoate System suitability— [189224-26-8] Test for required detectability: Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 200 mL, Ozagrel Sodium, when dried, contains not less than and use this solution as the solution for system suitability 98.0z and not more than 102.0z of C13H11N2NaO2. test. Pipet 2 mL of the solution for system suitability test, and add the mobile phase to make exactly 10 mL. Confirm Description Ozagrel Sodium occurs as white crystals or that the peak area of ozagrel obtained from 5 mL of this so- crystalline powder. lution is equivalent to 15 to 25z of that from 5 mLoftheso- It is freely soluble in water, soluble in methanol, and prac- lution for system suitability test. tically insoluble in ethanol (99.5). System performance: When the procedure is run with 5 mL Identification (1) Determine the absorption spectrum of a of the solution for system suitability test under the above op- solution of Ozagrel Sodium (1 in 200,000) as directed under erating conditions, the number of theoretical plates and the Ultraviolet-visible Spectrophotometry <2.24>,andcompare symmetry factor of the peak of ozagrel are not less than 6000 the spectrum with the Reference Spectrum or the spectrum and not more than 2.0, respectively. of a solution of Ozagrel Sodium RS prepared in the same System repeatability: When the test is repeated 6 times manner as the sample solution: both spectra exhibit similar with 5 mL of the solution for system suitability test under the intensities of absorption at the same wavelengths. above operating conditions, the relative standard deviation (2) Determine the infrared absorption spectrum of of the peak area of ozagrel is not more than 2.0z. Ozagrel Sodium as directed in the potassium bromide disk Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, method under Infrared Spectrophotometry <2.25>,andcom- 4 hours). pare the spectrum with the Reference Spectrum or the spectrum of Ozagrel Sodium RS: both spectra exhibit similar in- Assay Weigh accurately about 25 mg each of Ozagrel So- tensities of absorption at the same wave numbers. dium and Ozagrel Sodium RS, both previously dried, and (3) A solution of Ozagrel Sodium (1 in 20) responds to dissolve each in methanol to make exactly 25 mL. Pipet 5 the Qualitative Tests <1.09> for sodium salt. mL each of these solutions, add exactly 5 mL of the internal standard solution, and use these solutions as the sample so- pH <2.54> The pH of a solution prepared by dissolving lution and the standard solution, respectively. Perform the 0.5 g of Ozagrel Sodium in 10 mL of water is between 9.5 test with 1 mL each of the sample solution and standard solu- and 10.5. tion as directed under Liquid Chromatography <2.01> ac- Purity (1) Clarity and color of solution—Dissolve 0.5 g cording to the following conditions, and calculate the ratios, of Ozagrel Sodium in 10 mL of water: the solution is clear QT and QS, of the peak area of ozagrel to that of the internal and colorless. standard. (2) Chloride <1.03>—Dissolve 2.0 g of Ozagrel Sodium in Amount (mg) of C H N NaO ＝ M × Q /Q 30 mL of water, add 1 mL of acetic acid (100) and water to 13 11 2 2 S T S make 50 mL, shake, and allow to stand for 30 minutes. MS: Amount (mg) of Ozagrel Sodium RS Filter the solution, discard the first 5 mL of the filtrate, and Internal standard solution—A solution of benzoic acid in to 25 mL of the subsequent filtrate add 6 mL of dilute nitric methanol (1 in 100). acid and water to make 50 mL. Perform the test with this so- Operating conditions— lution as the test solution. Prepare the control solution as Detector: An ultraviolet absorption photometer (wave- follows: To 0.35 mL of 0.01 mol/L hydrochloric acid VS length: 272 nm). add 0.5 mL of acetic acid (100), 6 mL of dilute nitric acid Column: A stainless steel column 4.6 mm in inside diame- and water to make 50 mL (not more than 0.012z). ter and 15 cm in length, packed with octadecylsilanized silica (3) Heavy metals <1.07>—Proceed with 2.0 g of Ozagrel gel for liquid chromatography (5 mminparticlediameter). Sodium according to Method 2, and perform the test. Pre- Column temperature: A constant temperature of about pare the control solution with 2.0 mL of Standard Lead So- 259C. lution (not more than 10 ppm). Mobile phase: A mixture of a solution of ammonium ace- (4) Related substances—Dissolve 50 mg of Ozagrel So- tate (3 in 1000) and methanol (4:1). dium in 100 mL of the mobile phase, and use this solution as Flow rate: Adjust the flow rate so that the retention time the sample solution. Perform the test with 5 mLofthesam- of ozagrel is about 10 minutes. ple solution as directed under Liquid Chromatography System suitability— <2.01> according to the following conditions. Determine each System performance: When the procedure is run with 1 mL peak area by the automatic integration method, and calcu- of the standard solution under the above operating condi- late the amount of them by the area percentage method: each JP XVI Official Monographs / Pancreatin 1207 tions, the internal standard and ozagrel are eluted in this mg of Ozagrel Sodium RS, and dissolve in methanol to make order with the resolution between these peaks being not less exactly 25 mL. Pipet 5 mL of this solution, add exactly 5 mL than 2.0, and the symmetry factor of the peak of ozagrel is of the internal standard solution, and use this solution as the not more than 2.0. standard solution. Then, proceed as directed in the Assay System repeatability: When the test is repeated 6 times under Ozagrel Sodium. with 1 mL of the standard solution under the above operating Amount (mg) of ozagrel sodium (C H N NaO ) conditions, the relative standard deviation of the ratio of the 13 11 2 2 ＝ M × Q /Q × 16 peak area of ozagrel to that of the internal standard is not S T S more than 1.0z. MS: Amount (mg) of Ozagrel Sodium RS Containers and storage Containers—Tight containers. Internal standard solution—A solution of benzoic acid in Storage—Light-resistant. methanol (1 in 100). Containers and storage Containers—Hermetic containers. Ozagrel Sodium for Injection

注射用オザグレルナトリウム Pancreatin パンクレアチン Ozagrel Sodium for Injection is a preparation for injection, which is dissolved before use. Pancreatin is a substance containing enzymes pre- It contains not less than 95.0z and not more than pared from the pancreas of edible animals, mostly the 105.0z of the labeled amount of ozagrel sodium hog, and has amylolytic, proteolytic and lipolytic ac- (C H N NaO : 250.23). 13 11 2 2 tivities. Method of preparation Prepare as directed under Injec- It contains not less than 2800 starch saccharifying tions, with Ozagrel Sodium. activity units, not less than 28,000 proteolytic activity units, and not less than 960 lipolytic activity units Description Ozagrel Sodium for Injection occurs as white per g. masses or powder. It is usually diluted with suitable excipients. Identification Dissolve an amount of Ozagrel Sodium for Description Pancreatin occurs as a white to light yellow Injection, equivalent to 40 mg of Ozagrel Sodium according powder. It has a characteristic odor. to the labeled amount, in water to make 40 mL. To 1 mL of this solution add water to make 200 mL, and determine the Purity (1) Rancidity—Pancreatin has no unpleasant or absorption spectrum of this solution as directed under Ultra- rancid odor and is tasteless. violet-visible Spectrophotometry <2.24>: it exhibits a maxi- (2) Fat—Add 20 mL of diethyl ether to 1.0 g of Pancrea- mum between 269 nm and 273 nm. tin, extract with occasional shaking for 30 minutes, and filter. Wash the residue with 10 mL of diethyl ether, combine pH Being specified separately. the washing with the filtrate, evaporate the diethyl ether, and Purity Related substances—Dissolve an amount of Ozagrel dry the residue at 1059C for 2 hours: the mass of the residue Sodium for Injection, equivalent to 0.20 g of Ozagrel So- does not exceed 20 mg. dium according to the labeled amount, in the mobile phase Loss on drying <2.41> Not more than 4.0z (1 g, in vacu- to make 100 mL. To 5 mL of this solution add the mobile um, phosphorus (V) oxide, 24 hours). phase to make 20 mL, and use this solution as the sample solution. Then, proceed as directed in the Purity (4) under Residue on ignition <2.44> Not more than 5z (1 g). Ozagrel Sodium. Assay (1) Starch digestive activity <4.03> Bacterial endotoxins <4.01> Less than 3.7 EU/mg. (i) Substrate solution—Use potato starch TS for amylolytic activity test, prepared by adding 10 mL of phosphate Uniformity of dosage units <6.02> It meets the requirement buffer solution for pancreatin instead of 10 mL of 1 mol/L of the Mass variation test. acetic acid-sodium acetate buffer solution, pH 5.0. Foreign insoluble matter <6.06> Perform the test according (ii) Sample solution—Weigh accurately about 0.1 g of to Method 2: it meets the requirement. Pancreatin, add a suitable amount of ice-cold water, stir, and add ice-cold water to make exactly 100 mL. Pipet 10 mL Insoluble particulate matter <6.07> It meets the require- of this solution, and add ice-cold water to make exactly 100 ment. mL. Sterility <4.06> Perform the test according to the Mem- (iii) Procedure—Proceed as directed in 1.1. Measure- brane filtration method: it meets the requirement. ment of starch saccharifying activity of 1. Assay for starch digestive activity under Digestion Test. Assay Dissolve an amount of Ozagrel Sodium for Injec- (2) Protein digestive activity <4.03> tion, equivalent to about 0.4 g of ozagrel sodium (i) Substrate solution—Use the substrate solution 2 (C H N NaO ), in water to make exactly 200 mL. Pipet 5 13 11 2 2 described in (2) Assay for protein digestive activity under mL of this solution, add exactly 10 mL of the internal stand- Digestion Test after adjusting the pH to 8.5. ard solution and 5 mL of water, mix, and use this solution as (ii) Sample solution—Weigh accurately about 0.1 g of the sample solution. Separately, weigh accurately about 25 Pancreatin, add a suitable amount of ice-cold water, stir, 1208 Pancuronium Bromide / Official Monographs JP XVI and add ice-cold water to make exactly 200 mL. clear and colorless. (iii) Procedure—Proceed as directed in (2) Assay for (2) Related substances—Dissolve 50 mg of Pancuronium protein digestive activity under Digestion Test, using Bromide in 5 mL of ethanol (95), and use this solution as the trichloroacetic acid TS B as the precipitation reagent. sample solution. Pipet 1 mL of the sample solution, add (3) Fat digestive activity <4.03> ethanol (95) to make exactly 100 mL, and use this solution as (i) Emulsifier—Prepare with 18 g of polyvinyl alcohol I the standard solution (1). Separately, weigh exactly 5 mg of and 2 g of polyvinyl alcohol II as directed in (3) Assay for fat dacuronium bromide for thin-layer chromatography, add digestive activity under Digestion Test. ethanol (95) to make exactly 25 mL, and use this solution as (ii) Substrate solution—Use the substrate solution the standard solution (2). Perform the test with these solu- described in (3) Assay for fat digestive activity under the tions as directed under Thin-layer Chromatography <2.03>. Digestion Test. Spot 2 mL each of the sample solution and standard solutions (iii) Sample solution—Weigh accurately about 0.1 g of (1) and (2) on a plate of silica gel for thin-layer chromatogra- Pancreatin, add a suitable amount of ice-cold water, stir, phy. Develop the plate with a mixture of 2-propanol, aceto- and add ice-cold water to make exactly 100 mL. nitrile and a solution of sodium iodide (1 in 5) (17:2:1) to a (iv) Procedure—Proceed as directed in (3) Assay for fat distance of about 12 cm, and air-dry the plate. Spray evenly digestive activity under Digestion Test, using phosphate a solution of sodium nitrite in methanol (1 in 100) on the buffer solution, pH 8.0, as the buffer solution. plate, allow to stand for 2 minutes, and spray evenly potassium bismuth iodide TS on the plate: a spot from the sample Containers and storage Containers—Tight containers. solution, corresponding to that from the standard solution Storage—Not exceeding 309C. (2), has no more color than that from the standard solution (2), and the spots other than the principal spot and the above mentioned spot from the sample solution have no more color Pancuronium Bromide than the spot from the standard solution (1). パンクロニウム臭化物 Water <2.48> Not more than 8.0z (0.3 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.2 g of Pancuronium Bromide, dissolve in 50 mL of acetic anhydride by warming, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination, and make any necessary correction.

C35H60Br2N2O4:732.67 Each mL of 0.1 mol/L perchloric acid VS 1,1?-(3a,17b-Diacetoxy-5a-androstan-2b,16b-diyl)bis(1- ＝ 36.63 mg of C35H60Br2N2O4 methylpiperidinium) dibromide Containers and storage Containers—Tight containers. [15500-66-0] Storage—Light-resistant. Pancuronium Bromide contains not less than 98.0z and not more than 102.0z of C35H60Br2N2O4, calculated on the dehydrated basis. Panipenem Description Pancuronium Bromide occurs as a white crys- パニペネム talline powder. It is very soluble in water, and freely soluble in ethanol (95) and in acetic anhydride. It is hygroscopic. Identification (1) Determine the infrared absorption spectrum of Pancuronium Bromide as directed in the potassium C15H21N3O4S: 339.41 bromide disk method under Infrared Spectrophotometry (5R,6S)-6-[(1R)-1-Hydroxyethyl]-3-[(3S)-1-(1- <2.25>, and compare the spectrum with the Reference Spec- iminoethyl)pyrrolidin-3-ylsulfanyl]-7-oxo-1- trum: both spectra exhibit similar intensities of absorption at azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid the same wave numbers. [87726-17-8] (2) A solution of Pancuronium Bromide (1 in 100) responds to the Qualitative Tests <1.09> (1) for bromide. Panipenem contains not less than 900 mg (potency) and not more than 1010 mg (potency) per mg, calcu- Optical rotation <2.49> [a]20: ＋38 – ＋429(0.75 g calcu- D lated on the anhydrous basis and corrected on the lated on the dehydrated basis, water, 25 mL, 100 mm). amount of the residual solvent. The potency of pH <2.54> The pH of a solution of Pancuronium Bromide Panipenem is expressed as mass (potency) of pani- (1 in 100) is between 4.5 and 6.5. penem (C15H21N3O4S). Purity (1) Clarity and color of solution—Dissolve 1.0 g Description Panipenem occurs as a white to light yellow, of Pancuronium Bromide in 10 mL of water: the solution is crystalline powder or mass. JP XVI Official Monographs / Panipenem 1209

It is very soluble in water, freely soluble in methanol, nal standard from the standard solution (2). Calculate the slightly soluble in ethanol (99.5), and practically insoluble in amount of the ethanol and acetone by the following for- diethyl ether. mula: ethanol is not more than 5.0z and acetone is not It is hygroscopic. more than 1.0z. It deliquesces in the presence of moisture. Amount (z) of ethanol in Panipenem

Identification (1) Dissolve 0.02 g of Panipenem in 2 mL ＝ 15 × 0.79 × (QTa ＋ QSa2 － 2QSa1)/2(QSa2 － QSa1) of water, add 1 mL of hydroxylammonium chloride-ethanol × 1/1000 × 100/M TS, allow to stand for 3 minutes, add 1 mL of acidic ammo- M: Amount (g) of Panipenem nium iron (III) sulfate TS, and shake: a red-brown color develops. Amount (z) of acetone in Panipenem

(2) Determine the absorption spectrum of a solution of ＝ 3 × 0.79 × (QTb ＋ QSb2 － 2QSb1)/2(QSb2 － QSb1) Panipenem in 0.02 mol/L 3-(N-morpholino)propanesulfonic × 1/1000 × 100/M acid buffer solution, pH 7.0 (1 in 50,000) as directed under M: amount (g) of Panipenem Ultraviolet-visible Spectrophotometry <2.24>:itexhibitsa 0.79: Specific gravity (d 20 ) of ethanol (99.5) and acetone maximum between 296 nm and 300 nm. 20 (3) Determine the infrared absorption spectrum of Internal standard solution—A solution of 1-propanol (1 in Panipenem as directed in the potassium bromide disk 400). method under Infrared Spectrophotometry <2.25>:itexhibits Operating conditions— absorption at the wave numbers of about 1760 cm－1, 1676 Detector: Hydrogen flame-ionization detector. cm－1, 1632 cm－1, 1588 cm－1,1384cm－1 and 1249 cm－1. Column: A glass column 1 mm in inside diameter and 40 m in length, coated with porous polymer beads for gas Absorbance <2.24> E 1z (298 nm): 280 – 310 (50 mg calcu- 1cm chromatography. lated on the anhydrous and desolvent basis, 0.02 mol/L 3- Column temperature: A constant temperature of about (N-morpholino)propanesulfonic acid buffer solution, pH 1409C. 7.0, 2500 mL). Carrier gas: Helium. 20 Optical rotation <2.49> [a]D : ＋55 – ＋659(0.1 g, calcu- Flow rate: Adjust the flow rate so that the retention time lated on the anhydrous and corrected on the amount of the of 1-propanol is about 6 minutes. residual solvent, 0.1 mol/L 3-(N-morpholino)propane- System suitability— sulfonic acid buffer solution, pH 7.0, 10 mL, 100 mm). System performance: When the procedure is run with 1 mL of the gas of the standard solution (2) under the above pH <2.54> Dissolve 0.5 g of Panipenem in 10 mL of water: operating conditions, ethanol, acetone and the internal the pH of the solution is between 4.5 and 6.5. standard are eluted in this order with the resolution between Purity (1) Clarity and color of solution—Being specified ethanol and acetone being not less than 4. separately. System repeatability: When the test is repeated 6 times (2) Heavy metals <1.07>—Proceed with 1.0 g of Panipen- with 1 mL of the gas of the standard solution (2) under the em according to Method 4, and perform the test. Prepare the above operating conditions, the relative standard deviation control solution with 2.0 mL of Standard Lead Solution (not of the ratios of the peak area of ethanol to that of the inter- more than 20 ppm). nal standard is not more than 5.0z. (3) Residual solvents <2.46>—Weigh accurately about (4) Related substances—Being specified separately. 0.2 g of Panipenem, transfer to a 20-mL narrow-mouthed Water <2.48> Weigh accurately about 0.5 g of Panipenem, cylindrical glass bottle, add exactly 2 mL of the internal transfer to a 15-mL narrow-mouthed cylindrical glass bottle, standard solution and 2 mL of water to dissolve, seal tightly add exactly 2 mL of the internal standard solution to dis- a rubber stopper with aluminum cap, and use this solution as solve, seal tightly a rubber stopper with aluminum cap, and the sample solution. Separately, pipet 15 mL of ethanol use this solution as the sample solution. Separately, weigh (99.5) and 3 mL of acetone, add water to make exactly 200 accurately 2 g of water, and add the internal standard solu- mL. Pipet 1 mL and 2 mL of this solution, and add water to tion to make exactly 100 mL. Pipet 5 mL and 10 mL of this them to make exactly 20 mL. Transfer exactly 2 mL each of solution, add the internal standard solution to make exactly these solutions to a 20-mL narrow-mouthed cylindrical glass 20 mL, and use these solutions as the standard solution (1) bottle, add exactly 2 mL of the internal standard solution, and the standard solution (2). Perform the test with 1 mLof seal tightly a rubber stopper with aluminum cap, and use the sample solution and standard solutions (1) and (2) as di- these solutions as the standard solution (1) and the standard rected under Gas Chromatography <2.02> according to the solution (2). Shake gently the sample solution and the stan- following condition, and calculate the ratios, Q , Q and dard solutions (1) and (2) in a water bath at a constant room T S1 Q of the peak area of water to that of the internal stand- temperature, and allow to stand for 30 minutes. Perform the S2 ard. Calculate the amount of water by the following for- test with 1 mL of the gas in each container as directed under mula: water is not more than 5.0z. Gas Chromatography <2.02> according to the following conditions. Calculate the ratios, QTa and QTb, of the peak area Amount of water (z) of ethanol and acetone to that of the internal standard from ＝ MS/MT × (QT ＋ QS2 － 2QS1)/2(QS2 － QS1) the sample solution, the ratios, QSa1 and QSb1, of the peak × 1/100 × 100 area of ethanol and acetone to that of the internal standard M : Amount (g) of water from the standard solution (1), and the ratios, Q and Q , S Sa2 Sb2 M : Amount (g) of Panipenem of the peak area of ethanol and acetone to that of the inter- T 1210 Pantethine / Official Monographs JP XVI

Internal standard solution—A solution of acetonitrile in of the internal standard is about 12 minutes. methanol (1 in 100). System suitability— Operating conditions— System performance: When the procedure is run with 10 Detector: A thermal conductivity detector. mL of the standard solution under the above operating con- Column: A glass column 3 mm in inside diameter and 2 m ditions, panipenem and the internal standard are eluted in in length, packed with porous ethylvinylbenzene-divinylben- this order with the resolution between these peaks being not zene copolymer for gas chromatography (150 to 180 mmin less than 3. particle diameter). System repeatability: When the test is repeated 6 times Column temperature: A constant temperature of about with 10 mL of the standard solution under the above operat- 1259C. ing conditions, the relative standard deviation of the ratios Carrier gas: Helium. of the peak area of panipenem to that of the internal stand- Flow rate: Adjust the flow rate so that the retention time ard is not more than 2.0z. of acetonitrile is about 8 minutes. Containers and storage Containers—Tight containers. System suitability— Storage—Not exceeding －109C. System performance: When the procedure is run with 1 mL of the standard solution (2) under the above operating conditions, water, methanol, and the internal standard are eluted in this order with the resolution between water and internal Pantethine standard being not less than 10. パンテチン System repeatability: When the test is repeated 6 times with 1 mL of the standard solution (2) under the above operating conditions, the relative standard deviation of the ratios of the peak area of water to that of the internal standard is not more than 5.0z. Residue on ignition Being specified separately. Bacterial endotoxins <4.01> Less than 0.15 EU/mg (potency). C22H42N4O8S2:554.72 Bis(2-{3-[(2R)-2,4-dihydroxy-3,3- Assay Weigh accurately an amount of Panipenem and dimethylbutanoylamino]propanoylamino}ethyl) disulfide Panipenem RS, equivalent to about 0.1 g (potency), dissolve [16816-67-4] separately in 0.02 mol/L 3-(N-morpholino)propanesulfonic acid buffer solution, pH 7.0 to make exactly 100 mL. Pipet 5 Pantethine is an aqueous solution containing 80z mL each of these solutions, add exactly 5 mL of the internal of pantethine. standard solution, add 0.02 mol/L 3-(N-morpholino)pro- Pantethine contains not less than 98.0z of pan- panesulfonic acid buffer solution, pH 7.0 to make 20 mL, tethine (C H N O S ), calculated on the anhydrous and use these solutions as the sample solution and standard 22 42 4 8 2 basis. solution. Perform the test within 30 minutes after preparation of the solutions with 10 mL of the sample solution and Description Pantethine is a clear, colorless to pale yellow standard solution as directed under Liquid Chromatography viscous liquid. <2.01> according to the following conditions, and calculate It is miscible with water, with methanol and with ethanol the ratios, QT and QS, of the peak area of panipenem to that (95). of the internal standard. It is decomposed by light.

Amount [mg (potency)] of panipenem (C15H21N3O4S) Identification (1) To 0.7 g of Pantethine add 5 mL of so- ＝ MS × QT/QS × 1000 dium hydroxide TS, shake, and add 1 to 2 drops of copper (II) sulfate TS: a blue-purple color develops. M : Amount [mg (potency)] of Panipenem RS S (2) To 0.7 g of Pantethine add 3 mL of water, shake, Internal standard solution—A solution of sodium p- add 0.1 g of zinc powder and 2 mL of acetic acid (100), and styrenesulfonate in 0.02 mol/L 3-(N-morpholino)propane- boil for 2 to 3 minutes. After cooling, add 1 to 2 drops of so- sulfonic acid buffer solution, pH 7.0 (1 in 1000). dium pentacyanonitrosylferrate (III) TS: a red-purple color Operating conditions— develops. Detector: An ultraviolet absorption photometer (wave- (3) To 1.0 g of Pantethine add 500 mL of water, and length: 280 nm). shake. To 5 mL of this solution add 3 mL of 1 mol/L hydro- Column: A stainless steel column 4.6 mm in inside diame- chloric acid TS, and heat on a water bath for 30 minutes. ter and 25 cm in length, packed with octadecylsilanized sili- After cooling, add 7 mL of a solution of hydroxylammo- cone polymer coated silica gel for liquid chromatography (5 nium chloride in sodium hydroxide TS (3 in 140), and allow mm in particle diameter). to stand for 5 minutes. Add 3 drops of 2,4-dinitrophenol TS, Column temperature: A constant temperature of about and add 1 mol/L hydrochloric acid TS dropwise until the so- 409C. lution has no color, and then add 1 mL of iron (III) chloride Mobile phase: A mixture of 0.02 mol/L 3-(N-morpho- TS: a red-purple color develops. lino)propanesulfonic acid buffer solution, pH 8.0 and aceto- Optical rotation <2.49> [a]20: ＋15.0 – ＋18.09(1 g calcu- nitrile (50:1). D lated on the anhydrous basis, water, 25 mL, 100 mm). Flow rate: Adjust the flow rate so that the retention time JP XVI Official Monographs / Papaverine Hydrochloride 1211

Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Pantethine according to Method 1, and perform the test. Papaverine Hydrochloride Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). パパベリン塩酸塩 (2) Arsenic <1.11>—Prepare the test solution with 2.0 g of Pantethine according to Method 3, and perform the test (not more than 1 ppm). (3) Related substances—Dissolve 0.6 g of Pantethine in 10 mL of water, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin- layer Chromatography <2.03>. Spot 2 mLeachofthesample solution and standard solution on a plate of silica gel for C20H21NO4.HCl: 375.85 thin-layer chromatography. Develop the plate with 2-buta- 6,7-Dimethoxy-1-(3,4-dimethoxybenzyl)isoquinoline none saturated with water to a distance of about 10 cm, and monohydrochloride air-dry the plate. Allow the plate to stand for about 10 [61-25-6] minutes in iodide vapor: the spots other than the principal spot from the sample solution are not more intense than the Papaverine Hydrochloride, when dried, contains spot from the standard solution. not less than 98.5z of C20H21NO4.HCl. (4) Mercapto compounds—To 1.5 g of Pantethine add Description Papaverine Hydrochloride occurs as white 20 mL of water, shake, add 1 drop of ammonia TS and 1 to crystals or crystalline powder. 2 drops of sodium pentacyanonitrosylferrate (III) TS: a red It is sparingly soluble in water and in acetic acid (100), color is not developed. slightly soluble in ethanol (95), and practically insoluble in Water <2.48> 18–22z (0.2 g, volumetric titration, direct acetic anhydride and in diethyl ether. titration). The pH of a solution of Papaverine Hydrochloride (1 in 50) is between 3.0 and 4.0. Residue on Ignition <2.44> Not more than 0.1z (2 g). Identification (1) To 1 mg of Papaverine Hydrochloride Assay Weigh accurately about 0.3 g of Pantethine, add add 1 drops of formaldehyde-sulfuric acid TS: a colorless to water to make exactly 20 mL. Transfer exactly 5 mL of this light yellow-green color is produced, and it gradually solution in an iodine bottle, and add exactly 25 mL of 0.05 changestodeepred,thentobrown. mol/L bromine VS and 100 mL of water. Add 5 mL of di- (2) Dissolve 0.02 g of Papaverine Hydrochloride in 1 mL luted sulfuric acid (1 in 5) rapidly, stopper tightly immedi- of water, and add 3 drops of sodium acetate TS: a white pre- ately, and warm at 40 to 509C for 15 minutes with occa- cipitate is produced. sional shaking. After cooling, carefully add 5 mL of a solu- (3) Dissolve 1 mg of Papaverine Hydrochloride in 3 mL tion of potassium iodide (2 in 5), then immediately stopper of acetic anhydride and 5 drops of sulfuric acid, heat in a tightly, shake, add 100 mL of water and titrate <2.50> the lib- water bath for 1 minute, and examine under ultraviolet light erated iodine with 0.1 mol/L sodium thiosulfate VS (indica- (main wavelength: 365 nm): the solution shows a yellow- tor: 2 mL of starch TS). Perform a blank determination. green fluorescence. Each mL of 0.05 mol/L bromine VS (4) Dissolve 0.1 g of Papaverine Hydrochloride in 10 mL

＝ 5.547 mg of C22H42N4O8S2 of water, make alkaline with ammonia TS, and shake with 10 mL of diethyl ether. Draw off the diethyl ether layer, Containers and storage Containers—Tight containers. wash with 5 mL of water, and filter. Evaporate the filtrate Storage—Light-resistant, at a temperature not exceeding on a water bath, and dry the residue at 1059C for 3 hours: 109C. the residue so obtained melts <2.60> between 1459Cand 1489C. (5) Alkalify a solution of Papaverine Hydrochloride (1 in 50) with ammonia TS, and filter the precipitate. Acidify the filtrate with dilute nitric acid: the solution responds to Qualitative Tests <1.09> (2) for chloride. Purity (1) Clarity and color of solution—Dissolve 0.10 g of Papaverine Hydrochloride in 10 mL of water: the solution is clear and colorless. (2) Morphine—Dissolve 10 mg of Papaverine Hydro- chloride in 1 mL of water, add 5 mL of 1-nitroso-2-naphthol TS and 2 mL of a solution of potassium nitrate (1 in 10), and warm at 409C for 2 minutes. Add 1 mL of a solution of sodium nitrate (1 in 5000), and warm at 409C for 5 minutes. After cooling, shake the mixture with 10 mL of chloroform, centrifuge, and separate the aqueous layer: the solution so obtained has no more color than a pale red color. 1212 Papaverine Hydrochloride Injection / Official Monographs JP XVI

(3) Readily carbonizable substances <1.15>—Perform the ment. test with 0.12 g of Papaverine Hydrochloride: the solution Sterility <4.06> Perform the test according to the Mem- has no more color than Matching Fluid for Color S or P. brane filtration method: it meets the requirement. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, Assay Dilute an exactly measured volume of Papaverine 4 hours). Hydrochloride Injection, equivalent to about 0.2 g of

Residue on ignition <2.44> Not more than 0.2z (1 g). papaverine hydrochloride (C20H21NO4.HCl), with water to 10 mL, render the solution alkaline with ammonia TS, and Assay Weigh accurately about 0.5 g of Papaverine Hydro- extract with 20-mL, 15-mL, 10-mL and 10-mL portions of chloride, previously dried, dissolve in 100 mL of a mixture chloroform. Combine the extracts, wash with 10 mL of of acetic anhydride and acetic acid (100) (7:3) by warming, water, and re-extract the washings with two 5-mL portions cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS of chloroform. Combine all the chloroform extracts, and (potentiometric titration). Perform a blank determination, distil the chloroform on a water bath. Dissolve the residue in and make any necessary correction. 30 mL of acetic acid (100), and titrate <2.50> with 0.05 Each mL of 0.1 mol/L perchloric acid VS mol/L perchloric acid VS (indicator: 2 drops of crystal violet

＝ 37.59 mg of C20H21NO4.HCl TS). Perform a blank determination, and make any necessary correction. Containers and storage Containers—Tight containers. Storage—Light-resistant. Each mL of 0.05 mol/L perchloric acid VS

＝ 18.79 mg of C20H21NO4.HCl Containers and storage Containers—Hermetic containers. Papaverine Hydrochloride Injection Storage—Light-resistant. パパベリン塩酸塩注射液 Paraffin Papaverine Hydrochloride Injection is an aqueous solution for injection. パラフィン It contains not less than 95.0z and not more than 105.0z of the labeled amount of papaverine hydro- Paraffin is a mixture of solid hydrocarbons ob- chloride (C H NO .HCl: 375.85). 20 21 4 tained from petroleum. Method of preparation Prepare as directed under Injec- Description Paraffin occurs as a colorless or white, more tions, with Papaverine Hydrochloride. or less transparent, crystalline mass. It is odorless and taste- Description Papaverine Hydrochloride Injection is a clear, less. colorless liquid. It is sparingly soluble in diethyl ether and practically in- pH:3.0–5.0 soluble in water, in ethanol (95) and in ethanol (99.5). Specific gravity d 20: about 0.92 (proceed as directed in Identification (1) To 1 mL of Papaverine Hydrochloride 20 4.2. in 4. Specific gravity under Fats and Fatty Oils Test Injection add 3 drops of sodium acetate TS: a white precipi- <1.13>). tate is produced. (2) Dilute a volume of Papaverine Hydrochloride Injec- Identification (1) Heat Paraffin strongly in a porcelain tion, equivalent to 0.1 g of Papaverine Hydrochloride ac- dish, and ignite: it burns with a bright flame and the odor of cording to the labeled amount, with water to 10 mL, render paraffin vapor is perceptible. the solution alkaline with ammonia TS, and shake with 10 (2) Heat 0.5 g of Paraffin with 0.5 g of sulfur with shak- mL of diethyl ether. Draw off the diethyl ether layer, wash ing carefully: the odor of hydrogen sulfide is perceptible. with 5 mL of water, and filter. Evaporate the filtrate on a Melting point <2.60> 50–759C (Method 2). water bath to dryness, and dry the residue at 1059Cfor3 hours: the residue so obtained melts <2.60> between 1459C Purity (1) Acidity or alkalinity—Boil 10.0 g of Paraffin and 1489C. with 10 mL of hot water and 1 drop of phenolphthalein TS (3) Proceed with 1 mg each of the residue obtained in (2) in a water bath for 5 minutes, and shake vigorously: a red as directed in the Identification (1) and (3) under Papaverine color is not produced. Add 0.20 mL of 0.02 mol/L sodium Hydrochloride. hydroxide VS to this solution, and shake: a red color is pro- (4) Alkalify 2 mL of Papaverine Hydrochloride Injection duced. with ammonia TS, filter the precipitate off, and acidity the (2) Heavy metals <1.07>—Ignite2.0gofParaffinina filtrate with dilute nitric acid: the solution responds to crucible, first moderately until charred, then between 4509C Qualitative Tests <1.09> (2) for chloride. and 5509C to ash. Cool, add 2 mL of hydrochloric acid, and evaporate on a water bath to dryness. To the residue add 2 Bacterial endotoxins <4.01> Less than 6.0 EU/mg. mL of dilute acetic acid and water to make 50 mL, and per- Extractable volume <6.05> It meets the requirement. form the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Foreign insoluble matter <6.06> Perform the test according Solution add 2 mL of dilute acetic acid and water to make 50 to Method 1: it meets the requirement. mL (not more than 10 ppm). Insoluble particulate matter <6.07> It meets the require- (3) Arsenic <1.11>—Prepare the test solution with 1.0 g JP XVI Official Monographs / Liquid Paraffin 1213 of Paraffin according to Method 3, and perform the test (not acid, and evaporate on a water bath to dryness. To the more than 2 ppm). residue add 2 mL of dilute acetic acid and water to make 50 (4) Sulfur compounds—To 4.0 g of Paraffin add 2 mL mL, and perform the test using this solution as the test solu- of ethanol (99.5), further add 2 drops of a clear saturated so- tion. Prepare the control solution as follows: to 2.0 mL of lution of lead (II) oxide in a solution of sodium hydroxide (1 Standard Lead Solution add 2 mL of dilute acetic acid and in 5), and heat for 10 minutes at 709C with occasional shak- water to make 50 mL (not more than 10 ppm). ing: no dark brown color develops in the aqueous layer. (4) Arsenic <1.11>—Prepare the test solution with 1.0 g (5) Readily carbonizable substances—Melt 5.0 g of of Liquid Paraffin, according to Method 3 except that after Paraffin placed in a Nessler tube at a temperature near the addition of 10 mL of a solution of magnesium nitrate hexa- melting point. Add 5 mL of sulfuric acid for readily car- hydrate in ethanol (95) (1 in 50), add 1.5 mL of hydrogen bonizable substances, and warm at 709C for 5 minutes in a peroxide (30), fire to burn, and perform the test (not more water bath. Remove the tube from the water bath, immedi- than 2 ppm). ately shake vigorously and vertically for 3 seconds, and (5) Solid paraffin—Transfer 50 mL of Liquid Paraffin, warm for 1 minute in a water bath at 709C. Repeat this previously dried at 1059C for 2 hours, to a Nessler tube, and procedure five times: the color of the sulfuric acid layer is cool in ice water for 4 hours: the turbidity produced, if any, not darker than that of the following control solution. is not deeper than that of the following control solution. Control solution: Add 1.5 mL of Cobalt (II) Chloride CS, Control solution: To 1.5 mL of 0.01 mol/L hydrochloric 0.5 mL of Copper (II) Sulfate CS and 5 mL of liquid acid VS add 6 mL of dilute nitric acid and water to make 50 paraffin to 3.0 mL of Iron (III) Chloride CS, and shake mL, add 1 mL of silver nitrate TS, and allow to stand for 5 vigorously. minutes. (6) Sulfur compounds—Prepare a saturated solution of Containers and storage Containers—Well-closed contain- lead (II) oxide in a solution of sodium hydroxide (1 in 5), ers. and mix 2 drops of this clear solution with 4.0 mL of Liquid Paraffinand2mLofethanol(99.5).Heatat709Cfor10 minutes with frequent shaking, and cool: no dark brown Liquid Paraffin color develops. (7) Polycyclic aromatic hydrocarbons—Take 25 mL of 流動パラフィン Liquid Paraffin by a 25-mL measuring cylinder, transfer to a 100-mL separator, and wash out the cylinder with 25 mL of Liquid Paraffin is a mixture of liquid hydrocarbons hexane for ultraviolet-visible spectrophotometry. Combine obtained from petrolatum. the washings with the liquid in the separator, and shake vig- Tocopherols of a suitable form may by added at a orously. Shake this solution vigorously for 2 minutes with concentration not exceeding 0.001z as a stabilizer. 5.0 mL of dimethylsulfoxide for ultraviolet-visible spectrophotometry, and allow to stand for 15 minutes. Transfer the Description Liquid Paraffin is a colorless, transparent, oily lower layer to a 50-mL separator, add 2 mL of hexane for liquid, nearly free from fluorescence. It is odorless and taste- ultraviolet-visible spectrophotometry, shake vigorously for 2 less. minutes, and allow to stand for 2 minutes. Transfer the It is freely soluble in diethyl ether, very slightly soluble in lower layer to a 10-mL glass-stoppered centrifuge tube, and ethanol (99.5), and practically insoluble in water and in centrifuge between 2500 revolutions per minute and 3000 ethanol (95). revolutions per minute for about 10 minutes, and use the Boiling point: above 3009C. clear solution obtained as the sample solution. Transfer 25 Identification (1) Heat Liquid Paraffin strongly in a por- mL of hexane for ultraviolet-visible spectrophotometry to celain dish, and fire: it burns with a bright flame and the another 50-mL separator, shake vigorously for 2 minutes odor of paraffin vapor is perceptible. with 5.0 mL of dimethylsulfoxide for ultraviolet-visible spec- (2) Heat 0.5 of Liquid Paraffin with 0.5 g of sulfur with trophotometry, and allow to stand for 2 minutes. Transfer shaking carefully: the odor of hydrogen sulfide is percepti- the lower layer to a 10-mL glass-stoppered centrifuge tube, ble. centrifuge between 2500 revolutions per minute and 3000 revolutions per minute for about 10 minutes, and use the Specific gravity <2.56> d 20: 0.860 – 0.890 20 clear solution thus obtained as a control solution. Immedi- Viscosity <2.53> Not less than 37 mm2/s (Method 1, ately determine the absorbance of the sample solution using 37.89C). the control solution as the blank as directed under Ultravio- let-visible Spectrophotometry <2.24>: not more than 0.10 at Purity (1) Odor—Transfer a suitable amount of Liquid the wavelength region between 260 nm and 350 nm. Paraffin to a small beaker, and heat on a water bath: a for- (8) Readily carbonizable substances—Transfer 5 mL of eign odor is not perceptible. Liquid Paraffin to a Nessler tube, and add 5 mL of sulfuric (2) Acidity or alkalinity—Shake vigorously 10 mL of acid for readily carbonizable substances. After heating in a Liquid Paraffin with 10 mL of hot water and 1 drop of phe- water bath for 2 minutes, remove the tube from the water nolphthalein TS: no red color develops. Shake this solution bath, and immediately shake vigorously and vertically for 5 with 0.20 mL of 0.02 mol/L sodium hydroxide VS: a red seconds. Repeat this procedure four times: the Liquid color develops. Paraffin layer remains unchanged in color, and the sulfuric (3) Heavy metals <1.07>—Ignite 2.0 g of Liquid Paraffin acid layer has no more color than the following control solu- in a crucible, first moderately until charred, then between tion. 4509C and 5509C to ash. Cool, add 2 mL of hydrochloric Control solution: Mix 3.0 mL of Iron (III) Chloride CS 1214 Light Liquid Paraffin / Official Monographs JP XVI with 1.5 mL of Cobalt (II) Chloride CS and 0.50 mL of Liquid Paraffin and 2 mL of ethanol (99.5). Heat at 709C Copper (II) Sulfate CS. for 10 minutes with frequent shaking, and cool: no dark brown color develops. Containers and storage Containers—Tight containers. (7) Polycyclic aromatic hydrocarbons—Take 25 mL of Light Liquid Paraffin by a 25-mL measuring cylinder, transfer to a 100-mL separator, and wash out the cylinder with 25 Light Liquid Paraffin mL of hexane for ultraviolet-visible spectrophotometry. Combine the washings with the liquid in the separator, and 軽質流動パラフィン shake vigorously. Shake this solution vigorously for 2 minutes with 5.0 mL of dimethylsulfoxide for ultraviolet- Light Liquid Paraffin is a mixture of liquid visible spectrophotometry, and allow to stand for 15 hydrocarbons obtained from petroleum. minutes. Transfer the lower layer to a 50-mL separator, add Tocopherols of a suitable form may be added at a 2 mL of hexane for ultraviolet-visible spectrophotometry, concentration not exceeding 0.001z as a stabilizer. shake vigorously for 2 minutes, and allow to stand for 2 minutes. Transfer the lower layer to a glass-stoppered 10-mL Description Light Liquid Paraffin is a clear, colorless oily centrifuge tube, and centrifuge between 2500 revolutions per liquid, nearly free from fluorescence. It is odorless and taste- minute and 3000 revolutions per minute for about 10 less. minutes, and use the clear solution so obtained as the sample It is freely soluble in diethyl ether, and practically insolu- solution. Separately, transfer 25 mL of hexane for ultravio- bleinwaterandinethanol(95). let-visible spectrophotometry to a 50-mL separator, add 5.0 Boiling point: above 3009C. mL of dimethylsulfoxide for ultraviolet-visible spectropho- Identification (1) Heat Light Liquid Paraffin strongly in tometry, shake vigorously for 2 minutes, and allow to stand a porcelain dish, and fire: it burns with a bright flame and for 2 minutes. Transfer the lower layer to a glass-stoppered the odor of paraffin vapor is perceptible. 10-mL centrifuge tube, centrifuge between 2500 revolutions (2) Heat 0.5 of Light Liquid Paraffin with 0.5 g of sulfur per minute and 3000 revolutions per minute for about 10 with shaking carefully: the odor of hydrogen sulfide is per- minutes, and use the clear solution so obtained as a control ceptible. solution. Immediately determine the absorbance of the sample solution using the control solution as the blank as di- Specific gravity <2.56> d 20: 0.830 – 0.870 20 rected under Ultraviolet-visible Spectrophotometry <2.24>: Viscosity <2.53> Less than 37 mm2/s (Method 1, 37.89C). not more than 0.10 at the wavelength region between 260 nm and 350 nm. Purity (1) Odor—Transfer a suitable amount of Light (8) Readily carbonizable substances—Transfer 5 mL of LiquidParaffintoasmallbeaker,andheatonawaterbath: Light Liquid Paraffin to a Nessler tube, and add 5 mL of no foreign odor is perceptible. sulfuric acid for readily carbonizable substances. After heat- (2) Acidity or alkalinity—Shake vigorously 10 mL of ing in a water bath for 2 minutes, remove the tube from the Light Liquid Paraffin with 10 mL of hot water and 1 drop of water bath, and immediately shake vigorously and vertically phenolphthalein TS: no red color develops. Shake this solu- for 5 seconds. Repeat this procedure four times: the liquid tion with 0.20 mL of 0.02 mol/L sodium hydroxide VS: a paraffin layer remains unchanged in color, and sulfuric acid red color develops. layer has no more color than the following control solution. (3) Heavy metals <1.07>—Ignite 2.0 g of Light Liquid Control solution: Mix 3.0 mL of Iron (III) Chloride CS Paraffin in a crucible, first moderately until charred, then with 1.5 mL of Cobalt (II) Chloride CS and 0.50 mL of between 4509C and 5509C to ash. Cool, add 2 mL of hydro- Copper (II) Sulfate CS. chloric acid, and evaporate on a water bath to dryness. To the residue add 2 mL of dilute acetic acid and water to make Containers and storage Containers—Tight containers. 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and Paraformaldehyde water to make 50 mL (not more than 10 ppm). (4) Arsenic <1.11>—Prepare the test solution with 1.0 g パラホルムアルデヒド of Light Liquid Paraffin according to Method 3, and perform the test (not more than 2 ppm). (CH O) (5) Solid paraffin—Transfer 50 mL of Light Liquid 2 n Poly(oxymethylene) Paraffin, previously dried at 1059C for 2 hours, to a Nessler [30525-89-4] tube, and cool in ice water for 4 hours: the turbidity produced, if any, is not deeper than that of the following con- Paraformaldehyde contains not less than 95.0z of trol solution. CH O: 30.03. Control solution: To 1.5 mL of 0.01 mol/L hydrochloric 2 acid VS add 6 mL of dilute nitric acid and water to make 50 Description Paraformaldehyde occurs as a white powder. mL, add 1 mL of silver nitrate TS, and allow to stand for 5 It has a slight odor of formaldehyde, but a very strong minutes. irritating odor is perceptible when it is heated. (6) Sulfur compounds—Prepare a saturated solution of It is practically insoluble in water, in ethanol (95) and in lead (II) oxide in a solution of sodium hydroxide (1 in 5), diethyl ether. and mix 2 drops of this clear solution with 4.0 mL of Light It dissolves in hot water, in hot dilute hydrochloric acid, in JP XVI Official Monographs / Dental Paraformaldehyde Paste 1215 sodium hydroxide TS and in ammonia TS. It sublimes at about 1009C. Dental Paraformaldehyde Paste Identification (1) Dissolve 0.1 g of Paraformaldehyde in 歯科用パラホルムパスタ 5 mL of ammonia TS, add 5 mL of silver nitrate TS, shake, and add 3 mL of a solution of sodium hydroxide (1 in 10): a mirror of metallic silver is immediately formed on the sides Method of preparation of the container. Paraformaldehyde, finely powdered 35 g (2) Add a solution of 0.04 g of salicylic acid in 5 mL of Procaine Hydrochloride, finely sulfuric acid to 0.02 g of Paraformaldehyde, and warm powdered 35 g slowly: a persistent, dark red color is produced. Hydrous Lanolin a sufficient quantity Purity (1) Clarity and color of solution—Dissolve 0.20 g To make 100 g of Paraformaldehyde in 10 mL of ammonia TS: the solution is clear and colorless. Prepare as directed under Ointments, with the above in- (2) Acidity or alkalinity—To 0.5 g of Paraformaldehyde gredients. add 10 mL of water, shake vigorously for 1 minute, and Description Dental Paraformaldehyde Paste is yellowish filter: the filtrate is neutral. white in color. It has a characteristic odor. (3) Chloride <1.03>—Dissolve 1.5 g of Paraformalde- hyde in 75 mL of water and 7.5 mL of sodium carbonate TS, Identification (1) To 0.15 g of Dental Paraformaldehyde evaporate on a water bath to dryness, and ignite at about Paste add 20 mL of diethyl ether and 20 mL of 0.5 mol/L 5009C. Dissolve the residue in 15 mL of water, filter, if nec- sodium hydroxide TS, shake well, separate the water layer, essary, neutralize with diluted nitric acid (3 in 10), and add 6 and dilute with water to make 100 mL. To 1 mL of this solu- mL of dilute nitric acid and water to make 50 mL. Perform tion add 10 mL of acetylacetone TS, and heat on a water the test using this solution as the test solution. Prepare the bath for 10 minutes: a yellow color is produced (paraformal- control solution as follows: to 0.25 mL of 0.01 mol/L hydro- dehyde). chloric acid VS add 7.5 mL of sodium carbonate TS, a (2) To the diethyl ether layer obtained in (1) add 5 mL of volume of diluted nitric acid (3 in 10) required for neutraliza- dilute hydrochloric acid and 20 mL of water, shake well, and tion of the sample, 6 mL of dilute nitric acid and water to separate the water layer: the solution responds to Qualitative make 50 mL (not more than 0.006z). Tests <1.09> for primary aromatic amines (procaine hydro- (4) Sulfate <1.14>—Dissolve 1.5 g of Paraformaldehyde chloride). in 45 mL of water and 4.5 mL of sodium carbonate TS, (3) To 0.15 g of Dental Paraformaldehyde Paste add 25 evaporate on a water bath to dryness, and ignite at abut mL of diethyl ether and 25 mL of water, shake, separate the 5009C. Dissolve the residue in 15 mL of water, filter, if nec- water layer, filter, and use the filtrate as the sample solution. essary, neutralize the diluted hydrochloric acid (3 in 5), and Seperately, dissolve 0.01 g of procaine hydrochloride in 5 boil for 5 minutes. After cooling, add 1 mL of dilute hydro- mL of water, and use this solution as standard solution. Per- chloric acid and water to make 50 mL. Perform the test form the test with these solutions as directed under Thin- using this solution as the test solution. Prepare the control layer Chromatography <2.03>. Spot 5 mLeachofthesample solution as follows: to 4.5 mL of sodium carbonate TS add solution and standard solution on a plate of silica gel with an equal volume of diluted hydrochloric acid (3 in 5) for the fluorescent indicator for thin-layer chromatography. Devel- neutralization of the sample and 15 mL of water, and boil op the plate with a mixture of ethyl acetate, ethanol (99.5) for 5 minutes. After cooling, add 0.35 mL of 0.005 mol/L and ammonia solution (28) (50:5:1) to a distance of about 10 sulfuric acid VS, 1 mL of dilute hydrochloric acid and water cm, and air-dry the plate. Examine under ultraviolet light to make 50 mL (not more than 0.011z). (main wavelength: 254 nm): spots from the sample solution and standard solution show the same Rf value. Residue on ignition <2.44> Not more than 0.1z (1 g). Containers and storage Containers—Tight containers. Assay Dissolve about 50 mg of Paraformaldehyde, accurately weighed, in 10 mL of potassium hydroxide TS in an iodine flask. Add 40 mL of water and an exactly measured 50 mL of 0.05 mol/L iodine VS, stopper, and allow to stand for 5 minutes. Then add 5 mL of dilute hydrochloric acid, stopper immediately, allow to stand for 15 minutes, and titrate <2.50> the excess iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Perform a blank determination.

Each mL of 0.05 mol/L iodine VS ＝ 1.501 mg of CH2O Containers and storage Containers—Tight containers. 1216 Parnaparin Sodium / Official Monographs JP XVI

tion as directed under Liquid Chromatography <2.01> ac- Parnaparin Sodium cording to the following conditions. Determine the peak height, HUV, in chromatogram obtained by the ultraviolet パルナパリンナトリウム absorption photometer, and determine the peak height, HRI, in chromatogram obtained by the differential refractometer.

Calculate the ratio of HUV to HRI, HRI/HUV,ateachpeak. Assume the molecular mass in the 4th peak from the low molecular mass in chromatogram obtained by the ultraviolet absorption photometer as 2400, and make the calculation of

the standard coefficient from dividing 2400 by the HRI/HUV at the corresponding peak. Make the calculation to multiply

the HRI/HUV at each peak by the standard coefficient, and determine the molecular mass of each peak by the calculation. Prepare the calculation curve by plotting the logarithm of molecular masses at each peak on the vertical axis and the retention time on the chromatogram obtained by the differential refractometer on the horizontal axis. Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 234 nm) and a differential refractometer. Column: Connect two stainless steel columns which are Parnaparin Sodium is a low-molecular heparin 7.5 mm in inside diameter and 30 cm in length, and are sodium obtained by depolymerization, with hydrogen packed with porous silica gel for liquid chromatography; one peroxide and with copper (II) acetate, of heparins column, the molecular mass of limited size exclusion is about sodium from the healthy edible porcine intestinal 500,000; the other, the molecular mass of limited size exclu- mucosa. The mass-average molecular mass ranges be- sion is about 100,000. Connect a pump, the about 500,000- tween 4500 and 6400. molecular mass of limited size exclusion column, the about The potency is not less than 70 low-molecular-mass- 100,000-molecular mass of limited size exclusion column, heparin units and not more than 95 low-molecular- the ultraviolet absorption photometer and the differential mass-heparin units of anti-factor Xa activity per milli- refractometer in this order. gram calculated with reference of the dried substance. Column temperature; A constant temperature of about 409C. Description Parnaparin Sodium occurs as a white or light Mobile phase: Dissolve 28.4 g of sodium sulfate anhydride yellow powder. in 1000 mL of water, and 5.0 with 0.05 mol/L sulfuric acid It is freely soluble in water, and practically insoluble in TS. ethanol (99.5). Flow rate: 0.5 mL per minute. It is hygroscopic. System suitability— Identification (1) Mix 0.1 mL of a solution of Parnaparin System performance: When the procedure is run with 50 Sodium (1 in 20) and 10 mL of a solution of tritoluidine blue mL of the standard solution under the above operating con- O (1 in 100,000), and shake the mixture: the blue color of so- ditions, confirm that more than ten peaks in chromatogram lution immediately changes to purple. obtained as directed under either the Ultraviolet-visible Spec- (2) A solution of Parnaparin Sodium (1 in 20) responds trophotometry, or the Differential Refractometry are ob- to Qualitative Tests <1.09> for sodium salt. served. System repeatability: When the tests repeated 6 times with pH <2.54> Dissolve 0.1 g of Parnaparin Sodium in 10 mL 50 mL of the standard solution under the above operating of water: the pH of this solution is between 6.0 and 8.0. conditions, relative standard deviation of the 4th peak height

Purity (1) Clarity and color of solution—Dissolve 1.0 g in chromatogram (HUV and HRI) is not more than 3.0z. of Parnaparin Sodium in 10 mL of water: the solution is (ii) Determination of molecular mass—Dissolve the 20 clear and colorless or pale yellow. mg of Parnaparin Sodium with 2.0 mL of mobile phase, and (2) Heavy metals <1.07>—Proceedwith1.0gofPar- use this solution as the sample solution. Perform the test naparin Sodium according to Method 2, and perform the with 50 mL of the sample solution as directed under Liquid test. Prepare the control solution with 2.0 mL of Standard Chromatography <2.01> according to the following condi- Lead Solution (not more than 20 ppm). tions. Divide the main peak observed between 30 min and 45 min to 30 sec-interval fractions, and determine the strength Loss on drying <2.41> Not more than 8.0z (0.2 g, in vacu- of differential refractometer of each 30 sec-interval fraction. um, phosphorus (V) oxide, 609C, 3 hours). Determine the molecular mass of each fraction using the Molecular mass Calculate the molecular mass of Parnapa- calibration curve and the retention time of each fraction. De- rin Sodium by the following methods: The mass-average mo- termine the mean of molecular mass in the entire peak using lecular mass ranges between 4500 and 6400. the strength of differential refractometer and the molecular (i) Creation of calibration curve—Weigh 20 mg of low- mass in every fractions. molecularmassheparinforcalibrationofmolecularmass, Mean molecular mass of parnaparin sodium and dissolve it in 2.0 mL of the mobile phase as the standard ＝ S(n ･M )/Sn solution. Perform the test with 50 mL of the standard solu- i i i JP XVI Official Monographs / Parnaparin Sodium 1217

ni: The differential refractometer strength of fraction i in the mixing incubate accurately for 5 minutes at 37 ± 19C. the main peak of chromatogram Then, to each tube add 0.10 mL of sodium calcium solution

Mi: Molecular mass of fraction i in main peak (277 in 100,000) which is pre-warmed at 37 ± 19C, mix, start a stop watch simultaneously, and permit to stand at the Operating conditions— same temperature. Determine the time for the first appear- Detector: A differential refractometer. ance of fibrin clot. Column, column temperature, mobile phase, and flow (iv) Calculation Determine the low-molecular-mass- rate: Proceed as directed in the operating conditions in (i) heparin unit (anti-factor IIa activity) of the sample solution Creation of calibration curve. from calibration curve obtained plots of clotting times for System suitability— each standard solution; calculate the low-molecular-mass- Proceed as directed in (i) Creation of calibration curve. heparin unit (anti-factor IIa activity) for 1 mg of parnaparin Distribution of molecular mass Themolecularmassof sodium as following equation. Parnaparin Sodium is calculated as directed in the determi- The low-molecular-mass-heparin unit (anti-factor IIa nation of molecular mass and the distribution of molecular activity) for 1 mg of parnaparin sodium mass is calculated by the following equation: the molecular ＝ the low-molecular-mass-heparin unit (anti-factor IIa mass of not less than 80z parnaparin sodium is between activity) in 1 mL of sample solution × b/a 1500 and 10,000. a: Amount (mg) of Parnaparin Sodium Distribution of molecular mass (z) b: The total volume (mL) in which Parnaparin Sodium ＝ (Sn /Sn ) × 100 j i has been dissolved with isotonic sodium chloride solu-

ni: The differential refractometer strength of fraction i in tion for the preparation of sample solution the main peak of chromatogram The ratio of anti-factor Xa activity to anti-factor IIa activity Sn : Sum of differential refractometer strength in the each j Divide the anti-factor Xa activity, obtained in the Assay, by fraction between 1500 and 10,000 molecular mass in the anti-factor IIa activity which has been obtained from the the main peak test according to the method of anti-factor IIa activity; the The degree of sulfate ester Dissolve 0.5 g of Parnaparin ratio of anti-factor Xa activity to anti-factor IIa activity is Sodium with 10 mL water. Treat the solution with 5 mL of a between 1.5 and 2.5. strongly basic ion exchange resin, and subsequently with 10 Assay mL of a strongly acidic ion exchange resin. Dilute the solu- (i) Standard solution Dissolve Low-molecular Mass- tion with water to 50 mL, and titrate <2.50> with 0.1 mol/L Heparin RS in isotonic sodium chloride solution to make so- Sodium hydroxide VS (potentiometric titration). Calculate lutions which contain 0.4, 0.6 and 0.8 low-molecular-mass- the degree of sulfate ester of Parnaparin Sodium from the heparin units (anti-factor Xa activity) in 1 mL, respectively. equivalence point by the following equation; it is between (ii) Sample solution Weigh accurately about 50 mg of 2.0 and 2.4. Parnaparin Sodium, and dissolve it in isotonic sodium chlo- Thedegreeofsulfateester ride solution to make a solution which contains 7 mgpar- ＝ the first equivalence point (mL)/[the second naparin sodium in 1 mL. equivalence point (mL) – first equivalence point (mL)] (iii) Procedure To each plastic tube add 0.10 mL of either the sample solution or the standard solution, sepa- Total nitrogen Weigh accurately about 0.10 g of Parnapa- rately. Subsequently to the every tubes add 0.70 mL of Tris- rin Sodium which is dried, and perform the test as directed buffered solution (pH 8.4), 0.10 mL of anti-thrombin III under Nitrogen Determination <1.08>: it contains not less TS, and 0.10 mL of normal human plasma, and mix them. than 1.9z and not more than 2.3z of nitrogen (N:14.01). To another plastic tube transfer 0.20 mL of these solutions, Anti-factor IIa activity Determine the potency of anti- separately, and incubate for accurate 3 minutes at 37 ± 19C. factor IIa activity of Parnaparin Sodium according to the Next, to each tube add 0.10 mL of facter Xa TS and mix it, following method, it contains not less than 35 and not more permit to stand 37 ± 19C accurately for 30 seconds, and im- than 60 low-molecular-mass-heparin unit per milligram cal- mediately add 0.20 mL of chromogenic synthetic substrate culated with reference to the dried substance. solution (3 in 4000) and mix it, and subsequently incubate (i) Standard solution Dissolve Low-molecular Mass accurately for 3 min at 37 ± 19C. To each test tube add 0.30 Heparin RS with isotonic sodium chloride solution to make mL of diluted acetic acid (100) solution (1 in 2) to stop the solutions which contain 0.1, 0.2 and 0.3 low-molecular- reaction. Separately, to plastic tube add 0.10 mL of isotonic mass-heparin unit (anti-factor IIa activity) in 1 mL, respec- sodium chloride solution, 0.70 mL of Tris-buffered solution tively. (pH 8.4), 0.10 mL of anti-thrombin III TS, and 0.10 mL of (ii) Sample solution Weigh accurately about 50 mg of normal human plasma to every tubes, and mix well. To Parnaparin Sodium, and dissolve it with isotonic sodium another plastic tube transfer 0.2 mL of the solution, sepa- chloride solution to adjust the solution which contains 4 mg rately, and add both 0.30 mL of water and 0.30 mL of di- parnaparin sodium in 1 mL. luted acetic acid (100) (1 in 2). Determine the absorbance of (iii) Procedure To each plastic tube add 0.10 mL of the both the sample solution and the standard solution at 405 sample solution and the standard solution, separately. To nm as directed under Ultraviolet-visible Spectrophotometry each tube add 0.10 mL of human normal plasma and mix, <2.24> using a solution obtained from this solution as the and incubate at 37 ± 19C accurately for 1 minute. Next, to blank. each test tube add 0.10 mL of activated thromboplastin-time (iv) Calculation method Determine the low-molecular- assay solution, which is pre-warmed at 37 ± 19C, and after mass unit (anti-factor Xa activity) of the sample solution 1218 Peanut Oil / Official Monographs JP XVI using the calibration curve prepared from the absorbance of Iodine value <1.13> 84–103 the standard solutions and their logarithmic concentrations, Containers and storage Containers—Tight containers. and calculate the low-molecular-mass unit (anti-factor Xa activity) in 1 mg of Parnaparin Sodium. Low-molecular-mass-heparin unit (anti-factor Xa activity) in Pemirolast Potassium 1 mg of Parnaparin Sodium ＝ the low-molecular-mass-heparin unit (anti-factor Xa ペミロラストカリウム activity) in 1 mL of the sample solution × b/a a: Amount (mg) of Parnaparin Sodium b: The total volume (mL) in which Parnaparin Sodium has been dissolved with isotonic sodium chloride solution for the preparation of sample solution Container and Storage C H KN O: 266.30 Container—Well-closed containers. 10 7 6 Monopotassium 5-(9-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidin-3- yl)-1H-tetrazol-1-ide Peanut Oil [100299-08-9] Pemirolast Potassium contains not less than 98.5 Oleum Arachidis z and not more than 101.0z of C10H7KN6O, calculated on the anhydrous basis. ラッカセイ油 Description Pemirolast Potassium occurs as a light yellow crystalline powder. Peanut Oil is the fixed oil obtained from the seeds It is freely soluble in water, slightly soluble in methanol, of Arachis hypogaea Linne(á Leguminosae). and very slightly soluble in ethanol (99.5). Description Peanut Oil is a pale yellow, clear oil. It is odor- It dissolves in potassium hydroxide TS. less or has a slight odor. It has a mild taste. Melting point: about 3229C (with decomposition). It is miscible with diethyl ether and with petroleum ether. Identification (1) Determine the absorption spectrum of a It is slightly soluble in ethanol (95). solution of Pemirolast Potassium in diluted potassium hy- Specific gravity d 25: 0.909 – 0.916 25 droxide TS (1 in 10,000) (1 in 100,000) as directed under Ul- Congealing point of the fatty acids: 22 – 339C traviolet-visible Spectrophotometry <2.24>, and compare the Identification Saponify 5 g of Peanut Oil by boiling with spectrum with the Reference Spectrum or the spectrum of a 2.5 mL of sodium hydroxide solution (3 in 10) and 12.5 mL solution of Pemirolast Potassium RS prepared in the same of ethanol (95). Evaporate the ethanol, dissolve the residue manner as the sample solution: both spectra exhibit similar in 50 mL of hot water, and add dilute hydrochloric acid in intensities of absorption at the same wavelengths. excess until the free fatty acids separate as an oily layer. (2) Determine the infrared absorption spectrum of Cool the mixture, remove the separated fatty acids, and dis- Pemirolast Potassium as directed in the potassium bromide solve them in 75 mL of diethyl ether. To the diethyl ether so- disk method under Infrared Spectrophotometry <2.25>,and lution add a solution of 4 g of lead (II) acetate trihydrate in compare the spectrum with the Reference Spectrum or the 40 mL of ethanol (95), and allow the mixture to stand for 18 spectrum of Pemirolast Potassium RS: both spectra exhibit hours. Filter the supernatant liquid, transfer the precipitate similar intensities of absorption at the same wave numbers. to the filter with the aid of diethyl ether, and filter by suc- (3) Pemirolast Potassium responds to the Qualitative tion. Place the precipitate in a beaker, heat it with 40 mL of Tests <1.09> (1) for potassium salt. dilute hydrochloric acid and 20 mL of water until the oily Purity (1) Clarity and color of solution—A solution ob- layer is entirely clear, cool, and decant the water layer. Boil tained by dissolving 0.5 g of Pemirolast Potassium in 10 mL the fatty acids with 50 mL of diluted hydrochloric acid (1 in of water is clear and colorless. 100). When the solution prepared by dissolving 0.1 g of the (2) Heavy metals <1.07>—Proceedwith0.5gof fatty acids in 10 mL of ethanol (95) is not darkened by the Pemirolast Potassium according to Method 2, and perform addition of 2 drops of sodium sulfide TS, allow the fatty the test. Prepare the control solution with 1.0 mL of Stand- acids to solidify, and press them between dry filter papers to ard Lead Solution (not more than 20 ppm). exclude moisture. Dissolve the solid fatty acid in 25 mL of (3) Related substances—Dissolve 50 mg of Pemirolast diluted ethanol (9 in 10) with the aid of gentle heat, and then Potassium in 50 mL of a mixture of phosphate buffer solu- cool to 159C to crystallize the fatty acids. Recrystallize them tion, pH 8.0 and methanol (3:2), and use this solution as the from diluted ethanol (9 in 10) and dry in a desiccator sample solution. Pipet 2 mL of the sample solution, and add (phosphorus (V) oxide, in vacuum) for 4 hours: the melting a mixture of phosphate buffer solution, pH 8.0 and metha- point <1.13> of the dried crystals is between 739Cand769C. nol (3:2) to make exactly 100 mL. To exactly 2.5 mL of this Acid value <1.13> Not more than 0.2. solution add a mixture of phosphate buffer solution, pH 8.0 and methanol (3:2) to make exactly 50 mL, and use this solu- Saponification value <1.13> 188–196 tion as the standard solution. Perform the test with exactly Unsaponifiable matters <1.13> Not more than 1.5z. 10 mL each of the sample solution and standard solution as JP XVI Official Monographs / Pemirolast Potassium for Syrup 1219 directed under Liquid Chromatography <2.01> according to acid (100) (30:20:1). the following conditions. Determine each peak area by the Flow rate: Adjust the flow rate so that the retention time automatic integration method: the area of the peak other of pemirolast is about 5 minutes. than pemirolast obtained from the sample solution is not System suitability— larger than the peak area of pemirolast from the standard so- System performance: When the procedure is run with 10 lution. mL of the standard solution under the above operating con- Operating conditions— ditions, pemirolast and the internal standard are eluted in Detector, column, column temperature, mobile phase, and this order with the resolution between these peaks being not flow rate: Proceed as directed in the operating conditions in less than 5. the Assay. System repeatability: When the test is repeated 6 times Time span of measurement: About 9 times as long as the with 10 mL of the standard solution under the above operat- retention time of pemirolast. ing conditions, the relative standard deviation of the ratio of System suitability— the peak area of pemirolast to that of the internal standard is Test for required detectability: To exactly 5 mL of the not more than 1.0z. standard solution add a mixture of phosphate buffer solu- Containers and storage Containers—Tight containers. tion, pH 8.0 and methanol (3:2) to make exactly 25 mL. Storage—Light-resistant. Confirm that the peak area of pemirolast obtained with 10 mL of this solution is equivalent to 15 to 25z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 Pemirolast Potassium for Syrup mL of the standard solution under the above operating con- シロップ用ペミロラストカリウム ditions, the number of theoretical plates and the symmetry factor of the peak of pemirolast are not less than 3000 and not more than 1.7, respectively. Pemirolast Potassium for Syrup is a preparation for System repeatability: When the test is repeated 6 times syrup, which is dissolved before use. with 10 mL of the standard solution under the above operat- It contains not less than 95.0z and not more than ing conditions, the relative standard deviation of the peak 105.0z of the labeled amount of pemirolast potas- area of pemirolast is not more than 2.0z. sium (C10H7KN6O: 266.30). (4) Residual solvent—Being specified separately. Method of preparation Prepare as directed under Prepara- Water <2.48> Not more than 0.5z (0.1 g, coulometric tions for Syrups, with Pemirolast Potassium. titration). Identification Determine the absorption spectrum of the Assay Weigh accurately about 50 mg each of Pemirolast sample solution obtained in the Assay as directed under Ul- Potassium and Pemirolast Potassium RS (separately deter- traviolet-visible Spectrophotometry <2.24>: it exhibits maxi- mine the water <2.48> in the same manner as Pemirolast ma between 255 nm and 259 nm and between 355 nm and Potassium), dissolve in a mixture of phosphate buffer solu- 359 nm. tion, pH 8.0 and methanol (3:2) to make them exactly 50 pH Being specified separately. mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard solution to each, then add a mixture of Uniformity of dosage units <6.02> Perform the test accord- phosphate buffer solution, pH 8.0 and methanol (3:2) to ing to the following method: Pemirolast Potassium for make 50 mL, and use these solutions as the sample solution Syrup in single-unit containers meet the requirement of the and the standard solution, respectively. Perform the test Content uniformity test. with 10 mL each of the sample solution and standard solution Dissolve the total amount of the content of 1 container of as directed under Liquid Chromatography <2.01> according Pemirolast Potassium for Syrup in water to make exactly to the following conditions, and calculate the ratios, QT and V mL so that each mL contains about 50 mgofpemirolast QS, of the peak area of pemirolast to that of the internal potassium (C10H7KN6O). Pipet 10 mL of this solution, add standard. water to make exactly 50 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of C10H7KN6O ＝ MS × QT/QS Amount (mg) of pemirolast potassium (C H KN O) M : Amount (mg) of Pemirolast Potassium RS, calculated 10 7 6 S ＝ M × A /A × V/400 on the anhydrous basis S T S M : Amount (mg) of Pemirolast Potassium RS, calculated Internal standard solution—A solution of ethyl aminobenzo- S on the anhydrous basis ate in methanol (1 in 1000). Operating conditions— Assay Powder Pemirolast Potassium for Syrup. Weigh ac- Detector: An ultraviolet absorption photometer (wave- curately a portion of the powder, equivalent to about 5 mg length: 260 nm). of pemirolast potassium (C10H7KN6O), and dissolve in water Column: A stainless steel column 4.6 mm in inside diame- to make exactly 100 mL. Pipet 10 mL of this solution, add ter and 15 cm in length, packed with octadecylsilanized silica water to make exactly 50 mL, and use this solution as the gel for liquid chromatography (5 mm in particle diameter). sample solution. Separately, weigh accurately about 20 mg Column temperature: A constant temperature of about of Pemirolast Potassium RS (separately determine the water 259C. <2.48> inthesamemannerasPemirolast Potassium), and Mobile phase: A mixture of water, methanol and acetic dissolve in water to make exactly 100 mL. Pipet 5 mL of this 1220 Pemirolast Potassium Tablets / Official Monographs JP XVI solution, add water to make exactly 100 mL, and use this so- pemirolast potassium (C10H7KN6O) according to the labeled lution as the standard solution. Determine the absorbances, amount. Pipet 4 mL of this solution, add exactly 2 mL of

AT and AS, at 357 nm of the sample solution and standard diluted potassium hydroxide TS (1 in 10), and use this solu- solution as directed under Ultraviolet-visible Spectropho- tion as the sample solution. Separately, weigh accurately tometry <2.24>. about 28 mg of Pemirolast Potassium RS (separately determine the water <2.48> in the same manner as Pemirolast Amount (mg) of pemirolast potassium (C H KN O) 10 7 6 Potassium), dissolve in water to make exactly 100 mL. Pipet ＝ M × A /A × 1/4 S T S 5 mL of this solution, add water to make exactly 50 mL.

MS: Amount (mg) of Pemirolast Potassium RS, calculated Pipet 5 mL of this solution, add water to make exactly 25 on the anhydrous basis mL. Pipet 4 mL of this solution, add exactly 2 mL of diluted potassium hydroxide TS (1 in 10), and use this solution as Containers and storage Containers—Tight containers. the standard solution. Then, proceed as directed in the Storage—Light-resistant. Assay. Dissolution rate (z) with respect to the labeled amount Pemirolast Potassium Tablets of pemirolast potassium (C10H7KN6O) ＝ MS × AT/AS × V?/V × 1/C × 18 ペミロラストカリウム錠 MS: Amount (mg) of Pemirolast Potassium RS, calculated on the anhydrous basis Pemirolast Potassium Tablets contain not less than C: Labeled amount (mg) of pemirolast potassium 95.0z and not more than 105.0z of the labeled (C10H7KN6O) in 1 tablet amount of pemirolast potassium (C H KN O: 10 7 6 Assay Accurately weigh the mass of not less than 20 266.30). Pemirolast Potassium Tablets, and powder. Weigh accu- Method of preparation Prepare as directed under Tablets, rately a portion of the powder, equivalent to about 5 mg of with Pemirolast Potassium. pemirolast potassium (C10H7KN6O), add 50 mL of water, shake thoroughly for 20 minutes, then add water to make ex- Identification Determine the absorption spectrum of the actly 100 mL. Filter, discard the first 10 mL of the filtrate, sample solution obtained in the Assay as directed under Ul- pipet 10 mL of the subsequent filtrate, add 1 mL of diluted traviolet-visible Spectrophotometry <2.24>: it exhibits maxi- potassium hydroxide TS (1 in 100), add water to make ex- ma between 255 nm and 259 nm, and between 355 nm and actly 50 mL, and use this solution as the sample solution. 359 nm. Separately, weigh accurately about 20 mg of Pemirolast Uniformity of dosage units <6.02> Perform the test accord- Potassium RS (separately determine the water <2.48> in the ing to the following method: it meets the requirement of the same manner as Pemirolast Potassium), and dissolve in Content uniformity test. water to make exactly 100 mL. Pipet 5 mL of this solution, To 1 tablet of Pemirolast Potassium Tablets add 50 mL of add 1 mL of diluted potassium hydroxide TS (1 in 100), add water for 5 mg of pemirolast potassium (C10H7KN6O), and water to make exactly 100 mL, and use this solution as the shake to disintegrate the tablet completely. Then, add water standard solution. Determine the absorbances, AT sand AS, to make exactly V mL so that each mL contains about 50 mg at 357 nm of the sample solution and standard solution as di- of pemirolast potassium (C10H7KN6O),andfilter.Discard rected under Ultraviolet-visible Spectrophotometry <2.24>, the first 10 mL of the filtrate, pipet 10 mL of the subsequent using water as the blank. filtrate, add 1 mL of diluted potassium hydroxide TS (1 in Amount (mg) of pemirolast potassium (C H KN O) 100), add water to make exactly 50 mL, and use this solution 10 7 6 ＝ M × A /A × 1/4 as the sample solution. Then, proceed as directed in the S T S

Assay. MS: Amount (mg) of Pemirolast Potassium RS, calculated on the anhydrous basis Amount (mg) of pemirolast potassium (C10H7KN6O) ＝ MS × AT/AS × V/400 Containers and storage Containers—Tight containers. Storage—Light-resistant. MS: Amount (mg) of Pemirolast Potassium RS, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of disodium hydrogen phosphate-citric acid buffer solution, pH 5.0 as the dissolution medium, the dissolution rate in 45 minutes of a 5-mg tablet is not less than 75z,andthat in 60 minutes of a 10-mg tablet is not less than 70z. Start the test with 1 tablet of Pemirolast Potassium Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis- card the first 10 mL of the filtrate, pipet V mL of the subsequent filtrate, and add the dissolution medium to make exactly V?mL so that each mL contains about 5.6 mgof JP XVI Official Monographs / Pentazocine 1221

other than the principal spot from the sample solution are Penbutolol Sulfate not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, ペンブトロール硫酸塩 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g). Assay Weigh accurately about 0.8 g of Penbutolol Sulfate, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-

(C18H29NO2)2.H2SO4: 680.94 form a blank determination, and make any necessary correc- (2S)-3-(2-Cyclopentylphenoxy)-1- tion. (1,1-dimethylethyl)aminopropan-2-ol hemisulfate Each mL of 0.1 mol/L perchloric acid VS [38363-32-5] ＝ 68.09 mg of (C18H29NO2)2.H2SO4 Penbutolol Sulfate, when dried, contains not less Containers and storage Containers—Well-closed contain- than 98.5z of (C18H29NO2)2.H2SO4. ers. Description Penbutolol Sulfate occurs as a white crystalline powder. It is very soluble in acetic acid (100), freely soluble in Pentazocine methanol, sparingly soluble in ethanol (95), slightly soluble ペンタゾシン in water, and practically insoluble in acetic anhydride and in diethyl ether. Identification (1) Determine the absorption spectrum of a solution of Penbutolol Sulfate in methanol (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Pen- butolol Sulfate, previously dried, as directed in the paste C H NO: 285.42 method under Infrared Spectrophotometry <2.25>,andcom- 19 27 (2RS,6RS,11RS)-6,11-Dimethyl- pare the spectrum with the Reference Spectrum: both spectra 3-(3-methylbut-2-en-1-yl)-1,2,3,4,5,6-hexahydro- exhibit similar intensities of absorption at the same wave 2,6-methano-3-benzoazocin-8-ol numbers. [359-83-1] (3) Dissolve 0.1 g of Penbutolol Sulfate in 25 mL of water by warming, and cool: this solution responds to Pentazocine, when dried, contains not less than Qualitative Tests <1.09> for sulfate. 99.0z of C19H27NO. Optical rotation <2.49> [a]20: －23 – －259(after drying, D Description Pentazocine occurs as a white to pale yellowish 0.2g,methanol,20mL,100mm). white, crystalline powder. It is odorless. Melting point <2.60> 213–2179C It is freely soluble in acetic acid (100) and in chloroform, soluble in ethanol (95), sparingly soluble in diethyl ether and Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of practically insoluble in water. Penbutolol Sulfate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Identification (1) To 1 mg of Pentazocine add 0.5 mL of Lead Solution (not more than 10 ppm). formaldehyde-sulfuric acid TS: a deep red color is produced, (2) Arsenic <1.11>—Prepare the test solution with 1.0 g and it changes to grayish brown immediately. of Penbutolol Sulfate according to Method 4, and perform (2) Dissolve 5 mg of Pentazocine in 5 mL of sulfuric the test (not more than 2 ppm). acid, add 1 drop of iron (III) chloride TS, and heat in a (3) Related substances—Dissolve 0.8 g of Penbutolol water bath for 2 minutes: the color of the solution changes Sulfate in 10 mL of methanol, and use this solution as the from light yellow to deep yellow. Shake the solution with 1 sample solution. Pipet 1 mL of the sample solution, add drop of nitric acid: the solution remains yellow in color. methanol to make exactly 200 mL, and use this solution as (3) Determine the absorption spectrum of a solution of the standard solution. Perform the test with these solutions Pentazocine in 0.01 mol/L hydrochloric acid TS (1 in as directed under Thin-layer Chromatography <2.03>. Spot 10,000) as directed under Ultraviolet-visible Spectropho- 10 mL each of the sample solution and standard solution on a tometry <2.24>, and compare the spectrum with the Refer- plate of silica gel with fluorescent indicator for thin-layer ence Spectrum: both spectra exhibit similar intensities of ab- chromatography. Develop the plate with a mixture of 2- sorption at the same wavelengths. propanol, ethanol (95) and ammonia solution (28) (85:12:3) Absorbance <2.24> E 1z (278 nm): 67.5 – 71.5 (after dry- to a distance of about 10 cm, and air-dry the plate. Examine 1cm ing, 0.1 g, 0.01 mol/L hydrochloric acid TS, 1000 mL). under ultraviolet light (main wavelength: 254 nm): the spots 1222 Pentobarbital Calcium / Official Monographs JP XVI

Melting point <2.60> 150–1589C It is sparingly soluble in water, slightly soluble in ethanol (95), and practically insoluble in acetonitrile. Purity (1) Clarity and color of solution—Dissolve 0.10 g A solution of Pentobarbital Calcium (1 in 100) shows no of Pentazocine in 20 mL of 0.1 mol/L hydrochloric acid TS: optical rotation. the solution is clear and colorless. (2) Heavy metals <1.07>—Proceed with 1.0 g of Pentazo- Identification (1) Determine the infrared absorption spec- cine according to Method 2, and perform the test. Prepare trum of Pentobarbital Calcium as directed in the potassium the control solution with 2.0 mL of Standard Lead Solution bromide disk method under Infrared Spectrophotometry (not more than 20 ppm). <2.25>, and compare the spectrum with the Reference Spec- (3) Arsenic <1.11>—Prepare the test solution with 1.0 g trum: both spectra exhibit similar intensities of absorption at of Pentazocine according to Method 3, and perform the test the same wave numbers. with a solution of magnesium nitrate hexahydrate in ethanol (2) To 1 g of Pentobarbital Calcium add 5 mL of ethanol (95) (1 in 10) (not more than 2 ppm). (95) and 5 mL of dilute hydrochloric acid, dissolve by warm- (4) Related substances—Dissolve 0.20 g of Pentazocine ing with shaking, shake with 5 mL of dilute hydrochloric in 10 mL of chloroform, and use this solution as the sample acid and 10 mL of water, allow to cool, and filter. To the fil- solution. Pipet 1 mL of the sample solution, add chloroform trate add 1 drop of methyl red TS, and add ammonia TS to make exactly 100 mL, and use this solution as the stand- until a slight yellow color develops: the solution responds to ard solution. Perform the test with these solutions as di- Qualitative Tests <1.09> (1), (2) and (3) for calcium salt. rected under Thin-layer Chromatography <2.03>. Spot 10 mL Purity (1) Chloride <1.03>—To 1.0 g of Pentobarbital each of the sample solution and standard solution on a plate Calcium add 5 mL of ethanol (95) and 2.5 mL of dilute nitric of silica gel for thin-layer chromatography. Develop the acid, dissolve by warming with shaking, cool, add water to plate with a mixture of chloroform, methanol and isopro- make 50 mL, shake well, and filter. Discard the first 10 mL pylamine (94:3:3) to a distance of about 13 cm, and air-dry of the filtrate, and to 15 mL of the subsequent filtrate add 6 the plate. Allow to stand for 5 minutes in iodine vapor: any mL of dilute nitric acid and water to make 50 mL. Perform spot other than the principal spot from the sample solution is the test using this solution as the test solution. Prepare the not more intense than the spot from the standard solution. control solution as follows: To 0.30 mL of 0.01 mol/L hy- Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- drochloric acid VS add 1.5 mL of ethanol (95), 6 mL of dium, phosphorus (V) oxide, 609C, 5 hours). lute nitric acid and water to make 50 mL (not more than 0.035z). Residue on ignition <2.44> Not more than 0.2z (1 g). (2) Heavy metals <1.07>—To 2.0 g of Pentobarbital Cal- Assay Weigh accurately about 0.5 g of Pentazocine, previ- cium add 5 mL of ethanol (95) and 5 mL of dilute hydro- ously dried, dissolve in 50 mL of acetic acid (100), and titrate chloric acid, dissolve by warming with shaking, cool, add <2.50> with 0.1 mol/L perchloric acid VS (indicator: 2 drops water to make 80 mL, shake well, and filter. Discard the first of crystal violet TS). Perform a blank determination, and 10 mL of the filtrate, to 40 mL of the subsequent filtrate add make any necessary correction. 1 drop of phenolphthalein TS, add dropwise ammonia TS until a pale red color develops, and add 2 mL of dilute acetic Each mL of 0.1 mol/L perchloric acid VS acid and water to make 50 mL. Perform the test using this ＝ 28.54 mg of C H NO 19 27 solution as the test solution. Prepare the control solution as Containers and storage Containers—Well-closed contain- follows: To 2.5 mL of ethanol (95) add 2.5 mL of dilute hy- ers. drochloric acid and water to make 30 mL. Add 1 drop of phenolphthalein TS, add dropwise ammonia TS until a pale red color develops, then add 2.0 mL of Standard Lead Solu- Pentobarbital Calcium tion, 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). ペントバルビタールカルシウム (3) Related substances—Dissolve 10 mg of Pentobarbital Calcium in 100 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mLeach of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the areas of each peak by

C22H34CaN4O6: 490.61 the automatic integration method: the area of any peak Monocalcium bis[5-ethyl-5-[(1RS)-1-methylbutyl]-4,6- other than the peak of pentobarbital from the sample solu- dioxo-1,4,5,6-tetrahydropyrimidin-2-olate] tion is not larger than 3/10 times the peak area of pentobar- [76-74-4,Pentobarbital] bital from the standard solution, and the total of these peak area is not larger than the peak area of pentobarbital from Pentobarbital Calcium contains not less than 98.0z the standard solution. and not more than 102.0z of C22H34CaN4O6, calcu- Operating conditions— latedonthedriedbasis. Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in Description Pentobarbital Calcium occurs as a white pow- the Assay. der. Time span of measurement: About 3 times as long as the JP XVI Official Monographs / Pentoxyverine Citrate 1223 retention time of pentobarbital beginning after the solvent ing conditions, the relative standard deviation of the ratios peak. of the peak area of pentobarbital to that of the internal System suitability— standard is not more than 1.0z. Test for required detection: Pipet 2 mL of the standard so- Containers and storage Containers—Well-closed contain- lution, add water to make exactly 20 mL, and confirm that ers. the peak area of pentobarbital obtained from 20 mLofthis solution is equivalent to 5 to 15z of that from 20 mLofthe standard solution. System performance: Proceed as directed in the system Pentoxyverine Citrate performance in the Assay. System repeatability: When the test is repeated 6 times Carbetapentane Citrate with 20 mL of the standard solution under the above operat- Carbetapentene Citrate ing conditions, the relative standard deviation of the peak areas of pentobarbital is not more than 5z. ペントキシベリンクエン酸塩 Loss on drying <2.41> Not more than 7.0z (1 g, 1059C, 5 hours). Assay Weigh accurately about 20 mg of Pentobarbital Cal- cium, dissolve in 5 mL of water, add exactly 5 mL of the internal standard solution and water to make 50 mL. To 5 mL C20H31NO3.C6H8O7: 525.59 of this solution add water to make 20 mL. To 2 mL of this 2-[2-(Diethylamino)ethoxy]ethyl solution add water to make 20 mL, and use this solution as 1-phenylcyclopentanecarboxylate monocitrate the sample solution. Separately, weigh accurately about 18 [23142-01-0] mg of Pentobarbital RS, previously dried at 1059Cfor2 hours, dissolve in 10 mL of acetonitrile, add exactly 5 mL of Pentoxyverine Citrate, when dried, contains not less the internal standard solution and water to make 50 mL. To than 98.5z of C H NO .C H O . 5 mL of this solution add water to make 20 mL. To 2 mL of 20 31 3 6 8 7 this solution add water to make 20 mL, and use this solution Description Pentoxyverine Citrate occurs as a white, crys- as the standard solution. Perform the test with 20 mLeachof talline powder. the sample solution and standard solution as directed under It is very soluble in acetic acid (100), freely soluble in Liquid Chromatography <2.01> according to the following water and in ethanol (95), and practically insoluble in diethyl conditions, and calculate the ratios, QT and QS, of the peak ether. area of pentobarbital to that of the internal standard. Identification (1) Dissolve 0.1 g of Pentoxyverine Citrate

Amount (mg) of C22H34CaN4O6 in 10 mL of water, and add 10 mL of Reinecke salt TS: a ＝ MS × QT/QS × 1.084 light red precipitate is formed. (2) Determine the infrared absorption spectrum of Pen- MS: Amount (mg) of Pentobarbital RS toxyverine Citrate, previously dried, as directed in the paste Internal standard solution—Dissolve 0.2 g of isopropyl par- method under Infrared Spectrophotometry <2.25>,andcom- ahydroxybenzoate in 20 mL of acetonitorile, and add water pare the spectrum with the Reference Spectrum: both spectra to make 100 mL. exhibit similar intensities of absorption at the same wave Operating conditions— numbers. Detector: An ultraviolet absorption photometer (wave- (3) A solution of Pentoxyverine Citrate (1 in 10) re- length: 210 nm). sponds to Qualitative Tests <1.09> (1) and (2) for citrate. Column: A stainless steel column 4.6 mm in inside diame- Melting point <2.60> 92–959C ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Purity (1) Clarity and color of solution—Dissolve 1.0 g Column temperature: A constant temperature of about of Pentoxyverine Citrate in 10 mL of water: the solution is 409C. clear and colorless. Mobile phase: Dissolve 1.36 g of potassium dihydrogen- (2) Heavy metals <1.07>—Proceed with 2.0 g of Pentox- phosphate in 1000 mL of water, and adjust to pH 4.0 with yverine Citrate according to Method 2, and perform the test. diluted phosphoric acid (1 in 10). To 650 mL of this solution Prepare the control solution with 2.0 mL of Standard Lead add 350 mL of acetonitorile. Solution (not more than 10 ppm). Flow rate: Adjust the flow rate so that the retention time (3) Arsenic <1.11>—Prepare the test solution with 1.0 g of pentobarbital is about 7 minutes. of Pentoxyverine Citrate according to Method 3, and per- System suitability— form the test (not more than 2 ppm). System performance: When the procedure is run with 20 (4) Related substances—Dissolve 0.20 g of Pentoxyver- mL of the standard solution under the above operating con- ine Citrate in 10 mL of ethanol (95), and use this solution as ditions, pentobarbital and the internal standard are eluted in the sample solution. Pipet 1 mL of the sample solution, add this order with the resolution between these peaks being not ethanol (95) to make exactly 200 mL, and use this solution as less than 5. the standard solution. Perform the test with these solutions System repeatability: When the test is repeated 6 times as directed under Thin-layer Chromatography <2.03>. Spot with 20 mL of the standard solution under the above operat- 15 mL each of the sample solution and standard solution on a 1224 Peplomycin Sulfate / Official Monographs JP XVI plate of silica gel for thin-layer chromatography. Immedi- It is hygroscopic. ately after air-drying, develop the plate with a mixture of Identification (1) To 4 mg of Peplomycin Sulfate add 5 chloroform, methanol, ethyl acetate and ammonia solution mL of copper (II) sulfate TS, and dissolve in water to make (28) (25:10:10:1) to a distance of about 10 cm, and air-dry 100 mL. Determine the absorption spectrum of this solution the plate. Allow to stand in iodine vapor for 10 minutes: the as directed under Ultraviolet-visible Spectrophotometry spots other than the principal spot from the sample solution <2.24>, and compare the spectrum with the Reference Spec- are not more intense than the spot from the standard solu- trum or the spectrum of a solution of Peplomycin Sulfate RS tion. prepared in the same manner as the sample solution: both Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- spectra exhibit similar intensities of absorption at the same um, phosphorus (V) oxide, 609C, 4 hours). wavelengths. (2) Dissolve 10 mg each of Peplomycin Sulfate and Residue on ignition <2.44> Not more than 0.1z (1 g). Peplomycin Sulfate RS in 6 mL of water, add 0.5 mL of a Assay Weigh accurately about 0.5 g of Pentoxyverine solution of copper (II) sulfate pentahydrate (1 in 125), and Citrate, previously dried, dissolve in 30 mL of acetic acid use these solutions as the sample solution and the standard (100), add 30 mL of acetic anhydride, and titrate <2.50> with solution. Perform the test with 10 mL each of these solutions 0.1 mol/L of perchloric acid VS until the color of the solu- as directed under Liquid Chromatography <2.01> according tion changes from purple through blue-green to green (indi- to the following conditions: the retention time of the princi- cator: 3 drops of crystal violet TS). Perform a blank deter- pal peak obtained from the sample solution is the same as mination, and make any necessary correction. that from the standard soution. Operating conditions— Each mL of 0.1 mol/L perchloric acid VS Detector, column, column temperature, mobile phase ＝ 52.56 mg of C H NO .C H O 20 31 3 6 8 7 stock solution, mobile phase A, mobile phase B, flowing of Containers and storage Containers—Well-closed contain- the mobile phase, and flow rate: Proceed as directed in the ers. operating conditions in the Purity (3). (3) A solution of Peplomycin Sulfate (1 in 200) responds to Qualitative Tests <1.09> (1)and(2)forsulfate. Peplomycin Sulfate 20 Optical rotation <2.49> [a]D : －2–－59(0.1 g calculated on the dried basis, 0.1 mol/L phosphate buffer solution, pH ペプロマイシン硫酸塩 5.3, 10 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 0.10 g of Peplomycin Sulfate in 20 mL of water is between 4.5 and 6.0. Purity (1) Clarity and color of solution—Dissolve 80 mg of Peplomycin Sulfate in 4 mL of water: the solution is clear and colorless. (2) Copper—Dissolve exactly 75 mg of Peplomycin Sul- fate in exactly 10 mL of diluted nitric acid (1 in 100), and use this solution as the sample solution. Separately, to 5.0 mL of Standard Copper Stock Solution add diluted nitric acid (1 in 100) to make exactly 100 mL. To 3.0 mL of this solution add diluted nitric acid (1 in 100) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under

C61H88N18O21S2.H2SO4: 1571.67 Atomic Absorption Spectrophotometry <2.23> according to N 1-{3-[(1S)-(1-Phenylethyl)amino]propyl}bleomycinamide the following conditions: the absorbance of the sample solu- monosulfate tion is not more than that of the standard solution (not more [70384-29-1] than 200 ppm). Gas: Combustible gas—Acetylene. Peplomycin Sulfate is the sulfate of a substance Supporting gas—Air. having antitumor activity produced by the growth of Lamp: Copper hollow cathode lamp. Streptomyces verticillus. Wavelength: 324.8 nm. It contains not less than 865 mg (potency) and not (3) Related substances—Dissolve about 10 mg of more than 1010 mg (potency) per mg, calculated on Peplomycin Sulfate in 6 mL of water, add 0.5 mL of a solu- the dried basis. The potency of Peplomycin Sulfate tion of copper (II) sulfate pentahydrate (1 in 125), and use is expressed as mass (potency) of peplomycin this solution as the sample solution. Perform the test with 10 (C61H88N18O21S2: 1473.59). mL of the sample solution as directed under Liquid Chroma- tography <2.01> according to the following conditions. De- Description Peplomycin Sulfate occurs as a white to light termine the areas of the peaks, appeared after the peak of yellowish white powder. copper sulfate, by the automatic integration method, and It is freely soluble in water, and practically insoluble in calculate the amounts of them by the area percentage ethanol (95). method: the total amount of the peaks other than peplomy- JP XVI Official Monographs / Peplomycin Sulfate 1225 cin is not more than 7.0z. Glycerin 10.0 g Operating conditions— Peptone 10.0 g Detector: An ultraviolet absorption photometer (wave- Meat extract 10.0 g length: 254 nm). Sodium chloride 3.0 g Column: A stainless steel column 4.6 mm in inside diame- Agar 15.0 g ter and 25 cm in length, packed with octadecylsilanized silica Water 1000 mL gel for liquid chromatography (7 mm in particle diameter). Mix all the ingredients and adjust the pH of the solution Column temperature: A constant temperature of about with sodium hydroxide TS so that it will be 6.9 to 7.1 after 409C. sterilization. Mobile phase stock solution: Dissolve 0.96 g of sodium 1- (iii) Liquid medium for suspending test organism pentanesulfonate and 1.86 g of disodium dihydrogen ethyl- Glycerin 10.0 g enediamine tetraacetate dihydrate in 1000 mL of water and 5 Peptone 10.0 g mL of acetic acid (100), and adjust the pH to 4.3 with am- Meat extract 10.0 g monia TS. Sodium chloride 3.0 g Mobile phase A: A mixture of mobile phase stock solution Water 1000 mL and methanol (9:1). Mix all the ingredients and adjust the pH of the solution Mobile phase B: A mixture of mobile phase stock solution with sodium hydroxide TS so that it will be 6.9 to 7.1 after and methanol (3:2). sterilization. Flowing of the mobile phase: Control the gradient by mix- (iv) Preparation of agar medium of seeded ing the mobile phases A and B as directed in the following layer—Inoculate the test organism onto the slant of the agar table. medium for transferring test organism, and incubate the slantat279C for 40 to 48 hours. Inoculate the subcultured Time after injection Mobile phase A Mobile phase B test organism into 100 mL of the liquid medium for suspend- of sample (min) (volz) (volz) ing test organism, incubate at 25 to 279C for 5 days while shaking, and use this suspension as the suspension of the test 0–60 100→ 00→ 100 organism. Keep the suspension of the test organism at a tem- 60–75 0 100 perature of not exceeding 59C and use within 14 days. Add 0.5 mL of the suspension of the test organism in 100 mL of Flow rate: 1.2 mL per minute. the Agar medium for seed layer previously kept at 489C, mix Time span of measurement: As long as 20 minutes after thoroughly, and use this as the agar medium of seeded layer. elution of peplomycin beginning after the peak of copper (v) Preparation of cylinder-agar plate—Proceed as di- sulfate. rected in 1.7. Preparation of cylinder-agar plates with the ex- System suitability— ception of the amounts of the agar medium for base layer Test for required detectability: Measure exactly 1 mL of and the agar medium of seeded layer to put in the Petri dish, the sample solution, add water to make exactly 10 mL, and which are 5.0 mL and 8.0 mL, respectively. use this solution as the solution for system suitability test. (vi) Standard solutions—Weigh accurately an amount of Pipet 1 mL of the solution for system suitability test, and Peplomycin Sulfate RS, equivalent to about 20 mg (po- add water to make exactly 10 mL. Confirm that the peak tency), dissolve in 0.1 mol/L phosphate buffer solution, pH area of peplomycin obtained from 10 mL of this solution is 6.8 to make exactly 100 mL, and use this solution as the equivalent to 7 to 13z of that from 10 mL of the solution for standard stock solution. Keep the standard stock solution at system suitability test. a temperature not exceeding 59C, and use within 15 days. System performance: When the procedure is run with 10 Take exactly a suitable amount of the standard stock solu- mL of the sample solution under the above operating condition before use, add 0.1 mol/L phosphate buffer solution, tions, the number of theoretical plates and the symmetry fac- pH 6.8 to make solutions so that each mL contains 4 mg (po- tor of the peak of peplomycin are not less than 30,000 and tency) and 2 mg (potency), and use these solutions as the high not more than 2.0, respectively. concentration standard solution and the low concentration System repeatability: When the test is repeated 6 times standard solution, respectively. with 10 mL of the sample solution under the above operating (vii) Sample solutions—Weigh accurately an amount of conditions, the relative standard deviation of the peak area Peplomycin Sulfate, equivalent to about 20 mg (potency), of peplomycin is not more than 2.0z. and dissolve in 0.1 mol/L phosphate buffer solution, pH 6.8 to make exactly 100 mL. Take exactly a suitable amount of Loss on drying <2.41> Not more than 3.0z (60 mg, in this solution, add 0.1 mol/L phosphate buffer solution, pH vacuum, phosphorus (V) oxide, 609C, 3 hours). Handle the 6.8 to make solutions so that each mL contains 4 mg (po- sample avoiding absorption of moisture. tency) and 2 mg (potency), and use these solutions as the high Assay Perform the test according to the Cylinder-plate concentration sample solution and the low concentration method as directed under Microbial Assay for Antibiotics sample solution, respectively. <4.02> according to the following conditions. Containers and storage Containers—Tight containers. (i) Test organism—Mycobacterium smegmatis ATCC 607 (ii) Agar media for base and seed layer, and for transferring test organism 1226 Peplomycin Sulfate for Injection / Official Monographs JP XVI

(i) Test organism, culture medium, liquid medium for Peplomycin Sulfate for Injection suspending test organisms, preparation of seeded agar layer, preparation of cylinder-agar plate and the standard solutions 注射用ペプロマイシン硫酸塩 —Proceed as directed in the Assay under Peplomycin Sul- fate. (ii) Sample solutions—Weigh accurately the mass of the Peplomycin Sulfate for Injection is a preparation contents of not less than 10 containers of Peplomycin Sul- for injection which is dissolved before use. fate for Injection. Weigh accurately an amount of the con- It contains not less than 90.0z and not more tents, equivalent to about 10 mg (potency) of Peplomycin than 115.0z of the labeled amount of peplomycin Sulfate, dissolve in 0.1 mol/L phosphate buffer solution, pH (C H N O S : 1473.59). 61 88 18 21 2 6.8, to make exactly 100 mL. Measure exactly a suitable Method of preparation Prepare as directed under Injec- quantity of this solution, add 0.1 mol/L phosphate buffer tions, with Peplomycin Sulfate. solution, pH 6.8 so that each mL contains 4 mg (potency) and 2 mg (potency), and use these solutions as the high con- Description Peplomycin Sulfate for Injection occurs as centration sample solution and the low concentration sample white light masses or powder. solution, respectively. Identification Take an amount of Peplomycin Sulfate for Containers and storage Containers—Hermetic containers. Injection, equivalent to 10 mg (potency) of Peplomycin Sul- fate according to the labeled amount, and dissolve in 15 mL of Copper (II) sulfate TS and water to make 2 mL. Apply this solution to the column (prepared by filling a 15 mm in- Perphenazine side diameter and 15 cm long chromatography tube with 15 ペルフェナジン mL of strongly basic ion exchange resin (Cl type) for column chromatography (75 – 150 mm in particle diameter) and run off. Then wash the column using water at 2.5 mL per minute, collect about 30 mL of the effluent. Add water to the effluent to make 250 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet- visible Spectrophotometry <2.24>: it exhibits maxima between 242 nm and 246 nm, and between 291 nm and 295 nm. C H ClN OS: 403.97 Further determine the absorbances A and A , at 243 nm and 21 26 3 1 2 2-{4-[3-(2-Chloro-10H-phenothiazin- 293 nm, respectively: the ratio A /A is 1.20 to 1.30. 1 2 10-yl)propyl]piperazin-1-yl}ethanol Osmotic pressure ratio Being specified separately. [58-39-9] pH <2.54> The pH of a solution prepared by dissolving an Perphenazine, when dried, contains not less than amount of Peplomycin Sulfate for Injection, equivalent to 98.5z of C H ClN OS. 50 mg (potency) of Peplomycin Sulfate according to the 21 26 3 labeled amount, in 10 mL of water is 4.5 to 6.0. Description Perphenazine occurs as white to light yellow crystals or crystalline powder. It is odorless, and has a bitter Purity Clarity and color of solution—A solution prepared taste. by dissolving an amount of Peplomycin Sulfate for Injec- It is freely soluble in methanol and in ethanol (95), soluble tion, equivalent to 10 mg (potency) of Peplomycin Sulfate in acetic acid (100), sparingly soluble in diethyl ether, and according to the labeled amount, in 10 mL of water is clear practically insoluble in water. and colorless. It dissolves in dilute hydrochloric acid. Loss on drying <2.41> Not more than 4.0z (60 mg, in It is gradually colored by light. vacuum, phosphorus (V) oxide, 609C, 3 hours). Perform the Identification (1) Dissolve 5 mg of Perphenazine in 5 mL sampling preventing from moisture absorption. of sulfuric acid: a red color, changing to deep red-purple Bacterial endotoxins <4.01> Less than 1.5 EU/mg (po- upon warming, is produced. tency). (2) Dissolve 0.2 g of Perphenazine in 2 mL of methanol, add this solution to 10 mL of a warm solution of 2,4,6- Uniformity of dosage units <6.02> It meets the requirement trinitrophenol in methanol (1 in 25), and allow to stand for 4 of the Mass variation test. hours. Collect the crystals, wash with a small volume of Foreign insoluble matter <6.06> Perform the test according methanol, and dry at 1059C for 1 hour: the crystals so ob- to Method 2: it meets the requirement. tained melt <2.60> between 2379C and 2449C (with decomposition). Insoluble particulate matter <6.07> It meets the require- (3) Determine the absorption spectrum of a solution of ment. Perphenazine in 0.1 mol/L hydrochloric acid TS (1 in Sterility <4.06> Perform the test according to the Mem- 200,000) as directed under Ultraviolet-visible Spectropho- brane filtration method: it meets the requirement. tometry <2.24>, and compare the spectrum with the Refer- ence Spectrum 1 or the spectrum of a solution of Perphena- Assay Perform the test according to the Cylinder-plate zine RS prepared in the same manner as the sample solution: method as directed under Microbial Assay for Antibiotics both spectra exhibit similar intensities of absorption at the <4.02> according to the following conditions. JP XVI Official Monographs / Perphenazine Tablets 1227 same wavelengths. Separately, to 10 mL of the solution add phenazine Tablets, equivalent to 25 mg of Perphenazine ac- 10 mL of water. Determine the absorption spectrum of the cording to the labeled amount, with 10 mL of methanol, and solution as directed under Ultraviolet-visible Spectropho- filter. Evaporate 2 mL of the filtrate on a water bath to dry- tometry <2.24>, and compare the spectrum with the Refer- ness. With the residue, proceed as directed in the Identifica- ence Spectrum 2 or the spectrum of a solution of Perphena- tion (1) under Perphenazine. zine RS prepared in the same manner as the sample solution: (2) Add 5 mL of the filtrate obtained in the Identifica- both spectra exhibit similar intensities of absorption at the tion (1) to 10 mL of a warm solution of 2,4,6-trinitrophenol same wavelengths. inmethanol(1in25),andproceedasdirectedintheIdentifi- (4) Perform the test with Perphenazine as directed under cation (2) under Perphenazine. Flame Coloration Test <1.04> (2): a green color appears. (3) Determine the absorption spectrum of the filtrate obtained in the Assay as directed under Ultraviolet-visible Melting point <2.60> 95 – 1009C Spectrophotometry <2.24>: it exhibits a maximum between Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of 309 nm and 313 nm. Add 30 mL of methanol to another 10 Perphenazine according to Method 2, and perform the test. mL of the filtrate, and determine the absorption spectrum: it Prepare the control solution with 2.0 mL of Standard Lead exhibits a maximum between 256 nm and 260 nm. Solution (not more than 20 ppm). Uniformity of dosage units <6.02> Perform the test accord- (2) Related substances—Perform the test in the current ing to the following method: it meets the requirement of the of nitrogen in light-resistant containers under the protection Content uniformity test. from sunlight. Dissolve 0.10 g of Perphenazine in 10 mL of Disintegrate 1 Perphenazine Tablet by shaking with 5 mL ethanol (95), and use this solution as the sample solution. of water, shake well with 70 mL of methanol, and add Pipet 1 mL of the sample solution, and add ethanol (95) methanol to make exactly 100 mL. Centrifuge this solution, to make exactly 10 mL. Pipet 1 mL of this solution, add pipet V mL of the supernatant liquid, add methanol to make ethanol (95) to make exactly 20 mL, and use this solution as exactly V?mL of a solution containing about 4 mgofper- the standard solution. Perform the test with these solutions phenazine (C H ClN OS)ineachml,andusethissolution as directed under Thin-layer Chromatography <2.03>. Spot 21 26 3 as the sample solution. Separately, weigh accurately about 10 mL each of the sample solution and standard solution on a 10 mg of Perphenazine RS, previously dried in vacuum over plate of silica gel with fluorescent indicator for thin-layer phosphorus (V) oxide at 659C for 4 hours, dissolve in metha- chromatography. Develop the plate with a mixture of 1- nol to make exactly 250 mL. Pipet 5 mL of this solution, add butanol and 1 mol/L ammonia TS (5:1) to a distance of methanol to make exactly 50 mL, and use this solution as the about 12 cm, and air-dry the plate. Examine under ultravio- standard solution. Determine the absorbances, A and A , let light (main wavelength: 254 nm): any spot other than the T S of the sample solution and standard solution at 258 nm as di- principal spot from the sample solution is not more intense rected under Ultraviolet-visible Spectrophotometry <2.24>. than that from the standard solution. Amount (mg) of perphenazine (C H ClN OS) Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- 21 26 3 ＝ M × A /A × V?/V × 1/25 um, phosphorus (V) oxide, 659C, 4 hours). S T S M : Amount (mg) of Perphenazine RS Residue on ignition <2.44> Not more than 0.1z (1 g). S Dissolution <6.10> When the test is performed at 100 revo- Assay Weigh accurately about 0.4 g of Perphenazine, pre- lutions per minute according to the Paddle method, using viously dried, dissolve in 50 mL of acetic acid (100), and 900 mL of 2nd fluid for dissolution test as the dissolution titrate <2.50> with 0.1 mol/L perchloric acid VS until the medium, the dissolution rate in 90 minutes of Perphenazine color of the solution changes from purple through blue-pur- Tablets is not less than 70z. ple to blue-green (indicator: 3 drops of crystal violet TS). Start the test with 1 tablet of Perphenazine Tablets, Perform a blank determination, and make any necessary withdraw not less than 20 mL of the medium at the specified correction. minute after starting the test, and filter through a membrane Each mL of 0.1 mol/L perchloric acid VS filter with a pore size not exceeding 0.8 mm. Discard the first

＝ 20.20 mg of C21H26ClN3OS 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 10 mg Containers and storage Containers—Tight containers. of Perphenazine RS, previously dried in vacuum with phos- Storage—Light-resistant. phorus (V) oxide at 659C for 4 hours, dissolve in 5 mL of 0.1 mol/L hydrochloric acid TS, and add the dissolution medium to make exactly 250 mL. Pipet 5 mL of this solu- Perphenazine Tablets tion, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Determine the ペルフェナジン錠 absorbances, AT and AS, of the sample solution and standard solution at 255 nm as directed under Ultraviolet-visible Perphenazine Tablets contain not less than 90.0z Spectrophotometry <2.24>. The dissolution rate of Per- and not more than 110.0z of the labeled amount of phenazine Tablets in 90 minutes is not less than 70z. perphenazine (C H ClN OS: 403.97). 21 26 3 Dissolution rate (z) with respect to the labeled amount

Method of preparation Prepare as directed under Tablets, of perphenazine (C21H26ClN3OS) with Perphenazine. ＝ MS × AT/AS × 1/C × 36

Identification (1) Shake well a quantity of powdered Per- MS: Amount (mg) of Perphenazine RS 1228 Perphenazine Maleate / Official Monographs JP XVI

C: Labeled amount (mg) of perphenazine (C21H26ClN3OS) methanol, and pour into 10 mL of a warm solution of 2,4,6- in 1 tablet trinitrophenol in methanol (1 in 25). Allow to stand for 4 hours, collect the crystals, wash with a small amount of Assay Weigh accurately and powder not less than 20 methanol, and dry at 1059C for 1 hour: the crystals melt Perphenazine Tablets. Weigh accurately a portion of the <2.60> between 2379Cand2449C (with decomposition). powder, equivalent to about 4 mg of perphenazine (3) Determine the absorption spectrum of a solution of (C H ClN OS), add 70 mL of methanol, shake well, and 21 26 3 Perphenazine Maleate (1 in 20,000) as directed under Ultra- add methanol to make exactly 100 mL. Filter the solution, violet-visible Spectrophotometry <2.24>, and compare the and discard the first 20 mL of the filtrate. Pipet 5 mL of the spectrum with the Reference Spectrum 1: both spectra ex- subsequent filtrate, add methanol to make exactly 50 mL, hibit similar intensities of absorption at the same wave- and use this solution as the sample solution. Weigh accu- lengths. Separately, to 10 mL of the solution add 30 mL of rately about 10 mg of Perphenazine RS, previously dried in water. Determine the absorption spectrum of the solution as vacuum over phosphorus (V) oxide at 659C for 4 hours, and directed under Ultraviolet-visible Spectrophotometry <2.24>, dissolve in methanol to make exactly 250 mL. Pipet 5 mL of and compare the spectrum with the Reference Spectrum 2: this solution, add methanol to make exactly 50 mL, and use both spectra exhibit similar intensities of absorption at the this solution as the standard solution. Determine the absor- same wavelengths. bances, A and A , of the sample solution and the standard T S (4) Perform the test with Perphenazine Maleate as di- solution at 258 nm as directed under Ultraviolet-visible Spec- rected under Flame Coloration Test <1.04> (2): a green color trophotometry <2.24>. appears.

Amount (mg) of perphenazine (C21H26ClN3OS) (5) Evaporate the aqueous layer reserved in (2) to dry- ＝ MS × AT/AS × 2/5 ness. To the residue add 1 mL of dilute sulfuric acid and 5 mL of water, and extract with four 25-mL portions of M : Amount (mg) of Perphenazine RS S diethyl ether. Combine the diethyl ether extracts, and evapo- Containers and storage Containers—Tight containers. rate in a water bath at about 359C with the aid of a current Storage—Light-resistant. of air: the residue melts <2.60> between 1289Cand1369C. Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of perphenazine maleate according to Method 2, and perform Perphenazine Maleate the test. Prepare the control solution with 2.0 mL of Stand- ard Lead Solution (not more than 10 ppm). ペルフェナジンマレイン酸塩 (2) Arsenic <1.11>—Prepare the test solution with 1.0 g of Perphenazine Maleate according to Method 3, and perform the test (not more than 2 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.5 g of Perphenazine Male- C H ClN OS.2C H O :636.11 21 26 3 4 4 4 ate, previously dried, dissolve in 70 mL of acetic acid (100), 2-{4-[3-(2-Chloro-10H-phenothiazin-10-yl)propyl]piperazin- and titrate <2.50> with 0.1 mol/L perchloric acid VS until the 1-yl}ethanol dimaleate color of the solution changes from purple through blue to [58-39-9, Perphenazine] blue-green (indicator: 3 drops of crystal violet TS). Perform a blank determination, and make any necessary correction. Perphenazine Maleate, when dried, contains not less than 98.0z of C21H26ClN3OS.2C4H4O4. Each mL of 0.1 mol/L perchloric acid VS ＝ 31.81 mg of C H ClN OS.2C H O Description Perphenazine Maleate occurs as a white to 21 26 3 4 4 4 light yellow powder. It is odorless. Containers and storage Containers—Well-closed contain- It is sparingly soluble in acetic acid (100), slightly soluble ers. in water and in ethanol (95), and practically insoluble in Storage—Light-resistant. chloroform. It dissolves in dilute hydrochloric acid. It is gradually colored by light. Perphenazine Maleate Tablets Melting point: about 1759C (with decomposition). ペルフェナジンマレイン酸塩錠 Identification (1) Dissolve 8 mg of Perphenazine Maleate in 5 mL of sulfuric acid: a red color is produced, which becomes deep red-purple on warming. Perphenazine Maleate Tablets contain not less than (2) Dissolve 0.3 g of Perphenazine Maleate in 3 mL of 93.0z and not more than 107.0z of the labeled dilute hydrochloric acid, add 2 mL of water and 3 mL of am- amount of perphenazine maleate (C21H26ClN3OS.2C4 monia solution (28), shake, and extract with three 10-mL H4O4: 636.11). portions of chloroform. [Reserve the aqueous layer, and use Method of preparation Prepare as directed under Tablets, for test (5)]. Evaporate the combined chloroform extracts on with Perphenazine Maleate. a water bath to dryness, dissolve the residue in 20 mL of JP XVI Official Monographs / Perphenazine Maleate Tablets 1229

Identification (1) Shake a quantity of powdered Per- Maleate Tablets is not less than 70z. phenazine Maleate Tablets, equivalent to 0.04 g of Per- Conduct this procedure without exposure to light. Start phenazine Maleate according to the labeled amount, with 3 the test with 1 tablet of Perphenazine Maleate Tablets, with- mL of dilute hydrochloric acid and 30 mL of water, centri- draw not less than 20 mL of the medium at the specified fuge, filter the supernatant liquid, add 3 mL of ammonia so- minute after starting the test, and filter through a membrane lution (28) to the filtrate, and extract with three 10-mL por- filter with a pore size not exceeding 0.45 mm. Discard the tions of chloroform. [Reserve the aqueous layer, and use for first 10 mL of the filtrate, pipet V mL of the subsequent fil- test (4).] Wash the combined chloroform extracts with two trate, add the dissolution medium to make exactly V?mL so 5-mL portions of water, and separate the chloroform layer. that each mL contains about 3.5 mg of perphenazine maleate

Evaporate 6 mL of the chloroform solution on a water bath (C21H26ClN3OS.2C4H4O4) according to the labeled amount, to dryness. Proceed with the residue as directed in the Iden- and use this solution as the sample solution. Separately, tification (1) under Perphenazine Maleate. weigh accurately about 28 mg of perphenazine maleate for (2) Evaporate 20 mL of the chloroform solution ob- assay, previously dried at 1059C for 3 hours, dissolve in 10 tained in (1) on a water bath to dryness, dissolve the residue mL of 0.1 mol/L hydrochloric acid TS, and add the dissolu- in 20 mL of methanol, and filter, if necessary. Warm the fil- tion medium to make exactly 200 mL. Pipet 5 mL of this so- trate, add 5 mL of a warm solution of 2,4,6-trinitrophenol in lution, add the dissolution medium to make exactly 200 mL, methanol (1 in 25), allow to stand for 4 hours, and proceed and use this solution as the standard solution. Determine the as directed in the Identification (2) under Perphenazine absorbances, AT and AS, at 255 nm of the sample solution Maleate. and standard solution as directed under Ultraviolet-visible (3) To 2 mL of the filtrate obtained in the Assay add Spectrophotometry <2.24>. water to make 50 mL. Determine the absorption spectrum of Dissolution rate (z) with respect to the labeled amount the solution as directed under Ultraviolet-visible Spectropho- of perphenazine maleate (C H ClN OS.2C H O ) tometry <2.24>: it exhibits maxima between 253 nm and 257 21 26 3 4 4 4 ＝ M × A /A × V?/V × 1/C × 45/4 nm and between 303 nm and 313 nm. S T S

(4) Filter, if necessary, the aqueous layer reserved in (1), MS: Amount (mg) of perphenazine maleate for assay evaporate the filtrate to make about 5 mL, add 2 mL of di- C: Labeled amount (mg) of perphenazine maleate lute sulfuric acid, and extract with two 10-mL portions of (C21H26ClN3OS.2C4H4O4)in1tablet diethyl ether. Combine the diethyl ether extracts, evaporate Assay Weigh accurately and powder not less than 20 Per- on a water bath to dryness, dissolve the residue in 5 mL of phenazine Maleate Tablets. Weigh accurately a portion of sulfuric acid TS, and add 1 to 2 drops of potassium perman- the powder, equivalent to about 40 mg of perphenazine ganate TS: the red color of potassium permanganate TS maleate (C H ClN OS.2C H O ), shake well with 15 mL of fades immediately. 21 26 3 4 4 4 1 mol/L hydrochloric acid TS and 50 mL of methanol, add Uniformity of dosage units <6.02> Perform the test accord- water to make exactly 100 mL, and filter. Discard the first 20 ing to the following method: it meets the requirement of the mL of the filtrate, measure exactly 5 mL of the subsequent Content uniformity test. filtrate, add water to make exactly 250 mL, and use this so- Disintegrate 1 tablet of Perphenazine Maleate Tablets by lution as the sample solution. Separately, weigh accurately shaking with 15 mL of 0.1 mol/L hydrochloric acid TS, about 40 mg of perphenazine maleate for assay, previously shake vigorously with 50 mL of methanol, add water to dried at 1059C for 3 hours, dissolve in 15 mL of 1 mol/L hy- make exactly 100 mL, and centrifuge. Pipet V mL of the drochloric acid TS and 50 mL of methanol, and add water to supernatant liquid, add water to make exactly V?mL of a make exactly 100 mL. Pipet 5 mL of this solution, add water solution containing about 6 mg of perphenazine maleate to make exactly 250 mL, and use this solution as the stand-

(C21H26ClN3OS.2C4H4O4)ineachml,andusethissolution ard solution. Determine the absorbances, AT and AS,ofthe as the sample solution. Separately, weigh accurately 30 mg sample solution and the standard solution at 255 nm as di- of perphenazine maleate for assay, previously dried at 1059C rected under Ultraviolet-visible Spectrophotometry <2.24>, for 3 hours, dissolve in 15 mL of 0.1 mol/L hydrochloric using water as the blank. acid TS and 50 mL of methanol, and add water to make ex- Amount (mg) of perphenazine maleate actly 100 mL. Pipet 5 mL of this solution, add 3 mL of 0.1 (C H ClN OS.2C H O ) mol/L hydrochloric acid TS, 10 mL of methanol and water 21 26 3 4 4 4 ＝ M × A /A to make exactly 250 mL, and use this solution as the stand- S T S ard solution. Determine the absorbances, AT and AS,ofthe MS: Amount (mg) of perphenazine maleate for assay sample solution and the standard solution at 255 nm as di- Containers and storage Containers—Tight containers. rected under Ultraviolet-visible Spectrophotometry <2.24>, Storage—Light-resistant. using water as the blank. Amount (mg) of perphenazine maleate

(C21H26ClN3OS.2C4H4O4) ＝ MS × AT/AS × V?/V × 1/50

MS: Amount (mg) of perphenazine maleate for assay Dissolution <6.10> When the test is performed at 75 revolutions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 30 minutes of Perphenazine 1230 Adsorbed Purified Pertussis Vaccine / Official Monographs JP XVI

of Pethidine Hydrochloride in 10 mL of water: the solution Adsorbed Purified Pertussis is clear and colorless. (2) Sulfate <1.14>—Perform the test with 0.20 g of Vaccine Pethidine Hydrochloride. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 沈降精製百日せきワクチン 0.240z). (3) Related substances—Dissolve 0.05 g of Pethidine Hy- Adsorbed Purified Pertussis Vaccine is a liquid for drochloride in 20 mL of the mobile phase, and use this solu- injection prepared by adding an aluminum salt to a tion as the sample solution. Pipet 1 mL of the sample solu- liquid containing the protective antigen of Bordetella tion, add the mobile phase to make exactly 100 mL, and use pertussis to make the antigen insoluble. this solution as the standard solution. Perform the test with It conforms to the requirements of Adsorbed Puri- exactly 20 mL each of the sample solution and standard solu- fied Pertussis Vaccine in the Minimum Requirements tion as directed under Liquid Chromatography <2.01> ac- for Biological Products. cording to the following conditions. Determine each peak area obtained from both solutions by the automatic integra- Description Adsorbed Purified Pertussis Vaccine forms a tion method: the total area of the peaks other than pethidine homogeneous, white turbidity on shaking. from the sample solution is not larger than the peak area of perthidine from the standard solution. Operating conditions— Pethidine Hydrochloride Detector: An ultraviolet absorption photometer (wavelength: 257 nm). Operidine Column: A stainless steel column 4.6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica ペチジン塩酸塩 gel for liquid chromatography (5 mminparticlediameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2.0 g of sodium lauryl sulfate in 1000 mL of diluted phosphoric acid (1 in 1000), adjust the pH to 3.0 with sodium hydroxide TS, and to 550 mL of this solution add 450 mL of acetonitrile. Flow rate: Adjust the flow rate so that the retention time C H NO .HCl: 283.79 15 21 2 of pethidine is about 7 minutes. Ethyl 1-methyl-4-phenylpiperidine-4-carboxylate Time span of measurement: About 2 times as long as the monohydrochloride retention time of pethidine beginning after the solvent peak. [50-13-5] System suitability— Test for required detection: To exactly 2 mL of the stand- Pethidine Hydrochloride, when dried, contains not ard solution add the mobile phase to make exactly 20 mL. less than 98.0 of C H NO .HCl. z 15 21 2 Confirm that the peak area of pethidine obtained from 20 Description Pethidine Hydrochloride occurs as a white, mL of this solution is equivalent to 5 to 15z of that from 20 crystalline powder. mL of the standard solution. It is very soluble in water and in acetic acid (100), freely System performance: To 2 mL each of the sample solution soluble in ethanol (95), sparingly soluble in acetic anhydride, and a solution of isoamyl parahydroxybenzoate in the mo- and practically insoluble in diethyl ether. bile phase (1 in 50,000) add the mobile phase to make 10 mL. The pH of a solution dissolved 1.0 g of Pethidine Hydro- When the procedure is run with 20 mL of this solution ac- chloride in 20 mL of water is between 3.8 and 5.8. cording to the above operating conditions, pethidine and isoamyl parahydroxybenzoate are eluted in this order with Identification (1) Determine the absorption spectrum of a the resolution between these peaks being not less than 2.0. solution of Pethidine Hydrochloride (1 in 2000) as directed System repeatability: When the test is repeated 6 times under Ultraviolet-visible Spectrophotometry <2.24>,and with 20 mL of the standard solution under the above operat- compare the spectrum with the Reference Spectrum: both ing conditions, the relative standard deviation of the peak spectra exhibit similar intensities of absorption at the same area of pethidine is not more than 2.0z. wavelengths. (2) Determine the infrared absorption spectrum of Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, Pethidine Hydrochloride, previously dried, as directed in the 3 hours). potassium bromide disk method under Infrared Spectropho- Residue on ignition <2.44> Not more than 0.1z (0.5 g). tometry <2.25>, and compare the spectrum with the Refer- ence Spectrum: both spectra exhibit similar intensities of Assay Weigh accurately about 0.5 g of Pethidine Hydro- absorption at the same wave numbers. chloride, previously dried, dissolve in 50 mL of a mixture of (3) A solution of Pethidine Hydrochloride (1 in 50) re- acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> sponds to Qualitative Tests <1.09> (2) for chloride. with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination, and make any necessary Melting point <2.60> 187–1899C correction. Purity (1) Clarity and color of solution—Dissolve 1.0 g JP XVI Official Monographs / Hydrophilic Petrolatum 1231

Each mL of 0.1 mol/L perchloric acid VS MS: Amount (mg) of pethidine hydrochloride for assay ＝ 28.38 mg of C H NO .HCl 15 21 2 Internal standard solution—A solution of isoamyl parahy- Containers and storage Containers—Tight containers. droxybenzoate in the mobile phase (1 in 12,500). Storage—Light-resistant. Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 257 nm). Pethidine Hydrochloride Injection Column: A stainless steel column 4.6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica Operidine Injection gel for liquid chromatography (5 mminparticlediameter). Column temperature: A constant temperature of about ペチジン塩酸塩注射液 409C. Mobile phase: Dissolve 2.0 g of sodium lauryl sulfate in 1000 mL of diluted phosphoric acid (1 in 1000), adjust the Pethidine Hydrochloride Injection is an aqueous so- pH to 3.0 with sodium hydroxide TS, and to 550 mL of this lution for injection. solution add 450 mL of acetonitrile. It contains not less than 95.0z and not more than Flow rate: Adjust the flow rate so that the retention time 105.0z of the labeled amount of pethidine hydrochlo- of pethidine is about 7 minutes. ride (C H NO .HCl: 283.79). 15 21 2 System suitability— Method of preparation Prepare as directed under Injec- System performance: When the procedure is run with 20 tions, with Pethidine Hydrochloride. mL of the standard solution under the above operating conditions, pethidine and the internal standard are eluted in this Description Pethidine Hydrochloride Injection is a clear, order with the resolution between these peaks being not less colorless liquid. than 2.0. It is affected by light. System repeatability: When the test is repeated 6 times pH4.0–6.0 with 20 mL of the standard solution under the above operat- Identification Take a volume of Pethidine Hydrochloride ing conditions, the relative standard deviation of the ratios Injection equivalent to 0.1 g of Pethidine Hydrochloride ac- of the peak area of pethidine to that of the internal standard cording to the labeled amount, and add water to make 200 is not more than 1.0z. mL. Determine the absorption spectrum of this solution as Containers and storage Containers—Hermetic containers, directed under Ultraviolet-visible Spectrophotometry <2.24>: and colored containers may be used. it exhibits maxima between 250 nm and 254 nm, between 255 Storage—Light-resistant. nm and 259 nm, and between 261 nm and 265 nm. Bacterial endotoxins <4.01> Less than 6.0 EU/mg. Extractable volume <6.05> It meets the requirement. Hydrophilic Petrolatum Foreign insoluble matter <6.06> Perform the test according 親水ワセリン to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require- Method of preparation ment. White Beeswax 80 g Sterility <4.06> Perform the test according to the Mem- Stearyl Alcohol or Cetanol 30 g brane filtration method: it meets the requirement. Cholesterol 30 g White Petrolatum a sufficient quantity Assay Measure exactly a volume of Pethidine Hydrochlo- ride Injection, equivalent to about 0.1 g of pethidine hydro- To make 1000 g chloride (C15H21NO2.HCl), add exactly 10 mL of the internal Melt and mix Stearyl Alcohol or Cetanol, White Beeswax standard solution, and add the mobile phase to make 50 mL. and White Petrolatum on a water bath. Add Cholesterol, To 5 mL of this solution add the mobile phase to make 20 and melt completely by stirring. Stop warming, and stir until mL, and use this solution as the sample solution. Separately, the mixture congeals. weigh accurately about 0.1 g of pethidine hydrochloride for assay, previously dried at 1059C for 3 hours, add exactly 10 Description Hydrophilic Petrolatum is white in color. It mL of the internal standard solution, and add the mobile has a slight, characteristic odor. phase to make 50 mL. To 5 mL of this solution add the mo- When mixed with an equal volume of water, it retains the bile phase to make 20 mL, and use this solution as the stand- consistency of ointment. ard solution. Perform the test with 20 mLofthesamplesolu- Containers and storage Containers—Tight containers. tion and standard solution as directed under Liquid Chroma- tography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of pethidine to that of the internal standard.

Amount (mg) of pethidine hydrochloride (C15H21NO2.HCl) ＝ MS × QT/QS 1232 White Petrolatum / Official Monographs JP XVI

White Petrolatum Yellow Petrolatum

白色ワセリン黄色ワセリン

White Petrolatum is a decolorized and purified mix- Yellow Petrolatum is a purified mixture of ture of hydrocarbons obtained from petroleum. hydrocarbons obtained from petroleum. Description White Petrolatum is a white to pale yellow, ho- Description Yellow Petrolatum occurs as a yellow, homo- mogeneous, unctuous mass. It is odorless and tasteless. geneous, unctuous mass, It is odorless and tasteless. It is practically insoluble in water, in ethanol (95) and in It is slightly soluble in ethanol (95), and practically insolu- ethanol (99.5). ble in water. It dissolves in diethyl ether making a clear liquid or It dissolves in diethyl ether, in petroleum benzine and in producing slight insoluble substances. turpentine oil, making a clear liquid or producing slight in- It becomes a clear liquid when warmed. soluble substances. It becomes a yellow, clear liquid with slight fluorescence Melting point <2.60> 38–609C (Method 3). when warmed. Purity (1) Color—Melt White Petrolatum by warming, Melting point <2.60> 38–609C (Method 3). and pour 5 mL of it into a test tube, and keep the content in a liquid condition: the liquid has no more color than the fol- Purity (1) Color—Melt Yellow Petrolatum by warming, lowing control solution, when observed transversely from and pour 5 mL of it into a test tube, and keep the content in side against a white background. a liquid condition: the liquid has no more color than the fol- Control solution: Add 3.4 mL of water to 1.6 mL of Iron lowing control solution, when observed transversely from (III) Chloride CS. side against a white background. (2) Acidity or alkalinity—To 35.0 g of White Petrolatum Control solution: To 3.8 mL of Iron (III) Chloride CS add add 100 mL of hot water, shake vigorously for 5 minutes, 1.2 mL of Cobalt (II) Chloride CS. and then draw off the aqueous layer. Treat the White (2) Acidity or alkalinity—To 35.0 g of Yellow Petrola- Petrolatum layer in the same manner using two 50-mL por- tum add 100 mL of hot water, shake vigorously for 5 tions of hot water. To the combined aqueous layer add 1 minutes, and then draw off the aqueous layer. Treat the Yel- drop of phenolphthalein TS, and boil: no red color is pro- low Petrolatum layer in the same manner using two 50-mL duced. Further add 2 drops of methyl orange TS: no red portions of hot water. To the combined aqueous layer add 1 color is produced. drop of phenolphthalein TS, and boil: no red color is pro- (3) Heavy metals <1.07>—Proceedwith1.0gofWhite duced. Further add 2 drops of methyl orange TS: no red Petrolatum according to Method 2, and perform the test. color is produced. Prepare the control solution with 3.0 mL of Standard Lead (3) Heavy metals <1.07>—Proceed with 1.0 g of Yellow Solution (not more than 30 ppm). Petrolatum according to Method 2, and perform the test. (4) Arsenic <1.11>—Prepare the test solution with 1.0 g Prepare the control solution with 3.0 mL of Standard Lead of White Petrolatum, according to Method 3, and perform Solution (not more than 30 ppm). the test. Add 10 mL of a solution of magnesium nitrate hex- (4) Arsenic <1.11>—Prepare the test solution with 1.0 g ahydrate in ethanol (95) (1 in 50), then add 1.5 mL of hydro- of Yellow Petrolatum, according to Method 3, and perform gen peroxide (30), and fire to burn (not more than 2 ppm). the test. Add 10 mL of a solution of magnesium nitrate hex- (5) Sulfur compound—To 4.0 g of White Petrolatum ahydrate in ethanol (95) (1 in 50), then add 1.5 mL of hydro- add 2 mL of ethanol (99.5) and 2 drops of sodium hydroxide gen peroxide (30), and fire to burn (not more than 2 ppm). solution (1 in 5) saturated with lead (II) oxide, warm the (5) Sulfur compound—To 4.0 g of Yellow Petrolatum mixture for 10 minutes at about 709C with frequent shaking, add 2 mL of ethanol (99.5) and 2 drops of sodium hydroxide and allow to cool: no dark color is produced. solution (1 in 5) saturated with lead (II) oxide, warm the (6) Organic acids—To 100 mL of dilute ethanol add 1 mixture for 10 minutes at about 709C with frequent shaking, drop of phenolphthalein TS, and titrate with 0.01 mol/L so- and allow to cool: no dark color is produced. dium hydroxide VS, until the color of the solution changes (6) Organic acids—To 100 mL of dilute ethanol add 1 to light red. Mix this solution with 20.0 g of White Petrola- drop of phenolphthalein TS, and titrate with 0.01 mol/L so- tum, and boil for 10 minutes under a reflux condenser. Add dium hydroxide VS, until the color of the solution changes 2 to 3 drops of phenolphthalein TS to the mixture and 0.40 to light red. Mix this solution with 20.0 g of Yellow Petrola- mL of 0.1 mol/L sodium hydroxide VS with vigorous shak- tum, and boil for 10 minutes under a reflux condenser. Add ing: the color of the solution remains red. 2 to 3 drops of phenolphthalein TS to the mixture and 0.40 (7) Fats and fatty oils or resins—To 10.0 g of White mL of 0.1 mol/L sodium hydroxide VS with vigorous shak- Petrolatum add 50 mL of sodium hydroxide solution (1 in ing: the color of the solution remains red. 5), and boil for 30 minutes under a reflux condenser. Cool (7) Fats and fatty oils or resins—To 10.0 g of Yellow the mixture, separate the aqueous layer, and filter, if neces- Petrolatum add 50 mL of sodium hydroxide solution (1 in sary. To the aqueous layer add 200 mL of dilute sulfuric 5), and boil for 30 minutes under a reflux condenser. Cool acid: neither oily matter nor precipitate is produced. the mixture, separate the aqueous layer, and filter, if necessary. To the aqueous layer add 200 mL of dilute sulfuric Residue on ignition <2.44> Not more than 0.05z (2 g). acid: neither oily matter nor precipitate is produced. Containers and storage Containers—Tight containers. JP XVI Official Monographs / Phenethicillin Potassium 1233

Residue on ignition <2.44> Not more than 0.05z (2 g). Containers and storage Containers—Tight containers. Phenethicillin Potassium フェネチシリンカリウム Petroleum Benzin

石油ベンジン

Petroleum Benzin is a mixture of low-boiling point hydrocarbons from petroleum. C17H19KN2O5S: 402.51 Description Petroleum Benzin occurs as a colorless, clear, Monopotassium (2S,5R,6R)-3,3-dimethyl-7-oxo-6- volatile liquid. It shows no fluorescence. It has a chracteris- [(2RS)-2-phenoxypropanoylamino]-4-thia-1- tic odor. azabicyclo[3.2.0]heptane-2-carboxylate It is miscible with ethanol (99.5) and with diethyl ether. [132-93-4] It is practically insoluble in water. It is very flammable. Phenethicillin Potassium contains not less than 1400 20 Specific gravity d 20:0.65–0.71 units and not more than 1480 units per mg, calculated on the dried basis. The potency of Phenethicillin Po- Purity (1) Acid—Shake vigorously 10 mL of Petroleum tassium is expressed as unit based on the amount of Benzin with 5 mL of water for 2 minutes, and allow to stand: phenethicillin potassium (C H KN O S). One unit of the separated aqueous layer does not change moistened blue 17 19 2 5 Phenethicillin Potassium is equivalent to 0.68 mgof litmus paper to red. phenethicillin potassium (C H KN O S). (2) Sulfur compounds and reducing substances—To 10 17 19 2 5 mL of Petroleum Benzin add 2.5 mL of ammonia-ethanol Description Phenethicillin Potassium occurs as a white to TS and 2 to 3 drops of silver nitrate TS, and warm the mix- light yellowish white crystalline powder. ture at about 509C for 5 minutes, protected from light: no It is freely soluble in water, and slightly soluble in ethanol brown color develops. (99.5). (3) Fatty oil and sulfur compounds—Drop and evapo- Identification (1) Determine the absorption spectrum of a rate 10 mL of Petroleum Benzin in small portions on odor- solution of Phenethicillin Potassium (1 in 5000) as directed less filter paper spread on a previously warmed glass plate: under Ultraviolet-visible Spectrophotometry <2.24>,and no spot or no foreign odor is perceptible. compare the spectrum with the Reference Spectrum: both (4) Benzene—Warm 5 drops of Petroleum Benzin with 2 spectra exhibit similar intensities of absorption at the same mL of sulfuric acid and 0.5 mL of nitric acid for about 10 wavelengths. minutes, allow to stand for 30 minutes, transfer the mixture (2) Determine the infrared absorption spectrum of to a porcelain dish, and dilute with water: no odor of Phenethicillin Potassium as directed in the potassium bro- nitrobenzene is perceptible. mide disk method under Infrared Spectrophotometry <2.25>, (5) Residue on evaporation—Evaporate 140 mL of and compare the spectrum with the Reference Spectrum: PetroleumBenzinonawaterbathtodryness,andheatthe both spectra exhibit similar intensities of absorption at the residue at 1059C to constant mass: the mass is not more than same wave numbers. 1mg. (3) Phenethicillin Potassium responds to Qualitative (6) Readily carbonizable substances—Shake vigorously 5 Tests <1.09> (1) for potassium salt. mL of Petroleum Benzin with 5 mL of sulfuric acid for rea- 20 dily carbonizable substances for 5 minutes in a Nessler tube, Optical rotation <2.49> [a]D : ＋217 – ＋2449(1 g calculated and allow to stand: the sulfuric acid layer has no more color on the dried basis, phosphate TS, 100 mL, 100 mm). than Matching Fluid A. L-a-Phenethicillin potassium Dissolve about 50 mg of Distilling range <2.57> 50–809C, not less than 90 volz. Phenethicillin Potassium in the mobile phase to make 50 mL, and use this solution as the sample solution. Perform Containers and storage Containers—Tight containers. the test with 10 mL of the sample solution as directed under Storage—Remote from fire, and not exceeding 309C. Liquid Chromatography <2.01> according to the following

conditions, and determine the peak areas, AD and AL,ofD- a-phenethicillin and L-a-phenethicillin by the automatic inte-

gration method: AL/(AD ＋ AL) is between 0.50 and 0.70. Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 254 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Adjust the pH of a mixture of a solution of 1234 Phenobarbital / Official Monographs JP XVI diammonium hydrogen phosphate (1 in 150) and acetonitrile for exactly 15 minutes. Add 0.2 – 0.5 mL of starch TS, and (41:10) to 7.0 with phosphoric acid. titrate <2.50> with 0.01 mol/L sodium thiosulfate VS until Flow rate: Adjust the flow rate so that the retention time the color of the solution disappears. Separately, to exactly 2 of L-a-phenethicillin is about 25 minutes. mL each of the sample solution and standard solution add System suitability— exactly 10 mL of 0.005 mol/L iodine VS, then proceed in the System performance: When the procedure is run with 10 same manner as above without allowing to stand for 15 mL of the sample solution under the above operating condi- minutes as a blank determination, and make any necessary tions, D-a-phenethicillin and L-a-phenethicillin are eluted in correction. Determine the volumes, VT and VS,of0.005 this order with the resolution between these peaks being not mol/L iodine VS consumed in the sample solution and less than 1.5. standard solution. System repeatability: When the test is repeated 6 times Amount (unit) of C H KN O S ＝ M × V /V with 10 mL of the sample solution under the above operating 17 19 2 5 S T S conditions, the relative standard deviation of the peak area MS: Amount (unit) of Phenethicillin Potassium RS of L-a-phenethicillin is not more than 2.0z. Containers and storage Containers—Well-closed contain- Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of ers. Phenethicillin Potassium according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Phenobarbital (2) Arsenic <1.11>—Prepare the test solution with 1.0 g of Phenethicillin Potassium according to Method 4 and, per- フェノバルビタール form the test (not more than 2 ppm). (3) Related substances—Dissolve 50 mg of Phenethicillin Potassium in 50 mL of the mobile, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as C H N O : 232.24 directed under Liquid Chromatography <2.01> according to 12 12 2 3 5-Ethyl-5-phenylpyrimidine-2,4,6(1H,3H,5H)-trione the following conditions, and determine each peak area by [50-06-6] the automatic integration method: the total of the peak areas other than -a-phenethicillin and -a-phenethicillin obtained D L Phenobarbital, when dried, contains not less than from the sample solution is not larger than 5 times the total 99.0z and not more than 101.0z of C12H12N2O3. of the peak areas of D-a-phenethicillin and L-a-phenethicillin from the standard solution. Description Phenobarbital occurs as white crystals or crys- Operating conditions— talline powder. Detector, column, column temperature, mobile phase, and It is very soluble in N,N-dimethylformamide, freely solu- flow rate: Proceed as directed in the operating conditions in ble in ethanol (95) and in acetone, sparingly soluble in aceto- the L-a-Phenethicillin potassium. nitrile, and very slightly soluble in water. Time span of measurement: About 1.5 times as long as the It dissolves in sodium hydroxide TS. retention time of L-a-phenethicillin. The pH of a saturated solution of Phenobarbital is be- System suitability— tween 5.0 and 6.0. Test for required detectability: Measure exactly 2 mL of Identification (1) Determine the absorption spectrum of a the standard solution, and add the mobile phase to make ex- solution of Phenobarbital in boric acid-potassium chloride- actly 10 mL. Confirm that the peak area of L-a-phenethicil- sodium hydroxide buffer solution, pH 9.6 (1 in 100,000) as lin obtained from 10 mL of this solution is equivalent to 14 to directed under Ultraviolet-visible Spectrophotometry <2.24>, 26z of that from 10 mL of the standard solution. and compare the spectrum with the Reference Spectrum: System performance, and system repeatability: Proceed as both spectra exhibit similar intensities of absorption at the directed in the system suitability in the L-a-Phenethicillin po- same wavelengths. tassium. (2) Determine the infrared absorption spectrum of Loss on drying <2.41> Not more than 1.0z (0.1 g, in vacu- Phenobarbital as directed in the potassium bromide disk um, 609C, 3 hours). method under Infrared Spectrophotometry <2.25>,andcompare the spectrum with the Reference Spectrum: both spectra Assay Weigh accurately an amount of Phenethicillin Po- exhibit similar intensities of absorption at the same wave tassium and dried Phenethicillin Potassium RS, equivalent numbers. to about 40,000 units, dissolve each in phosphate buffer solution, pH 6.0 to make exactly 20 mL, and use these solu- Melting point <2.60> 175 – 1799C tions as the sample solution and the standard solution, Purity (1) Clarity and color of solution—Dissolve 0.5 g respectively. Pipet 2 mL each of these solutions in 100-mL of Phenobarbital in 5 mL of sodium hydroxide TS: the solu- glass-stoppered flasks, add 2.0 mL of sodium hydroxide TS tion is clear and colorless. to them, and allow to stand for exactly 15 minutes. To them (2) Chloride <1.03>—Dissolve 0.30 g of Phenobarbital in add 2.0 mL of diluted hydrochloric acid (1 in 10) and exactly 20 mL of acetone, and add 6 mL of dilute nitric acid and 10 mL of 0.005 mol/L iodine VS, and allow them to stand water to make 50 mL. Perform the test using this solution as JP XVI Official Monographs / 10 Phenobarbital Powder 1235 the test solution. Prepare the control solution as follows: mixture of 50 mL of N,N-dimethylformamide and 22 mL of take 0.30 mL of 0.01 mol/L hydrochloric acid VS, 20 mL of ethanol (95), and make any necessary correction. acetone and 6 mL of dilute nitric acid, and add water to Each mL of 0.1 mol/L potassium hydroxide-ethanol VS make 50 mL (not more than 0.035z). ＝ 23.22 mg of C H N O (3) Heavy metals <1.07>—Proceed with 1.0 g of 12 12 2 3 Phenobarbital according to Method 2, and perform the test. Containers and storage Containers—Well-closed contain- Prepare the control solution with 2.0 mL of Standard Lead ers. solution (not more than 20 ppm). (4) Phenylbarbituric acid—Boil 1.0 g of Phenobarbital with 5 mL of ethanol (95) for 3 minutes: the solution is clear. 10 Phenobarbital Powder (5) Related substances—Dissolve 0.10 g of Phenobar- bital in 100 mL of acetonitrile, and use this solution as the Phenobarbital Powder sample solution. Pipet 2 mL of the sample solution, add acetonitrile to make exactly 100 mL. Pipet 5 mL of this solu- フェノバルビタール散 10 tion, add acetonitrile to make exactly 100 mL, and use this solution as the standard solution. Perform the test with 10z Phenobarbital Powder contains not less than exactly 10 mL each of the sample solution and standard 9.3z and not more than 10.7z of phenobarbital solution as directed under Liquid Chromatography <2.01> (C H N O : 232.24). according to the following conditions, and determine each 12 12 2 3 peak area of both solutions by the automatic integration Method of preparation method: the area of the peak other than phenobarbital ob- Phenobarbital 100 g tained from the sample solution is not larger than the peak Starch, Lactose Hydrate or area of phenobarbital from the standard solution. their mixture a sufficient quantity Operating conditions— Detector: An ultraviolet absorption photometer (wave- To make 1000 g length: 210 nm). Prepare as directed under Granules or Powders, with the Column: A stainless steel column 4.6 mm in inside diame- above ingredients. ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Identification (1) Determine the absorption spectrum of Column temperature: A constant temperature of about the sample solution obtained in the Assay as directed under 459C. Ultraviolet-visible Spectrophotometry <2.24>:itexhibitsa Mobile phase: A mixture of water and acetonitrile (11:9). maximum between 238 nm and 242 nm. Flow rate: Adjust the flow rate so that the retention time (2) To 6 g of 10z Phenobarbital Powder add 150 mL of of phenobarbital is about 5 minutes. ethanol, shake well, and filter. Condense the filtrate on a Time span of measurement: About 12 times as long as the water bath to about 5 mL, add about 50 mL of water, filter retention time of phenobarbital, beginning after the solvent to collect the formed crystals, and dry them at 1059Cfor2 peak. hours. Determine the infrared absorption spectrum of the System suitability— crystals as directed in the potassium bromide disk method Test for required detectability: Pipet 5 mL of the standard under Infrared Spectrophotometry <2.25>, and compare the solution, and add acetonitrile to make exactly 20 mL. Con- spectrum with the Reference Spectrum: both spectra exhibit firm that the peak area of phenobarbital obtained with 10 mL similar intensities of absorption at the same wave numbers. of this solution is equivalent to 20 to 30z of that with 10 mL Dissolution <6.10> When the test is performed at 50 revolu- of the standard solution. tions per minute according to the Paddle method, using 900 System performance: When the procedure is run with 10 mL of water as the dissolution medium, the dissolution rate mL of the standard solution under the above operating con- in 30 minutes of 10z Phenobarbital Powder is not less than ditions, the number of theoretical plates and the symmetry 80z. factor of the peak of phenobarbital are not less than 3000 Start the test with an accurately weighted about 0.3 g of and not more than 2.0, respectively. 10z Phenobarbital Powder, withdraw not less than 20 mL System repeatability: When the test is repeated 6 times of the medium at the specified minute after starting the test, with 10 mL of the standard solution under the above operat- and filter through a membrane filter with a pore size not ing conditions, the relative standard deviation of the peak exceeding 0.45 mm. Discard the first 10 mL of the filtrate, area of phenobarbital is not more than 3.0z. pipet 5 mL of the subsequent filtrate, add exactly 10 mL of Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, boric acid-potassium chloride-sodium hydroxide buffer solu- 2 hours). tion, pH 9.6, and use this solution as the sample solution. Separately, weigh accurately about 17 mg of phenobarbital Residue on ignition <2.44> Not more than 0.1z (1 g). for assay, previously dried at 1059C for 2 hours, and dis- Assay Weigh accurately about 0.5 g of Phenobarbital, pre- solve in water to make exactly 100 mL. Pipet 5 mL of this viously dried, dissolve in 50 mL of N,N-dimethylformamide, solution, and add water to make exactly 25 mL. Pipet 5 mL and titrate <2.50> with 0.1 mol/L potassium hydroxide- of this solution, add exactly 10 mL of boric acid-potassium ethanol VS until the color of the solution change from yel- chloride-sodium hydroxide buffer solution, pH 9.6, and use low to yellow-green (indicator: 1 mL of alizarin yellow GG- this solution as the standard solution. Perform the test with thymolphthalein TS). Perform a blank determination using a the sample solution and standard solution as directed under 1236 Phenol / Official Monographs JP XVI

Ultraviolet-visible Spectrophotometry <2.24>,usingamix- Identification (1) Add 1 drop of iron (III) chloride TS to ture of boric acid-potassium chloride-sodium hydroxide 10 mL of a solution of Phenol (1 in 100): a blue-purple color buffer solution, pH 9.6, and water (2:1) as the blank, and develops. determine the absorbances, AT and AS,at240nm. (2) Add bromine TS dropwise to 5 mL of a solution of Phenol (1 in 10,000): a white precipitate is produced, which Dissolution rate (z) with respect to the labeled amount at first dissolves with shaking, but becomes permanent as of phenobarbital (C H N O ) 12 12 2 3 excess of the reagent is added. ＝ MS/MT × AT/AS × 1/C × 180 Purity (1) Clarity and color of solution and acidity or M : Amount (mg) of phenobarbital for assay S alkalinity—Dissolve 1.0 g of Phenol in 15 mL of water: the M : Amount (g) of 10z Phenobarbital Powder T solution is clear, and neutral or only faintly acid. Add 2 C: Labeled amount (mg) of phenobarbital (C H N O )in 12 12 2 3 drops of methyl orange TS: no red color develops. 1g (2) Residue on evaporation—Weigh accurately about 5 g Assay Weigh accurately about 0.2 g of 10z Phenobarbital of Phenol, evaporate on a water bath, and dry the residue at Powder, dissolve in a boric acid-potassium chloride-sodium 1059C for 1 hour: the mass is not more than 0.05z of the hydroxide buffer solution, pH 9.6 to make exactly 100 mL. mass of the sample. Pipet 5 mL of this solution, add a boric acid-potassium chlo- Assay Dissolve about 1.5 g of Phenol, accurately weighed, ride-sodium hydroxide buffer solution, pH 9.6 to make ex- in water to make exactly 1000 mL. Transfer exactly 25 mL of actly 100 mL, and use this solution as the sample solution. this solution to an iodine flask, add exactly 30 mL of 0.05 Separately, weigh accurately about 20 mg of phenobarbital mol/L bromine VS, then 5 mL of hydrochloric acid, and im- for assay, previously dried at 1059C for 2 hours, and add a mediately stopper the flask. Shake the flask repeatedly for boric acid-potassium chloride-sodium hydroxide buffer solu- 30 minutes, allow to stand for 15 minutes, then add 7 mL of tion, pH 9.6 to make exactly 100 mL. Pipet 5 mL of this so- potassium iodide TS, at once stopper the flask, and shake lution, add a boric acid-potassium chloride-sodium hydrox- well. Add 1 mL of chloroform, stopper the flask, and shake ide buffer solution, pH 9.6 to make exactly 100 mL, and use thoroughly. Titrate <2.50> the liberated iodine with 0.1 this solution as the standard solution. Perform the test with mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). the sample solution and standard solution as directed under Perform a blank determination. Ultraviolet-visible Spectrophotometry <2.24>, using a boric acid-potassium chloride-sodium hydroxide buffer solution, Each mL of 0.05 mol/L bromine VS pH 9.6 as the blank, and determine the absorbances, AT and ＝ 1.569 mg of C6H6O A , at 240 nm. S Containers and storage Containers—Tight containers.

Amount (mg) of phenobarbital (C12H12N2O3) Storage—Light-resistant. ＝ MS × AT/AS MS: Amount (mg) of phenobarbital for assay Phenol for Disinfection Containers and storage Containers—Well-closed containers. Carbolic Acid for Disinfection

消毒用フェノール Phenol Phenol for Disinfection contains not less than Carbolic Acid 95.0z of phenol (C6H6O: 94.11). フェノール Description Phenol for Disinfection occurs as colorless to slightly red crystals, crystalline masses, or liquid containing these crystals. It has a characteristic odor. It is very soluble in ethanol (95) and in diethyl ether, and freely soluble in water.

C6H6O: 94.11 Phenol for Disinfection (10 g) is liquefied by addition of 1 Phenol mL of water. [108-95-2] It cauterizes the skin, turning it white. Congealing point: about 309C. Phenol contains not less than 98.0z of C H O. 6 6 Identification (1) To 10 mL of a solution of Phenol for Description Phenol occurs as colorless to slightly red crys- Disinfection (1 in 100) add 1 drop of iron (III) chloride TS: a tals or crystalline masses. It has a characteristic odor. blue-purple color is produced. It is very soluble in ethanol (95) and in diethyl ether, and (2) To 5 mL of a solution of Phenol for Disinfection (1 soluble in water. in 10,000) add bromine TS dropwise: a white precipitate is Phenol (10 g) is liquefied by addition of 1 mL of water. formed, and it dissolves at first upon shaking but becomes The color changes gradually through red to dark red by permanent as excess of the reagent is added. light or air. Purity (1) Clarity of solution—Dissolve 1.0 g of Phenol It cauterizes the skin, turning it white. for Disinfection in 15 mL of water: the solution is clear. Congealing point: about 409C (2) Residue on evaporation—Weigh accurately about 5 g JP XVI Official Monographs / Phenol and Zinc Oxide Liniment 1237 of Phenol for Disinfection, evaporate on a water bath, and mL of 0.05 mol/L bromine VS, then 5 mL of hydrochloric dry the residue at 1059C for 1 hour: the mass is not more acid, and immediately stopper the flask. Shake the flask than 0.10z of the mass of the sample. repeatedly for 30 minutes, allow to stand for 15 minutes, then add 7 mL of potassium iodide TS, at one stopper the Assay Dissolve about 1 g of Phenol for Disinfection, accu- flask tightly, and shake well. Add 1 mL of chloroform, stop- rately weighed, in water to make exactly 1000 mL. Pipet 25 per the flask, and shake thoroughly. Titrate <2.50> the liber- mL of the solution into an iodine flask, add exactly 30 mL of ated iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 0.05 mol/L bromine VS and 5 mL of hydrochloric acid, 1 mL of starch TS). Perform a blank determination. stopper immediately, shake for 30 minutes and allow to stand for 15 minutes. Add 7 mL of potassium iodide TS, Each mL of 0.05 mol/L bromine VS stopper immediately, shake well, and titrate <2.50> the liber- ＝ 1.569 mg of C6H6O ated iodine with 0.1 mol/L sodium thiosulfate VS (indicator: Containers and storage Containers—Tight containers. 1 mL of starch TS). Perform a blank determination. Storage—Light-resistant. Each mL of 0.05 mol/L bromine VS

＝ 1.569 mg of C6H6O Containers and storage Containers—Tight containers. Dental Phenol with Camphor Storage—Light-resistant. 歯科用フェノール・カンフル

Liquefied Phenol Method of preparation Phenol 35 g Liquefied Carbolic Acid d-ordl-Camphor 65 g 液状フェノール To make 100 g Melt Phenol by warming, add d-Camphor or dl-Cam- Liquefied Phenol is Phenol maintained in a liquid phor, and mix. condition by the presence of 10z of Water, Purified Description Dental Phenol with Camphor is a colorless or Water or Purified Water in Containers. light red liquid. It has a characteristic odor. It contains not less than 88.0z of phenol (C6H6O: 94.11) Containers and storage Containers—Tight containers. Storage—Light-resistant. Description Liquefied Phenol is a colorless or slightly reddish liquid. It has a characteristic odor. It is miscible with ethanol (95), with diethyl ether and with glycerin. Phenol and Zinc Oxide Liniment A mixture of equal volumes of Liquefied Phenol and glyc- フェノール・亜鉛華リニメント erin is miscible with water. The color changes gradually to dark red on exposure to light or air. Method of preparation It cauterizes the skin, turning it white. 20 Liquefied Phenol 22 mL Specific gravity d 20: about 1.065 Powdered Tragacanth 20 g Identification (1) Add 1 drop of iron (III) chloride TS to Carmellose Sodium 30 g 10 mL of a solution of Liquefied Phenol (1 in 100): a blue- Glycerin 30 mL purple color develops. Zinc Oxide 100 g (2) Add bromine TS dropwise to 5 mL of a solution of Purified Water or Purified Liquefied Phenol (1 in 10,000): a white precipitate is pro- Water in Containers a sufficient quantity duced, which at first dissolves with shaking, but becomes To make 1000 g permanent as excess of the reagent is added. Mix Liquefied Phenol, Glycerin and Purified Water or Boiling point <2.57> Not more than 1829C. Purified Water in Containers, add Powdered Tragacanth in Purity (1) Clarity and color of solution and acidity or small portions by stirring, and allow the mixture to stand alkalinity—Dissolve 1.0 g of Liquefied Phenol in 15 mL of overnight. To the mixture add Carmellose Sodium in small water: the solution is clear, and neutral or only faintly acid. portions by stirring to make a pasty mass, add Zinc Oxide in Add 2 drops of methyl orange TS: no red color develops. small portions, and mix. Less than 5 g of Powdered (2) Residue on evaporation—Weigh accurately about 5 g Tragacanth or Carmellose Sodium can be replaced by each of Liquefied Phenol, evaporate on a water bath, and dry the other to make 50 g in total. residue at 1059C for 1 hour: the mass is not more than Description Phenol and Zinc Oxide Liniment is a white, 0.05z of the mass of the sample. pasty mass. It has a slight odor of phenol. Assay Dissolve about 1.7 g of Liquefied Phenol, accurately Identification (1) Shake well 1 g of Phenol and Zinc weighed, in a water to make exactly 1000 mL. Transfer ex- Oxide Liniment with 10 mL of diethyl ether, and filter. To actly 25 mL of this solution to an iodine flask, add exactly 30 the filtrate add 10 mL of dilute sodium hydroxide TS, shake 1238 Phenolated Water / Official Monographs JP XVI well, and separate the water layer. To 1 mL of the water Each mL of 0.05 mol/L bromine VS layer add 1 mL of sodium nitrite TS and 1 mL of dilute hy- ＝ 1.569 mg of C6H6O drochloric acid, shake, and add 3 mL of sodium hydroxide Containers and storage Containers—Tight containers. TS: a yellow color develops (phenol). (2) Place 1 g of Phenol and Zinc Oxide Liniment in a porcelain crucible, heat gradually raising the temperature until the content is charred, and then ignite it strongly: a Phenolated Water for Disinfection yellow color develops, and disappears on cooling. To the 消毒用フェノール水 residue add 10 mL of water and 5 mL of dilute hydrochloric acid, shake well, and filter. To the filtrate add 2 to 3 drops of potassium hexacyanoferrate (II) TS: a white precipitate is Phenolated Water for Disinfection contains not less produced (zinc oxide). than 2.8 w/vz and not more than 3.3 w/vz of (3) Shake 0.5 g of Phenol and Zinc Oxide Liniment with phenol (C6H6O: 94.11). 1 mL of water and 5 mL of chloroform, separate the chlo- Method of preparation roform layer, and use this solution as the sample solution. Separately, dissolve 0.01 g of phenol in 5 mL of chloroform, Phenol for Disinfection 31 g and use this solution as the standard solution. Perform the Water, Purified Water or Purified test with these solutions as directed under Thin-layer Chro- Water in Containers a sufficient quantity matography <2.03>. Spot 5 mL each of the sample solution To make 1000 mL and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl Mix the above ingredients. acetate, ethanol (99.5) and ammonia solution (28) (50:5:1) to Description Phenolated Water for Disinfection is a clear, a distance of about 10 cm, and air-dry the plate. Allow the colorless liquid, having the odor of phenol. plate to stand in iodine vapor: the spots obtained from the sample solution and the standard solution show the same Rf Identification (1) Add 1 drop of iron (III) chloride TS to value. 10 mL of Phenolated Water for Disinfection: a blue-purple color develops. Containers and storage Containers—Tight containers. (2) Proceed with 5 mL of a solution of Phenolated Water for Disinfection (1 in 200) as directed in the Identifi- Phenolated Water cation (2) under Phenol for Disinfection. Assay Take exactly 5 mL of Phenolated Water for Disin- フェノール水 fection, add water to make exactly 100 mL, then pipet 25 mL of the solution into an iodine flask, and proceed as directed in the Assay under Phenol for Disinfection. Phenolated Water contains not less than 1.8 w/vz and not more than 2.3 w/vz of phenol (C6H6O: Each mL of 0.05 mol/L bromine VS 94.11). ＝ 1.569 mg of C6H6O Method of preparation Containers and storage Containers—Tight containers. Liquefied Phenol 22 mL Water, Purified Water or Purified Water in Containers a sufficient quantity Phenolsulfonphthalein To make 1000 mL フェノールスルホンフタレイン Mix the above ingredients. Description Phenolated Water is a colorless, clear liquid, having the odor of phenol. Identification (1) Add 1 drop of iron (III) chloride TS to 10 mL of Phenolated Water: a blue-purple color develops. (2) To 5 mL of a solution of Phenolated Water (1 in 200) add bromine TS dropwise: a white precipitate is formed, and C19H14O5S: 354.38 it dissolves at first upon shaking but becomes permanent as 2-[Bis(4-hydroxyphenyl)methyliumyl]benzenesulfonate excess of the reagent is added. [143-74-8] Assay Take exactly 2 mL of Phenolated Water into an Phenolsulfonphthalein, when dried, contains not iodine flask, add 25 mL of water, then add exactly 40 mL of less than 98.0z of C H O S. 0.05 mol/L bromine VS and 5 mL of hydrochloric acid, 19 14 5 stopper immediately, shake for 30 minutes, and allow to Description Phenolsulfonphthalein occurs as a vivid red to stand for 15 minutes. Add 7 mL of potassium iodide TS, dark red, crystalline powder. stopper tightly at once, shake well, and titrate <2.50> the lib- It is very slightly soluble in water and in ethanol (95). erated iodine with 0.1 mol/L sodium thiosulfate VS (indica- It dissolves in sodium hydroxide TS. tor: 1 mL of starch TS). Perform a blank determination. Identification (1) Dissolve 5 mg of Phenolsulfonphtha- JP XVI Official Monographs / Phenolsulfonphthalein Injection 1239 lein in 2 to 3 drops of sodium hydroxide TS, add 2 mL of 0.05 mol/L bromine VS and 1 mL of dilute sulfuric acid, Phenolsulfonphthalein Injection shake well, and allow to stand for 5 minutes. Render the solution alkaline with sodium hydroxide TS: a deep blue-pur- フェノールスルホンフタレイン注射液 ple color develops. (2) Dissolve 0.01 g of Phenolsulfonphthalein in diluted Phenolsulfonphthalein Injection is an aqueous solu- sodium carbonate TS (1 in 10) to make 200 mL. To 5 mL of tion for injection. this solution add diluted sodium carbonate TS (1 in 10) to It contains not less than 0.54 w/vz and not more make 100 mL. Perform the test with this solution as directed than 0.63 w/vz of phenolsulfonphthalein (C H O S: under Ultraviolet-visible Spectrophotometry <2.24>,and 19 14 5 354.38). compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same Method of preparation wavelengths. Phenolsulfonphthalein 6 g Purity (1) Insoluble substances—To about 1 g of Phenol- Sodium Chloride 9 g sulfonphthalein, accurately weighed, add 20 mL of a solu- Sodium Bicarbonate 1.43 g tion of sodium hydrogen carbonate (1 in 40). Allow the mix- (or Sodium Hydroxide 0.68 g) ture to stand for 1 hour with frequent shaking, dilute with Water for Injection or Sterile Water water to 100 mL, and allow to stand for 24 hours. Collect for Injection in Containers a sufficient quantity the insoluble substances using a tared glass filter (G4), wash To make 1000 mL with 25 mL of a solution of sodium hydrogen carbonate (1 in 100) and with five 5-mL portions of water, and dry at 1059C Prepare as directed under Injections, with the above ingre- for 1 hour: the mass of the residue is not more than 0.2z. dients. (2) Related substances—Dissolve 0.10 g of Phenolsul- Description Phenolsulfonphthalein Injection is a clear, fonphthalein in 5 mL of dilute sodium hydroxide TS, and orange-yellow to red liquid. use this solution as the sample solution. Pipet 0.5 mL of the sample solution, add dilute sodium hydroxide TS to make Identification To 1 mL of Phenolsulfonphthalein Injection exactly 100 mL, and use this solution as the standard solu- add 2 to 3 drops of sodium hydroxide TS, and proceed as dition. Perform the test with these solutions as directed under rected in the Identification (1) under Phenolsulfonphthalein. 2.03 Thin-layer Chromatography < >. Spot 10 mLeachofthe pH <2.54> 6.0–7.6 sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Bacterial endotoxins <4.01> Less than 7.5 EU/mg. Develop the plate with a mixture of t-amyl alcohol, acetic Extractable volume <6.05> It meets the requirement. acid (100) and water (4:1:1) to a distance of about 15 cm, and air-dry the plate. After allowing the plate to stand in an Foreign insoluble matter <6.06> Perform the test according ammonia vapor, examine under ultraviolet light (main wave- to Method 1: it meets the requirement. length: 254 nm): the spots other than the principal spot from Insoluble particulate matter <6.07> Perform the test ac- the sample solution are not more intense than the spot from cording to Method 2: it meets the requirement. the standard solution. Sterility <4.06> Perform the test according to the Mem- Loss on drying <2.41> Not more than 1.0z (1 g, silica gel, brane filtration method: it meets the requirement. 4 hours). Sensitivity To 1.0 mL of Phenolsulfonphthalein Injection Residue on ignition <2.44> Not more than 0.2z (1 g). add 5 mL of water. To 0.20 mL of this solution add 50 mL Assay Weigh accurately about 0.15 g of Phenolsul- of freshly boiled and cooled water and 0.40 mL of 0.01 fonphthalein, previously dried, transfer to an iodine flask, mol/L sodium hydroxide VS: a deep red-purple color devel- dissolve in 30 mL of a solution of sodium hydroxide (1 in ops, and it changes to light yellow on the addition of 0.40 250), and add water to make 200 mL. Add exactly measured mL of 0.005 mol/L sulfuric acid VS. 50 mL of 0.05 mol/L bromine VS, add 10 mL of hydrochlo- Assay Pipet 5 mL of Phenolsulfonphthalein Injection, and ric acid to the solution quickly, and stopper immediately. add a solution of anhydrous sodium carbonate (1 in 100) to Allow the mixture to stand for 5 minutes with occasional make exactly 250 mL. Pipet 5 mL of this solution, add a so- shaking, add 7 mL of potassium iodide TS, stopper again lution of anhydrous sodium carbonate (1 in 100) to make ex- immediately, and shake gently for 1 minute. Titrate <2.50> actly 200 mL, and use this solution as the sample solution. the liberated iodine with 0.1 mol/L sodium thiosulfate VS Separately, weigh accurately about 30 mg of phenolsul- (indicator: 1 mL of starch TS). Perform a blank determina- fonphthalein for assay, previously dried in a desiccator (sili- tion. ca gel) for 4 hours, and dissolve in a solution of anhydrous Each mL of 0.05 mol/L bromine VS sodium carbonate (1 in 100) to make exactly 250 mL. Pipet 5

＝ 4.430 mg of C19H14O5S mL of this solution, add a solution of anhydrous sodium carbonate (1 in 100) to make exactly 200 mL, and use this so- Containers and storage Containers—Well-closed contain- lution as the standard solution. Determine the absorbances, ers. AT and AS, of the sample solution and standard solution at 559 nm as directed under Ultraviolet-visible Spectropho- tometry <2.24>. 1240 L-Phenylalanine / Official Monographs JP XVI

Amount (mg) of phenolsulfonphthalein (C19H14O5S) nine in 25 mL of water, and use this solution as the sample ＝ MS × AT/AS solution. Pipet 1 mL of the sample solution, and add water to make exactly 50 mL. Pipet 5 mL of this solution, add M : Amount (mg) of phenolsulfonphthalein for assay S water to make exactly 20 mL, and use this solution as the Containers and storage Containers—Hermetic containers. standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a L-Phenylalanine plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid L-フェニルアラニン (100) (3:1:1) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat at 809Cfor5 minutes: the spots other than the principal spot from the sample solution are not more intense than the spot from the

C9H11NO2:165.19 standard solution. (2S)-2-Amino-3-phenylpropanoic acid Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, [63-91-2] 3 hours).

L-Phenylalanine, when dried, contains not less than Residue on ignition <2.44> Not more than 0.1z (1 g). 98.5z of C9H11NO2. Assay Weigh accurately about 0.17 g of L-Phenylalanine, Description L-Phenylalanine occurs as white crystals or previously dried, and dissolve in 3 mL of formic acid, add 50 crystalline powder. It is odorless or has a faint characteristic mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L odor, and has a slightly bitter taste. perchloric acid VS (potentiometric titration). Perform a It is freely soluble in formic acid, sparingly soluble in blank determination, and make any necessary correction. water, and practically insoluble in ethanol (95). Each mL of 0.1 mol/L perchloric acid VS It dissolves in dilute hydrochloric acid. ＝ 16.52 mg of C9H11NO2 Identification Determine the infrared absorption spectrum Containers and storage Containers—Tight containers. of L-Phenylalanine, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho- tometry <2.25>, and compare the spectrum with the Refer- ence Spectrum: both spectra exhibit similar intensities of Phenylbutazone absorption at the same wave numbers. フェニルブタゾン 20 Optical rotation <2.49> [a]D : －33.0 – －35.59(after drying, 0.5 g, water, 25 mL, 100 mm). pH <2.54> Dissolve 0.20 g of L-Phenylalanine in 20 mL of water: the pH of this solution is between 5.3 and 6.3. Purity (1) Clarity and color of solution—Dissolve 0.5 g of L-Phenylalanine in 10 mL of 1 mol/L hydrochloric acid TS: the solution is clear and colorless.

(2) Chloride <1.03>—Perform the test with 0.5 g of L- C19H20N2O2: 308.37 Phenylalanine. Prepare the control solution with 0.30 mL of 4-Butyl-1,2-diphenylpyrazolidine-3,5-dione 0.01 mol/L hydrochloric acid VS (not more than 0.021z). [50-33-9] (3) Sulfate <1.14>—Perform the test with 0.6 g of L- Phenylalanine. Prepare the control solution with 0.35 mL of Phenylbutazone, when dried, contains not less than 0.005 mol/L sulfuric acid VS (not more than 0.028z). 99.0z of C19H20N2O2. (4) Ammonium <1.02>—Perform the test with 0.25 g of Description Phenylbutazone occurs as a white to slightly L-Phenylalanine. Prepare the control solution with 5.0 mL yellowish white, crystalline powder. It is odorless, and is at of Standard Ammonium Solution (not more than 0.02z). first tasteless but leaves a slightly bitter aftertaste. (5) Heavy metals <1.07>—Dissolve 1.0 g of L-Phenylala- It is freely soluble in acetone, soluble in ethanol (95) and nine in 40 mL of water and 2 mL of dilute acetic acid by in diethyl ether, and practically insoluble in water. warming, cool, and add water to make 50 mL. Perform the It dissolves in sodium hydroxide TS. test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solu- Identification (1) To 0.1 g of Phenylbutazone add 1 mL tion add 2 mL of dilute acetic acid and water to make 50 mL of acetic acid (100) and 1 mL of hydrochloric acid, and heat (not more than 20 ppm). on a water bath under a reflux condenser for 30 minutes. (6) Arsenic <1.11>—Dissolve 1.0 g of L-Phenylalanine in Add 10 mL of water, and cool with ice water. Filter, and to 5 mL of dilute hydrochloric acid and 15 mL of water, and the filtrate add 3 to 4 drops of sodium nitrite TS. To 1 mL of perform the test with this solution as the test solution (not this solution add 1 mL of 2-naphthol TS and 3 mL of chlo- more than 2 ppm). roform, and shake: a deep red color develops in the chlo- (7) Related substances—Dissolve 0.10 g of L-Phenylala- roform layer. JP XVI Official Monographs / Phenylephrine Hydrochloride 1241

(2) Dissolve 1 mg of Phenylbutazone in 10 mL of dilute Description Phenylephrine Hydrochloride occurs as white sodium hydroxide TS, and dilute with water to make 100 crystals or crystalline powder. It is odorless, and has a bitter mL. Determine the absorption spectrum of the solution as taste. directed under Ultraviolet-visible Spectrophotometry <2.24>, It is very soluble in water, freely soluble in ethanol (95), and compare the spectrum with the Reference Spectrum: and practically insoluble in diethyl ether. both spectra exhibit similar intensities of absorption at the The pH of a solution of Phenylephrine Hydrochloride (1 same wavelengths. in 100) is between 4.5 and 5.5. Melting point <2.60> 104–1079C Identification (1) To 1 mL of a solution of Phenylephrine Hydrochloride (1 in 100) add 1 drop of copper (II) sulfate TS Purity (1) Clarity of solution—Dissolve 1.0 g of Phenyl- and 1 mL of a solution of sodium hydroxide (1 in 5): a blue butazone in 20 mL of sodium hydroxide solution (2 in 25), color is produced. To the solution so obtained add 1 mL of and allow to stand at 25 ± 19C for 3 hours: the solution is diethyl ether, and shake vigorously: no blue color develops clear. Determine the absorbance of this solution at 420 nm as in the diethyl ether layer. directed under Ultraviolet-visible Spectrophotometry <2.24>: (2) To 1 mL of a solution of Phenylephrine Hydrochlo- it is not more than 0.05. ride (1 in 100) add 1 drop of iron (III) chloride TS: a persis- (2) Heavy metals <1.07>—Proceed with 2.0 g of Phenyl- tent purple color is produced. butazone according to Method 2, and perform the test. Pre- (3) Dissolve 0.3 g of Phenylephrine Hydrochloride in 3 pare the control solution with 2.0 mL of Standard Lead So- mL of water, add 1 mL of ammonia TS, and rub the inner lution (not more than 10 ppm). side of the test tube with a glass rod: a precipitate is pro- (3) Arsenic <1.11>—Prepare the test solution with 1.0 g duced. Collect the precipitate, wash with a few drops of ice- of phenylbutazone, according to Method 3, and perform the cold water, and dry at 1059C for 2 hours: it melts <2.60> be- test (not more than 2 ppm). tween 1709C and 1779C. (4) Readily carbonizable substances—Dissolve 1.0 g of (4) A solution of Phenylephrine Hydrochloride (1 in 100) Phenylbutazone in 20 mL of sulfuric acid, and allow to responds to Qualitative Tests <1.09> (2) for chloride. stand at 25 ± 19C for exactly 30 minutes: the solution is 20 clear. Determine the absorbance of this solution at 420 nm as Optical rotation <2.49> [a]D : －42.0 – －47.59(after dry- directed under Ultraviolet-visible Spectrophotometry <2.24>: ing, 0.5 g, water, 10 mL, 100 mm). it is not more than 0.10. Melting point <2.60> 140 – 1459C Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- Purity (1) Clarity and color of solution—Dissolve 1.0 g um, silica gel, 4 hours). of Phenylephrine Hydrochloride in 10 mL of water: the solu- Residue on ignition <2.44> Not more than 0.1z (1 g). tion is clear and colorless. (2) Sulfate <1.14>—Take 0.5 g of Phenylephrine Hydro- Assay Weigh accurately about 0.5 g of Phenylbutazone, chloride, and perform the test. Prepare the control solution previously dried, dissolve in 25 mL of acetone, and titrate with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more <2.50> with 0.1 mol/L sodium hydroxide VS until the solu- than 0.048z). tion shows a blue color which persists for 15 seconds (indica- (3) Ketone—Dissolve 0.20 g of Phenylephrine Hydro- tor: 5 drops of bromothymol blue TS). Perform a blank de- chloride in 1 mL of water, and add 2 drops of sodium penta- termination with a mixture of 25 mL of acetone and 16 mL cyanonitrosylferrate (III) TS, 1 mL of sodium hydroxide TS of water, and make any necessary correction. and then 0.6 mL of acetic acid (100): the solution has no Each mL of 0.1 mol/L sodium hydroxide VS more color than the following control solution.

＝ 30.84 mg of C19H20N2O2 Control solution: Prepare as directed above without Phenylephrine Hydrochloride. Containers and storage Containers—Tight containers. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Phenylephrine Hydrochloride Residue on ignition <2.44> Not more than 0.2z (1 g). フェニレフリン塩酸塩 Assay Weigh accurately about 0.1 g of Phenylephrine Hy- drochloride, previously dried, dissolve in 40 mL of water contained in an iodine flask, add exactly measured 50 mL of 0.05 mol/L bromine VS, then add 5 mL of hydrochloric acid, and immediately stopper tightly. Shake the mixture, and allow to stand for 15 minutes. To this solution add 10 mL of potassium iodide TS carefully, stopper tightly imme-

C9H13NO2.HCl: 203.67 diately, shake thoroughly, allow to stand for 5 minutes, and (1R)-1-(3-Hydroxyphenyl)-2-methylaminoethanol titrate <2.50> with 0.1 mol/L sodium thiosulfate VS (indica- monohydrochloride tor: 1 mL of starch TS). Perform a blank determination. [61-76-7] Each mL of 0.05 mol/L bromine VS ＝ 3.395 mg of C H NO .HCl Phenylephrine Hydrochloride, when dried, contains 9 13 2 not less than 98.0z and not more than 102.0z of Containers and storage Containers—Tight containers. C9H13NO2.HCl. Storage—Light-resistant. 1242 Phenytoin / Official Monographs JP XVI

(3) Chloride <1.03>—Dissolve 0.30 g of Phenytoin in 30 Phenytoin mL of acetone, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the Diphenylhydantoin test solution. Prepare the control solution from 0.60 mL of 0.01 mol/L hydrochloric acid VS, 30 mL of acetone and 6 フェニトイン mL of dilute nitric acid, and add water to 50 mL (not more than 0.071z). (4) Heavy metals <1.07>—Proceedwith1.0gof Phenytoin according to Method 2, and perform the test. Pre- pare the control solution with 2.0 mL of Standard Lead So- lution (not more than 20 ppm). Loss on drying <2.41> Not more than 0.5z (2 g, 1059C,

C15H12N2O2: 252.27 2 hours). 5,5-Diphenylimidazolidine-2,4-dione Residue on ignition <2.44> Not more than 0.1z (1 g). [57-41-0] Assay Weigh accurately about 0.5 g of Phenytoin, previ- Phenytoin, when dried, contains not less than ously dried, dissolve in 40 mL of ethanol (95) with the aid of 99.0z of C15H12N2O2. gentle heating, add 0.5 mL of thymolphthalein TS immediately, and titrate with 0.1 mol/L sodium hydroxide VS until Description Phenytoin occurs as a white, crystalline pow- a light blue color develops. Then add 1 mL of pyridine, 5 der or granules. It is odorless and tasteless. drops of phenolphthalein TS and 25 mL of silver nitrate TS, It is sparingly soluble in ethanol (95) and in acetone, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS until slightly soluble in diethyl ether, and practically insoluble in a light red color, which persists for 1 minute, develops. water. It dissolves in sodium hydroxide TS. Each mL of 0.1 mol/L sodium hydroxide VS

Melting point: about 2969C (with decomposition). ＝ 25.23 mg of C15H12N2O2 Identification (1) Dissolve 0.02 g of Phenytoin in 2 mL of Containers and storage Containers—Well-closed contain- ammonia TS, and add 5 mL of silver nitrate TS: a white pre- ers. cipitate is produced. (2) Boil a mixture of 0.01 g of Phenytoin, 1 mL of ammonia TS and 1 mL of water, and add dropwise 2 mL of a Phenytoin Powder mixture prepared from 50 mL of a solution of copper (II) sulfate pentahydrate (1 in 20) and 10 mL of ammonia TS: a Diphenylhydantoin Powder red, crystalline precipitate is produced. (3) Heat 0.1 g of Phenytoin with 0.2 g of sodium hydrox- フェニトイン散 ide, and fuse: the gas evolved turns moistened red litmus paper blue. Phenytoin Powder contains not less than 95.0z and (4) Add 3 mL of chlorinated lime TS to 0.1 g of not more than 105.0z of the labeled amount of Phenytoin, shake for 5 minutes, and dissolve the oily pre- phenytoin (C H N O : 252.27). cipitate in 15 mL of hot water. After cooling, add 1 mL of 15 12 2 2 dilute hydrochloric acid dropwise, then add 4 mL of water. Method of preparation Prepare as directed under Granules Filter the white precipitate thus obtained, wash with water, or Powders, with Phenytoin. and press it with dry filter paper to remove the accom- Identification Weigh a portion of Phenytoin Powder, panying water. Dissolve the precipitate with 1 mL of chlo- equivalent to 0.3 g of Phenytoin according to the labeled roform, add 5 mL of diluted ethanol (9 in 10), and rub the amount, stir well with two 100-mL portions of diethyl ether, inner surface of the flask to produce a white, crystalline pre- and extract. Combine the diethyl ether extracts, and filter. cipitate. Collect the precipitate, wash with ethanol (95), and Evaporate the filtrate on a water bath to dryness, and dry: the melting point <2.60> is between 1659C and 1699C. proceed with the residue as directed in the Identification Purity (1) Clarity and color of solution—Dissolve 0.20 g under Phenytoin. of Phenytoin in 10 mL of 0.2 mol/L sodium hydroxide VS: Dissolution Being specified separately. the solution is clear and colorless. Then heat the solution: no turbidity is produced. Cool, and mix the solution with 5 mL Assay Weigh accurately an amount of Phenytoin Powder, of acetone: the solution is clear and colorless. equivalent to about 50 mg of phenytoin (C15H12N2O2), add (2) Acidity or alkalinity—Shake 2.0 g of Phenytoin with 30 mL of methanol, treat with ultrasonic waves for 15 40 mL of water for 1 minute, filter, and perform the follow- minutes with occasional shaking, shake for another 10 ing tests using this filtrate as the sample solution. minutes, and add methanol to make exactly 50 mL. Centri- (i) To 10 mL of the sample solution add 2 drops of phe- fuge this solution, pipet 5 mL of the supernatant liquid, add nolphthalein TS: no color develops. Then add 0.15 mL of exactly 5 mL of the internal standard solution, and use this 0.01 mol/L sodium hydroxide VS: a red color develops. solution as the sample solution. Separately, weigh accurately (ii) To 10 mL of the sample solution add 0.30 mL of 0.01 about 25 mg of phenytoin for assay, previously dried at mol/L hydrochloric acid VS and 5 drops of methyl red TS: a 1059C for 2 hours, and dissolve in methanol to make exactly red to orange color develops. 25 mL. Pipet 5 mL of this solution, add exactly 5 mL of the JP XVI Official Monographs / Phenytoin Tablets 1243 internal standard solution, and use this solution as the standing to the following method: it meets the requirement of the ard solution. Perform the test with 10 mLeachofthesample Content uniformity test. solution and standard solution as directed under Liquid To 1 tablet of Phenytoin Tablets add 3V/5 mL of a mix- Chromatography <2.01> according to the following condi- ture of water and acetonitrile (1:1), treat with ultrasonic tions, and calculate the ratios, QT and QS, of the peak area waves for 15 minutes with occasional shaking, shake for of phenytoin to that of the internal standard. another 10 minutes, and add a mixture of water and acetonitrile (1:1) to make exactly V mL so that each mL contains Amount (mg) of phenytoin (C H N O ) 15 12 2 2 about 1 mg of phenytoin (C H N O ). Centrifuge this solu- ＝ M × Q /Q × 2 15 12 2 2 S T S tion, pipet 5 mL of the supernatant liquid, add exactly 5 mL

MS: Amount (mg) of phenytoin for assay of the internal standard solution, and use this solution as the sample solution. Proceed as directed in the Assay. Internal standard solution—A solution of propyl parahydroxybenzoate in the mobile phase (1 in 25,000). Amount (mg) of phenytoin (C15H12N2O2) Operating conditions— ＝ MS × QT/QS × V/25 Detector: An ultraviolet absorption photometer (wave- M : Amount (mg) of phenytoin for assay length: 258 nm). S Column: A stainless steel column 4.6 mm in inside diame- Internal standard solution—A solution of propyl parahy- ter and 15 cm in length, packed with octadecylsilanized silica droxybenzoate in the mobile phase (1 in 25,000). gel for liquid chromatography (5 mm in particle diameter). Dissolution Being specified separately. Column temperature: A constant temperature of about 409C. Assay Weigh accurately the mass of not less than 20 Mobile phase: A mixture of methanol and 0.02 mol/L Phenytoin Tablets, and powder in an agate mortar. Weigh phosphate buffer solution, pH 3.5 (11:9). accurately a portion of the powder, equivalent to about 50

Flow rate: Adjust the flow rate so that the retention time mg of phenytoin (C15H12N2O2), add 30 mL of a mixture of of phenytoin is about 5 minutes. water and acetonitrile (1:1), treat with ultrasound waves for System suitability— 15 minutes with occasional shaking, shake for another 10 System performance: When the procedure is run with 10 minutes, and add a mixture of water and acetonitrile (1:1) to mL of the standard solution under the above operating con- make exactly 50 mL. Centrifuge this solution, pipet 5 mL of ditions, phenytoin and the internal standard are eluted in the supernatant liquid, add exactly 5 mL of the internal this order with the resolution between these peaks being not standard solution, and use this solution as the sample solu- less than 8. tion. Separately, weigh accurately about 25 mg of phenytoin System repeatability: When the test is repeated 6 times for assay, previously dried at 1059C for 2 hours, and dis- with 10 mL of the standard solution under the above operat- solve in a mixture of water and acetonitrile (1:1) to make ex- ing conditions, the relative standard deviation of the ratio of actly 25 mL. Pipet 5 mL of this solution, add exactly 5 mL the peak area of phenytoin to that of the internal standard is of the internal standard solution, and use this solution as the not more than 1.0z. standard solution. Perform the test with 10 mLeachofthe sample solution and standard solution as directed under Liq- Containers and storage Containers—Well-closed contain- uid Chromatography <2.01> according to the following con- ers. ditions, and calculate the ratios, QT and QS, of the peak area of phenytoin to that of the internal standard.

Phenytoin Tablets Amount (mg) of phenytoin (C15H12N2O2) ＝ MS × QT/QS × 2 Diphenylhydantoin Tablets MS: Amount (mg) of phenytoin for assay フェニトイン錠 Internal standard solution—A solution of propyl parahydroxybenzoate in the mobile phase (1 in 25,000). Operating conditions— Phenytoin Tablets contain not less than 95.0z and Detector: An ultraviolet absorption photometer (wave- not more than 105.0z of the labeled amount of length: 258 nm). phenytoin (C H N O : 252.27). 15 12 2 2 Column: A stainless steel column 4.6 mm in inside diame- Method of preparation Prepare as directed under Tablets, ter and 15 cm in length, packed with octadecylsilanized silica with Phenytoin. gel for liquid chromatography (5 mminparticlediameter). Column temperature: A constant temperature of about Identification Weigh a portion of powdered Phenytoin 409C. Tablets, equivalent to about 0.3 g of Phenytoin according to Mobile phase: A mixture of methanol and 0.02 mol/L the labeled amount, transfer to a separator, and add 1 mL of phosphate buffer solution, pH 3.5 (11:9). dilute hydrochloric acid and 10 mL of water. Extract with Flow rate: Adjust the flow rate so that the retention time 100 mL of diethyl ether, then with four 25-mL potions of of phenytoin is about 5 minutes. diethyl ether. Combine the extracts, evaporate the diethyl System suitability— ether on a water bath, and dry the residue at 1059Cfor2 System performance: When the procedure is run with 10 hours. Proceed with the residue as directed in the Identifica- mL of the standard solution under the above operating con- tion under Phenytoin. ditions, phenytoin and the internal standard are eluted in Uniformity of dosage units <6.02> Perform the test accord- this order with the resolution between these peaks being not 1244 Phenytoin Sodium for Injection / Official Monographs JP XVI less than 8. Loss on drying <2.41> Not more than 2.5z (1 g, 1059C, System repeatability: When the test is repeated 6 times 4 hours). with 10 mL of the standard solution under the above operat- Assay Weigh accurately the content of not less than 10 ing conditions, the relative standard deviation of the ratio of preparations of Phenytoin Sodium for Injection, transfer the peak area of phenytoin to that of the internal standard is about 0.3 g of the content, previously dried and accurately not more than 1.0z. weighed, to a separator, dissolve in 50 mL of water, add 10 Containers and storage Containers—Well-closed contain- mL of dilute hydrochloric acid,andextractwith100mLof ers. diethyl ether, then with four 25-mL portions of diethyl ether. Combine the diethyl ether extracts, and evaporate on a water bath. Dry the residue at 1059C for 2 hours, and weigh it as Phenytoin Sodium for Injection the mass of phenytoin (C15H12N2O2: 252.27). Amount (mg) of phenytoin sodium (C H N NaO ) Diphenylhydantoin Sodium for Injection 15 11 2 2 ＝ amount (mg) of phenytoin (C15H12N2O2) × 1.087 注射用フェニトインナトリウム Containers and storage Containers—Hermetic containers.

Phytonadione Phytomenadione

Vitamin K1 C15H11N2NaO2: 274.25 Monosodium 5,5-diphenyl-4-oxoimidazolidin-2-olate フィトナジオン [630-93-3]

Phenytoin Sodium for Injection is a preparation for injection which is dissolved before use. When dried, it contains not less than 98.5z of phenytoin sodium (C15H11N2NaO2), and contains not less than 92.5z and not more than 107.5z of the C31H46O2:450.70 labeled amount of phenytoin sodium (C15H11N2NaO2). 2-Methyl-3-[(2E,7R,11R)-3,7,11,15-tetramethylhexadec- Method of preparation Prepare as directed under Injec- 2-en-1-yl]-1,4-naphthoquinone tions. [84-80-0]

Description Phenytoin Sodium for Injection occurs as Phytonadione contains not less than 97.0z and not white crystals or crystalline powder. It is odorless. more than 102.0z of C H O . It is soluble in water and in ethanol (95), and practically 31 46 2 insoluble in chloroform and in diethyl ether. Description Phytonadione is a clear yellow to orange-yel- The pH of a solution of Phenytoin Sodium for Injection low, viscous liquid. (1 in 20) is about 12. It is miscible with isooctane. It is hygroscopic. It is soluble in ethanol (99.5), and practically insoluble in A solution of Phenytoin Sodium for Injection absorbs car- water. bon dioxide gradually when exposed to air, and a crystalline It decomposes gradually and changes to a red-brown by precipitate of phenytoin is produced. light. 20 Specific gravity d 20: about 0.967 Identification (1) With the residue obtained in the Assay, proceed as directed in the Identification under Phenytoin. Identification (1) Determine the absorption spectrum of a (2) Ignite 0.5 g of Phenytoin Sodium for Injection, cool, solution of Phytonadione in isooctane (1 in 100,000) as di- and dissolve the residue in 10 mL of water: the solution rected under Ultraviolet-visible Spectrophotometry <2.24>, changes red litmus paper to blue, and responds to Qualita- and compare the spectrum with the Reference Spectrum 1: tive Tests <1.09> (1) for sodium salt. both spectra exhibit similar intensities of absorption at the same wavelengths. Separately, determine the absorption Purity (1) Clarity and color of solution—Dissolve 1.0 g spectrum of a solution of Phytonadione in isooctane (1 in of Phenytoin Sodium for Injection in 20 mL of freshly 10,000) as directed under Ultraviolet-visible Spectropho- boiled and cooled water in a glass-stoppered test tube: the tometry <2.24>, and compare the spectrum with the Refer- solution is clear and colorless. If any turbidity is produced, ence Spectrum 2: both spectra exhibit similar intensities of add 4.0 mL of 0.1 mol/L sodium hydroxide VS: the solution absorption at the same wavelengths. becomes clear and colorless. (2) Determine the infrared absorption spectrum of (2) Heavy metals <1.07>—Proceed with 1.0 g of Phytonadione as directed in the liquid film method under In- Phenytoin Sodium for Injection according to Method 2, and frared Spectrophotometry <2.25>, and compare the spectrum perform the test. Prepare the control solution with 2.0 mL of with the Reference Spectrum: both spectra exhibit similar in- Standard Lead Solution (not more than 20 ppm). tensities of absorption at the same wave numbers. JP XVI Official Monographs / Pilocarpine Hydrochloride 1245

20 Refractive index <2.45> nD : 1.525 – 1.529 MS: Amount (mg) of Phytonadione RS Purity (1) Ratio of absorbances—Determine the absor- Internal standard solution—A solution of cholesterol benzo- bances, A1, A2 and A3, of a solution of Phytonadione in ate in the mobile phase (1 in 400). isooctane (1 in 100,000) at 248.5 nm, 253.5 nm and 269.5 Operating conditions— nm, respectively, as directed under Ultraviolet-visible Spec- Detector: An ultraviolet absorption photometer (wave- trophotometry <2.24>:theratioA2/A1 is between 0.69 and length: 254 nm). 0.73, and the ratio A2/A3 is between 0.74 and 0.78. Deter- Column: A stainless steel column 4.6 mm in inside diame- mine the absorbances, A4 and A5, of a solution of ter and 25 cm in length, packed with porous silica gel for liq- Phytonadione in isooctane (1 in 10,000) at 284.5 nm and 326 uid chromatography (5 mm in particle diameter). nm, respectively: the ratio A4/A5 is between 0.28 and 0.34. Column temperature: A constant temperature of about (2) Heavy metals <1.07>—Carbonize 1.0 g of Phytona- 309C. dione by gentle heating. Cool, add 10 mL of a solution of Mobile phase: A mixture of hexane and n-amyl alcohol magnesium nitrate hexahydrate in ethanol (95) (1 in 10), and (4000 : 3). ignite the ethanol to burn. Cool, add 1 mL of sulfuric acid, Flow rate: Adjust the flow rate so that the retention time proceed according to Method 4, and perform the test. Pre- of the peak of E-isomer of phytonadione is about 25 pare the control solution with 2.0 mL of Standard Lead So- minutes. lution (not more than 20 ppm). System suitability— (3) Menadione—Dissolve 20 mg of Phytonadione in 0.5 System performance: When the procedure is run with 50 mL of a mixture of water and ethanol (95) (1:1), add 1 drop mL of the standard solution under the above operating con- of a solution of 3-methyl-1-phenyl-5-pyrazolone in ethanol ditions, the internal standard, Z-isomer and E-isomer are (95) (1 in 20) and 1 drop of ammonia solution (28), and al- eluted in this order with the resolution between the peaks of low to stand for 2 hours: no blue-purple color develops. Z-isomer and E-isomer being not less than 1.5. System repeatability: When the test is repeated 6 times Isomer ratio Conduct this procedure rapidly and without with 50 mL of the standard solution under the above operat- exposure to light. Dissolve 30 mg of Phytonadione in 50 mL ing conditions, the relative standard deviation of the ratio of of the mobile phase. To 4 mL of this solution add the mobile the total area of the peaks of Z-isomer and E-isomer to the phase to make 25 mL. To 10 mL of this solution add the peak area of the internal standard is not more than 1.0z. mobile phase to make 25 mL, and use this solution as the sample solution. Perform the test with 50 mLofthesample Containers and storage Containers—Tight containers. solution as directed under Liquid Chromatography <2.01> Storage—Light-resistant, at a cold place or in containers according to the following conditions, and determine the in which air has been displaced by Nitrogen. peak areas of Z-isomer and E-isomer, ATZ and ATE: ATZ/ (ATZ ＋ ATE) is between 0.05 and 0.18. Operating conditions— Pilocarpine Hydrochloride Proceed as directed in the operating conditions in the Assay. ピロカルピン塩酸塩 System suitability— System performance: When the procedure is run with 50 mL of the sample solution under the above operating conditions, Z-isomer and E-isomer are eluted in this order with the resolution between these peaks being not less than 1.5.

System repeatability: When the test is repeated 6 times C11H16N2O2.HCl: 244.72 with 50 mL of the sample solution under the above operating (3S,4R)-3-Ethyl-4-(1-methyl-1H-imidazol-5-ylmethyl)- conditions, the relative standard deviation of the total area 4,5-dihydrofuran-2(3H)-one monohydrochloride of the peaks of Z-isomer and E-isomer is not more than [54-71-7] 2.0z. Pilocarpine Hydrochloride, when dried, contains Assay Conduct this procedure rapidly and without expo- not less than 99.0z of C H N O .HCl. sure to light. Weigh accurately about 30 mg each of 11 16 2 2 Phytonadione and Phytonadione RS, and dissolve each in Description Pilocarpine Hydrochloride occurs as colorless the mobile phase to make exactly 50 mL. Pipet 4 mL each of crystals or white powder. It is odorless, and has a slightly these solutions, and add the mobile phase to make exactly 25 bitter taste. mL. To exactly 10 mL each of these solutions add exactly 7 It is very soluble in acetic acid (100), freely soluble in mL of the internal standard solution and the mobile phase to water, in methanol and in ethanol (95), soluble in acetic an- make 25 mL, and use these as the sample solution and the hydride, and practically insoluble in diethyl ether. standard solution, respectively. Perform the test with 50 mL The pH of a solution of Pilocarpine Hydrochloride (1 in each of the sample solution and standard solution as directed 10) is between 3.5 and 4.5. under Liquid Chromatography <2.01> according to the fol- It is hygroscopic. lowing conditions, and calculate the ratios, QT and QS,of It is affected by light. the total area of the peaks of Z-isomer and E-isomer to the Identification (1) Dissolve 0.1 g of Pilocarpine Hydro- peak area of the internal standard. chloride in 5 mL of water, add 1 drop of dilute nitric acid, 1

Amount (mg) of C31H46O2 ＝ MS × QT/QS mL of hydrogen peroxide TS, 1 mL of chloroform and 1 drop of a potassium dichromate solution (1 in 300), and 1246 Pimaricin / Official Monographs JP XVI shake the mixture vigorously: a violet color develops in the chloroform layer while no color or a light yellow color is Pimaricin produced in the aqueous layer. (2) To 1 mL of a solution of Pilocarpine Hydrochloride Natamycin (1 in 20) add 1 mL of dilute nitric acid and 2 to 3 drops of silver nitrate TS: a white precipitate or opalescence is pro- ピマリシン duced. Melting point <2.60> 200–2039C Purity (1) Sulfate—Dissolve 0.5 g of Pilocarpine Hydro- chloride in 20 mL of water, and use this solution as the sample solution. To 5.0 mL of the sample solution add 1 mL of dilute hydrochloric acid and 0.5 mL of barium chloride TS: no turbidity is produced. (2) Nitrate—To 2.0 mL of the sample solution obtained in (1) add 2 mL of iron (II) sulfate TS, and superimpose the mixture upon 4 mL of sulfuric acid: no dark brown color develops at the zone of contact. C33H47NO13: 665.73 (3) Related substances—Dissolve 0.3 g of Pilocarpine (1R*,3S*,5R*,7R*,8E,12R*,14E,16E,18E,20E,22R*, Hydrochloride in 10 mL of methanol, and use this solution 24S*,25R*,26S*)-22-(3-Amino-3,6-dideoxy-b-D- as the sample solution. Pipet 1 mL of the sample solution, mannopyranosyloxy)-1,3,26-trihydroxy-12-methyl-10-oxo- add methanol to make exactly 100 mL, and use this solution 6,11,28-trioxatricyclo[22.3.1.05,7]octacosa-8,14,16,18,20- as the standard solution. Perform the test with these solu- pentaene-25-carboxylic acid tions as directed under Thin-layer Chromatography <2.03>. [7681-93-8] Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Pimaricin is a polyene macrolide substance having Develop the plate with a mixture of chloroform, methanol antifungal activity produced by the growth of Strep- and ammonia TS (85:14:2) to a distance of about 13 cm, and tomyces natalensis. dry the plate at 1059C for 10 minutes. Cool, and spray It contains not less than 900 mg (potency) and not evenly bismuth potassium iodide TS on the plate: the spots more than 1020 mg (potency) per mg, calculated on the other than the principal spot from the sample solution are anhydrous basis. The potency of Pimaricin is ex- not more intense than the spot from the standard solution. pressedasmass(potency)ofpimaricin(C33H47NO13). (4) Readily carbonizable substances <1.15>—Take 0.25 g Description Pimaricinoccursaswhite to yellowish white of Pilocarpine Hydrochloride, and perform the test: the so- crystalline powder. lution has no more color than Matching Fluid B. It is slightly soluble in methanol and in acetic acid (100), Loss on drying <2.41> Not more than 3.0z (1 g, 1059C, and practically insoluble in water and in ethanol (99.5). 2 hours). Identification (1) To 3 mg of Pimaricin add 1 mL of hy- Residue on ignition <2.44> Not more than 0.5z (0.1 g). drochloric acid, and mix: a blue-purple color appears. (2) Dissolve 5 mg of Pimaricin in a solution of acetic Assay Weigh accurately about 0.5 g of Pilocarpine Hydro- acid (100) in methanol (1 in 100) to make 1000 mL. Deter- chloride, previously dried, dissolvein50mLofamixtureof mine the absorption spectrum of this solution as directed acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> under Ultraviolet-visible Spectrophotometry <2.24>,and with 0.1 mol/L perchloric acid VS (potentiometric titration). compare the spectrum with the Reference Spectrum or the Perform a blank determination, and make any necessary spectrum of a solution of Pimaricin RS prepared in the same correction. manner as the sample solution: both spectra exhibit similar Each mL of 0.1 mol/L perchloric acid VS intensities of absorption at the same wavelengths.

＝ 24.47 mg of C11H16N2O2.HCl 20 Optical rotation <2.49> [a]D : ＋243 – ＋2599(0.1 g, acetic Containers and storage Containers—Tight containers. acid (100), 25 mL, 100 mm). Storage—Light-resistant. Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Pimaricin according to Method 4, and perform the test. Pre- pare the control solution with 3.0 mL of Standard Lead So- lution (not more than 30 ppm). (2) Related substances—Dissolve 20 mg of Pimaricin in methanol to make 100 mL, and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the total area of the peaks other than pimaricin by the automatic integration method: not more than 4.0z. Operating conditions— Detector: An ultraviolet absorption photometer (wave- JP XVI Official Monographs / Pimozide 1247 length: 303 nm). Column: A stainless steel column 3.9 mm in inside diame- Pimozide ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mminparticlediameter). ピモジド Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.0 g of ammonium acetate in 1000 mL of a mixture of water, methanol and tetrahydrofuran (47:44:2). Flow rate: Adjust the flow rate so that the retention time of pimaricin is about 10 minutes. Time span of measurement: About 3 times as long as the retention time of pimaricin.

System suitability— C28H29F2N3O: 461.55 Test for required detectability: Measure exactly 1 mL of 1-{1-[4,4-Bis(4-fluorophenyl)butyl]piperidin-4-yl}- the sample solution, add methanol to make exactly 100 mL, 1,3-dihydro-2H-benzoimidazol-2-one and use this solution as the solution for system suitability [2062-78-4] test. Pipet 1 mL of the solution for system suitability test, and add methanol to make exactly 10 mL. Confirm that the Pimozide contains not less than 98.5z and not peak area of pimaricin obtained from 10 mL of this solution more than 101.0z of C28H29F2N3O. is equivalent to 7 to 13z of that from 10 mL of the solution Description Pimozide occurs as a white to pale yellowish for system suitability test. white powder. System performance: When the procedure is run with 10 It is freely soluble in acetic acid (100), slightly soluble in mL of the solution for system suitability test under the above methanol and in ethanol (99.5), and practically insoluble in operating conditions, the number of theoretical plates and water. the symmetry factor of the peak of pimaricin are not less than 1500 and not more than 2.0, respectively. Identification (1) Determine the absorption spectrum of a System repeatability: When the test is repeated 6 times solution of Pimozide in methanol (1 in 25,000) as directed with 10 mL of the solution for system suitability test under under Ultraviolet-visible Spectrophotometry <2.24>,and the above operating conditions, the relative standard devia- compare the spectrum with the Reference Spectrum: both tion of the peak area of pimaricin is not more than 2.0z. spectra exhibit similar intensities of absorption at the same wavelengths. Water <2.48> Between 6.0z and 9.0z (0.2 g, volumetric (2) Determine the infrared absorption spectrum of titration, direct titration). Pimozide as directed in the potassium bromide disk method Assay Weigh accurately an amount of Pimaricin and under Infrared Spectrophotometry <2.25>, and compare the Pimaricin RS, equivalent to about 25 mg (potency), and dis- spectrum with the Reference Spectrum: both spectra exhibit solve each in methanol to make exactly 100 mL. Pipet 2 mL similar intensities of absorption at the same wave numbers. each of these solutions, add a solution of acetic acid (100) in Melting point <2.60> 216 – 2209C methanol (1 in 100) to make exactly 100 mL, and use these solutions as the sample solution and standard solution. De- Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of termine the absorbances at 295.5 nm, AT1 and AS1, at 303 Pimozide according to Method 2, and perform the test. Pre- nm, AT2 and AS2, and at 311 nm, AT3 and AS3,ofthesample pare the control solution with 2.0 mL of Standard Lead So- solution and standard solution as directed under Ultraviolet- lution by using 5 mL of sulfuric acid (not more than 10 visible Spectrophotometry <2.24>. ppm). (2) Arsenic <1.11>—Prepare the test solution with 1.0 g Amount [mg (potency)] of C H NO 33 47 13 of Pimozide according to Method 3, and perform the test AT1 ＋ AT3 (not more than 2 ppm). AT2 － 2 (3) Related substances—Dissolve 0.10 g of Pimozide in ＝ MS × × 1000 A ＋ A 10 mL of methanol, and use this solution as the sample solu- A － S1 S3 S2 2 tion. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard M : Amount [mg (potency)] of Pimaricin RS S solution. Perform the test with exactly 10 mLeachofthe Containers and storage Containers—Tight containers. sample solution and standard solution as directed under Liq- Storage—Light resistant. uid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of the peak other than the peak of pimozide from the sample solution is not lager than the peak area of pimozide from the standard solution, and the total area of the peaks other than the peak of pimozide from the sample solution is not larger than 1.5 times of the peak area of pimozide from the standard solution. Operating conditions— 1248 Pindolol / Official Monographs JP XVI

Detector: An ultraviolet absorption photometer (wavelength: 280 nm). Pindolol Column: A stainless steel column 4.6 mm in inside diameter and 10 cm in length, packed with octadecylsilanized silica ピンドロール gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: Dissolve 2.5 g of ammonium acetate and 8.5 g of tetrabutylammonium hydrogensulfate in water to make 1000 mL. C H N O : 248.32 Mobile phase B: Acetonitrile. 14 20 2 2 (2RS)-1-(1H-Indol-4-yloxy)- Flowing of the mobile phase: Control the gradient by mix- 3-(1-methylethyl)aminopropan-2-ol ing the mobile phases A and B as directed in the following [13523-86-9] table. Pindolol, when dried, contains not less than 98.5z Time after injection Mobile phase A Mobile phase B of C14H20N2O2. of sample (min) (volz) (volz) Description Pindolol occurs as a white, crystalline powder. 0–10 80→ 70 20 → 30 It has a slight, characteristic odor. 10–15 70 30 It is sparingly soluble in methanol, slightly soluble in ethanol (95), and practically insoluble in water and in diethyl Flow rate: 2.0 mL per minute. ether. Time span of measurement: 1.5 times as long as the reten- It dissolves in dilute sulfuric acid and in acetic acid (100). tion time of pimozide. Identification (1) To 1 mL of a solution of Pindolol in System suitability— methanol (1 in 10,000) add 1 mL of a solution of 1-(4- Test for required detectability: Pipet 1 mL of the standard pyridyl)-pyridinium chloride hydrochloride (1 in 1000) and 1 solution, and add methanol to make exactly 10 mL. Confirm mL of sodium hydroxide TS, then add 1 mL of hydrochloric that the peak area of pimozide obtained from 10 mLofthis acid: a blue to blue-purple color, changing to red-purple, is solution is equivalent to 8 to 12z of that of pimozide from produced. 10 mL of the standard solution. (2) Dissolve 0.05 g of Pindolol in 1 mL of dilute sulfuric System performance: Dissolve 5 mg of Pimozide and 2 mg acid, and add 1 mL of Reinecke salt TS: a light red precipi- of mebendazole in methanol to make 100 mL. When the tate is produced. procedure is run with 10 mL of this solution under the above (3) Determine the absorption spectrum of a solution of operating conditions, mebendazole and pimozide are eluted Pindolol in methanol (1 in 50,000) as directed under Ultra- in this order with the resolution between these peaks being violet-visible Spectrophotometry <2.24>, and compare the not less than 5. spectrum with the Reference Spectrum: both spectra exhibit System repeatability: When the test is repeated 6 times similar intensities of absorption at the same wavelengths. with 10 mL of the standard solution under the above operat- (4) Determine the infrared absorption spectrum of Pin- ing conditions, the relative standard deviation of the peak dolol, previously dried, as directed in the potassium bromide area of pimozide is not more than 2.0z. disk method under Infrared Spectrophotometry <2.25>,and (4) Residual solvent—Being specified separately. compare the spectrum with the Reference Spectrum: both Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, spectra exhibit similar intensities of absorption at the same 3 hours). wave numbers. 1z Residue on ignition <2.44> Not more than 0.1z (1 g). Absorbance <2.24> E1cm (264 nm): 333 – 350 (10 mg, methanol, 500 mL). Assay Weigh accurately about 70 mg of Pimozide, previously dried, dissolve in 25 mL of acetic acid for nonaqueous Melting point <2.60> 169 – 1739C titration, and titrate <2.50> with 0.02 mol/L perchloric acid Purity (1) Clarity and color of solution—Dissolve 0.5 g VS (indicator: 2 drops of crystal violet TS). Perform a blank of Pindolol in 10 mL of acetic acid (100), and observe imme- determination in the same manner, and make any necessary diately: the solution is clear, and has no more color than the correction. following control solution. Each mL of 0.02 mol/L perchloric acid VS Control solution: Measure accurately 4 mL of Matching Fluid A, add exactly 6 mL of water, and mix. ＝ 9.231 mg of C28H29F2N3O (2) Heavy metals <1.07>—Proceed with 1.0 g of Pindolol Containers and storage Containers—Well-closed contain- according to Method 2, and perform the test. Prepare the ers. control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (3) Arsenic <1.11>—Prepare the test solution with 1.0 g of Pindolol according to Method 3, and perform the test (not more than 2 ppm). (4) Related substances—Dissolve 0.10 g of Pindolol in 10 mL of methanol, and use this solution as the sample solu- JP XVI Official Monographs / Pioglitazone Hydrochloride 1249 tion. Pipet 2 mL of the sample solution, and add methanol as the sample solution: both spectra exhibit similar intensi- to make exactly 100 mL. Pipet 5 mL of this solution, add ties of absorption at the same wavelengths. methanol to make exactly 20 mL, and use this solution as the (2) Determine the infrared absorption spectrum of standard solution. Perform the test with these solutions as Pioglitazone Hydrochloride as directed in the potassium bro- directed under Thin-layer Chromatography <2.03>. Spot 5 mide disk method under Infrared Spectrophotometry <2.25>, mL each of the sample solution and standard solution on a and compare the spectrum with the Reference Spectrum or plate of silica gel for thin-layer chromatography. Develop the spectrum of Pioglitazone Hydrochloride RS: both spec- the plate with a mixture of chloroform, acetone and tra exhibit similar intensities of absorption at the same wave isopropylamine (5:4:1) to a distance of about 12 cm, and air- numbers. dry the plate. Spray evenly diluted sulfuric acid (3 in 5) and a (3) Dissolve 50 mg of Pioglitazone Hydrochloride in 1 sodium nitrite solution (1 in 50) on the plate: the spots other mL of nitric acid, and add 4 mL of dilute nitric acid: the so- than the principal spot from the sample solution are not lution responds to the Qualitative Tests <1.09> (2) for chlo- more intense than the spot from the standard solution. ride. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of 4 hours). Pioglitazone Hydrochloride according to Method 4, and perform the test. After incineration, use 3 mL of hydrobromic Residue on ignition <2.44> Not more than 0.1z (1 g). acid instead of 3 mL of hydrochloric acid. Prepare the con- Assay Weigh accurately about 0.5 g of Pindolol, previ- trol solution with 1.0 mL of Standard Lead Solution (not ously dried, dissolve in 80 mL of methanol, and titrate more than 10 ppm). <2.50> with 0.1 mol/L hydrochloric acid VS (potentiometric (2) Related substances—Dissolve 20 mg of Pioglitazone titration). Perform a blank determination, and make any Hydrochloride in 20 mL of methanol, add the mobile phase necessary correction. to make 100 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile Each mL of 0.1 mol/L hydrochloric acid VS phase to make exactly 200 mL, and use this solution as the ＝ 24.83 mg of C H N O 14 20 2 2 standard solution. Perform the test with exactly 40 mLeach Containers and storage Containers—Tight containers. of the sample solution and standard solution as directed Storage—Light-resistant. under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of the Pioglitazone Hydrochloride peaks, having the relative retention times of about 0.7, about 1.4 and about 3.0 with respect to pioglitazone from the sam- ピオグリタゾン塩酸塩 ple solution, is not larger than 2/5 times the peak area of pioglitazone from the standard solution, and the area of each peak other than the peak of pioglitazone and other than those mentioned above is smaller than 1/5 times the peak area of pioglitazone from the standard solution. Further- more, the total area of the peaks other than the peak of pioglitazone is not larger than the peak area of pioglitazone

C19H20N2O3S.HCl: 392.90 from the standard solution. (5RS)-5-{4-[2-(5-Ethylpyridin- Operating conditions— 2-yl)ethoxy]benzyl}thiazolidine-2,4-dione Detector, column, column temperature, mobile phase and monohydrochloride flow rate: Proceed as directed in the operating conditions in [112529-15-4] the Assay. Time span of measurement: About 4 times as long as the Pioglitazone Hydrochloride contains not less than retention time of pioglitazone, beginning after the solvent 99.0z and not more than 101.0z of C19H20N2O3S. peak. HCl, calculated on the anhydrous basis. System suitability— Test for required detectability: Pipet 1 mL of the standard Description Pioglitazone Hydrochloride occurs as white solution, and add the mobile phase to make exactly 10 mL. crystals or crystalline powder. Confirm that the peak area of pioglitazone obtained from 40 It is soluble in N,N-dimethylformamide and in methanol, mL of this solution is equivalent to 7 to 13z of that of slightly soluble in ethanol (99.5), and practically insoluble in pioglitazone from 40 mL of the standard solution. water. System performance: Dissolve 50 mg of Pioglitazone Hy- It dissolves in 0.1 mol/L hydrochloric acid TS. drochloride in 10 mL of a solution of benzophenone in A solution of Pioglitazone Hydrochloride in N,N- methanol (1 in 750), and add methanol to make 100 mL. To dimethylformamide (1 in 20) shows no optical rotation. 1 mL of this solution add the mobile phase to make 20 mL. Identification (1) Determine the absorption spectrum of a When the procedure is run with 40 mL of this solution under solution of Pioglitazone Hydrochloride in 0.1 mol/L hydro- the above operating conditions, pioglitazone and benzophe- chloric acid TS (1 in 50,000) as directed under Ultraviolet- none are eluted in this order with the resolution between visible Spectrophotometry <2.24>, and compare the spectrum these peaks being not less than 10. with the Reference Spectrum or the spectrum of a solution of System repeatability: When the test is repeated 6 times Pioglitazone Hydrochloride RS prepared in the same manner with 40 mL of the standard solution under the above operat- 1250 Pioglitazone Hydrochloride Tablets / Official Monographs JP XVI ing conditions, the relative standard deviation of the peak area of pioglitazone is not more than 2.0z. Pioglitazone Hydrochloride Tablets (3) Residual solvent—Being specified separately. ピオグリタゾン塩酸塩錠 Water <2.48> Not more than 0.2z (0.5 g, coulometric titration). For anolyte solution, use anolyte solution for water determination A. Pioglitazone Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled Residue on ignition <2.44> Not more than 0.1z (1 g). amount of pioglitazone hydrochloride (C19H20N2O3S. Assay Weigh accurately about 50 mg each of Pioglitazone HCl: 392.90). Hydrochloride and Pioglitazone Hydrochloride RS (sepa- Method of preparation Prepare as directed under Tablets, rately, determine the water <2.48> inthesamemanneras with Pioglitazone Hydrochloride. Pioglitazone Hydrochloride), add exactly 10 mL of the internal standard solution and methanol to make 100 mL. Pipet 2 Identification To an amount of powdered Pioglitazone Hy- mL each of these solutions, add the mobile phase to make 20 drochloride Tablets, equivalent to 2.8 mg of Pioglitazone mL, and use these solutions as the sample solution and the Hydrochloride according to the labeled amount, add 100 mL standard solution, respectively. Perform the test with 20 mL of 0.1 mol/L hydrochloric acid TS, shake, and filter through each of the sample solution and standard solution as directed a membrane filter with a pore size not exceeding 0.45 mm. under Liquid Chromatography <2.01> according to the fol- Determine the absorption spectrum of the filtrate as directed lowing conditions, and calculate the ratios, QT and QS,of under Ultraviolet-visible Spectrophotometry <2.24>:itexhib- the peak area of pioglitazone to that of the internal stand- its a maximum between 267 nm and 271 nm. ard. Uniformity of dosage units <6.02> Perform the test accord- Amount (mg) of pioglitazone hydrochloride ing to the following method: it meets the requirement of the

(C19H20N2O3S.HCl) Content uniformity test. ＝ MS × QT/QS Disintegrate 1 tablet of Pioglitazone Hydrochloride Tablets with 10 mL of 0.1 mol/L hydrochloric acid TS, add M : Amount (mg) of Pioglitazone Hydrochloride RS, S 70 mL of methanol, shake vigorously for 10 minutes, then calculated on the anhydrous basis add methanol to make exactly 100 mL, and centrifuge. Take Internal standard solution—A solution of benzophenone in exactly V mL of the supernatant liquid, add a mixture of methanol (1 in 750). methanol and 0.1 mol/L hydrochloric acid TS (9:1) to make Operating conditions— exactly V?mL so that each mL contains about 26 mgof

Detector: An ultraviolet absorption photometer (wave- pioglitazone hydrochloride (C19H20N2O3S.HCl), and use this length: 269 nm). solution as the sample solution. Separately, weigh accurately Column: A stainless steel column 4.6 mm in inside diame- about 33 mg of Pioglitazone Hydrochloride RS (separately, ter and 15 cm in length, packed with octadecylsilanized silica determine the water <2.48> in the same manner as Pioglita- gel for liquid chromatography (5 mm in particle diameter). zone Hydrochloride), dissolve in 10 mL of 0.1 mol/L hydro- Column temperature: A constant temperature of about chloric acid TS, and add methanol to make exactly 100 mL. 259C. Pipet 4 mL of this solution, add a mixture of methanol and Mobile phase: A mixture of ammonium acetate solution 0.1 mol/L hydrochloric acid TS (9:1) to make exactly 50 mL, (77 in 10,000), acetonitrile and acetic acid (100) (25:25:1). and use this solution as the standard solution. Determine the

Flow rate: Adjust the flow rate so that the retention time absorbances, AT and AS, of the sample solution and stand- of pioglitazone is about 7 minutes. ard solution at 269 nm as directed under Ultraviolet-visible System suitability— Spectrophotometry <2.24> using a mixture of methanol and System performance: When the procedure is run with 20 0.1 mol/L hydrochloric acid TS (9:1) as the blank. mL of the standard solution under the above operating con- Amount (mg) of pioglitazone hydrochloride ditions, pioglitazone and the internal standard are eluted in (C H N O S.HCl) this order with the resolution between these peaks being not 19 20 2 3 ＝ M × A /A × V?/V × 2/25 less than 10. S T S

System repeatability: When the test is repeated 6 times MS: Amount (mg) of Pioglitazone Hydrochloride RS, with 20 mL of the standard solution under the above operat- calculated on the anhydrous basis ing conditions, the relative standard deviation of the peak Dissolution <6.10> When the test is performed at 50 revolu- area of pioglitazone is not more than 1.0z. tions per minute according to the Paddle method, using 900 Containers and storage Containers—Well-closed contain- mL of a solution, which is prepared by mixing 50 mL of 0.2 ers. mol/L hydrochloric acid TS and 150 mL of potassium chloride solution (3 in 20), adding water to make 1000 mL and adjusting to pH 2.0 with 5 mol/L hydrochloric acid TS, as the dissolution medium, the dissolution rate in 45 minutes of Pioglitazone Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Pioglitazone Hydrochloride Tablets, withdraw 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the JP XVI Official Monographs / Pipemidic Acid Hydrate 1251 first 5 mL of the filtrate, pipet V mL of the subsequent fil- of pioglitazone is about 7 minutes. trate, add the dissolution medium to make exactly V?mL so System suitability— that each mL contains about 18 mg of pioglitazone hydro- System performance: When the procedure is run with 20 chloride (C19H20N2O3S.HCl) according to the labeled mL of the standard solution under the above operating con- amount, and use this solution as the sample solution. Sepa- ditions, pioglitazone and the internal standard are eluted in rately, weigh accurately about 23 mg of Pioglitazone Hydro- this order with the resolution between these peaks being not chloride RS (separately determine the water <2.48> in the less than 10. same manner as Pioglitazone Hydrochloride), dissolve in 10 System repeatability: When the test is repeated 6 times mL of methanol, and add the dissolution medium to make with 20 mL of the standard solution under the above operat- exactly 50 mL. Pipet 2 mL of this solution, add the dissoluing conditions, the relative standard deviation of the ratio of tion medium to make exactly 50 mL, and use this solution as the peak area of pioglitazone to that of the internal standard the standard solution. Determine the absorbances, AT and is not more than 1.0z. A , of the sample solution and standard solution at 269 nm S Containers and storage Containers—Tight containers. as directed under Ultraviolet-visible Spectrophotometry <2.24> using the dissolution medium as the blank. Dissolution rate (z) with respect to the labeled amount Pipemidic Acid Hydrate of pioglitazone hydrochloride (C19H20N2O3S.HCl) ＝ MS × AT/AS × V?/V × 1/C × 72 ピペミド酸水和物

MS: Amount (mg) of Pioglitazone Hydrochloride RS, calculated on the anhydrous basis C Labeled amount (mg) of pioglitazone hydrochloride

(C19H20N2O3S.HCl) in 1 tablet Assay Accurately weigh the mass of not less than 20 Pioglitazone Hydrochloride Tablets, and powder. Weigh ac- C H N O .3H O: 357.36 curately a portion of the powder, equivalent to about 25 mg 14 17 5 3 2 8-Ethyl-5-oxo-2-(piperazin-1-yl)- of pioglitazone hydrochloride (C H N O S.HCl), add 45 19 20 2 3 5,8-dihydropyrido[2,3-d]pyrimidine- mL of methanol and exactly 5 mL of the internal standard 6-carboxylic acid trihydrate solution, agitate with the aid of ultrasonic waves, and centri- [51940-44-4, anhydride] fuge. To 2 mL of the supernatant liquid add the mobile phase to make 20 mL, and use this solution as the sample so- Pipemidic Acid Hydrate contains not less than lution. Separately, weigh accurately about 25 mg of Pioglita- 98.5z and not more than 101.0z of pipemidic acid zone Hydrochloride RS (separately, determine the water (C H N O : 303.32), calculated on the anhydrous <2.48> in the same manner as Pioglitazone Hydrochloride), 14 17 5 3 basis. dissolve in 45 mL of methanol, and add exactly 5 mL of the internal standard solution. Pipet 2 mL of this solution, add Description Pipemidic Acid Hydrate occurs as a pale yel- the mobile phase to make 20 mL, and use this solution as the low, crystalline powder. standard solution. Perform the test with exactly 20 mLeach It is freely soluble in acetic acid (100), very slightly soluble of the sample solution and standard solution as directed in water and in ethanol (99.5), and practically insoluble in under Liquid Chromatography <2.01> according to the fol- methanol. lowing conditions, and calculate the ratios, QT and QS,of It dissolves in sodium hydroxide TS. the peak area of pioglitazone to that of the internal stand- It is gradually colored on exposure to light. ard. Melting point: about 2509C (with decomposition). Amount (mg) of pioglitazone hydrochloride Identification (1) Dissolve 0.1 g of Pipemidic Acid Hy-

(C19H20N2O3S.HCl) drate in 20 mL of sodium hydroxide TS, and dilute with ＝ MS × QT/QS water to make 200 mL. To 1 mL of the solution add water to make 100 mL. Determine the absorption spectrum of the so- M : Amount (mg) of Pioglitazone Hydrochloride RS, S lution as directed under Ultraviolet-visible Spectrophotome- calculated on the anhydrous basis try <2.24>, and compare the spectrum with the Reference Internal standard solution—A solution of benzophenone in Spectrum: both spectra exhibit similar intensities of absorp- methanol (1 in 750). tion at the same wavelengths. Operating conditions— (2) Determine the infrared absorption spectrum of Detector: An ultraviolet absorption photometer (wave- Pipemidic Acid Hydrate as directed in the potassium bro- length: 269 nm). mide disk method under Infrared Spectrophotometry <2.25>, Column: A stainless steel column 4.6 mm in inside diame- and compare the spectrum with the Reference Spectrum: ter and 15 cm in length, packed with octadecylsilanized silica both spectra exhibit similar intensities of absorption at the gel for liquid chromatography (5 mm in particle diameter). same wave numbers. Column temperature: A constant temperature of about Purity (1) Chloride <1.03>—Dissolve 1.0 g of Pipemidic 259C. Acid Hydrate in 35 mL of water and 10 mL of sodium hy- Mobile phase: A mixture of ammonium acetate solution droxide TS, then add 15 mL of dilute nitric acid, shake well, (77 in 10,000), acetonitrile and acetic acid (100) (25:25:1). and filter through a glass filter (G3). To 30 mL of the filtrate Flow rate: Adjust the flow rate so that the retention time 1252 Piperacillin Hydrate / Official Monographs JP XVI add 6 mL of dilute nitric acid and water to make 50 mL. Per- form the test using this solution as the test solution. Prepare Piperacillin Hydrate the control solution as follows: to 0.30 mL of 0.01 mol/L hydrochloric acid VS add 5 mL of sodium hydroxide TS, ピペラシリン水和物 13.5 mL of dilute nitric acid and water to make 50 mL (not more than 0.021z). (2) Sulfate <1.14>—Dissolve 1.0 g of Pipemidic Acid Hy- drate in 35 mL of water and 10 mL of sodium hydroxide TS, then add 15 mL of dilute hydrochloric acid, shake well, and filter through a glass filter (G3). To 30 mL of the filtrate add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to C H N O S.H O: 535.57 0.50 mL of 0.005 mol/L sulfuric acid VS add 5 mL of sodi- 23 27 5 7 2 (2S,5R,6R)-6-{(2R)-2-[(4-Ethyl-2,3-dioxopiperazine- um hydroxide TS, 7.5 mL of dilute hydrochloric acid and 1-carbonyl)amino]-2-phenylacetylamino}-3,3-dimethyl- water to make 50 mL (not more than 0.048z). 7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid (3) Heavy metals <1.07>—Proceed with 2.0 g of Pipe- monohydrate midic Acid Hydrate according to Method 2, and perform the [66258-76-2] test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Piperacillin Hydrate contains not less than 970 mg (4) Arsenic <1.11>—Prepare the test solution with 1.0 g (potency) and not more than 1020 mg (potency) per of Pipemidic Acid Hydrate according to Method 3, and per- mg, calculated on the anhydrous basis. The potency of form the test (not more than 2 ppm). Piperacillin Hydrate is expressed as mass (potency) of (5) Related substances—Dissolve 0.10 g of Pipemidic piperacillin (C H N O S: 517.55). Acid Hydrate in 10 mL of diluted acetic acid (100) (1 in 20), 23 27 5 7 and use this solution as the sample solution. Pipet 1 mL of Description Piperacillin Hydrate occurs as a white crystal- the sample solution, add diluted acetic acid (100) (1 in 20) to line powder. make exactly 200 mL, and use this solution as the standard It is freely soluble in methanol, soluble in ethanol (99.5) solution. Perform the test with these solutions as directed and in dimethylsulfoxide, and very slightly soluble in water. under Thin-layer Chromatography <2.03>. Spot 5 mLeachof Identification (1) Determine the infrared absorption spec- the sample solution and standard solution on a plate of silica trum of Piperacillin Hydrate as directed in the potassium gel with fluorescent indicator for thin-layer chromatogra- bromide disk method under Infrared Spectrophotometry phy. Develop the plate with a mixture of chloroform, metha- <2.25>, and compare the spectrum with the Reference Spec- nol, formic acid and triethylamine (25:15:5:1) to a distance trum or the spectrum of Piperacillin RS: both spectra exhibit of about 10 cm, and air-dry the plate. Examine under ultra- similar intensities of absorption at the same wave numbers. violet light (main wavelength: 254 nm): the spots other than (2) Determine the 1H spectrum of a solution of Piperacil- the principal spot from the sample solution are not more lin Hydrate in deuterated dimethylsulfoxide for nuclear mag- intense than the spot from the standard solution. netic resonance spectroscopy (1 in 3) as directed under Water <2.48> 14.5 – 16.0z (20 mg, coulometric titration). Nuclear Magnetic Resonance Spectroscopy <2.21>, using tet- ramethylsilane for nuclear magnetic resonance spectroscopy Residue on ignition <2.44> Not more than 0.1z (1 g). as an internal reference compound: it exhibits a triple signal Assay Weigh accurately about 0.35 g of Pipemidic Acid A at about d 1.1 ppm, a single signal B at about d 4.2 ppm, Hydrate, dissolve in 40 mL of acetic acid (100), and titrate and a multiple signal C at about d 7.4 ppm, and the ratio of <2.50> with 0.1 mol/L perchloric acid VS (potentiometric the integrated intensity of each signal, A:B:C, is about 3:1:5. titration). Perform a blank determination in the same man- Optical rotation <2.49> [a]20: ＋162 – ＋1729(0.2 g, metha- ner, and make any necessary correction. D nol,20mL,100mm). Each mL of 0.1 mol/L perchloric acid VS Purity (1) Heavy metal <1.07>—Proceed with 2.0 g of ＝ 30.33 mg of C H N O 14 17 5 3 Piperacillin Hydrate according to Method 2, and perform Containers and storage Containers—Well-closed contain- the test. Prepare the control solution with 2.0 mL of Stand- ers. ard Lead Solution (not more than 10 ppm). Storage—Light-resistant. (2) Related substances 1—Conduct this procedure rapidly after the preparation of the sample solution and standard solution. Dissolve 20 mg of Piperacillin Hydrate in 20 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution (1). Pipet 2 mL of the standard solution (1), add the mobile phase to make exactly 10 mL, and use this solution as the standard solution (2). Perform the test with exactly 20 mL each of the sample solution and the standard solutions (1) and (2) as directed under Liquid Chroma- tography <2.01> according to the following conditions, and JP XVI Official Monographs / Piperacillin Hydrate 1253 determine each peak area by the automatic integration about 6.6 with respect to piperacillin, after multiplying by method: the total area of the peaks, having the relative reten- the relative response factor, 2.0. tion time of about 0.38 and about 0.50 with respect to piper- Operating conditions— acillin, obtained from the sample solution is not larger than Detector, column and column temperature: Proceed as di- 2 times the peak area of piperacillin from the standard solu- rected in the operating conditions in the Assay. tion (2), the total area of the peaks, having the relative reten- Mobile phase: Take 60.1 g of acetic acid (100) and 101.0 g tion time of about 0.82 and about 0.86 with respect to piper- of triethylamine, add water to make 1000 mL. To 25 mL of acillin, obtained from the sample solution is not larger than this solution add 300 mL of acetonitrile and 25 mL of dilute the peak area of piperacillin from the standard solution (2), acetic acid, and add water to make 1000 mL. and the area of the peak other than piperacillin and other Flow rate: Adjust the flow rate so that the retention time than the peaks having the relative retention time of about of piperacillin is about 1.2 minutes. 0.38, about 0.50, about 0.82 and about 0.86 with respect to Time span of measurement: About 8 times as long as the piperacillin, obtained from the sample solution, is not larger retention time of piperacillin, beginning after the piperacillin than the peak area of piperacillin from the standard solution peak. (2). Furthermore, the total area of the peaks other than System suitability— piperacillin obtained from the sample solution is not larger Test for required detectability: Confirm that the peak area than the peak area of piperacillin from the standard solution of piperacillin obtained from 20 mL of the standard solution (1). (2) is equivalent to 15 to 25z of that from 20 mLofthe Operating conditions— standard solution (1). Detector, column, column temperature, mobile phase, and System performance: When the procedure is run with 20 flow rate: Proceed as directed in the operating conditions in mL of the standard solution (1) under the above operating the Assay. conditions, the number of theoretical plates and the symme- Time span of measurement: About 3 times as long as the try factor of the peak of piperacillin are not less than 1500 retention time of piperacillin, beginning after the solvent and not more than 2.0, respectively. peak. System repeatability: When the test is repeated 6 times System suitability— with 20 mL of the standard solution (2) under the above op- Test for required detectability: Confirm that the peak area erating conditions, the relative standard deviation of the of piperacillin obtained from 20 mL of the standard solution peak area of piperacillin is not more than 4.0z. (2) is equivalent to 15 to 25z of that from 20 mLofthe (4) Residual solvents <2.46>—Transferexactly10mgof standard solution (1). Piperacillin Hydrate to an about 3 mL-vial, add exactly 1 System performance: When the procedure is run with 20 mL of saturated sodium hydrogen carbonate solution to dis- mL of the standard solution (1) under the above operating solve and stop the vial tightly. After heating this at 909Cfor conditions, the number of theoretical plates and the symme- 10 minutes, use the gas inside the container as the sample try factor of the peak of piperacillin are not less than 3000 gas. Separately, measure exactly 1 mL of ethyl acetate, dis- and not more than 1.5, respectively. solve in water to make exactly 200 mL. Pipet 10 mL of this System repeatability: When the test is repeated 6 times solution, add water to make exactly 20 mL. Pipet 2 mLof with 20 mL of the standard solution (2) under the above op- this solution in an about 3-mL vial containing exactly 1 mL erating conditions, the relative standard deviation of the of saturated sodium hydrogen carbonate solution, and stop peak area of piperacillin is not more than 3.0z. the vial tightly. Run the procedure similarly to the sample, (3) Related substances 2—Dissolve 20 mg of Piperacillin and use the gas as the standard gas. Perform the test with ex- Hydrate in 20 mL of the mobile phase, and use this solution actly 0.5 mL each of the sample gas and standard gas as di- as the sample solution. Pipet 1 mL of the sample solution, rected under Gas Chromatography <2.02> according to the add the mobile phase to make exactly 200 mL, and use this following conditions, and determine the peak area of ethyl solution as the standard solution (1). Pipet 2 mL of the acetate by the automatic integration method: the peak area standard solution (1), add the mobile phase to make exactly of ethyl acetate obtained from the sample gas is not larger 10 mL, and use this solution as the standard solution (2). than that from the standard gas. Perform the test with exactly 20 mLeachofthesamplesolu- Operating conditions— tion, and the standard solutions (1) and (2) as directed under Detector: A hydrogen flame-ionization detector. Liquid Chromatography <2.01> according to the following Column: A glass column 3 mm in inside diameter and 1 m conditions, and determine each peak area by the automatic in length, packed with porous stylene-divinyl benzene co- integration method: the area of the peak, having the relative polymer for gas chromatography (average pore diameter of retention time of about 6.6 with respect to piperacillin, ob- 0.0085 mm, 300 – 400 m2/g) with the particle size of 125 to tained from the sample solution is not larger than 3 times the 150 mm. peak area of piperacillin from the standard solution (2), and Column temperature: A constant temperature of about the area of the peaks other than the peak of piperacillin and 1459C. the peak having the relative retention time of about 6.6 with Carrier gas: Nitrogen. respect to piperacillin from the sample solution are not Flow rate: Adjust the flow rate so that the retention time larger than 1.4 times the peak area of piperacillin from the of ethyl acetate is about 4 minutes. standard solution (2). Furthermore, the total area of the System suitability— peaks other than the peak of piperacillin from the sample so- System performance: Take 1 mL of saturated sodium lution is not larger than the area of the peak of piperacillin hydrogen carbonate solution in an about 3 mL-vial, add 2 from the standard solution (1). For these calculations, use mL each of ethyl acetate solution (1 in 400) and acetone solu- the area of the peak, having the relative retention time of tion (1 in 400), and stop the vial tightly. When the procedure 1254 Piperacillin Sodium / Official Monographs JP XVI is run under the above operating conditions, acetone and ethyl acetate are eluted in this order with the resolution be- Piperacillin Sodium tween these peaks being not less than 2.0. System repeatability: Take 1 mL of saturated sodium ピペラシリンナトリウム hydrogen carbonate solution in an about 3 mL-vial, add 2 mL of ethyl acetate solution (1 in 400), stop the vial tightly, and perform the test under the above operating conditions. When the procedure is repeated 6 times, the relative standard deviation of the peak area of ethyl acetate is not more than 10z. Water <2.48> Not less than 3.2z and not more than 3.8z

(0.5 g, volumetric titration, direct titration). C23H26N5NaO7S: 539.54 Monosodium (2S,5R,6R)-6-{(2R)-2-[(4-ethyl-2,3- Residue on ignition <2.44> Not more than 0.1z (1 g). dioxopiperazine-1-carbonyl)amino]-2-phenylacetylamino}- Bacterial endotoxins <4.01> Less than 0.07 EU/mg (po- 3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2- tency). carboxylate [59703-84-3] Assay Weigh accurately an amount of Piperacillin Hydrate and Piperacillin RS, equivalent to about 50 mg (potency), Piperacillin Sodium contains not less than 863 mg dissolve each in the mobile phase to make exactly 50 mL. (potency) per mg, calculated on the anhydrous basis. Pipet 5 mL each of these solutions, add exactly 5 mL of the The potency of Piperacillin Sodium is expressed as internal standard solution, and use these solutions as the mass (potency) of piperacillin (C H N O S: 517.55). sample solution and the standard solution, respectively. Per- 23 27 5 7 form the test with 5 mL each of the sample solution and Description Piperacillin Sodium occurs as a white powder standard solution as directed under Liquid Chromatography or mass. <2.01> according to the following conditions, and calculate It is very soluble in water, freely soluble in methanol and the ratios, HT and HS, of the peak height of piperacillin to in ethanol (95), and practically insoluble in acetonitrile. that of the internal standard. Identification (1) Determine the infrared absorption spec-

Amount [mg (potency)] of piperacillin (C23H27N5O7S) trum of Piperacillin Sodium as directed in the potassium ＝ MS × HT/HS × 1000 bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- M : Amount [mg (potency)] of Piperacillin RS S trum: both spectra exhibit similar intensities of absorption at Internal standard solution—A solution of acetanilide in the the same wave numbers. mobile phase (1 in 5000). (2) Piperacillin Sodium responds to Qualitative Tests Operating conditions— <1.09> (1) for sodium salt. Detector: An ultraviolet absorption photometer (wave- Optical rotation <2.49> [a]20: ＋175 – ＋1909(0.8 g calcu- length: 254 nm). D lated on the anhydrous basis, water, 20 mL, 100 mm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel pH <2.54> Dissolve 1.0 g of Piperacillin Sodium in 4 mL of for liquid chromatography (5 mminparticlediameter). water: the pH of the solution is between 5.0 and 7.0. Column temperature: A constant temperature of about Purity (1) Clarity and color of solution—Dissolve 1.0 g 259C. of Piperacillin Sodium in 10 mL of water: the solution is Mobile phase: Take 60.1 g of acetic acid (100) and 101.0 g clear and colorless. of triethylamine, add water to make 1000 mL. To 25 mL of (2) Heavy metals <1.07>—Proceed with 2.0 g of Piper- this solution add 210 mL of acetonitrile and 25 mL of dilute acillin Sodium according to Method 4, and perform the test. acetic acid, and add water to make 1000 mL. Prepare the control solution with 2.0 mL of Standard Lead Flow rate: Adjust the flow rate so that the retention time Solution (not more than 10 ppm). of piperacillin is about 5 minutes. (3) Arsenic <1.11>—Prepare the test solution with 2.0 g System suitability— of Piperacillin Sodium according to Method 4, and perform System performance: When the procedure is run with 5 mL the test (not more than 1 ppm). of the standard solution under the above operating condi- (4) Related substances—Dissolve 0.1 g of Piperacillin tions, the internal standard and piperacillin are eluted in this Sodium in 50 mL of the mobile phase A, and use this solu- order with the resolution between these peaks being not less tion as the sample solution. Pipet 1 mL of the sample solu- than 3. tion, add the mobile phase A to make exactly 100 mL, and System repeatability: When the test is repeated 6 times use this solution as the standard solution. Perform the test with 5 mL of the standard solution under the above operating with exactly 20 mL each of the sample solution and standard conditions, the relative standard deviation of the ratio of the solution as directed under Liquid Chromatography <2.01> peak height of piperacillin to that of the internal standard is according to the following conditions, and determine the not more than 1.0z. areas of each peak by the automatic integration method: the Containers and storage Containers—Tight containers. area of the peak of ampicillin appeared at the retention time of about 7 minutes from the sample solution is not larger than 1/2 times that of piperacillin from the standard solu- JP XVI Official Monographs / Piperacillin Sodium for Injection 1255 tion, the total area of related compounds 1 appeared at the mL. Pipet 5 mL of this solution, add exactly 5 mL of the in- retention times of about 17 minutes and about 21 minutes is ternal standard solution, and use this solution as the stand- not larger than 2 times of the peak area of piperacillin from ard solution. Perform the test with 5 mLeachofthesample the standard solution, the peak area of related compound 2 solution and standard solution as directed under Liquid appeared at the retention time of about 56 minutes is not Chromatography <2.01> according to the following condi- larger than that of piperacillin from the standard solution, tions, and calculate the ratios, QT and QS, of the peak height and the total area of the peaks other than piperacillin is not of piperacillin to that of the internal standard. larger than 5 times of the peak area of piperacillin from the Amount [mg (potency)] of piperacillin (C H N O S) standard solution. The peak areas of ampicillin, related com- 23 27 5 7 ＝ M × Q /Q × 1000 pounds 1 and related compound 2 are used after multiplying S T S by their relative response factors, 1.39, 1.32 and 1.11, MS: Amount [mg (potency)] of Piperacillin RS respectively. Internal standard solution—A solution of acetanilide in the Operating conditions— mobile phase (1 in 5000). Detector: An ultraviolet absorption photometer (wave- Operating conditions— length: 220 nm). Detector: An ultraviolet absorption photometer (wave- Column: A stainless steel column 4.6 mm in inside diame- length: 254 nm). ter and 15 cm in length, packed with octadecylsilanized silica Column: A stainless steel column 4.6 mm in inside diame- gel for liquid chromatography (5 mm in particle diameter). ter and 15 cm in length, packed with octadecylsilanized silica Column temperature: A constant temperature of about gel for liquid chromatography (5 mminparticlediameter). 259C. Column temperature: A constant temperature of about Mobile phase A: A mixture of water, acetonitrile and 0.2 259C. mol/L potassium dihydrogenphosphate (45:4:1). Mobile phase: To 60.1 g of acetic acid (100) and 101.0 g of Mobile phase B: A mixture of acetonitrile, water and 0.2 triethylamine add water to make exactly 1000 mL. To 25 mL mol/L potassium dihydrogenphosphate (25:24:1). of this solution add 25 mL of dilute acetic acid and 210 mL Flowing of the mobile phase: Control the gradient by mix- of acetonitrile, and add water to make exactly 1000 mL. ing the mobile phases A and B as directed in the following Flow rate: Adjust the flow rate so that the retention time table. of piperacillin is about 5 minutes. System suitability— Time after injection Mobile phase A Mobile phase B System performance: When the procedure is run with 5 mL of sample (min) (volz) (volz) of the standard solution under the above operating conditions, the internal standard and piperacillin are eluted in this 0 – 7 100 0 order with the resolution between these peaks being not less 7–13 100→ 83 0 → 17 than 3. 13–41 83 17 System repeatability: When the test is repeated 6 times 41–56 83→ 20 17 → 80 with 5 mL of the standard solution under the above operating 56–60 20 80 conditions, the relative standard deviation of the ratios of the peak height of piperacillin to that of the internal stand- Flow rate: 1.0 mL per minute. The retention time of piper- ard is not more than 1.0z. acillin is about 33 minutes. Time span of measurement: About 1.8 times as long as the Containers and storage Containers—Hermetic containers. retention time of piperacillin beginning after the solvent peak. System suitability— Piperacillin Sodium for Injection System performance: When the procedure is run with 20 mL of the standard solution under the above operating con- 注射用ピペラシリンナトリウム ditions, the number of theoretical plates and the symmetry factor of the peak of piperacillin are not less than 15,000 and Piperacillin Sodium for Injection is a preparation not more than 1.5, respectively. for injection which is dissolved before use. System repeatability: When the test is repeated 3 times It contains not less than 93.0z and not more with 20 mL of the standard solution under the above operat- than 107.0z of the labeled amount of piperacillin ing conditions, the relative standard deviation of the peak (C23H27N5O7S: 517.55). areas of piperacillin is not more than 2.0z. Method of preparation Prepare as directed under Injec- Water <2.48> Not more than 1.0z (3 g, volumetric titrations, with Piperacillin Sodium. tion, direct titration). Description Piperacillin Sodium for Injection is a white Assay Weigh accurately an amount of Piperacillin Sodium, powder or masses. equivalent to about 0.1 g (potency), and dissolve in water to make exactly 100 mL. To exactly 5 mL of this solution add Identification Proceed as directed in the Identification exactly 5 mL of the internal standard solution, and use this under Piperacillin Sodium. solution as the sample solution. Separately, weigh accurately pH <2.54> The pH of a solution prepared by dissolving an an amount of Piperacillin RS, equivalent to about 0.1 g (po- amount of Piperacillin Sodium for Injection, equivalent to tency), and dissolve in the mobile phase to make exactly 100 1.0 g (potency) of Piperacillin Sodium according to the la- 1256 Piperazine Adipate / Official Monographs JP XVI beled amount, in 4 mL of water is 5.0 – 7.0. line powder. It is odorless, and has a slightly acid taste. It is soluble in water and in acetic acid (100), and practi- Purity (1) Clarity and color of solution—Dissolve an cally insoluble in ethanol (95), in acetone and in diethyl amount of Piperacillin Sodium for Injection, equivalent to ether. 4.0 g (potency) of Piperacillin Sodium according to the Melting point: about 2509C (with decomposition). labeled amount, in 17 mL of water: the solution is clear and colorless. Identification (1) Dissolve 0.5 g of Piperazine Adipate in (2) Related substances—Proceed as directed in the Purity 10 mL of water, add 1 mL of hydrochloric acid, and extract (4) under Piperacillin Sodium. with two 20-mL portions of diethyl ether. Combine the diethyl ether extracts, evaporate to dryness on a water bath, Water <2.48> Not more than 1.0z (3 g, volumetric titra- and dry the residue at 1059C for 1 hour: the melting point tion, direct titration). <2.60> is between 1529Cand1559C. Bacterial endotoxins <4.01> Less than 0.04 EU/mg (po- (2) To 3 mL of a solution of Piperazine Adipate (1 in tency). 100) add 3 drops of Reinecke salt TS: a light red precipitate is formed. Uniformity of dosage units <6.02> It meets the requirement (3) Determine the infrared absorption spectrum of Piper- of the Mass variation test. azine Adipate, previously dried, as directed in the potassium Foreign insoluble matter <6.06> Perform the test according bromide disk method under Infrared Spectrophotometry to Method 2: it meets the requirement. <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at Insoluble particulate matter <6.07> It meets the require- the same wave numbers. ment. pH <2.54> The pH of a solution of Piperazine Adipate (1 in Sterility <4.06> Perform the test according to the Mem- 20) is between 5.0 and 6.0. brane filtration method: it meets the requirement. Purity (1) Clarity and color of solution—Dissolve 1.0 g Assay Weigh accurately the mass of the contents of not less of Piperazine Adipate in 30 mL of water: the solution is than 10 Piperacillin Sodium for Injection. Weigh accurately clear and colorless. an amount of the contents, equivalent to about 20 mg (po- (2) Heavy metals <1.07>—Proceed with 2.0 g of Pipera- tency) of Piperacillin Sodium, dissolve in water to make ex- zine Adipate according to Method 2, and perform the test. actly 20 mL. Pipet 5 mL of this solution, add exactly 5 mL Prepare the control solution with 2.0 mL of Standard Lead of the internal standard solution, and use this solution as the Solution (not more than 10 ppm). sample solution. Separately, weigh accurately about 20 mg (potency) of Piperacillin RS, and dissolve in the mobile Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, phase to make exactly 20 mL. Pipet 5 mL of this solution, 4 hours). add exactly 5 mL of the internal standard solution, and use Residue on ignition <2.44> Not more than 0.1z (1 g). this solution as the standard solution. Proceed as directed in the Assay under Piperacillin Sodium. Assay Weigh accurately about 0.2 g of Piperazine Adipate, previously dried, dissolve in a mixture of 20 mL of acetic Amount [mg (potency)] of piperacillin (C H N O S) 23 27 5 7 acid for nonaqueous titration and 40 mL of acetone for ＝ M × Q /Q S T S nonaqueous titration, and titrate <2.50> with 0.1 mol/L per-

MS: Amount [mg (potency)] of Piperacillin RS chloric acid VS until the red-purple color of the solution changes to blue-purple (indicator: 6 drops of bromocresol Internal standard solution—A solution of acetanilide in the green-methylrosaniline chloride TS). Perform a blank deter- mobile phase (1 in 5000). mination, and make any necessary correction. Containers and storage Containers—Hermetic containers. Each mL of 0.1 mol/L perchloric acid VS Plastic containers for aqueous injections may be used. ＝ 11.61 mg of C4H10N2.C6H10O4 Containers and storage Containers—Well-closed contain- Piperazine Adipate ers.

ピペラジンアジピン酸塩 Piperazine Phosphate Hydrate

ピペラジンリン酸塩水和物

C4H10N2.C6H10O4: 232.28 Piperazine hexanedioate [142-88-1]

C4H10N2.H3PO4.H2O: 202.15 Piperazine Adipate, when dried, contains not less Piperazine monophosphate monohydrate than 98.5z of C4H10N2.C6H10O4. [18534-18-4] Description Piperazine Adipate occurs as a white, crystal- Piperazine Phosphate Hydrate contains not less JP XVI Official Monographs / Piperazine Phosphate Tablets 1257 than 98.5z of piperazine phosphate (C4H10N2.H3PO4: phate Hydrate, dissolve in 10 mL of formic acid, add 60 mL 184.13), calculated on the anhydrous basis. of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank Description Piperazine Phosphate Hydrate occurs as white determination, and make any necessary correction. crystals or crystalline powder. It is odorless, and has a slightly acid taste. Each mL of 0.1 mol/L perchloric acid VS

It is soluble in formic acid, sparingly soluble in water, very ＝ 9.207 mg of C4H10N2.H3PO4 slightly soluble in acetic acid (100), and practically insoluble Containers and storage Containers—Well-closed contain- in methanol, in ethanol (95) and in diethyl ether. ers. It dissolves in dilute hydrochloric acid. Melting point: about 2229C (with decomposition). Identification (1) To 3 mL of a solution of Piperazine Piperazine Phosphate Tablets Phosphate Hydrate (1 in 100) add 3 drops of Reinecke salt TS:a light red precipitate is formed. ピペラジンリン酸塩錠 (2) Determine the infrared absorption spectrum of Piperazine Phosphate Hydrate as directed in the potassium Piperazine Phosphate Tablets contain not less than bromide disk method under Infrared Spectrophotometry 95.0z and not more than 105.0z of the labeled <2.25>, and compare the spectrum with the Reference Spec- amount of piperazine phosphate hydrate (C H N . trum: both spectra exhibit similar intensities of absorption at 4 10 2 H PO .H O: 202.15). the same wave numbers. 3 4 2 (3) A solution of Piperazine Phosphate Hydrate (1 in Method of preparation Prepare as directed under Tablets, 100) responds to Qualitative Tests <1.09> (1) and (3) for with Piperazine Phosphate Hydrate. phosphate. Identification Take a quantity of Piperazine Phosphate pH <2.54> Dissolve 1.0 g of Piperazine Phosphate Hydrate Tablets equivalent to 0.1 g of Piperazine Phosphate Hydrate in 100 mL of water: the pH of the solution is between 6.0 according to the labeled amount, previously powdered, add and 6.5. 10 mL of water, shake while warming for 10 minutes, allow to cool, and filter. To 3 mL of the filtrate add 3 drops of Purity (1) Chloride <1.03>—To 0.5 g of Piperazine Phos- Reinecke salt TS: a light red precipitate is formed. phate Hydrate add 6 mL of dilute nitric acid and water to make 50 mL. Use this solution as the test solution, and per- Disintegration <6.09> It meets the requirement. The time form the test. Prepare the control solution with 0.25 mL of limit of the test is 10 minutes. 0.01 mol/L hydrochloric acid VS (not more than 0.018z). Assay Weigh accurately not less than 20 Piperazine Phos- (2) Heavy metals <1.07>—To 2.0 g of Piperazine Phos- phate Tablets, and powder. Weigh accurately a quantity of phate Hydrate add 5 mL of dilute hydrochloric acid, 30 mL the powder, equivalent to about 0.15 g of piperazine phos- of water and 2 mL of dilute acetic acid, and dissolve. Add phate hydrate (C H N .H PO .H O). Add 5 mL of formic sodium hydroxide TS, adjust the pH of the solution to 3.3, 4 10 2 3 4 2 acid, shake for 5 minutes, centrifuge, and collect the super- and add water to make 50 mL. Perform the test using this natant liquid. To the residue add 5 mL of formic acid, shake solution as the test solution. Prepare the control solution for 5 minutes, centrifuge, and collect the supernatant liquid. with 2.0 mL of Standard Lead Solution (not more than 10 Repeat twice the same procedure with 5 mL each of acetic ppm). acid (100), combine all the supernatant liquids, add 50 mL of (3) Arsenic <1.11>—Dissolve 2.0 g of Piperazine Phos- acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric phate Hydrate in 5 mL of dilute hydrochloric acid, and use acid VS (potentiometric titration). Perform a blank determi- this solution as the test solution. Perform the test (not more nation, and make any necessary correction. than 1 ppm). (4) Related substances—Dissolve 50 mg of Piperazine Each mL of 0.1 mol/L perchloric acid VS

Phosphate Hydrate in 10 mL of water, and use this solution ＝ 10.11 mg of C4H10N2.H3PO4.H2O as the sample solution. Pipet 1 mL of the sample solution, Containers and storage Containers—Tight containers. add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ammonia solution (28), acetone and ethanol (99.5) (8:3:3:2) to a distance of about 13 cm, and air-dry the plate. Spray evenly 4- dimethylaminocinnamaldehyde TS, and allow to stand for 15 minutes: the spots other than the principal spot and the spot on the starting line from the sample solution are not more intense than the spot from the standard solution. Water <2.48> 8.0 – 9.5z (0.3 g, volumetric titration, direct titration). Assay Weigh accurately about 0.15 g of Piperazine Phos- 1258 Pirarubicin / Official Monographs JP XVI

cin according to Method 2, and perform the test. Prepare the Pirarubicin control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). ピラルビシン (3) Related substances—Dissolve 10 mg of Pirarubicin in 20 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the peak area of doxorubicin, having the relative retention time of about 0.45 with respect to pirarubicin, and the area of the peak, having the relative retention time of about 1.2 with respect to pirarubicin, obtained from the sample solution are not larger than the peak C H NO : 627.64 32 37 12 area of pirarubicin from the standard solution, respectively, (2S,4S)-4-{3-Amino-2,3,6-trideoxy-4-O-[(2R)-3,4,5,6- and the sum of the areas of the peaks, having the relative tetrahydro-2H-pyran-2-yl]-a-L-lyxo-hexopyranosyloxy}- retention times of about 1.9 and about 2.0 with respect to 2,5,12-trihydroxy-2-hydroxyacetyl-7-methoxy-1,2,3,4- pirarubicin, from the sample solution is not larger than 5 tetrahydrotetracene-6,11-dione times the peak area of pirarubicin from the standard solu- [72496-41-4] tion. For these calculations, use the peak area for doxorubicin after multiplying by the relative response factor 0.94 Pirarubicin is a derivative of daunorubicin. and the area for the two peaks, having the relative retention It contains not less than 950 mg (potency) per mg, times of about 1.9 and about 2.0, after multiplying by their calculated on the anhydrous basis. The potency of relative response factors, 1.09, respectively. Pirarubicin is expressed as mass (potency) of pirarubi- Operating conditions— cin (C H NO ). 32 37 12 Detector, column, column temperature, mobile phase, and Description Pirarubicin occurs as a red-orange crystalline flow rate: Proceed as directed in the operating conditions in powder. the Assay. It is soluble in chloroform, very slightly soluble in aceto- Time span of measurement: About 4 times as long as the nitrile, in methanol and in ethanol (99.5), and practically in- retention time of pirarubicin. soluble in water. System suitability— Test for required detectability: Measure exactly 2 mL of Identification (1) Dissolve 10 mg of Pirarubicin in 80 mL the standard solution, and add the mobile phase to make ex- of methanol and 6 mL of diluted hydrochloric acid (1 in actly 10 mL. Confirm that the peak area of pirarubicin ob- 5000), and add water to make 100 mL. To 10 mL of this so- tained from 20 mL of this solution is equivalent to 14 to 26z lution add diluted methanol (4 in 5) to make 100 mL. Deter- of that from 20 mL of the standard solution. mine the absorption spectrum of this solution as directed System performance, and system repeatability: Proceed as under Ultraviolet-visible Spectrophotometry <2.24>,and directed in the system suitability in the Assay. compare the spectrum with the Reference Spectrum or the spectrum of a solution of Pirarubicin RS prepared in the Water <2.48> Not more than 2.0z (0.1 g, volumetric titra- same manner as the sample solution: both spectra exhibit tion, direct titration). similar intensities of absorption at the same wavelengths. Assay Weigh accurately an amount of Pirarubicin and (2) Dissolve 5 mg each of Pirarubicin and Pirarubicin RS Pirarubicin RS, equivalent to about 10 mg (potency), and in 5 mL of chloroform, and use these solutions as the sample dissolve in the mobile phase to make exactly 10 mL. Pipet 5 solution and standard solution. Perform the test with these mL of these solutions, add exactly 5 mL of the internal solutions as directed under Thin-layer Chromatography standard solution, and use these solutions as the sample so- <2.03>. Spot 5 mL each of the sample solution and standard lution and standard solution. Perform the test with 20 mL solution on a plate of silica gel for thin-layer chromatogra- each of the sample solution and standard solution as directed phy. Develop the plate with a mixture of chloroform and under Liquid Chromatography <2.01> according to the fol- methanol (5:1) to a distance of about 10 cm, and air-dry the lowing conditions, and calculate the ratios, Q and Q ,of plate. Examine the spots with the necked eye: the principal T S thepeakareaofpirarubicintothat of the internal standard. spot obtained from the sample solution and the spot from the standard solution show a red-orange color and the same Amount [mg (potency)] of C32H37NO12 Rf value. ＝ MS × QT/QS × 1000 20 Optical rotation <2.49> [a]D : ＋195 – ＋2159(10 mg, chlo- MS: Amount [mg (potency)] of Pirarubicin RS roform, 10 mL, 100 mm). Internal standard solution—A solution of 2-naphthol in the Purity (1) Clarity and color of solution—Dissolve 10 mg mobile phase (1 in 1000). of Pirarubicin in 10 mL of 0.01 mol/L hydrochloric acid TS: Operating conditions— the solution is clear and red. Detector: An ultraviolet absorption photometer (wave- (2) Heavy metals <1.07>—Proceed with 1.0 g of Pirarubi- length: 254 nm). JP XVI Official Monographs / Pirenoxine 1259

Column: A stainless steel column 6 mm in inside diameter Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of and 15 cm in length, packed with octadecylsilanized silica gel Pirenoxine according to Method 2, and perform the test. for liquid chromatography (5 mminparticlediameter). Prepare the control solution with 2.0 mL of Standard Lead Column temperature: A constant temperature of about Solution (not more than 20 ppm). 259C. (2) Related substances—Dissolve 10 mg of Pirenoxine in Mobile phase: A mixture of 0.05 mol/L ammonium for- 50 mL of the mobile phase, and use this solution as the sam- mate buffer solution, pH 4.0 and acetonitrile (3:2). ple solution. Pipet 3 mL of the sample solution, add the mo- Flow rate: Adjust the flow rate so that the retention time bile phase to make exactly 200 mL, and use this solution as of pirarubicin is about 7 minutes. the standard solution. Perform the test with exactly 5 mL System suitability— each of the sample solution and standard solution as directed System performance: When the procedure is run with 20 under Liquid Chromatography <2.01> according to the fol- mL of the standard solution under the above operating con- lowing conditions. Determine each peak area of both solu- ditions, pirarubicin and the internal standard are eluted in tions by the automatic integration method: the total area of this order with the resolution between these peaks being not the peaks other than pirenoxine is not larger than the peak less than 9. area of pirenoxine from the standard solution. System repeatability: When the test is repeated 6 times Operating conditions— with 20 mL of the standard solution under the above operat- Detector: An ultraviolet absorption photometer (wave- ing conditions, the relative standard deviation of the ratios length: 230 nm). of the peak area of pirarubicin to that of the internal stand- Column: A stainless steel column 4 mm in inside diameter ard is not more than 1.0z. and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Containers and storage Containers—Hermetic containers. Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 1.39 g of tetra n-butylammonium Pirenoxine chloride and 4.5 g of disodium hydrogen phosphate dodecahydrate in 1000 mL of water, and adjust the pH to 6.5 with ピレノキシン phosphoric acid. To 700 mL of this solution add 200 mL of acetonitrile and 30 mL of tetrahydrofuran, and mix. Flow rate: Adjust the flow rate so that the retention time of pirenoxine is about 10 minutes. Time span of measurement: About 3 times as long as the retention time of pirenoxine.

C16H8N2O5308.25 System suitability— 1-Hydroxy-5-oxo-5H-pyrido[3,2-a]phenoxazine-3- Test for required detectability: To exactly 2 mL of the carboxylic acid standard solution add the mobile phase to make exactly 30 [1043-21-6] mL. Confirm that the peak area of pirenoxine obtained from 5 mL of this solution is equivalent to 5 to 8z of that of Pirenoxine, when dried, contains not less than pirenoxine obtained from 5 mL of the standard solution. 98.0z of C16H8N2O5. System performance: Dissolve 3 mg of Pirenoxine and 16 mg of methyl parahydroxybenzoate in 100 mL of the mobile Description Pirenoxine occurs as a yellow-brown powder. phase. When the procedure is run with 5 mL of this solution It is odorless, and has a slightly bitter taste. under the above operating conditions, pirenoxine and methyl It is very slightly soluble in dimethylsulfoxide, and practi- parahydroxybenzoate are eluted in this order with the resolu- cally insoluble in water, in acetonitrile, in ethanol (95), in tion between these peaks being not less than 2.0. tetrahydrofuran and in diethyl ether. System repeatability: When the test is repeated 6 times Melting point: about 2509C (with decomposition). with 5 mL of the standard solution under the above operating Identification (1) Dissolve 2 mg of Pirenoxine in 10 mL conditions, the relative standard deviation of the peak area of phosphate buffer solution, pH 6.5, add 5 mL of a solu- of pirenoxine is not more than 1.0z. tion of L-ascorbic acid (1 in 50), and shake vigorously: a Loss on drying <2.41> Not more than 1.5z (0.5 g, in vacu- dark purple precipitate is formed. um, 809C, 3 hours). (2) Determine the absorption spectrum of a solution of Pirenoxine in phosphate buffer solution, pH 6.5 (1 in Residue on ignition <2.44> Not more than 0.1z (1 g). 200,000) as directed under Ultraviolet-visible Spectropho- Assay Weigh accurately about 0.1 g of Pirenoxine, previ- tometry <2.24>, and compare the spectrum with the Refer- ously dried, dissolve in 140 mL of dimethylsulfoxide by heat- ence Spectrum: both spectra exhibit similar intensities of ab- ing on a water bath. After cooling, add 30 mL of water, and sorption at the same wavelengths. titrate <2.50> immediately with 0.02 mol/L sodium hydrox- (3) Determine the infrared absorption spectrum of ide VS (potentiometric titration). Perform a blank determi- Pirenoxine, previously dried, as directed in the potassium nation, and make any necessary correction. bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- Each mL of 0.02 mol/L sodium hydroxide VS trum: both spectra exhibit similar intensities of absorption at ＝ 6.165 mg of C16H8N2O5 the same wave numbers. Containers and storage Containers—Tight containers. 1260 Pirenzepine Hydrochloride Hydrate / Official Monographs JP XVI

solution add 5 mL of methanol and the mobile phase A to Pirenzepine Hydrochloride Hydrate make 10 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add 5 mL of metha- ピレンゼピン塩酸塩水和物 nol and the mobile phase A to make exactly 10 mL. Pipet 1 mL of this solution, add 5 mL of methanol and the mobile phase A to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mLeach of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than pirenzepine is not larger than 3/10 times the peak area of pirenzepine from the standard solution, and the total area of the peaks other than pirenzepine is not larger than 3/5 times the peak area of pirenzepine from the standard solution.

C19H21N5O2.2HCl.H2O: 442.34 Operating conditions— 11-[(4-Methylpiperazin-1-yl)acetyl]-5,11-dihydro-6H- Detector: An ultraviolet absorption photometer (wave- pyrido[2,3-b][1,4]benzodiazepin-6-one dihydrochloride length: 283 nm). monohydrate Column: A stainless steel column 4.6 mm in inside diame- [29868-97-1, anhydride] ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticlediameter). Pirenzepine Hydrochloride Hydrate contains not Column temperature: A constant temperature of about less than 98.5z and not more than 101.0z of pirenze- 409C. pine hydrochloride (C19H21N5O2.2HCl: 424.32), calcu- Mobile phase A: Dissolve 2 g of sodium lauryl sulfate in lated on the anhydrous basis. 900 mL of water, adjust the pH to 3.2 with acetic acid (100), andaddwatertomake1000mL. Description Pirenzepine Hydrochloride Hydrate occurs as Mobile phase B: Methanol. a white to pale yellow crystalline powder. Mobile phase C: Acetonitrile. It is freely soluble in water and in formic acid, slightly Flowing of the mobile phase: Control the gradient by mix- soluble in methanol, and very slightly soluble in ethanol ing the mobile phases A, B and C as directed in the following (99.5). table. The pH of a solution by dissolving 1 g of Pirenzepine Hy- drochloride Hydrate in 10 mL of water is between 1.0 and 2.0. Time after injection Mobile phase Mobile phase Mobile phase Melting point: about 2459C (with decomposition). of sample (min) A(volz) B(volz) C(volz) It is gradually colored by light. 0 – 15 55 → 25 30 15 → 45 Identification (1) Determine the absorption spectrum of a 15–253045 solution of Pirenzepine Hydrochloride Hydrate (1 in 40,000) as directed under Ultraviolet-visible Spectrophotometry Flow rate: Adjust the flow rate so that the retention time <2.24>, and compare the spectrum with the Reference Spec- of pirenzepine is about 8 minutes. trum: both spectra exhibit similar intensities of absorption at Time span of measurement: About 2 times as long as the thesamewavelengths. retention time of pirenzepine beginning after the solvent (2) Determine the infrared absorption spectrum of Piren- peak. zepine Hydrochloride Hydrate as directed in the potassium System suitability— chloride disk method under Infrared Spectrophotometry Test for required detectability: Pipet 1 mL of the standard <2.25>, and compare the spectrum with the Reference Spec- solution, and add 5 mL of methanol and the mobile phase A trum: both spectra exhibit similar intensities of absorption at to make exactly 10 mL. Confirm that the peak area of piren- the same wave numbers. zepine obtained from 10 mL of this solution is equivalent to 7 (3) A solution of Pirenzepine Hydrochloride Hydrate to 13z of that from 10 mL of the standard solution. (1 in 50) responds to Qualitative Tests <1.09> for chloride. System performance: Dissolve 0.1 g of phenylpiperazine Purity (1) Clarity and color of solution—A solution ob- hydrochloride in 10 mL of methanol. Mix 1 mL of this solu- tained by dissolving 1.0 g of Pirenzepine Hydrochloride Hy- tion and 1 mL of the sample solution, and add 5 mL of drate in 10 mL of water is clear and not more color than that methanol and the mobile phase A to make 10 mL. When the of the following control solution. procedure is run with 10 mL of this solution under the above Control solution: To 1.2 mL of Matching fluid for color F operating conditions, pirenzepine and phenylpiperazine are add8.8mLofdilutedhydrochloricacid(1in40). eluted in this order with the resolution between these peaks (2) Heavy metals <1.07>—Proceed with 2.0 g of Pirenze- being not less than 5. pine Hydrochloride Hydrate according to Method 2, and System repeatability: When the test is repeated 6 times perform the test. Prepare the control solution with 2.0 mL of with 10 mL of the standard solution under the above operat- Standard Lead Solution (not more than 10 ppm). ing conditions, the relative standard deviation of the peak (3) Related substances—Dissolve 0.3 g of Pirenzepine area of pirenzepine is not more than 2.0z. Hydrochloride Hydrate in 10 mL of water. To 1 mL of this Water <2.48> Not less than 3.5z and not more than 5.0z JP XVI Official Monographs / Piroxicam 1261

(0.3 g, volumetric titration, direct titration). lution (not more than 20 ppm). (2) Related substances—Dissolve 75 mg of Piroxicam in Residue on ignition <2.44> Not more than 0.1z (1 g). 50 mL of acetonitrile for liquid chromatography, and use Assay Weigh accurately about 0.2 g of Pirenzepine Hydro- this solution as the sample solution. Pipet 1 mL of the sam- chloride Hydrate, dissolve in 2 mL of formic acid, add 60 ple solution, add acetonitrile for liquid chromatography to mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L make exactly 10 mL. Pipet 1 mL of this solution, add aceto- perchloric acid VS (potentiometric titration). Perform a nitrile for liquid chromatography to make exactly 50 mL, blank determination in the same manner, and make any nec- and use this solution as the standard solution. Perform the essary correction. test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography Each mL of 0.1 mol/L perchloric acid VS <2.01> according to the following conditions, and determine ＝ 14.14 mg of C H N O .2HCl 19 21 5 2 each peak area by the automatic integration method: the Containers and storage Containers—Well-closed contain- area of the peak other than piroxicam obtained with the ers. sample solution is not larger than the peak area of piroxicam Storage—Light-resistant. with the standard solution, and the total area of the peaks other than piroxicam is not larger than 2 times the peak area of piroxicam with the standard solution. Piroxicam Operating conditions— Detector: An ultraviolet absorption photometer (wave- ピロキシカム length: 230 nm). Column: A stainless steel column 4.6 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticlediameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.05 mol/L potassium dihydrogen phosphate TS, pH 3.0 and acetonitrile for liquid C H N O S: 331.35 15 13 3 4 chromatography (3:2). 4-Hydroxy-2-methyl-N-(pyridin-2-yl)-2H-1,2- Flow rate: Adjust the flow rate so that the retention time benzothiazine-3-carboxamide 1,1-dioxide of piroxicam is about 10 minutes. [36322-90-4] Time span of measurement: About 5 times as long as the retention time of piroxicam beginning after the solvent peak. Piroxicam contains not less than 98.5z and not System suitability— more than 101.0z of C H N O S, calculated on the 15 13 3 4 Test for required detectability: To exactly 5 mL of the dried basis. standard solution add acetonitrile for liquid chromatography Description Piroxicamoccursasawhitetopaleyellow to make exactly 20 mL. Confirm that the peak area of crystalline powder. piroxicam obtained with 20 mL of this solution is equivalent It is sparingly soluble in acetic anhydride, slightly soluble to 17.5 to 32.5z of that with 20 mL of the standard solution. in acetonitrile, in methanol and in ethanol (99.5), very System performance: When the procedure is run with 20 slightly soluble in acetic acid (100), and practically insoluble mL of the standard solution under the above operating con- in water. ditions, the number of theoretical plates and the symmetry Melting point: about 2009C (with decomposition). factor of the peak of piroxicam are not less than 6000 and not more than 1.5, respectively. Identification (1) Dissolve 0.1 g of Piroxicam in a mix- System repeatability: When the test is repeated 6 times ture of methanol and 0.5 mol/L hydrochloric acid TS with 20 mL of the standard solution under the above operat- (490:1) to make 200 mL. To 1 mL of this solution add the ing conditions, the relative standard deviation of the peak mixture of methanol and 0.5 mol/L hydrochloric acid TS area of piroxicam is not more than 2.0z. (490:1) to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro- Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, photometry <2.24>, and compare the spectrum with the Ref- 3 hours). erence Spectrum: both spectra exhibit similar intensities of Residue on ignition <2.44> Not more than 0.2z (1 g). absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Assay Weigh accurately about 0.25 g of Piroxicam, dis- Piroxicam as directed in the potassium bromide disk method solve in 60 mL of a mixture of acetic anhydride and acetic under Infrared Spectrophotometry <2.25>, and compare the acid (100) (1:1), and titrate <2.50> with 0.1 mol/L perchloric spectrum with the Reference Spectrum: both spectra exhibit acid VS (potentiometric titration). Perform a blank determi- similar intensities of absorption at the same wave numbers. nation in the same manner, and make any necessary correc- If any difference appears between the spectra, dissolve the tion. sample with dichloromethane, evaporate the solvent, dry the Each mL of 0.1 mol/L perchloric acid VS residue on a water bath, and perform the test. ＝ 33.14 mg of C15H13N3O4S Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Containers and storage Containers—Tight containers. Piroxicam according to Method 2, and perform the test. Pre- pare the control solution with 2.0 mL of Standard Lead So- 1262 Pivmecillinam Hydrochloride / Official Monographs JP XVI

solution. Separately, dissolve 2.0 mg of Pivmecillinam Hy- Pivmecillinam Hydrochloride drochloride RS in 4.0 mL of water, and use this solution as the standard solution. Perform the test with these solutions ピブメシリナム塩酸塩 as directed under Thin-layer Chromatography <2.03>. Spot 2 mL of the standard solution on a plate of silica gel for thin- layer chromatography, allow to stand for 30 minutes, then spot 2 mL of the sample solution on the plate. Immediately, develop the plate with a mixture of acetone, water and acetic acid (100) (10:1:1) to a distance of about 12 cm, and air-dry the plate. Allow the plate to stand for 10 minutes in iodine vapor: the spot from the sample solution appeared at the po- C H N O S.HCl: 476.03 21 33 3 5 sition corresponding to the spot obtained from the standard 2,2-Dimethylpropanoyloxymethyl (2S,5R,6R)-6-[(azepan- solution is not larger and not more intense than the spot 1-ylmethylene)amino]-3,3-dimethyl-7-oxo-4-thia-1- from the standard solution, and any spot other than the azabicyclo[3.2.0]heptane-2-carboxylate monohydrochloride principal spot and the above spot is not observable. [32887-03-9] Water <2.48> Not more than 1.0z (0.25 g, coulometric Pivmecillinam Hydrochloride contains not less than titration). 630 mg (potency) and not more than 710 mg (potency) Assay Weigh accurately an amount of Pivmecillinam Hy- per mg, calculated on the anhydrous basis. The po- drochloride and Pivmecillinam Hydrochloride RS, equiva- tency of Pivmecillinam Hydrochloride is expressed as lent to about 20 mg (potency), dissolve in a suitable amount mass (potency) of mecillinam (C H N O S: 325.43). 15 23 3 3 of the mobile phase, add exactly 10 mL of the internal stand- Description Pivmecillinam Hydrochloride occurs as a ard solution and the mobile phase to make 100 mL, and use white to yellowish white crystalline powder. these solutions as the sample solution and the standard solu- It is very soluble in methanol and in acetic acid (100), tion, respectively. Perform the test with 10 mLeachofthe freely soluble in water and in ethanol (99.5), and soluble in sample solution and standard solution as directed under Liq- acetonitrile. uid Chromatography <2.01> according to the following conditions, and calculate the ratios, Q and Q , of the peak area Identification (1) Determine the infrared absorption spec- T S of pivmecillinam to that of the internal standard. trum of Pivmecillinam Hydrochloride as directed in the potassium bromide disk method under Infrared Spectro- Amount [mg (potency)] of mecillinam (C15H23N3O3S) photometry <2.25>, and compare the spectrum with the ＝ MS × QT/QS × 1000 Reference Spectrum or the spectrum of Pivmecillinam Hy- M : Amount [mg (potency)] of Pivmecillinam Hydrochlo- drochloride RS: both spectra exhibit similar intensities of S ride RS absorption at the same wave numbers. (2) Dissolve 0.5 g of Pivmecillinam Hydrochloride in 10 Internal standard solution—A solution of diphenyl in the mL of water, and add 1 mL of dilute nitric acid and 1 drop mobile phase (1 in 12,500). of silver nitrate TS: a white precipitate is formed. Operating conditions— Detector: An ultraviolet absorption photometer (wave- Optical rotation <2.49> [a]20: ＋200 – ＋2209(1 g calculated D length: 254 nm). on the anhydrous basis, water, 100 mL, 100 mm). Column: A stainless steel column 4 mm in inside diameter Purity (1) Heavy metals <1.07>—To 1.0 g of Pivmecillin- and 30 cm in length, packed with octadecylsilanized silica gel am Hydrochloride in a crucible add 10 mL of a solution of for liquid chromatography (10 mminparticlediameter). magnesium nitrate hexahydrate in ethanol (95) (1 in 10), fire Column temperature: A constant temperature of about the ethanol to burn, and heat gradually to incinerate. If a 259C. carbonized substance remains, moisten with a small amount Mobile phase: Dissolve 0.771 g of ammonium acetate in of nitric acid, and ignite to incinerate. Cool, add 3 mL of about 900 mL of water, adjust the pH to 3.5 with acetic acid hydrochloric acid to the residue, dissolve by warming on a (100), and add water to make 1000 mL. To 400 mL of this water bath, and heat to dryness. To the residue add 10 mL of solution add 600 mL of acetonitrile. water, and dissolve by warming on a water bath. After cool- Flow rate: Adjust the flow rate so that the retention time ing, adjust the pH to 3 to 4 with ammonia TS, add 2 mL of of pivmecillinam is about 6.5 minutes. dilute acetic acid, filter if necessary, and wash the crucible System suitability— and the filter with 10 mL of water. Put the filtrate and the System performance: When the procedure is run with 10 washings to a Nessler tube, add water to make 50 mL, and mL of the standard solution under the above operating con- use this solution as the test solution. Prepare the control so- ditions, pivmecillinam and the internal standard are eluted in lution in the same manner as the test solution with 2.0 mL of this order with the resolution between these peaks being not Standard Lead Solution (not more than 20 ppm). less than 4. (2) Arsenic <1.11>—Prepare the test solution with 1.0 g System repeatability: When the test is repeated 6 times of Pivmecillinam Hydrochloride according to Method 4, and with 10 mL of the standard solution under the above operat- perform the test (not more than 2 ppm). ing conditions, the relative standard deviation of the ratios (3) Related substances—Dissolve 50 mg of Pivmecillin- of the peak area of pivmecillinam to that of the internal am Hydrochloride in 4.0 mL of a mixture of acetonitrile and standard is not more than 1.0z. acetic acid (100) (97:3), and use this solution as the sample Containers and storage Containers—Tight containers. JP XVI Official Monographs / Polymixin B Sulfate 1263

Disintegration <6.09> Perform the test using the disk: it Pivmecillinam Hydrochloride meets the requirement. Tablets Assay Weigh accurately the mass of not less than 20 Piv- mecillinam Hydrochloride Tablets, and powder. Weigh ac- ピブメシリナム塩酸塩錠 curately a portion of the powder, equivalent to about 0.1 g (potency) of Pivmecillinam Hydrochloride, add 50 mL of the mobile phase, shake vigorously for 10 minutes, and add the Pivmecillinam Hydrochloride Tablets contains not mobile phase to make exactly 100 mL. Pipet 10 mL of this less than 93.0z and not more than 107.0z of the solution, add exactly 5 mL of the internal standard solution labeled potency of mecillinam (C H N O S: 325.43). 15 23 3 3 and the mobile phase to make 50 mL, filter through a mem- Method of preparation Prepare as directed under Tablets, brane filter with a pore size not exceeding 0.45 mm, discard with Pivmecillinam Hydrochloride. the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately an Identification Powder Pivmecillinam Hydrochloride amount of Pivmecillinam Hydrochloride RS, equivalent to Tablets, dissolve a portion of the powder, equivalent to 35 about 20 mg (potency), dissolve in the mobile phase, add mg (potency) of Pivmecillinam Hydrochloride according to exactly 10 mL of the internal standard solution, add the the labeled amount, in 4 mL of a mixture of acetonitrile and mobile phase to make 100 mL, and use this solution as the acetic acid (100) (97:3), and filter through a membrane filter standard solution. Then, proceed as directed in the Assay with a pore size not exceeding 0.45 mm. Discard the first 2 under Pivmecillinam Hydrochloride. mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately dissolve 25 mg of Pivmecillinam Amount [mg (potency)] of mecillinam (C15H23N3O3S) Hydrochloride RS in 2 mL of a mixture of acetonitrile and ＝ MS × QT/QS × 5 acetic acid (100) (97:3), and use this solution as the standard M : Amount [mg (potency)] of Pivmecillinam Hydrochlo- solution. Perform the test with these solutions as directed S ride RS under Thin-layer Chromatography <2.03>. Spot 2 mLeachof the sample solution and standard solution on a plate of silica Internal standard solution—A solution of diphenyl in the gel for thin-layer chromatography, and immediately develop mobile phase (1 in 12,500). the plate with a mixture of acetone, water and acetic acid Containers and storage Containers—Tight containers. (100) (10:1:1) to a distance of about 12 cm, and air-dry the plate. Allow the plate to stand in iodine vapor for 10 minutes: the principal spot obtained from the sample solution has the same Rf value as the spot from the standard Live Oral Poliomyelitis Vaccine solution. 経口生ポリオワクチン Water <2.48> Not more than 3.0z (1 g of powdered Piv- mecillinam Hydrochloride Tablets, volumetric titration, Live Oral Poliomyelitis Vaccine contains live at- direct titration). tenuated poliovirus of type I, II and III. Uniformity of dosage units <6.02> Perform the test accord- Monovalent or bivalent product may be prepared, if ing to the following method: it meets the requirement of the necessary. Content uniformity test. Live Oral Poliomyelitis Vaccine conforms to the re- To 1 tablet of Pivmecillinam Hydrochloride Tablets add quirements of Live Oral Poliomyelitis Vaccine in the 40 mL of the mobile phase, shake vigorously for 10 minutes, Minimum Requirements for Biological Products. and add the mobile phase to make exactly 50 mL. Pipet Description Live Oral Poliomyelitis Vaccine is a light yel- V mL, equivalent to about 10 mg (potency) of Pivmecillinam low-red to light red, clear liquid. Hydrochloride, add exactly 5 mL of the internal standard solution and the mobile phase to make 50 mL, filter through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 10 mL of the filtrate, and use the subsequent Polymixin B Sulfate filtrate as the sample solution. Separately, weigh accurately ポリミキシン B 硫酸塩 an amount of Pivmecillinam Hydrochloride RS, equivalent to about 20 mg (potency), dissolve in the mobile phase, add exactly 10 mL of the internal standard solution, add the mobile phase to make 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Pivmecillinam Hydrochloride.

Amount [mg (potency)] of mecillinam (C15H23N3O3S) ＝ MS × QT/QS × 25/V Polymixin B Sulfate is the sulfate of a mixture of M : Amount [mg (potency)] of Pivmecillinam Hydrochlo- S peptide substances having antibacterial activity pro- ride RS duced by the growth of Bacillus polymyxa. Internal standard solution—A solution of diphenyl in the It contains not less than 6500 units per mg, calcu- mobile phase (1 in 12,500). lated on the dried basis. The potency of Polymixin B 1264 Polyoxyl 40 Stearate / Official Monographs JP XVI

Sulfate is expressed as mass unit of polymixin B Purity Heavy metals <1.07>—Proceed with 1.0 g of Poly- (C55-56H96-98N16O13). One unit of Polymixin B Sulfate mixin B Sulfate according to Method 2, and perform the is equivalent to 0.129 mg of polymixin B sulfate test. Prepare the control solution with 2.0 mL of Standard (C55-56H96-98N16O13.1-2H2SO4). Lead Solution (not more than 20 ppm). Description Polymixin B Sulfate occurs as a white to yel- Loss on drying <2.41> Not more than 6.0z (1 g, in vacu- low-brown powder. um, 609C, 3 hours). It is freely soluble in water, and practically insoluble in Residue on ignition <2.44> Not more than 0.75z (1 g). ethanol (99.5). Assay Perform the test according to the Cylinder-plate Identification (1) To 5 mL of a solution of Polymixin B method as directed under Microbial Assay for Antibiotics Sulfate (1 in 10) add 5 mL of a solution of sodium hydroxide <4.02> according to the following conditions. (1 in 10), add 5 drops of a solution of copper (II) sulfate pen- (i) Test organism—Escherichia coli NIHJ tahydrate (1 in 100) while shaking: a purple color develops. (ii) Agar media for seed and base layer (2) Transfer 5 mg each of Polymixin B Sulfate and Poly- Peptone 10.0 g mixin B Sulfate RS separately into two glass stoppered test Meat extract 3.0 g tubes, add 1 mL of diluted hydrochloric acid (1 in 2), Sodium chloride 30.0 g stopper the tube, heat at 1359C for 5 hours, then heat to Agar 20.0 g dryness on a water bath, and keep the heating until no more Water 1000 mL hydrochloric acid odor is evolved. Dissolve the residue in 0.5 Mix all the ingredients, and sterilize. Adjust the pH <2.54> mL of water, and use these solutions as the sample solution of the solution so that it will be 6.5 to 6.6 after sterilization. and standard solution (1). Separately, dissolve 20 mg each of (iii) Standard solutions—Weigh accurately an amount of L-leucine, L-threonine, phenylalanine and L-serine separately Polymixin B Sulfate RS, equivalent to about 200,000 units, in 10 mL of water, and use these solutions as the standard dissolve in phosphate buffer solution, pH 6.0 to make ex- solutions (2), (3), (4) and (5), respectively. Perform the test actly 20 mL, and use this solution as the standard stock solu- with these solutions as directed under Thin-layer Chroma- tion. Keep the standard stock solution at not exceeding 59C tography <2.03>. Spot 3 mL each of the sample solution, the and use within 14 days. Take exactly a suitable amount of standard solutions (1), (2), (3), (4) and (5) on a plate of silica the standard stock solution before use, add phosphate buffer gel for thin-layer chromatography, and expose the plate to a solution, pH 6.0 to make solutions so that each mL contains saturated vapor of the developing solvent for 15 hours. De- 4000 units and 1000 units, and use these solutions as the high velop the plate with a mixture of phenol and water (3:1) to a concentration standard solution and the low concentration distance of about 13 cm while without exposure to light, and standard solution, respectively. dry the plate at 1109C for 5 minutes. Spray evenly nin- (iv) Sample solutions—Weigh accurately an amount of hydrin-acetic acid TS on the plate, and heat at 1109Cfor5 Polymixin B Sulfate, equivalent to about 200,000 units, and minutes: Rf value of each spot obtained from the sample so- dissolve in phosphate buffer solution, pH 6.0 to make ex- lution is the same with Rf value of the corresponding spots actly 20 mL. Take exactly a suitable amount of this solution, from the standard solution (1). Each of the spots from the add phosphate buffer solution, pH 6.0 to make solutions so sample solution appears at the position corresponding to that each mL contains 4000 units and 1000 units, and use each of the spots from the standard solutions (2), (3) and (4), these solutions as the high concentration sample solution and but not appears at the position corresponding to the spot the low concentration sample solution, respectively. from the standard solution (5). (3) A solution of Polymixin B Sulfate (1 in 20) responds Containers and storage Containers—Tight containers. to the Qualitative Tests <1.09> for sulfate. Storage—Light-resistant.

20 Optical rotation <2.49> [a]D : －78 – －909(0.5 g calculated on the dried basis, water, 25 mL, 100 mm). Polyoxyl 40 Stearate pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Polymixin B Sulfate in 50 mL of water is between ステアリン酸ポリオキシル 40 5.0 and 7.0. Phenylalanine Weigh accurately about 0.375 g of Polymix- Polyoxyl 40 Stearate is the monostearate of conden- in B Sulfate, dissolve in 0.1 mol/L hydrochloric acid TS to sation polymers of ethylene oxide represented by the make exactly 100 mL. Determine absorbances, A1, A2, A3, formula H(OCH2CH2)nOCOC17H35,inwhichn is ap- A4 and A5, of this solution at 252 nm, at 258 nm, at 264 nm, proximately 40. at 280 nm and at 300 nm, respectively, as directed under Description Polyoxyl 40 Stearate occurs as a white to light Ultraviolet-visible Spectrophotometry <2.24>, and calculate yellow, waxy solid or powder. It is odorless or has a faint the amount of phenylalanine by the following equation: the fat-like odor. amount of phenylalanine calculated on the dried basis is not It is soluble in water, in ethanol (95) and in diethyl ether. less than 9.0z and not more than 12.0z. Congealing point <2.42> 39.0 – 44.09C Amount (z) of phenylalanine

＝ (A2 － 0.5A1 ＋ 0.5A3 － 1.8A4 ＋ 0.8A5)/MT × 9.4787 Congealing point of the fatty acid <1.13> Not below 539C.

MT: Amount (g) of the sample, calculated on the dried Acid value <1.13> Not more than 1. basis Saponification value <1.13> 25–35 JP XVI Official Monographs / Potassium Bromide 1265

Purity (1) Clarity and color of solution—Dissolve 1.0 g Water <2.48> Not more than 3.0z (1 g, volumetric titra of Polyoxyl 40 Stearate in 20 mL of water: the solution is tion, back titration). clear and colorless. Residue on ignition <2.44> Not more than 0.1z (2 g). (2) Heavy metals <1.07>—Proceed with 2.0 g of Polyoxyl 40 Stearate according to Method 2, and perform the test. Containers and storage Containers—Tight containers. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (3) Arsenic <1.11>—Prepare the test solution with 0.67 g Potash Soap of Polyoxyl 40 Stearate, according to Method 3, and perform the test (not more than 3 ppm). カリ石ケン Residue on ignition <2.44> Not more than 0.1z (1 g). Potash Soap contains not less than 40.0z as fatty Containers and storage Containers—Tight containers. acids. Method of preparation Polysorbate 80 Fixed oil 470 mL Potassium Hydroxide a sufficient quantity ポリソルベート 80 Water, Purified Water or Purified Water in Containers a sufficient quantity Polysorbate 80 is a polyoxyethylene ether of anhy- To make 1000 g drous sorbitol, partially esterified with oleic acid. Dissolve Potassium Hydroxide, in required quantity for Description Polysorbate 80 is a colorless or orange-yellow, saponification, in Water, Purified Water or Purified Water viscous liquid, having a faint, characteristic odor and a in Containers, add this solution to fixed oil, previously warm, slightly bitter taste. warmed, add a sufficient quantity of Ethanol if necessary, It is miscible with methanol, with ethanol (95), with warm stir thoroughly, heat in a water bath, and continue the ethanol (95), with pyridine and with chloroform. saponification. After complete saponification, add Water, It is freely soluble in water and slightly soluble in diethyl Purified Water or Purified Water in Containers to make ether. 1000 g. The pH of a solution of Polysorbate 80 (1 in 20) is between 5.5 and 7.5. Description Potash Soap occurs as a yellow-brown, transparent, unctuous, soft mass, having a characteristic odor. Identification (1) To 5 mL of a solution of Polysorbate It is freely soluble in water and in ethanol (95). 80 (1 in 20) add 5 mL of sodium hydroxide TS, boil for 5 minutes, cool, and acidify with dilute hydrochloric acid: the Purity Silicic acid and alkalinity—Dissolve 10 g of Potash solution is opalescent. Soap in 30 mL of ethanol (95), and add 0.50 mL of 1 mol/L (2) To 5 mL of a solution of polysorbate 80 (1 in 20) add hydrochloric acid VS: no turbidity is produced. Add 1 drop 2 to 3 drops of bromine TS: the color of the test solution is of phenolphthalein TS to this solution: no red color devel- discharged. ops. (3) Mix 6 mL of Polysorbate 80 with 4 mL of water at an Assay Weigh accurately about 5 g of Potash Soap, dissolve ordinary, or lower than ordinary, temperature: a jelly-like in 100 mL of hot water, and transfer to a separator. Acidify mass is produced. the mixture with dilute sulfuric acid, and cool. Extract the (4) To 10 mL of a solution of Polysorbate 80 (1 in 20) solution with 50-mL, 40-mL, and 30-mL portions of diethyl add 5 mL of ammonium thiocyanate-cobalt (II) nitrate TS, ether. Wash the combined diethyl ether extracts with 10-mL shake well, add 5 mL of chloroform, shake, and allow to portions of water until the washing contains no acid. Trans- stand: a blue color develops in the chloroform layer. fer the diethyl ether solution to a tared flask, evaporate Viscosity <2.53> 345 – 445 mm2/s (Method 1, 259C). diethyl ether on a water bath at a temperature as low as pos-

20 sible. Dry the residue at 809C to constant mass, and weigh as Specific gravity <1.13> d 20: 1.065 – 1.095 fatty acids. Acid value <1.13> Not more than 2.0. Containers and storage Containers—Tight containers. Saponification value <1.13> 45–55 Iodine value <1.13> 19 – 24 Use chloroform instead of cyclohexane, and titrate <2.50> without using an indicator, Potassium Bromide until the yellow color of iodine disappears. 臭化カリウム Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Polysorbate 80 according to Method 2, and perform the test. KBr: 119.00 Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Potassium Bromide, when dried, contains not less (2) Arsenic <1.11>—Prepare the test solution with 1.0 g than 99.0z of KBr. of Polysorbate 80 according to Method 3, and perform the test (not more than 2 ppm). Description Potassium Bromide occurs as colorless or 1266 Potassium Canrenoate / Official Monographs JP XVI white crystals, granules or crystalline powder. It is odorless. It is freely soluble in water and in glycerin, soluble in hot Potassium Canrenoate ethanol (95), and slightly soluble in ethanol (95). カンレノ酸カリウム Identification A solution of Potassium Bromide (1 in 10) responds to Qualitative Tests <1.09> for potassium salt and for bromide. Purity (1) Clarity and color of solution—Dissolve 1.0 g of Potassium Bromide in 3 mL of water: the solution is clear and colorless. (2) Alkalinity—Dissolve 1.0 g of Potassium Bromide in

10 mL of water, add 0.10 mL of 0.05 mol/L sulfuric acid VS C22H29KO4: 396.56 and 1 drop of phenolphthalein TS, heat to boiling, and cool: Monopotassium 17-hydroxy-3-oxo-17a-pregna-4,6-diene- no color develops. 21-carboxylate (3) Chloride—Make a calculation from the result ob- [2181-04-6] tained in the Assay: not more than 84.5 mL of 0.1 mol/L silver nitrate VS is consumed for 1 g of Potassium Bromide. Potassium Canrenoate, when dried, contains not (4) Sulfate <1.14>—Proceed with 2.0 g of Potassium less than 98.0z and not more than 102.0z of Bromide, and perform the test. Prepare the control solution C22H29KO4. with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than Description Potassium Canrenoate occurs as a pale yellow- 0.024z). ish white to pale yellow-brown, crystalline powder. (5) Iodide—Dissolve 0.5 g of Potassium Bromide in 10 It is freely soluble in water, soluble in methanol, sparingly mL of water, add 2 to 3 drops of iron (III) chloride TS and 1 soluble in ethanol (95), and practically insoluble in chlo- mL of chloroform, and shake: no red-purple to purple color roform and in diethyl ether. develops in the chloroform layer. (6) Bromate—Dissolve 1.0 g of Potassium Bromide in 10 Identification (1) Dissolve 2 mg of Potassium Canrenoate mL of freshly boiled and cooled water, and add 0.1 mL of in 2 drops of sulfuric acid: an orange color develops. Ob- potassium iodide TS, 1 mL of starch TS and 3 drops of serve under ultraviolet light (main wavelength: 365 nm): the dilute sulfuric acid. Shake the mixture gently, and allow to solution shows a yellow-green fluorescence. Add 1 drop of stand for 5 minutes: no blue color develops. acetic anhydride to this solution: the color of the solution (7) Heavy metals <1.07>—Proceed with 2.0 g of Potas- changes to red. sium Bromide according to Method 1, and perform the test. (2) Determine the absorption spectrum of a solution of Prepare the control solution with 2.0 mL of Standard Lead Potassium Canrenoate in methanol (1 in 100,000) as directed Solution (not more than 10 ppm). under Ultraviolet-visible Spectrophotometry <2.24>,and (8) Barium—Dissolve 0.5 g of Potassium Bromide in 10 compare the spectrum with the Reference Spectrum: both mL of water, add 0.5 mL of dilute hydrochloric acid and 1 spectra exhibit similar intensities of absorption at the same mL of potassium sulfate TS, and allow to stand for 10 wavelengths. minutes: no turbidity is produced. (3) Determine the infrared absorption spectrum of Po- (9) Arsenic <1.11>—Prepare the test solution with 1.0 g tassium Canrenoate, previously dried, as directed in the of Potassium Bromide according to Method 1, and perform potassium bromide disk method under Infrared Spectropho- the test (not more than 2 ppm). tometry <2.25>, and compare the spectrum with the Refer- ence Spectrum: both spectra exhibit similar intensities of ab- Loss on drying <2.41> Not more than 1.0z (1 g, 1109C, sorption at the same wave numbers. 4 hours). (4) The solution of Potassium Canrenoate (1 in 10) re- Assay Weigh accurately about 0.4 g of Potassium sponds to Qualitative Tests <1.09> (1) for potassium salt. Bromide, previously dried, and dissolve in 50 mL of water. Optical rotation <2.49> [a]20: －71 – －769(after drying, Add 10 mL of dilute nitric acid and exactly measured 50 mL D 0.2 g, methanol, 20 mL, 100 mm). of 0.1 mol/L silver nitrate VS, and titrate <2.50> the excess silver nitrate with 0.1 mol/L ammonium thiocyanate VS (in- pH <2.54> Dissolve 1.0 g of Potassium Canrenoate in 20 dicator: 2 mL of ammonium iron (III) sulfate TS). Perform mL of water: the pH of this solution is between 8.4 and 9.4. a blank determination. Purity (1) Clarity and color of solution—Dissolve 0.5 g Each mL of 0.1 mol/L silver nitrate VS of Potassium Canrenoate in 5 mL of water: the solution is ＝ 11.90 mg of KBr clear, and shows a pale yellow to light yellow color. (2) Heavy metals <1.07>—Proceed with 2.0 g of Potas- Containers and storage Containers—Tight containers. sium Canrenoate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (3) Arsenic <1.11>—Prepare the test solution with 1.0 g of Potassium Canrenoate according to Method 3, and perform the test (not more than 2 ppm). (4) Canrenone—Place 0.40 g of Potassium Canrenoate in a glass-stoppered centrifuge tube, cool in ice-water to a JP XVI Official Monographs / Potassium Chloride 1267 temperature not higher than 59C, add 6 mL of boric acid- water bath to dryness, add 2 mL of dilute acetic acid and 2.0 potassium chloride-sodium hydroxide buffer solution, pH mL of Standard Lead Solution to dryness, and dilute with 10.0, being cooled to a temperature not higher than 59Cto water to 50 mL (not more than 20 ppm). dissolve, and add 8 mL of water being cooled to a tempera- (3) Sodium—Dissolve 1.0 g of Potassium Carbonate in ture not higher than 59C. Add exactly 10 mL of chloroform, 20 mL of water, and perform the test as directed under allow to stand for 3 minutes at a temperature not higher than Flame Coloration Test <1.04> (1): no persisting yellow color 59C, shake vigorously for 2 minutes, and centrifuge. Drain is produced. off the water layer, collect 5 mL of the chloroform layer, (4) Arsenic <1.11>—Prepare the test solution with 0.5 g transfer to a glass-stoppered centrifuge tube containing 3 mL of Potassium Carbonate, according to Method 1, and per- of boric acid-potassium chloride-sodium hydroxide buffer form the test (not more than 4 ppm). solution, pH 10.0, cooled to a temperature not higher than Loss on drying <2.41> Not more than 1.0z (3 g, 1809C, 59C, and 4 mL of water cooled to a temperature not higher 4 hours). than 59C, shake for 1 minute, and centrifuge. Drain off the water layer, pipet 2 mL of the chloroform layer, and add Assay Dissolve about 1.5 g of Potassium Carbonate, previ- chloroform to make exactly 10 mL. Determine the absor- ously dried and accurately weighed, in 25 mL of water, bance of this solution at 283 nm as directed under Ultravio- titrate with 0.5 mol/L sulfuric acid VS until the blue color of let-visible Spectrophotometry <2.24>: it is not more than the solution changes to yellow-green, boil cautiously, then 0.67. cool, and titrate <2.50> until a greenish yellow color develops (indicator: 2 drops of bromocresol green TS). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Each mL of 0.5 mol/L sulfuric acid VS ＝ 69.11 mg of K CO Assay Weigh accurately about 0.2 g of Potassium Can- 2 3 renoate, previously dried, dissolve in 75 mL of acetic acid Containers and storage Containers—Tight containers. (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Use a solution of saturated potassium chloride-acetic acid (100) as the internal liquid.). Per- Potassium Chloride form a blank determination, and make any necessary correction. 塩化カリウム Each mL of 0.1 mol/L perchloric acid VS

＝ 39.66 mg of C22H29KO4 KCl: 74.55 Containers and storage Containers—Tight containers. Potassium Chloride, when dried, contains not less than 99z of KCl. Potassium Carbonate Description Potassium Chloride occurs as colorless or white crystals or crystalline powder. It is odorless, and has a 炭酸カリウム saline taste. It is freely soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. K CO :138.21 2 3 A solution of Potassium Chloride (1 in 10) is neutral. Potassium Carbonate, when dried, contains not less Identification A solution of Potassium Chloride (1 in 50) than 99.0z of K2CO3. responds to Qualitative Tests <1.09> for potassium salt and for chloride. Description Potassium Carbonate occurs as white granules or powder. It is odorless. Purity (1) Clarity and color of solution—Dissolve 1.0 g It is very soluble in water, and practically insoluble in of Potassium Chloride in 5 mL of water: the solution is clear ethanol (95). and colorless. A solution of Potassium Carbonate (1 in 10) is alkaline. (2) Acidity and alkalinity—Dissolve 5.0 g of Potassium It is hygroscopic. Chloride in 50 mL of freshly boiled and cooled water, and add 3 drops of phenolphthalein TS: no red color develops. Identification A solution of Potassium Carbonate (1 in 10) Then add 0.50 mL of 0.01 mol/L sodium hydroxide VS: a responds to Qualitative Tests <1.09> for potassium salt and red color develops. for carbonate. (3) Bromide—Dissolve 1.0 g of Potassium Chloride in Purity (1) Clarity and color of solution—Dissolve 1.0 g water to make 100 mL. To 5 mL of the solution add 3 drops of Potassium Carbonate in 20 mL of water: the solution is of dilute hydrochloric acid and 1 mL of chloroform, and add clear and colorless. 3 drops of sodium toluensulfonchloramide TS dropwise (2) Heavy metals <1.07>—Dissolve 1.0 g of Potassium while shaking: no yellow to yellow-red color develops in the Carbonate in 2 mL of water and 6 mL of dilute hydrochloric chloroform layer. acid, and evaporate to dryness on a water bath. Dissolve the (4) Iodide—Dissolve 0.5 g of Potassium Chloride in 10 residue in 35 mL of water and 2 mL of dilute acetic acid, di- mL of water, add 3 drops of iron (III) chloride TS and 1 mL lute with water to 50 mL, and perform the test using this so- of chloroform, shake, allow to stand for 30 minutes, and lution as the test solution. Prepare the control solution as shake again: no red-purple to purple color develops in the follows: evaporate 6 mL of dilute hydrochloric acid on a chloroform layer. 1268 Potassium Clavulanate / Official Monographs JP XVI

(5) Heavy metals <1.07>—Proceed with 4.0 g of Potas- (2) Determine the infrared absorption spectrum of Po- sium Chloride according to Method 1, and perform the test. tassium Clavulanate as directed in the potassium bromide Prepare the control solution with 2.0 mL of Standard Lead disk method under Infrared Spectrophotometry <2.25>,and Solution (not more than 5 ppm). compare the spectrum with the Reference Spectrum: both (6) Calcium and magnesium—Dissolve 0.20 g of Potas- spectra exhibit similar intensities of absorption at the same sium Chloride in 20 mL of water, add 2 mL of ammonia TS, wave numbers. 2 mL of ammonium oxalate TS and 2 mL of disodium (3) Potassium Clavulanate responds to Qualitative Tests hydrogenphosphate TS, and then allow to stand for 5 <1.09> (1) for potassium salt. minutes: no turbidity is produced. Optical rotation <2.49> [a]20: ＋53 – ＋639(0.5 g calculated (7) Sodium—Dissolve 1.0 g of Potassium Chloride in 20 D on the anhydrous basis, water, 50 mL, 100 mm). mL of water, and perform the Flame Coloration Test <1.04> (1): no persistent, yellow color develops. Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of (8) Arsenic <1.11>—Prepare the test solution with 1.0 g Potassium Clavulanate according to Method 2, and perform of Potassium Chloride according to Method 1, and perform the test. Prepare the control solution with 4.0 mL of Stand- the test (not more than 2 ppm). ard Lead Solution (not more than 20 ppm). (2) Arsenic <1.11>—Prepare the test solution with 1.0 g Loss on drying <2.41> Not more than 0.5z (1 g, 1309C, of Potassium Clavulanate according to Method 3, and per- 2 hours). form the test (not more than 2 ppm). Assay Weigh accurately about 0.2 g of Potassium Chlo- (3) Related substances—Dissolve 0.10 g of Potassium ride, previously dried, dissolve in 50 mL of water, and titrate Clavulanate in 10 mL of the mobile phase A, and use this so- <2.50> with 0.1 mol/L silver nitrate VS while shaking vigor- lution as the sample solution. Pipet 1 mL of the sample solu- ously (indicator: 3 drops of fluorescein sodium TS). tion, add the mobile phase A to make exactly 100 mL, and use this solution as the standard solution. Perform the test Each mL of 0.1 mol/L silver nitrate VS ＝ 7.455 mg of KCl with exactly 20 mL each of the sample solution and standard Containers and storage Containers—Tight containers. solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of Potassium Clavulanate each peak other than clavulanic acid from the sample solution is not larger than the peak area of clavulanic acid from クラブラン酸カリウム the standard solution, and the total area of the peaks other than clavulanic acid from the sample solution is not larger than 2 times of the peak area of clavulanic acid from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 230 nm). C H KNO :237.25 8 8 5 Column: A stainless steel column 4.6 mm in inside diame- Monopotassium (2R,5R)-3-[(1Z)-2-hydroxyethylidene]-7- ter and 10 cm in length, packed with octadecylsilanized silica oxo-4-oxa-1-azabicyclo[3.2.0]heptane-2-carboxylate gel for liquid chromatography (5 mminparticlediameter). [61177-45-5] Column temperature: A constant temperature of about 409C. Potassium Clavulanate is the potassium salt of a Mobile phase A: Adjust the pH of 0.05 mol/L sodium substance having b-lactamase inhibiting activity pro- dihydrogen phosphate TS to 4.0 with phosphoric acid. duced by the growth of Streptomyces clavuligerus. Mobile phase B: A mixture of the mobile phase A and It contains not less than 810 mg (potency) and not methanol (1:1). more than 860 mg (potency) per mg, calculated on the Flowing of the mobile phase: Control the gradient by mix- anhydrous basis. The potency of Potassium Clavu- ing the mobile phases A and B as directed in the following lanate is expressed as mass (potency) of clavularic acid table. (C8H9NO5: 199.16).

Description Potassium Clavulanate occurs as a white to Time after injection Mobile phase A Mobile phase B light yellowish white, crystalline powder. of sample (min) (volz) (volz) It is very soluble in water, soluble in methanol, and slightly soluble in ethanol (95). 0– 4 100 0 It is hygroscopic. 4–15 100→ 00→ 100 15–25 0 100 Identification (1) To 1 mL of a solution of Potassium Clavulanate (1 in 50,000) add 5 mL of imidazole TS, and warm in a water bath at 309C for 12 minutes. After cooling, Flow rate: 1.0 mL per minute. determine the absorption spectrum of this solution as di- Time span of measurement: About 6 times as long as the rected under Ultraviolet-visible Spectrophotometry <2.24>, retention time of clavulanic acid. and compare the spectrum with the Reference Spectrum: System suitability— both spectra exhibit similar intensities of absorption at the Test for required detectability: Pipet 1 mL of the standard same wavelengths. solution, and add the mobile phase A to make exactly 10 JP XVI Official Monographs / Potassium Guaiacolsulfonate 1269 mL. Confirm that the peak area of clavulanic acid obtained from 20 mL of this solution is equivalent to 7 to 13z of that Potassium Guaiacolsulfonate from 20 mL of the standard solution. System performance: Dissolve 10 mg each of Potassium グアヤコールスルホン酸カリウム Clavulanate and Amoxycillin in 100 mL of the mobile phase A. When the procedure is run with 20 mL of this solution under the above operating conditions, clavulanic acid and amoxycillin are eluted in this order with the resolution between these peaks being not less than 8 and the number of theoretical plates of the peak of clavulanic acid is not less than 2500. C7H7KO5S: 242.29 System repeatability: When the test is repeated 3 times Monopotassium 4-hydroxy-3-methoxybenzenesulfonate with 20 mL of the standard solution under the above operat- [1321-14-8] ing conditions, the relative standard deviation of the peak area of clavulanic acid is not more than 2.0z. Potassium Guaiacolsulfonate contains not less than 98.5z of C H KO S, calculated on the anhydrous Water <2.48> Not more than 1.5z (5 g, volumetric titra- 7 7 5 basis. tion, direct titration). Description Potassium Guaiacolsulfonate occurs as white Assay Weigh accurately an amount of Potassium Clavu- crystals or crystalline powder. It is odorless or has a slight, lanate and Lithium Clavulanate RS, equivalent to about 12.5 characteristic odor and a slightly bitter taste. mg (potency), dissolve each in 30 mL of water, add exactly 5 It is freely soluble in water and in formic acid, soluble in mL of the internal standard solution and water to make 50 methanol, and practically insoluble in ethanol (95), in acetic mL, and use these solutions as the sample solution and the anhydride and in diethyl ether. standard solution, respectively. Perform the test with 5 mL each of the sample solution and standard solution as directed Identification (1) To 10 mL of a solution of Potassium under Liquid Chromatography <2.01> according to the fol- Guaiacolsulfonate (1 in 100) add 2 drops of iron (III) chlo- lowing conditions, and calculate the ratios, QT and QS,of ride TS: a blue-purple color develops. the peak area of clavularic acid to that of the internal stand- (2) Dissolve 0.25 g of Potassium Guaiacolsulfonate in ard. water to make 500 mL, and to 10 mL of this solution add phosphate buffer solution, pH 7.0, to make 100 mL. Deter- Amount [mg (potency)] of clavularic acid (C H NO ) 8 9 5 mine the absorption spectrum of this solution as directed ＝ M × Q /Q × 1000 S T S under Ultraviolet-visible Spectrophotometry <2.24>,and

MS: Amount [mg (potency)] of Lithium Clavulanate RS compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same Internal standard solution—Dissolve 0.3 g of sulfanilamide wavelengths. in 30 mL of methanol, and add water to make 100 mL. (3) A solution of Potassium Guaiacolsulfonate (1 in 10) Operating conditions— responds to Qualitative Tests <1.09> for potassium salt. Detector: An ultraviolet absorption photometer (wavelength: 230 nm). pH <2.54> Dissolve 1.0 g of Potassium Guaiacolsulfonate Column: A stainless steel column 4.6 mm in inside diame- in 20 mL of water: the pH of the solution is between 4.0 and ter and 25 cm in length, packed with octadecylsilanized silica 5.5. gel for liquid chromatography (5 mm in particle diameter). Purity (1) Clarity and color of solution—Dissolve 1.0 g Column temperature: A constant temperature of about of Potassium Guaiacolsulfonate in 20 mL of water: the solu- 259C. tion is clear and colorless. Mobile phase: Dissolve 1.36 g of sodium acetate trihydrate (2) Sulfate <1.14>—Perform the test with 0.8 g of Potas- in 900 mL of water, adjust to pH 4.5 with diluted acetic acid sium Guaiacolsulfonate. Prepare the control solution with (31) (2 in 5), and add 30 mL of methanol and water to make 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 1000 mL. 0.030z). Flow rate: Adjust the flow rate so that the retention time (3) Heavy metals <1.07>—Proceed with 1.0 g of Potas- of clavularic acid is about 6 minutes. sium Guaiacolsulfonate according to Method 1, and perform System suitability— the test. Prepare the control solution with 2.0 mL of Stand- System performance: When the procedure is run with 5 mL ard Lead Solution (not more than 20 ppm). of the standard solution under the above operating condi- (4) Arsenic <1.11>—Prepare the test solution with 1.0 g tions, clavularic acid and the internal standard are eluted in of Potassium Guaiacolsulfonate according to Method 1, and this order with the resolution between these peaks being not perform the test (not more than 2 ppm). less than 4. (5) Related substances—Dissolve 0.20 g of Potassium System repeatability: When the test is repeated 6 times Guaiacolsulfonate in 200 mL of mobile phase, and use this with 5 mL of the standard solution under the above operating solution as the sample solution. Pipet 1 mL of the sample so- conditions, the relative standard deviation of the ratios of lution, add the mobile phase to make exactly 100 mL, and the peak area of clavularic acid to that of the internal stand- use this solution as the standard solution. Perform the test ard is not more than 1.0z. with exactly 5 mL each of the sample solution and standard Containers and storage Containers—Tight containers. solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak 1270 Potassium Hydroxide / Official Monographs JP XVI area obtained from these solutions by the automatic integra- Identification (1) A solution of Potassium Hydroxide (1 tion method: the total area of peaks other than the peak of in 500) is alkaline. potassium guaiacolsulfonate from the sample solution is not (2) A solution of Potassium Hydroxide (1 in 25) re- larger than the peak area of potassium guaiacolsulfonate sponds to Qualitative Tests <1.09> for potassium salt. from the standard solution. Purity (1) Clarity and color of solution—Dissolve 1.0 g Operating conditions— of Potassium Hydroxide in 20 mL of water: the solution is Detector: An ultraviolet absorption photometer (wave- clear and colorless. length: 279 nm). (2) Chloride <1.03>—Dissolve 2.0 g of Potassium Hy- Column: A stainless steel column 4 mm in inside diameter droxide in water, and add water to make 100 mL. To 25 mL and 20 to 25 cm in length, packed with dimethylamino- of the solution add 8 mL of dilute nitric acid and water to propylsilanized silica gel for liquid chromatography (5 to 10 make 50 mL. Perform the test using this solution as the test mm in particle diameter). solution. Prepare the control solution with 0.7 mL of 0.01 Column temperature: A constant temperature of about mol/L hydrochloric acid VS (not more than 0.050z). 309C. (3) Heavy metals <1.07>—Dissolve 1.0 g of Potassium Mobile phase: A mixture of 0.05 mol/L potassium dihy- Hydroxide in 5 mL of water, add 7 mL of dilute hydrochlo- drogenphosphate TS and methanol (20:1). ric acid, and evaporate on a water bath to dryness. Dissolve Flow rate: Adjust the flow rate so that the retention time the residue in 35 mL of water, 2 mL of dilute acetic acid and of potassium guaiacolsulfonate is about 10 minutes. 1 drop of ammonia TS, add water to make 50 mL, and per- Selection of column: Weigh 50 mg each of potassium form the test using this solution as the test solution. Prepare guaiacolsulfonate and guaiacol, and dissolve in 50 mL of the the control solution as follows: evaporate 7 mL of dilute hy- mobile phase. Proceed with 5 mL of this solution under the drochloric acid on a water bath to dryness, dissolve the above operating conditions, and calculate the resolution. residue in 2 mL of dilute acetic acid and 3.0 mL of Standard Use a column giving elution of guaiacol and potassium Lead Solution, and add water to make 50 mL (not more than guaiacolsulfonate in this order with the resolution of these 30 ppm). peaks being not less than 4. (4) Sodium—Dissolve 0.10 g of Potassium Hydroxide in Detection sensitivity: Adjust the sensitivity so that the 10 mL of dilute hydrochloric acid, and perform the test as peak height of potassium guaiacolsulfonate from 5 mLofthe directed under Flame Coloration Test <1.04> (1): no persis- standard solution is not less than 10 mm. tent yellow color develops. Time span of measurement: About twice as long as the (5) Potassium carbonate—The amount of potassium car- retention time of potassium guaiacolsulfonate. bonate (K2CO3: 138.21) is not more than 2.0z when calcu- Water <2.48> 3.0 – 4.5z (0.3 g, volumetric titration, direct lated by the following equation using B (mL) obtained in the titration). Assay. Assay Weigh accurately about 0.3 g of Potassium Guaia- Amount of potassium carbonate (mg) ＝ 138.21 × B colsulfonate, dissolve in 2.0 mL of formic acid, add 50 mL Assay Weigh accurately about 1.5 g of Potassium Hydrox- of acetic anhydride, and titrate <2.50> with 0.1 mol/L per- ide, and dissolve in 40 mL of freshly boiled and cooled chloric acid VS (potentiometric titration). Perform a blank water. Cool the solution to 159C, add 2 drops of phenol- determination, and make any necessary correction. phthalein TS, and titrate <2.50> with 0.5 mol/L sulfuric acid Each mL of 0.1 mol/L perchloric acid VS VS until the red color of the solution disappears. Record the

＝ 24.23 mg of C7H7KO5S amount A (mL) of 0.5 mol/L sulfuric acid VS consumed, then add 2 drops of methyl orange TS, and titrate <2.50> Containers and storage Containers—Well-closed contain- again with 0.5 mol/L sulfuric acid VS until the solution ers. changes to a persistent light red color. Record the amount B Storage—Light-resistant. (mL) of 0.5 mol/L sulfuric acid VS consumed. Calculate the amount KOH from the amount, A (mL) － Potassium Hydroxide B (mL). Each mL of 0.5 mol/L sulfuric acid VS 水酸化カリウム＝ 56.11 mg of KOH Containers and storage Containers—Tight containers. KOH: 56.11

Potassium Hydroxide contains not less than 85.0z Potassium Iodide of KOH. ヨウ化カリウム Description Potassium Hydroxide occurs as white fused masses, in small pellets, in flakes, in sticks and in other forms. It is hard and brittle, and shows a crystalline fracture. KI: 166.00 It is freely soluble in water and in ethanol (95), and practically insoluble in diethyl ether. Potassium Iodide, when dried, contains not less It rapidly absorbs carbon dioxide in air. than 99.0z of KI. It deliquesces in the presence of moisture. Description Potassium Iodide occurs as colorless or white JP XVI Official Monographs / Potassium Permanganate 1271 crystals, or a white crystalline powder. Each mL of 0.05 mol/L potassium iodate VS It is very soluble in water, soluble in ethanol (95), and ＝ 16.60 mg of KI practically insoluble in diethyl ether. Containers and storage Containers—Tight containers. It is slightly deliquescent in moist air. Storage—Light-resistant. Identification A solution of Potassium Iodide (1 in 20) responds to Qualitative Tests <1.09> for potassium salt and for iodide. Potassium Permanganate Purity (1) Clarity and color of solution—Dissolve 1.0 g 過マンガン酸カリウム of Potassium Iodide in 2 mL of water: the solution is clear and colorless.

(2) Alkalinity—Dissolve 1.0 g of Potassium Iodide in 10 KMnO4: 158.03 mL of freshly boiled and cooled water, and add 0.50 mL of 0.005 mol/L sulfuric acid VS and 1 drop of phenolphthalein Potassium Permanganate, when dried, contains not TS: no color develops. less than 99.0z of KMnO4. (3) Chloride, bromide and thiosulfate—Dissolve 0.20 g Description Potassium Permanganate occurs as dark pur- of Potassium Iodide in 5 mL of ammonia TS, add 15.0 mL ple crystals and has a metallic luster. of 0.1 mol/L silver nitrate VS, shake for 2 to 3 minutes, and It is soluble in water. filter. To 10 mL of the filtrate, add 15 mL of dilute nitric A solution of Potassium Permanganate (1 in 1000) has a acid: no brown color develops. The solution has no more slightly sweet, astringent taste. turbidity than that of the following control solution. Control solution: To 0.30 mL of 0.01 mol/L hydrochloric Identification A solution of Potassium Permanganate (1 in acid VS add 2.5 mL of ammonia TS, and 7.5 mL of 0.1 100) responds to Qualitative Tests <1.09> for permanganate. mol/L silver nitrate VS and 15 mL of dilute nitric acid. Purity (1) Water-insoluble substances—Dissolve 2.0 g of (4) Nitrate, nitrite and ammonium—Place 1.0 g of Po- Potassium Permanganate, previously powdered, in 200 mL tassium Iodide in a 40-mL test tube, and add 5 mL of water, of water. Filter the insoluble substances through a tared 5 mL of sodium hydroxide TS and 0.2 g of aluminum wire. glass filter (G4), wash with water until the last washing Insert the absorbent cotton in the mouth of the test tube, shows no color, and dry at 1059C for 2 hours: the mass of and place a piece of moistened red litmus paper on it. Heat the residue is not more than 4 mg. the test tube carefully on a water bath for 15 minutes: the gas (2) Arsenic <1.11>—Dissolve 0.40 g of Potassium Per- evolved does not turn red litmus paper to blue. manganate in 10 mL of water, add 1 mL of sulfuric acid, (5) Cyanide—Dissolve 0.5 g of Potassium Iodide in 10 add hydrogen peroxide (30) dropwise until the solution mL of water. To 5 mL of this solution add 1 drop of iron (II) remains colorless, and evaporate on a sand bath nearly to sulfate TS and 2 mL of sodium hydroxide TS, warm, then dryness. Dissolve the residue in 5 mL of water, and perform add 4 mL of hydrochloric acid: no green color develops. the test with this solution as the test solution: the color pro- (6) Iodate—Dissolve 0.5 g of Potassium Iodide in 10 mL duced is not more intense than the following standard color. of freshly boiled and cooled water, and add 2 drops of dilute Standard color: To 10 mL of water add 1 mL of sulfuric sulfuric acid and 1 drop of starch TS: no blue color develops acid and the same volume of hydrogen peroxide (30) as used immediately. for the preparation of the test solution. Evaporate the solu- (7) Heavy metals <1.07>—Proceed with 2.0 g of Potas- tion on a sand bath nearly to dryness, add 2.0 mL of Stand- sium Iodide according to Method 1, and perform the test. ard Arsenic Solution and water to make 5 mL, and carry out Prepare the control solution with 2.0 mL of Standard Lead the test with this solution in the same manner as the test solu- Solution (not more than 10 ppm). tion (not more than 5 ppm). (8) Barium—Dissolve 0.5 g of Potassium Iodide in 10 mL of water, add 1 mL of dilute sulfuric acid, and allow to Loss on drying <2.41> Not more than 0.5z (1 g, silica gel, stand for 5 minutes: no turbidity is produced. 18 hours). (9) Sodium—Dissolve 1.0 g of Potassium Iodide in 10 Assay Weigh accurately about 0.6 g of Potassium Perman- mL of water, and perform the Flame Coloration Test (1) ganate, previously dried, dissolve in water to make exactly <1.04>: a yellow color develops, but does not persist. 200 mL, and use this solution as the sample solution. Pipet (10) Arsenic <1.11>—Prepare the test solution with 25 mL of 0.05 mol/L oxalic acid VS into a 500-mL conical 0.40 g of Potassium Iodide according to Method 1, and flask, add 200 mL of diluted sulfuric acid (1 in 20), and keep perform the test (not more than 5 ppm). at a temperature between 309Cand359C. Transfer the sam- Loss on drying <2.41> Not more than 1.0z (2 g, 1059C, ple solution to a buret. Add quickly 23 mL of the sample so- 4 hours). lution from the buret to the flask while shaking gently, and then allow the flask to stand until the red color disappears. Assay Weigh accurately about 0.5 g of Potassium Iodide, Warm the mixture to a temperature between 559Cand609C, previously dried, in an iodine flask, dissolve in 10 mL of and continue the titration <2.50> slowly until the red color water, add 35 mL of hydrochloric acid and 5 mL of chlo- persists for 30 seconds. roform, and titrate <2.50> with 0.05 mol/L potassium iodate VS with shaking until the red-purple color of the chloroform Each mL of 0.05 mol/L oxalic acid VS layer disappears. The end point is reached when the red-pur- ＝ 3.161 mg of KMnO4 ple color does not reappear in the chloroform layer within 5 Containers and storage Containers—Tight containers. minutes after the layer has been decolorized. 1272 Potassium Sulfate / Official Monographs JP XVI

Potassium Sulfate Potato Starch

硫酸カリウム Amylum Solani

バレイショデンプン K2SO4: 174.26 This monograph is harmonized with the European Phar- Potassium Sulfate, when dried, contains not less macopoeia and the U.S. Pharmacopeia. The parts of the text than 99.0z of K2SO4.  that are not harmonized are marked with symbols ( ). Description Potassium Sulfate occurs as colorless crystals or a white, crystalline powder. It has a slightly saline, some- Potato Starch consists of starch granules derived what bitter taste. from the tuber of Solanum tuberosum Linneá It is soluble in water and practically insoluble in ethanol (Solanaceae). (95). Description Potato Starch occurs as a white powder. Identification A solution of Potassium Sulfate (1 in 20) re- It is practically insoluble in water and in ethanol (99.5). sponds to Qualitative Tests <1.09> for potassium salt and for Identification (1) Examined under a microscope <5.01> sulfate. using a mixture of water and glycerin (1:1), Potato Starch Purity (1) Clarity and color of solution, and acid or presents granules, either irregularly shaped, ovoid or pear- alkali—Dissolve 1.0 g of Potassium Sulfate in 20 mL of shaped, usually 30 – 100 mm in size but occasionally water: the solution is clear, colorless and neutral. exceeding 100 mm, or rounded, 10 – 35 mm in size. There are (2) Chloride <1.03>—Perform the test with 0.5 g of occasional compound granules having two to four compo- Potassium Sulfate. Prepare the control solution with 0.40 nents. The ovoid and pear-shaped granules have an eccentric mL of 0.01 mol/L hydrochloric acid VS (not more than hilum and the rounded granules acentric or slightly eccentric 0.028z). hilum. All granules show clearly visible concentric striations. (3) Heavy metals <1.07>—Proceed with 2.0 g of Potas- Between orthogonally oriented polarizing plates or prisms, sium Sulfate according to Method 1, and perform the test. the granules show a distinct black cross intersecting at the hi- Prepare the control solution with 2.0 mL of Standard Lead lum. Solution (not more than 10 ppm). (2) To 1 g of Potato Starch add 50 mL of water, boil for (4) Sodium—Dissolve 1.0 g of Potassium Sulfate in 20 1 minute, and allow to cool: a subtle white-turbid, pasty liq- mL of water, and perform the test as directed under Flame uid is formed. Coloration Test <1.04> (1): no persistent yellow color devel- (3) To 1 mL of the pasty liquid obtained in (2) add 0.05 ops. mL of diluted iodine TS (1 in 10): an orange-red to deep blue (5) Arsenic <1.11>—Prepare the test solution with 0.40 g color is formed, and the color disappears by heating. of Potassium Sulfate according to Method 1, and perform pH <2.54> Put 5.0 g of Potato Starch in a non-metal vessel, the test (not more than 5 ppm). add 25.0 mL of freshly boiled and cooled water, mix gently Loss on drying <2.41> Not more than 1.0z (1 g, 1109C, for 1 minute, and allow to stand for 15 minutes: the pH of 4 hours). the solution is between 5.0 and 8.0. Assay Weigh accurately about 0.5 g of Potassium Sulfate, Purity (1) Iron—To 1.5 g of Potato Starch add 15 mL of previously dried, boil with 200 mL of water and 1.0 mL of 2 mol/L hydrochloric acid TS, mix, filter, and use the fil- hydrochloric acid, and add gradually 8 mL of boiling barium trate as the test solution. To 2.0 mL of Standard Iron Solu- chloride TS. Heat the mixture on a water bath for 1 hour, tion add water to make 20 mL, and use as the control solu- collect the precipitate, and wash the precipitate with water tion. Put 10 mL each of the test solution and the control until the last washing shows no opalescence on the addition solution in test tubes, add 2 mL of a solution of citric acid (1 of silver nitrate TS. Dry, heat strongly to constant mass be- in 5) and 0.1 mL of mercapto acetic acid, and mix. Alkalize tween 5009Cand6009C by raising the temperature gradu- with ammonia solution (28) to litmus paper, add water to ally, and weigh as barium sulfate (BaSO4:233.39). make 20 mL, and mix. Transfer 10 mL each of these solutions into test tubes, allow to stand for 5 minutes, and com- Amount (mg) of K SO 2 4 pare the color of these solutions against a white background: ＝ amount (mg) of barium sulfate (BaSO ) × 0.747 4 the color of the test solution is not darker than that of the Containers and storage Containers—Well-closed contain- control solution (not more than 10 ppm). ers. (2) Oxidizing substances—To 4.0 g of Potato Starch add 50.0 mL of water, shake for 5 minutes, and centrifuge. To 30.0 mL of the supernatant liquid add 1 mL of acetic acid (100) and 0.5 to 1.0 g of potassium iodide, shake, and allow to stand for 25 to 30 minutes at a dark place. Add 1 mL of starch TS, and titrate <2.50> with 0.002 mol/L sodium thiosulfate VS until the color of the solution disappears. Per- form a blank determination and make any necessary correction: the volume of 0.002 mol/L sodium thiosulfate VS consumed is not more than 1.4 mL (not more than 20 ppm, JP XVI Official Monographs / Povidone 1273 calculated as hydrogen peroxide). ments of the tissue of the original plant. (3) Sulfur dioxide— Loss on drying <2.41> Not more than 20.0z (1 g, 1309C, (i) Apparatus Use as shown in the figure. 90 minutes). Residue on ignition <2.44> Not more than 0.6z (1 g). Containers and storage Containers—Well-closed contain-

ers.

Povidone Polyvidone Polyvinylpyrrolidone

ポビドン

(C6H9NO)n Poly[(2-oxopyrrolidin-1-yl)ethylene] A: Boiling flask (500 mL) [9003-39-8] B: Funnel (100 mL) C: Condenser Povidone is a chain polymer of 1-vinyl-2-pyrroli- D: Test-tube done. E: Tap It contains not less than 11.5z and not more than 12.8z of nitrogen (N: 14.01), calculated on the anhy- (ii) Procedure Introduce 150 mL of water into the boil- drous basis. ing flask, close the tap of the funnel, and pass carbon diox- It has a nominal K-value of not less than 25 and not ide through the whole system at a rate of 100 ± 5mLper more than 90. minute. Pass cooling water through the condenser, and place The nominal K-value is shown on the label. 10 mL of hydrogen peroxide-sodium hydroxide TS in the test-tube. After 15 minutes, remove the funnel without inter- Description Povidone occurs as a white to slightly yellow- rupting the stream of carbon dioxide, and introduce through ish fine powder. It is odorless or has a faint, characteristic the opening into the flask about 25 g of Potato Starch, accu- odor. rately weighed, with the aid of 100 mL of water. Apply tap It is freely soluble in water, in methanol and in ethanol grease to the outside of the connection part of the funnel, (95), slightly soluble in acetone, and practically insoluble in and load the funnel. Close the tap of the funnel, pour 80 mL diethyl ether. of 2 mol/L hydrochloric acid TS into the funnel, open the It is hygroscopic. tap to introduce the hydrochloric acid into the flask, and Identification Determine the infrared absorption spectrum close the tap while several mL of the hydrochloric acid of Povidone, previously dried at 1059C for 6 hours, as di- remains, in order to avoid losing sulfur dioxide. Place the rected in the potassium bromide disk method under Infrared flask in a water bath, and heat the mixture for 1 hour. Spectrophotometry <2.25>, and compare the spectrum with Transfer the contents of the test-tube with the aid of a little the Reference Spectrum or the spectrum of Povidone RS water to a wide-necked conical flask. Heat in a water bath previously dried at 1059C for 6 hours: both spectra exhibit for 15 minutes, and cool. Add 0.1 mL of bromophenol blue similar intensities of absorption at the same wave numbers. TS, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS until the color changes from yellow to violet-blue lasting for pH <2.54> Dissolve 1.0 g of Povidone in 20 mL of water: at least 20 seconds. Perform a blank determination and the pH of this solution is between 3.0 and 5.0 for Povidone make any necessary correction. Calculate the amount of sul- having the nominal K-value of 30 or less, and between 4.0 fur dioxide by applying the following formula: it is not more and 7.0 for Povidone having the nominal K-value exceeding than 50 ppm. 30. Amount (ppm) of sulfur dioxide ＝ V/M × 1000 × 3.203 Purity (1) Clarity and color of solution—Dissolve 1.0 g of Povidone in 20 mL of water: the solution is clear and col- M: Amount (g) of the sample orless to pale yellow, or pale red. V: Amount (mL) of 0.1 mol/L sodium hydroxide VS con- (2) Heavy metals <1.07>—Proceed with 2.0 g of Povi- sumed done according to Method 2, and perform the test. Prepare (4) Foreign matter—Under a microscope <5.01>, the control solution with 2.0 mL of Standard Lead Solution Potato Starch does not contain starch granules of any other (not more than 10 ppm). origin. It may contain a minute quantity, if any, of frag- (3) Aldehydes—Weigh accurately about 1.0 g of Povi- 1274 Povidone / Official Monographs JP XVI done and dissolve in 0.05 mol/L pyrophosphate buffer solu- Mobile phase: A mixture of water and methanol (4:1). tion, pH 9.0 to make exactly 100 mL. Stopper, heat at 609C Flow rate: Adjust the flow rate so that the retention time for 60 minutes, allow to cool to room temperature, and use of 1-vinyl-2-pyrrolidone is about 10 minutes. this solution as the sample solution. Separately, dissolve Selection of column: Dissolve 0.01 g of 1-vinyl-2-pyrroli- 0.100 g of freshly distilled acetaldehyde in water previously done and 0.5 g of vinyl acetate in 100 mL of methanol. To 1 cooled to 49C to make exactly 100 mL. Allow to stand at mL of this solution add diluted methanol (1 in 5) to make 49C for about 20 hours, pipet 1 mL of this solution, add 100 mL. Proceed with 50 mL of this solution under the above 0.05 mol/L pyrophosphate buffer solution, pH 9.0 to make operating conditions, and calculate the resolution. Use a exactly 100 mL, and use this solution as the standard solu- column giving elution of 1-vinyl-2-pyrrolidone and vinyl ace- tion. Measure 0.5 mL each of the sample solution, standard tate in this order with the resolution between these peaks solution and water (for blank test), transfer to separate cells, being not less than 2.0. add 2.5 mL of 0.05 mol/L pyrophosphate buffer solution, Detection sensitivity: Adjust the detection sensitivity so pH 9.0, and 0.2 mL of b-nicotinamide adenine dinucleotide that the peak height of 1-vinyl-2-pyrrolidone obtained from TS to each of these cells, mix and stopper tightly. Allow to 50 mL of the standard solution is between 10 mm and 15 mm. stand for 2 to 3 minutes at 22 ± 29C, and perform the test System repeatability: When the test is repeated 6 times with these solutions as directed under Ultraviolet-visible with the standard solution under the above operating condi- Spectrophotometry <2.24> using water as the control solutions, the relative standard deviation of obtained peak areas tion. Determine the absorbances, AT1, AS1 and AB1 of the of 1-vinyl-2-pyrrolidone is not more than 2z. subsequent solutions of the sample solution, the standard so- Washing of the guard column: After each test with the lution and water at 340 nm. Add 0.05 mL of aldehyde de- sample solution, wash away the polymeric material of Povi- hydrogenase solution to each of the cells, mix and stopper done from the guard column by passing the mobile phase tightly. Allow to stand for 5 minutes at 22 ± 29C. Deter- through the column backwards for about 30 minutes at the mine the absorbances, AT2, AS2 and AB2 of these solutions in same flow rate as applied in the test. the same manner as above: the content of aldehydes is not (5) Peroxides—Weigh exactly an amount of Povidone, more than 500 ppm (expressed as acetaldehyde). equivalent to 4.0 g calculated on the anhydrous basis, dissolve in water to make exactly 100 mL, and use this solution Content (ppm) of aldehydes expressed as acetaldehyde as the sample solution. To 25 mL of the sample solution add 1000 (A － A ) － (A － A ) 2 mL of titanium (III) chloride-sulfuric acid TS, and mix. ＝ × T2 T1 B2 B1 M (AS2 － AS1) － (AB2 － AB1) Allow to stand for 30 minutes, and perform the test with this solution as directed under Ultraviolet-visible Spectropho- M: Amount (g) of Povidone, calculated on the anhydrous tometry <2.24>, using a solution prepared by adding 2 mL of basis 13z sulfuric acid to 25 mL of the sample solution as a (4) 1-Vinyl-2-pyrrolidone—Weigh accurately about blank: the absorbance of the subsequent solution of the sam- 0.25 g of Povidone, dissolve in diluted methanol (1 in 5) to ple solution at 405 nm is not more than 0.35 (not more than make exactly 10 mL, and use this solution as the sample so- 400 ppm, expressed as hydrogen peroxide). lution. Separately, dissolve 50 mg of 1-vinyl-2-pyrrolidone in (6) Hydrazine—Transfer 2.5 g of Povidone to a 50-mL methanol to make exactly 100 mL. Pipet 1 mL of this solu- centrifuge tube, add 25 mL of water, and stir to dissolve. tion and add methanol to make exactly 100 mL. Pipet 5 mL Add 500 mL of a solution of salicylaldehyde in methanol (1 of this solution, add diluted methanol (1 in 5) to make ex- in 20), stir and warm at 609C for 15 minutes in a water bath. actly 100 mL, and use this solution as the standard solution. Allow to cool, add 2.0 mL of toluene, stopper tightly, shake Perform the test with exactly 50 mLeachofthesamplesolu- vigorously for 2 minutes, centrifuge, and use the upper layer tion and standard solution as directed under Liquid Chroma- of the mixture as the sample solution. Separately, dissolve tography <2.01> according to the following conditions, and 0.09 g of salicylaldazine in toluene to make exactly 100 mL. determine the peak areas, AT and AS, of 1-vinyl-2-pyrroli- Pipet 1 mL of this solution, add toluene to make exactly 100 done in each solution: the content of 1-vinyl-2-pyrrolidone is mL, and use this solution as the standard solution. Perform not more than 10 ppm. the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solu- Content (ppm) of 1-vinyl-2-pyrrolidone tion and standard solution on a plate coated with a 0.25-mm ＝ 2.5/M × A /A T S layer of dimethylsilanized silica gel with fluorescent indica- M: Amount (g) of Povidone, calculated on the anhydrous tor for thin-layer chromatography. Develop the plate with a basis mixture of methanol and water (2:1) to a distance of about three-fourths of the length of the plate, and air-dry the plate. Operating conditions— Examine under ultraviolet light (main wavelength: 365 nm): Detector: An ultraviolet spectrophotometer (detection the Rf value of the fluorescent spot from the standard solu- wavelength: 254 nm). tion is about 0.3, and the fluorescence of the spot from the Column: Stainless steel columns about 4 mm in inside di- sample solution corresponding to the spot from the standard ameter and about 25 mm in length, and about 4 mm in inside solution is not more intense than that of the spot from the diameter and about 250 mm in length, packed with octyl- standard solution (not more than 1 ppm). silanized silica gel for liquid chromatography (5 mminparticle diameter), and use them as a guard column and a separa- Water <2.48> Not more than 5.0z (0.5 g, volumetric titration column, respectively. tion, direct titration). Column temperature: A constant temperature of about Residue on ignition <2.44> Not more than 0.1z (1 g). 409C. JP XVI Official Monographs / Povidone-Iodine 1275

K-value Weigh accurately an amount of Povidone, equivalent to 1.00 g calculated on the anhydrous basis, and dissolve Povidone-Iodine in water to make exactly 100 mL, allow to stand for 60 minutes, and use this solution as the sample solution. Per- ポビドンヨード form the test with the sample solution and with water at 259C as directed in Method 1 under Viscosity Determination <2.53>, and calculate the K-value by the following formula. 1.5 log h － 1 300 c log h ＋ (c ＋ 1.5 c log h )2 K ＝ rel ＋ rel rel 0.15 ＋ 0.003 c 0.15 c ＋ 0.003 c2

c: Mass (g) of Povidone in 100 mL of the solution, calcu- (C6H9NO)n.xI lated on the anhydrous basis Poly[(2-oxopyrrolidin-1-yl)ethylene] iodine

hrel: Kinematic viscosity of the sample solution relative to [25655-41-8] that of water Povidone-Iodine is a complex of iodine with 1-vinyl- The K-value of Povidone is not less than 90z and not 2-pyrrolidone polymer. more than 108z of the nominal K-value. It contains not less than 9.0z and not more than Assay Weigh accurately about 0.1 g of Povidone, and 12.0z of available iodine (I: 126.90), and not less than place in a Kjeldahl flask. Add 5 g of a powdered mixture of 9.5z and not more than 11.5z of nitrogen (N: 33 g of potassium slfate, 1 g of copper (II) sulfate pentahy- 14.01), calculated on the dried basis. drate and 1 g of titanium (IV) oxide, and wash down any ad- Description Povidone-Iodine occurs as a dark red-brown hering sample from the neck of the flask with a small powder. It has a faint, characteristic odor. amount of water. Add 7 mL of sulfuric acid allowing to flow It is freely soluble in water and in ethanol (99.5). down the inside wall of the flask. Heat the flask gradually The pH of a solution obtained by dissolving 1.0 g of over a free flame until the solution has a clear, yellow-green Povidone-Iodine in 100 mL of water is between 1.5 and 3.5. color and the inside wall of the flask is free from a car- bonaceous material, and then heat for further 45 minutes. Identification (1) To10mLofdilutedstarchTS(1in10) After cooling, add cautiously 20 mL of water, cool the solu- add 1 drop of a solution of Povidone-Iodine (1 in 10): a deep tion, and connect the flask to the distillation apparatus pre- blue color develops. viously washed by passing steam through it. To the absorp- (2) To 1 mL of a solution of Povidone-Iodine (1 in 100) tion flask add 30 mL of a solution of boric acid (1 in 25), 3 add 1 mL of sodium thiosulfate TS, and add 1 mL of ammo- drops of bromocresol green-methyl red TS and sufficient nium thiocyanate-cobalt (II) nitrate TS and 2 drops of 1 water to immerse the lower end of the condenser tube. Add mol/L hydrochloric acid TS: a blue color develops, and a 30 mL of a solution of sodium hydroxide (2 in 5) through the blue precipitate is gradually formed. funnel, rinse cautiously the funnel with 10 ml of water, im- Purity (1) Clarity and color of solution—Dissolve 0.30 g mediately close the clamp attached to the rubber tube, then of Povidone-Iodine in 100 mL of water: the solution is clear start the distillation with steam to get 80 to 100 mL of the and brown. distillate. Remove the absorption flask from the lower end (2) Heavy metals <1.07>—Proceedwith1.0gof of the condenser tube, rinsing the end part with a small Povidone-Iodine according to Method 2, and perform the quantity of water, and titrate <2.50> the distillate with 0.025 test. Prepare the control solution with 2.0 mL of Standard mol/L sulfuric acid VS until the color of the solution Lead Solution (not more than 20 ppm). changes from green through pale grayish blue to pale grayish (3) Arsenic <1.11>—Prepare the test solution with 1.0 g red-purple. Perform a blank determination in the same man- of Povidone-Iodine according to Method 4, and perform the ner, and make any necessary correction. test (not more than 2 ppm). Each mL of 0.025 mol/L sulfuric acid VS (4) Iodide ion—Weigh accurately about 0.5 g of ＝ 0.700 mg of N Povidone-Iodine, dissolve in 100 mL of water, and add sodium hydrogensulfite TS until the color of iodine completely Containers and storage Containers—Tight containers. disappears. To this solution add exactly 25 mL of 0.1 mol/L silver nitrate VS, shake well with 10 mL of nitric acid, titrate <2.50> the excess silver nitrate with 0.1 mol/L ammonium thiocyanate VS until the solution develops a red-brown color, and calculate the total amount of iodine (indicator: 1 mL of ammonium iron (III) sulfate TS). Perform a blank determination. Each mL of 0.1 mol/L ammonium thiocyanate VS ＝ 12.69 mg of I Obtain the amount of iodide ion, calculated on the dried basis, by deducting the amount (z) of available iodine from the total amount (z) of iodine: it is not more than 6.6z. Loss on drying <2.41> Not more than 8.0z (1 g, 1009C, 3 hours). 1276 Pranoprofen / Official Monographs JP XVI

Residue on ignition <2.44> Not more than 0.05z (5 g). profen according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of the Standard Lead Solu- Assay (1) Available iodine—Weigh accurately about tion (not more than 10 ppm). 0.5 g of Povidone-Iodine, dissolve in 30 mL of water, and (3) Related Substances—Dissolve 50 mg of Pranoprofen titrate <2.50> with 0.02 mol/L sodium thiosulfate VS (indica- in 50 mL of the mobile phase, and use this solution as the tor: 2 mL of starch TS). sample solution. Pipet 1 mL of the sample solution, add the Each mL of 0.02 mol/L sodium thiosulfate VS mobile phase to make exactly 200 mL, and use this solution ＝ 2.538 mg of I as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed (2) Nitrogen—Weigh accurately about 20 mg of under Liquid Chromatography <2.01> according to the fol- Povidone-Iodine, and perform the test as directed under lowing conditions. Determine each peak area from both so- Nitrogen Determination <1.08>. lutions by the automatic integration method: the each area Containers and storage Containers—Tight containers. of the peaks other than the peak of pranoprofen from the sample solution is not larger than the peak area of pranoprofen from the standard solution, and the total peak Pranoprofen area of them is not larger than 2 times the peak area of pranoprofen from the standard solution. プラノプロフェン Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 275 nm). Column: A stainless steel column about 6 mm in inside diameter and about 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparti-

C15H13NO3: 255.27 cle diameter). (2RS)-2-(10H-9-Oxa-1-azaanthracen-6-yl)propanoic acid Column temperature: A constant temperature of about [52549-17-4] 259C. Mobile phase: Dissolve 7.02 g of sodium perchlorate Pranoprofen, when dried, contains not less than monohydrate in 1000 mL of water, and adjust the pH to 2.5 98.5z of C15H13NO3. with perchloric acid. To 2 volumes of this solution add 1 volume of acetonitrile. Description Pranoprofen occurs as a white to pale yellow- Flow rate: Adjust the flow rate so that the retention time ish white crystalline powder. of pranoprofen is about 10 minutes. It is freely soluble in N,N-dimethylformamide, soluble in Selection of column: Dissolve 4 mg each of Pranoprofen acetic acid (100), sparingly soluble in methanol, slightly solu- and ethyl parahydroxybenzoate in 200 mL of the mobile ble in acetonitrile, in ethanol (95) and in acetic anhydride, phase. Proceed with 10 mL of this solution under the above very slightly soluble in diethyl ether, and practically insolu- operating conditions, and calculate the resolution. Use a ble in water. column giving elution of pranoprofen and ethyl parahydrox- A solution of Pranoprofen in N,N-dimethylformamide ybenzoate in this order with the resolution between these (1 in 30) shows no optical rotation. peaks being not less than 2.1. Identification (1) Dissolve 0.02 g of Pranoprofen in 1 Detection sensitivity: Adjust the detection sensitivity so mol/L hydrochloric acid TS to make 100 mL, and dilute 10 that the peak height of pranoprofen from 10 mLofthe mL of the solution with water to make 100 mL. Determine standard solution is between 10 mm and 20 mm. the absorption spectrum of the solution as directed under Time span of measurement: About three times as long as Ultraviolet-visible Spectrophotometry <2.24>,andcompare the retention time of pranoprofen. the spectrum with the Reference Spectrum: both spectra Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- exhibit similar intensities of absorption at the same wave- um, phosphorus (V) oxide, 4 hours). lengths. (2) Determine the infrared absorption spectrum of Residue on ignition <2.44> Not more than 0.1z (1 g). Pranoprofen as directed in the potassium bromide disk Assay Weigh accurately about 0.4 g of Pranoprofen, previ- method under Infrared Spectrophotometry <2.25>,andcom- ously dried, dissolve in 70 mL of a mixture of acetic anhy- pare the spectrum with the Reference Spectrum: both spectra dride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 exhibit similar intensities of absorption at the same wave mol/L perchloric acid VS (potentiometric titration). Per- numbers. form a blank determination, and make any necessary correc- Melting point <2.60> 186–1909C tion. Purity (1) Chloride <1.03>—Dissolve 0.5 g of Prano- Each mL of 0.1 mol/L perchloric acid VS profen in 40 mL of methanol, and 6 mL of dilute nitric acid, ＝ 25.53 mg of C15H13NO3 and add water to make 50 mL. Perform the test using this Containers and storage Containers—Tight containers. solution as the test solution. Prepare the control solution as Storage—Light-resistant. follows. To 0.30 mL of 0.01 mol/L hydrochloric acid VS add 40 mL of methanol, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.021z). (2) Heavy metals <1.07>—Proceed with 2.0 g of Prano- JP XVI Official Monographs / Pravastatin Sodium 1277

cooled water is between 7.2 and 8.2. Pravastatin Sodium Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Pravastatin Sodium according to Method 2, and perform the プラバスタチンナトリウム test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 0.10 g of Pravastatin Sodium in 100 mL of a mixture of water and methanol (11:9), and use this solution as the sample solution. Pipet 10 mL of the sample solution, add the mixture of water and methanol (11:9) to make exactly 100 mL. Pipet 5 mL of this solution, add the mixture of water and methanol (11:9) to make exactly 100 mL, and use this solution as the standard C H NaO :446.51 23 35 7 solution. Perform the test with exactly 10 mLeachofthe Monosodium (3R,5R)-3,5-dihydroxy- sample solution and standard solution as directed under Liq- 7-{(1S,2S,6S,8S,8aR)-6-hydroxy-2-methyl- uid Chromatography <2.01> according to the following con- 8-[(2S)-2-methylbutanoyloxy]- ditions, and determine each peak area by the automatic inte- 1,2,6,7,8,8a-hexahydronaphthalen-1-yl}heptanoate gration method: the area of the peak other than pravastatin [81131-70-6] is not larger than 1/5 times the peak area of pravastatin from the standard solution, and the total area of the peaks Pravastatin Sodium contains not less than 98.5z other than pravastatin is not larger than the peak area of and not more than 101.0z of C H NaO , calculated 23 35 7 pravastatin from the standard solution. Keep the sample soon the anhydrous basis and corrected on the amount lution and standard solution at not over than159C. of the residual solvent. Operating conditions— Description Pravastatin Sodium occurs as a white to yel- Detector, column, column temperature, mobile phase, and lowish white, powder or crystalline powder. flow rate: Proceed as directed in the operating conditions in It is freely soluble in water and in methanol, and soluble in the Assay. ethanol (99.5). Time span of measurement: About 2.5 times as long as the It is hygroscopic. retention time of pravastatin beginning after the solvent peak. Identification (1) Determine the absorption spectrum of a System suitability— solution of Pravastatin Sodium (1 in 100,000) as directed Test for required detectability: To exactly 5 mL of the under Ultraviolet-visible Spectrophotometry <2.24>,and standard solution add a mixture of water and methanol compare the spectrum with the Reference Spectrum: both (11:9) to make exactly 50 mL. Confirm that the peak area of spectra exhibit similar intensities of absorption at the same pravastatin obtained with 10 mL of this solution is equivalent wavelengths. to 7 to 13z of that with 10 mL of the standard solution. (2) Determine the infrared absorption spectrum of System performance: Dissolve 5 mg of pravastatin sodium Pravastatin Sodium as directed in the potassium bromide in 50 mL of the mixture of water and methanol (11:9). When disk method under Infrared Spectrophotometry <2.25>:itex- the procedure is run with 10 mL of this solution under the hibits absorption at the wave numbers of about 2970 cm－1, above operating conditions, the number of theoretical plates 2880 cm－1,1727cm－1 and 1578 cm－1. and the symmetry factor of the peak of pravastatin are not (3) Dissolve 50 mg of Pravastatin Sodium in 5 mL of less than 3500 and not more than 1.6, respectively. methanol, and use this solution as the sample solution. System repeatability: When the test is repeated 6 times Separately, dissolve 24 mg of Pravastatin 1,1,3,3-Tetra- with 10 mL of the standard solution under the above operat- methylbutylammonium RS in 2 mL of methanol, and use ing conditions, the relative standard deviation of the peak this solution as the standard solution. Perform the test with area of pravastatin is not more than 2.0z. these solutions as directed under Thin-layer Chromatogra- (3) Residual solvent—Being specified separately. phy <2.03>. Spot 2 mL each of the sample solution and stand- ardsolutiononaplateofsilicagel with fluorescent indicator Water <2.48> Not more than 4.0z (0.5 g, volumetric titra- for thin-layer chromatography. Develop the plate with a tion, direct titration). mixture of ethyl acetate, ethanol (99.5) and acetic acid (100) Assay Weigh accurately about 0.1 g of Pravastatin So- (80:16:1) to a distance of about 8 cm, and air-dry the plate. dium, and dissolve in a mixture of water and methanol (11:9) Examine under ultraviolet light (main wavelength: 254 nm): to make exactly 100 mL. Pipet 10 mL of this solution, add the color tone and the Rf value of the principal spot with the exactly 10 mL of the internal standard solution and the sample solution are not different with them of the spot with mixture of water and methanol (11:9) to make 100 mL, and the standard solution. use this solution as the sample solution. Separately, weigh (4) A solution of Pravastatin Sodium (1 in 10) responds accurately about 30 mg of Pravastatin 1,1,3,3-Tetramethyl- to Qualitative Tests <1.09> (1) for sodium salt. butylammonium RS (previously determine the water <2.48> Optical rotation <2.49>: ＋153 – ＋1599(0.1 g calculated on with 0.5 g by direct titration in volumetric titration) dissolve the anhydrous basis and corrected on the amount of residual in the mixture of water and methanol (11:9) to make exactly solvent, water, 20 mL, 100 mm). 25 mL. Proceed with exactly 10 mL of this solution in the same manner for the preparation of the sample solution, and pH <2.54> The pH of a solution obtained by dissolving use the solution so obtained as the standard solution. Per- 1.0 g of Pravastatin Sodium in 20 mL of freshly boiled and 1278 Pravastatin Sodium Fine Granules / Official Monographs JP XVI form the test with 10 mL each of the sample solution and standard solution are stored at not exceeding 59Cafter standard solution as directed under Liquid Chromatography preparation. To an amount of Pravastatin Sodium Fine <2.01> according to the following conditions, and calculate Granules, equivalent to 25 mg of Pravastatin Sodium ac- the ratios, QT and QS, of the peak area of pravastatin to that cording to the labeled amount, add 25 mL of a mixture of of the internal standard. water and methanol (1:1), agitate for 15 minutes with the aid of ultrasonic waves, and centrifuge. Filter the supernatant Amount (mg) of C H NaO 23 35 7 liquid, discard the first 5 mL of the filtrate, and use the sub- ＝ M × Q /Q × 4 × 1.052 S T S sequent filtrate as the sample solution. Pipet 1 mL of the

MS: Amount (mg) of Pravastatin 1,1,3,3-Tetramethyl- sample solution, add a mixture of water and methanol (1:1) butylammonium RS, calculated on the anhydrous to make exactly 100 mL, and use this solution as the stand- basis ard solution. Perform the test with exactly 20 mLeachofthe sample solution and standard solution as directed under Liq- Internal standard solution—A solution of ethyl parahydrox- uid Chromatography <2.01> according to the following con- ybenzoate in the mixture of water and methanol (11:9) (3 in ditions. Determine each peak area of both solutions by the 4000). automatic integration method: the area of the peaks, having Operating conditions— the relative retention time of about 0.36 and about 1.9 to Detector: An ultraviolet absorption photometer (wave- pravastatin, obtained from the sample solution is not larger length: 238 nm). than 1/2 times and 3 times the peak area of pravastatin from Column: A stainless steel column 4.6 mm in inside diame- the standard solution, respectively, the area of the peak ter and 15 cm in length, packed with octadecylsilanized silica other than pravastatin and the peaks mentioned above gel for liquid chromatography (5 mm in particle diameter). obtained from the sample solution is not larger than 1/5 Column temperature: A constant temperature of about times the peak area of pravastatin from the standard solu- 259C. tion, and the total area of the peaks other than pravastatin Mobile phase: A mixture of water, methanol, acetic acid obtained from the sample solution is not larger than 4.5 (100) and triethylamine (550:450:1:1). times the peak area of pravastatin from the standard solu- Flow rate: Adjust the flow rate so that the retention time tion. For this calculation, use the area of peaks, obtained by of pravastatin is about 21 minutes. automatic integration method of related substances having System suitability— the relative retention time of about 0.36, about 0.28 and System performance: When the procedure is run with 10 about 0.88 to pravastatin, after multiplying by their relative mL of the standard solution under the above operating con- response factors, 0.58, 0.86 and 0.82, respectively. ditions, the internal standard and pravastatin are eluted in Operating conditions— this order with the resolution between these peaks being not Detector: An ultraviolet spectrophotometer (wavelength: less than 10. 238 nm). System repeatability: When the test is repeated 6 times Column: A stainless steel column 4.6 mm in inside diame- with 10 mL of the standard solution under the above operat- ter and 15 cm in length, packed with octadecylsilanized silica ing conditions, the relative standard deviation of the ratio of gel for liquid chromatography (5 mminparticlediameter). the peak area of pravastatin to that of the internal standard Column temperature: A constant temperature of about is not more than 1.0z. 259C. Containers and storage Containers—Tight containers. Mobile phase A: A mixture of water, methanol, acetic acid (100) and triethylamine (750:250:1:1). Mobile phase B: A mixture of methanol, water, acetic acid Pravastatin Sodium Fine Granules (100) and triethylamine (650:350:1:1). Flowing of mobile phase: Control the gradient by mixing プラバスタチンナトリウム細粒 the mobile phases A and B as directed in the following table.

Pravastatin Sodium Fine Granules contain not less Time after injection Mobile phase A Mobile phase B than 95.0z and not more than 105.0z of the labeled of sample (min) (volz) (volz) amount of pravastatin sodium (C H NaO : 446.51). 23 35 7 0–50 50 50 Method of preparation Prepare fine particles as directed 50–75 50→ 050→ 100 under Granules, with Pravastatin Sodium. Identification To an amount of Pravastatin Sodium Fine Flow rate: 1.3 mL per minute. Granules, equivalent to 10 mg of Pravastatin Sodium ac- Time span of measurement: For 75 minutes after injec- cording to the labeled amount, add 20 mL of water, agitate tion, beginning after the solvent peak. for 15 minutes with the aid of ultrasonic waves, and centri- System suitability— fuge. Filter the supernatant liquid, discard the first 5 mL of Test for required detectability: To exactly 1 mL of the the filtrate, and add water to 1 mL of the subsequent filtrate standard solution add a mixture of water and methanol (1:1) to make 50 mL. Determine the absorption spectrum of this to make exactly 10 mL. Confirm that the peak area of solution as directed under Ultraviolet-visible Spectropho- pravastatin obtained with 20 mL of this solution is equivalent tometry <2.24>: it exhibits a maximum between 237 nm and to 7 to 13z of that with 20 mL of the standard solution. 241 nm. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con- Purity Related substances—The sample solution and the JP XVI Official Monographs / Pravastatin Sodium Solution 1279 ditions, the number of theoretical plates and the symmetry C: Labeled amount (mg) of pravastatin sodium factor of the peak of pravastatin are not less than 3500 and (C23H35NaO7)in1g not more than 1.6, respectively. Particle size <6.03> It meets the requirements of Fine gran- System repeatability: When the test is repeated 6 times ules. with 20 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak Assay Weigh accurately an amount of Pravastatin Sodium area of pravastatin is not more than 1.5z. Fine Granules, equivalent to about 5 mg of pravastatin sodium (C H NaO ), add exactly 20 mL of the internal stand- Uniformity of dosage units <6.02> Perform the test accord- 23 35 7 ard solution, agitate for 15 minute with the aid of ultrasonic ing to the following method: the Pravastatin Sodium Fine waves, and centrifuge. Filter the supernatant liquid, discard Granules in single-unit container meets the requirement of the first 5 mL of the filtrate, to 2 mL of the subsequent fil- the Content uniformity test. trate add a mixture of water and methanol (1:1) to make 20 To the total amount of the content of 1 container of mL, and use this solution as the sample solution. Separately, Pravastatin Sodium Fine Granules add exactly V mL of the weigh accurately about 32 mg of Pravastatin 1,1,3,3- internal standard solution so that each mL contains 0.25 mg Tetramethylbutylammonium RS (separately determine the of pravastatin sodium (C H NaO ), agitate for 15 minutes 23 35 7 water <2.48> in the same manner as Pravastatin Sodium), with the aid of ultrasonic waves, and centrifuge. Filter the and dissolve in a mixture of water and methanol (1:1) to supernatant liquid, discard the first 5 mL of the filtrate, make exactly 100 mL. Pipet 5 mL of this solution, add ex- pipet 2 mL of the subsequent filtrate add a mixture of water actly 5 mL of the internal standard solution, then add a mix- and methanol (1 in 1) to make 20 mL, and use this solution ture of water and methanol (1:1) to make 50 mL, and use as the sample solution. Then, proceed as directed in the this solution as the standard solution. Perform the test with Assay. 10 mL each of the sample solution and standard solution as

Amount (mg) of pravastatin sodium (C23H35NaO7) directed under Liquid Chromatography <2.01> according to ＝ MS × QT/QS × V/100 × 1.052 the following conditions, and calculate the ratios, QT and Q , of the peak area of pravastatin to that of the internal M : Amount (mg) of pravastatin in taken Pravastatin S S standard. 1,1,3,3-Tetramethylbutylammonium RS, calculated

on the anhydrous basis Amount (mg) of pravastatin sodium (C23H35NaO7) ＝ M × Q /Q × 1/5 × 1.052 Internal standard solution—A solution of propyl parahy- S T S droxybenzoate in a mixture of water and methanol (1:1) (3 in MS: Amount (mg) of pravastatin in taken Pravastatin 10,000). 1,1,3,3-Tetramethylbutylammonium RS, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 Internal standard solution—A solution of propyl parahy- mL of water as the dissolution medium, the dissolution rate droxybenzoate in a mixture of water and methanol (1:1) (3 in in 15 minutes of Pravastatin Sodium Fine Granules is not 10,000). less than 80z. Operating conditions— Start the test with an accurately weighed amount of Proceed as directed in the operating conditions in the Pravastatin Sodium Fine Granules, equivalent to about 5 mg Assay under Pravastatin Sodium. of pravastatin sodium (C23H35NaO7) according to the labeled System suitability— amount, withdraw not less than 20 mL of the medium at the System performance: When the procedure is run with 10 specified minute after starting the test, and filter through a mL of the standard solution under the above operating con- membrane filter with a pore size not exceeding 0.45 mm. Dis- ditions, the internal standard and pravastatin are eluted in card the first 10 mL of the filtrate, and use the subsequent this order with the resolution between these peaks being not filtrate as the sample solution. Separately, weigh accurately less than 4. about 23 mg of Pravastatin 1,1,3,3-Tetramethylbutylammo- System repeatability: When the test is repeated 6 times nium RS (separately determine the water <2.48> in the same with 10 mL of the standard solution under the above operat- manner as Pravastatin Sodium), and dissolve in water to ing conditions, the relative standard deviation of the ratio of make exactly 100 mL. Pipet 3 mL of this solution, add water the peak area of pravastatin to that of the internal standard to make exactly 100 mL, and use this solution as the stand- is not more than 1.0z. ard solution. Determine the absorbances, A and A ,at T1 S1 Containers and storage Container—Well-closed contain- 238 nm and A and A at 265 nm of the sample solution T2 S2 ers. and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of Pravastatin Sodium Solution pravastatin sodium (C23H35NaO7) ＝ MS/MT × (AT1 － AT2)/(AS1 － AS2) プラバスタチンナトリウム液 × 1/C × 27 × 0.806

MS: Amount (mg) of Pravastatin 1,1,3,3-Tetramethyl- Pravastatin Sodium Solution contains not less than butylammonium RS, calculated on the anhydrous 95.0z and not more than 105.0z of the labeled basis amount of pravastatin sodium (C23H35NaO7: 446.51). M : Amount (g) of sample T Method of preparation Prepare as directed under Liquids 1280 Pravastatin Sodium Solution / Official Monographs JP XVI and Solutions for Oral Administration, with Pravastatin test. Sodium. Microbial limit <4.05> The acceptance criteria of TAMC Identification Pass a volume of Pravastatin Sodium Solu- and TYMC are 102 CFU/mL and 101 CFU/mL, respectively. tion, equivalent to 1 mg of Pravastatin Sodium according to Escherichia coli is not observed. the labeled amount, through a column [5.5 mm in inside di- Assay To a volume of Pravastatin Sodium Solution, ameter, packed with 30 mg of divinylbenzene-N-vinyl pyr- equivalent to 2 mg of pravastatin sodium (C H NaO ), add rolidone copolymer for column chromatography (30 mmin 23 35 7 exactly 5 mL of the internal standard solution, add water to particle size), and washed with 1 mL of methanol and 1 mL make 100 mL, and use this solution as the sample solution. ofwater].Thenwashwith1mLofwater,andelutewith1 Separately, weigh accurately about 20 mg of Pravastatin mL of methanol. To 0.1 mL of the eluate add water to make 1,1,3,3-Tetramethylbutylammonium RS (separately deter- 10 mL, and determine the absorption spectrum of this solu- mine the water <2.48> in the same manner as Pravastatin tion as directed under Ultraviolet-visible Spectrophotometry Sodium), and dissolve in a solution of disodium hydrogen <2.24>: it exhibits a maximum between 237 nm and 241 nm. phosphate dodecahydrate (1 in 200) to make exactly 50 mL. pH Being specified separately. Pipet 6 mL of this solution, add exactly 5 mL of the internal standard solution, add water to make 100 mL, and use this Purity Related substances—The sample solution and the solution as the standard solution. Perform the test with 10 standard solution are stored at not exceeding 159Cafter mL each of the sample solution and standard solution as preparation. To a volume of Pravastatin Sodium Solution, directed under Liquid Chromatography <2.01> according to equivalent to 2 mg of Pravastatin Sodium according to the the following conditions. Calculate the ratios, Q and Q ,of labeled amount, add a mixture of methanol and water (5:3) T S the peak area of pravastatin to that of the internal standard. to make 10 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of water Amount (mg) of pravastatin sodium and methanol (1:1) to make exactly 100 mL, and use this so- ＝ MS × QT/QS × 3/25 × 1.052 lution as the standard solution. Perform the test with exactly M : Amount (mg) of pravastatin in Pravastatin 1,1,3,3- 10 mL each of the sample solution and standard solution as S Tetramethylbutylammonium RS, calculated on the directed under Liquid Chromatography <2.01> according to anhydrous basis the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of Internal standard solution—A solution of ethyl parahy- the peaks, having the relative retention time about 0.24 and droxybenzoate in methanol (3 in 10,000). about 0.85 to pravastatin, obtained from the sample solution Operating conditions— is not larger than 2 times the peak area of pravastatin from Detector: An ultraviolet spectrophotometer (wavelength: the standard solution, the area of the peak other than 238 nm). pravastatin and the peaks mentioned above obtained from Column: A stainless steel column 3.9 mm in inside diame- the sample solution is not larger than 3/10 times the peak ter and 30 cm in length, packed with octadecylsilanized silica area of pravastatin from the standard solution, and the total gel for liquid chromatography (10 mm in particle diameter). area of the peaks other than pravastatin obtained from the Column temperature: A constant temperature of about sample solution is not larger than 3.5 times the peak area of 309C. pravastatin from the standard solution. Mobile phase: A mixture of water, methanol, acetic acid Operating conditions— (100) and triethylamine (500:500:1:1). Detector, column, column temperature, mobile phase, and Flow rate: Adjust the flow rate so that the retention time flow rate: Proceed as directed in the operating conditions in of pravastatin is about 20 minutes. the Assay. System suitability— Time span of measurement: About 2 times as long as the System performance: When the procedure is run with 10 retention time of pravasatin, beginning after the solvent mL of the standard solution under the above operating con- peak. ditions, the internal standard and pravastatin are eluted in System suitability— this order with the resolution between these peaks being not Test for required detectability: Pipet 2 mL of the standard less than 8. solution, and add a mixture of water and methanol (1:1) to System repeatability: When the test is repeated 6 times make exactly 10 mL. Confirm that the peak area of with 10 mL of the standard solution under the above operat- pravastatin obtained with 10 mL of this solution is equivalent ing conditions, the relative standard deviation of the ratio of to 15 to 25z of that with 10 mL of the standard solution. the peak area of pravastatin to that of the internal standard System performance: When the procedure is run with 10 is not more than 1.0z. mL of the standard solution under the above operating con- Containers and storage Container—Tight containers. ditions, the number of theoretical plates and the symmetry factor of the peak of pravastatin are not less than 3400 and not more than 1.6, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of pravastatin is not more than 2.5z. Uniformity of dosage units <6.02> The solution in single- unit container meets the requirement of the Mass variation JP XVI Official Monographs / Pravastatin Sodium Tablets 1281

Flowing of mobile phase: Control the gradient by mixing Pravastatin Sodium Tablets the mobile phases A and B as directed in the following table.

プラバスタチンナトリウム錠 Time after injection Mobile phase A Mobile phase B of sample (min) (volz) (volz) Pravastatin Sodium Tablets contain not less than 0–50 50 50 95.0 and not more than 105.0 of the labeled z z 50–75 50→ 050→ 100 amount of pravastatin sodium (C23H35NaO7: 446.51).

Method of preparation Prepare as directed under Tablets, Flow rate: 1.3 mL per minute. with Pravastatin Sodium. Time span of measurement: For 75 minutes after injec- Identification To a quantity of powdered Pravastatin So- tion, beginning after the solvent peak. dium Tablets, equivalent to 10 mg of Pravastatin Sodium ac- System suitability— cording to the labeled amount, add 20 mL of water, agitate Test for required detectability: Pipet 1 mL of the standard for 15 minutes with the aid of ultrasonic waves, and centri- solution, and add a mixture of water and methanol (1:1) fuge. Filter the supernatant liquid, discard the first 5 mL of to make exactly 10 mL. Confirm that the peak area of the filtrate, and add water to 1 mL of the subsequent filtrate pravastatin obtained with 20 mL of this solution is equivalent to make 50 mL. Determine the absorption spectrum of this to 7 to 13z of that with 20 mL of the standard solution. solution as directed under Ultraviolet-visible Spectropho- System performance: When the procedure is run with 20 tometry <2.24>: it exhibits a maximum between 237 nm and mL of the standard solution under the above operating con- 241 nm. ditions, the number of theoretical plates and the symmetry factor of the peak of pravastatin are not less than 3500 and Purity Related substances—The sample solution and the not more than 1.6, respectively. standard solution are stored at not exceeding 159Cafter System repeatability: When the test is repeated 6 times preparation. To a quantity of powdered Pravastatin Sodium with 20 mL of the standard solution under the above operat- Tablets, equivalent to 50 mg of Pravastatin Sodium according conditions, the relative standard deviation of the peak ing to the labeled amount, add 40 mL of a mixture of water area of pravastatin is not more than 1.5z. and methanol (1:1), agitate with the aid of ultrasonic waves, then add a mixture of water and methanol (1:1) to make 50 Uniformity of dosage units <6.02> Perform the test accord- mL, centrifuge, and use the supernatant liquid as the sample ing to the following method: it meets the requirement of the solution. Pipet 1 mL of the sample solution, add a mixture Content uniformity test. of water and methanol (1:1) to make exactly 100 mL, and To 1 tablet of Pravastatin Sodium Tablets add exactly use this solution as the standard solution. Perform the test V mL of the internal standard solution so that each mL con- with exactly 20 mL each of the sample solution and standard tains 0.25 mg of pravastatin sodium (C23H35NaO7), agitate solution as directed under Liquid Chromatography <2.01> for 15 minutes with the aid of ultrasonic waves, and centri- according to the following conditions. Determine each peak fuge. To 2 mL of the supernatant liquid add a mixture of area of both solutions by the automatic integration method: water and methanol (1:1) to make 20 mL, and use this solu- the area of the peaks, having the relative retention time tion as the sample solution. Then, proceed as directed in the about 0.36 and about 1.9 to pravastatin obtained from the Assay. sample solution is not larger than 3/10 times and 2 times the Amount (mg) of pravastatin sodium (C23H35NaO7) peak area of pravastatin from the standard solution, respec- ＝ MS × QT/QS × V/100 × 1.052 tively, the area of the peak other than pravastatin and the peaks mentioned above obtained from the sample solution is MS: Amount (mg) of pravastatin in Pravastatin 1,1,3,3- not larger than 1/5 times the peak area of pravastatin from Tetramethylbutylammonium RS, calculated on the the standard solution, and the total area of the peaks other anhydrous basis than pravastatin obtained from the sample solution is not Internal standard solution—A solution of propyl parahy- larger than 3 times the peak area of pravastatin from the droxybenzoate in a mixture of water and methanol (1:1) (3 in standard solution. For this calculation, use the area of the 10,000). peaks, having the relative retention time about 0.36, about 0.28 and about 0.88 to pravastatin, after multiplying by their Dissolution <6.10> When the test is performed at 50 revolu- relative response factors, 0.58, 0.86 and 0.82, respectively. tions per minute according to the Paddle method, using 900 Operating conditions— mL of water as the dissolution medium, the dissolution rate Detector: An ultraviolet spectrophotometer (wavelength: in 30 minutes of Pravastatin Sodium Tablets is not less than 238 nm). 85z. Column: A stainless steel column 4.6 mm in inside diame- Start the test with 1 tablet of Pravastatin Sodium Tablets, ter and 15 cm in length, packed with octadecylsilanized silica withdraw not less than 20 mL of the medium at the specified gel for liquid chromatography (5 mm in particle diameter). minute after starting the test, and filter through a membrane Column temperature: A constant temperature of about filter with a pore size not exceeding 0.45 mm. Discard the 259C. first 10 mL of the filtrate, pipet V mL of the subsequent fil- Mobile phase A: A mixture of water, methanol, acetic trate, add water to make exactly V?mL so that each mL con- acid (100) and triethylamine (750:250:1:1). tains about 5.5 mg of pravastatin (C23H36O7) according to the Mobile phase B: A mixture of methanol,water,aceticacid labeled amount, and use this solution as the sample solution. (100) and triethylamine (650:350:1:1). Separately, weigh accurately about 23 mg of Pravastatin 1282 Prazepam / Official Monographs JP XVI

1,1,3,3-Tetramethylbutylammonium RS (separately deter- is not more than 1.0z. mine the water <2.48> in the same manner as Pravastatin So- Containers and storage Container—Well-closed contain- dium), and dissolve in water to make exactly 100 mL. Pipet 3 ers. mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT1 and AS1, at 238 nm and AT2 and AS2 at 256 nm of the sample solution and standard solution as directed Prazepam under Ultraviolet-visible Spectrophotometry <2.24>. プラゼパム Dissolution rate (z) with respect to the labeled amount of

pravastatin sodium (C23H35NaO7) ＝ MS × (AT1 － AT2)/(AS1 － AS2) × V?/V × 1/C × 27 × 0.806

MS: Amount (mg) of Pravastatin 1,1,3,3-Tetramethyl- butylammonium RS, calculated on the anhydrous basis C: Labeled amount (mg) of pravastatin sodium

(C23H35NaO7)in1tablet C19H17ClN2O: 324.80 7-Chloro-1-(cyclopropylmethyl)-5-phenyl-1,3-dihydro- Assay Weigh accurately and powder not less than 20 2H-1,4-benzodiazepin-2-one Pravastatin Sodium Tablets. Weigh accurately a portion of [2955-38-6] the powder, equivalent to about 10 mg of pravastatin sodium (C H NaO ), add exactly 40 mL of the internal stand- 23 35 7 Prazepam, when dried, contains not less than 98.5z ard solution, agitate for 15 minutes with the aid of ultrasonic of C H ClN O. waves, and centrifuge. Filter the supernatant liquid, discard 19 17 2 the first 5 mL of the filtrate, to 2 mL of the subsequent fil- Description Prazepam occurs as white to light yellow crys- trate add a mixture of water and methanol (1:1) to make tals or crystalline powder. It is odorless. exactly 20 mL, and use this solution as the sample solution. It is freely soluble in acetone, soluble in acetic anhydride, Separately, weigh accurately about 32 mg of Pravastatin sparingly soluble in ethanol (99.5) and in diethyl ether, and 1,1,3,3-Tetramethylbutylammonium RS (separately deter- practically insoluble in water. mine the water <2.48> in the same manner as Pravastatin Identification (1) Dissolve0.01gofPrazepamin3mLof Sodium), and dissolve in a mixture of water and methanol sulfuric acid, and observe under ultraviolet light (main wave- (1:1) to make exactly 100 mL. Pipet 5 mL of this solution, length: 365 nm): the solution shows a grayish blue fluores- add exactly 5 mL of the internal standard solution, then add cence. a mixture of water and methanol (1:1) to make 50 mL, and (2) Dissolve 0.01 g of Prazepam in 1000 mL of a solution use this solution as the standard solution. Perform the test of sulfuric acid in ethanol (99.5) (3 in 1000). Determine the with 10 mL each of the sample solution and standard solution absorption spectrum of the solution as directed under Ultra- as directed under Liquid Chromatography <2.01> according violet-visible Spectrophotometry <2.24>, and compare the to the following conditions. Calculate the ratios, Q and T spectrum with the Reference Spectrum: both spectra exhibit Q , of the peak area of pravastatin to that of the internal S similar intensities of absorption at the same wavelengths. standard. (3) Determine the infrared absorption spectrum of

Amount (mg) of pravastatin sodium (C23H35NaO7) Prazepam, previously dried, as directed in the potassium ＝ MS × QT/QS × 2/5 × 1.052 bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- M : Amount (mg) of pravastatin in Pravastatin 1,1,3,3- S trum: both spectra exhibit similar intensities of absorption at Tetramethylbutylammonium RS, calculated on the the same wave numbers. anhydrous basis (4) Perform the Flame Coloration Tests <1.04> (2) with Internal standard solution—A solution of propyl parahy- Prazepam: a green color appears. droxybenzoate in a mixture of water and methanol (1:1) (3 in Melting point <2.60> 145 – 1489C 10,000). Operating conditions— Purity (1) Chloride <1.03>—To 1.0 g of Prazepam add 50 Proceed as directed in the operating conditions in the mL of water, allow to stand for 1 hour with occasional shak- Assay under Pravastatin Sodium. ing, and filter. To 20 mL of the filtrate add 6 mL of dilute System suitability— nitric acid and water to make 50 mL. Perform the test using System performance: When the procedure is run with 10 this solution as the test solution. Prepare the control solution mL of the standard solution under the above operating con- with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more ditions, the internal standard and pravastatin are eluted in than 0.036z). this order with the resolution between these peaks being not (2) Sulfate <1.14>—To 20 mL of the filtrate obtained in less than 4. (1) add 1 mL of dilute hydrochloric acid and water to make System repeatability: When the test is repeated 6 times 50 mL. Perform the test using this solution as the test solu- with 10 mL of the standard solution under the above operat- tion. Prepare the control solution with 0.40 mL of 0.005 ing conditions, the relative standard deviation of the ratio of mol/L sulfuric acid VS (not more than 0.048z). the peak area of pravastatin to that of the internal standard (3) Heavy metals <1.07>—Proceed with 2.0 g of Praze- JP XVI Official Monographs / Prazepam Tablets 1283 pam according to Method 2, and perform the test. Prepare nm, and minima between 263 nm and 267 nm and between the control solution with 2.0 mL of Standard Lead Solution 334 nm and 338 nm. (not more than 10 ppm). Dissolution <6.10> When the test is performed at 100 revo- (4) Arsenic <1.11>—Prepare the test solution with 1.0 g lutions per minute according to the Basket method, using of Prazepam according to Method 3, and perform the test 900 mL of 0.1 mol/L hydrochloric acid TS as the dissolution (not more than 2 ppm). medium, the dissolution rate in 30 minutes of Prazepam (5) Related substances—Dissolve 0.40 g of Prazepam in Tablets is not less than 80z. 10 mL of acetone, and use this solution as the sample solu- Start the test with 1 tablet of Prazepam Tablets, withdraw tion. Pipet 1 mL of the sample solution, and add acetone to not less than 20 mL of the medium at the specified minute make exactly 20 mL. Pipet 1 mL of this solution, add ace- after starting the test, and filter through a membrane filter tone to make exactly 25 mL, and use this solution as the with a pore size not exceeding 0.8 mm. Discard the first 10 standard solution. Perform the test with these solutions as mL of the filtrate, measure exactly the subsequent V mL of directed under Thin-layer Chromatography <2.03>. Spot 5 the filtrate, add the dissolution medium to make exactly mL each of the sample solution and standard solution on a V?mL so that each mL contains about 5 mg of prazepam plate of silica gel with fluorescent indicator for thin-layer (C H ClN O) according to the labeled amount, and use this chromatography. Develop the plate with a mixture of chlo- 19 17 2 solution as the sample solution. Separately, weigh accurately roform and acetone (9:1) to a distance of about 10 cm, and about 5 mg of prazepam for assay, previously dried at 1059C air-dry the plate. Examine under ultraviolet light (main for 2 hours, add 200 mL of the dissolution medium and dis- wavelength: 254 nm): the spots other than the principal spot solve with shaking, or by ultrasonication if necessary, add from the sample solution are not more intense than the spot the dissolution medium to make exactly 1000 mL, and use from the standard solution. this solution as the standard solution. Determine the absor-

Loss on drying <2.41> Not more than 0.20z (1 g, 1059C, bances, AT and AS, of the sample solution and the standard 2 hours). solution at 240 nm as directed under Ultraviolet-visible Spec- trophotometry <2.24>. Residue on ignition <2.44> Not more than 0.1z (1 g). Dissolution rate (z) with respect to the labeled amount Assay Weigh accurately about 0.4 g of Prazepam, previ- of prazepam (C H ClN O) ously dried, dissolve in 60 mL of acetic anhydride, and 19 17 2 ＝ M × A /A × V?/V × 1/C × 90 titrate <2.50> with 0.1 mol/L perchloric acid VS (potenti- S T S ometric titration). Perform a blank determination, and make MS: Amount (mg) of prazepam for assay any necessary correction. C: Labeled amount (mg) of prazepam (C19H17ClN2O) in 1 tablet Each mL of 0.1 mol/L perchloric acid VS

＝ 32.48 mg of C19H17ClN2O Assay Weigh accurately not less than 20 Prazepam Tablets, and powder. Weigh accurately a quantity of the powder, Containers and storage Containers—Tight containers. equivalent to about 50 mg of prazepam (C19H17ClN2O), add 30 mL of acetone, shake well, centrifuge, and separate the supernatant liquid. Repeat the same procedure twice with 30 Prazepam Tablets mL each of acetone, combine all the supernatants liquid, andevaporateonawaterbathtodryness.Dissolvethe プラゼパム錠 residue in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.02 mol/L perchlo- Prazepam Tablets contain not less than 93.0z and ric acid VS (potentiometric titration). Perform a blank deter- not more than 107.0z of the labeled amount of mination, and make any necessary correction. prazepam (C H ClN O: 324.80). 19 17 2 Each mL of 0.02 mol/L perchloric acid VS

Method of preparation Prepare as directed under Tablets, ＝ 6.496 mg of C19H17ClN2O with Prazepam. Containers and storage Containers—Tight containers. Identification (1) To a quantity of powdered Prazepam Tablets, equivalent to 0.05 g of Prazepam according to the labeled amount, add 25 mL of acetone, shake well, and filter. Take 5 mL of the filtrate, evaporate on a water bath to dryness, and dissolve the residue in 3 mL of sulfuric acid. With this solution, proceed as directed in the Identification (1) under Prazepam. (2) To a quantity of powdered Prazepam Tablets, equivalent to 0.02 g of Prazepam according to the labeled amount, add 200 mL of a solution of sulfuric acid in ethanol (99.5) (3 in 1000), shake well, and filter. To 5 mL of the filtrate add a solution of sulfuric acid in ethanol (99.5) (3 in 1000) to make 50 mL, and determine the absorption spectrum as directed under Ultraviolet-visible Spectrophotome- try <2.24>: it exhibits maxima between 241 nm and 245 nm, between 283 nm and 287 nm and between 363 nm and 367 1284 Prazosin Hydrochloride / Official Monographs JP XVI

integration method: the area of each peak other than the Prazosin Hydrochloride peak of prazosin from the sample solution is not larger than 2 times the peak area of prazosin from the standard solution, プラゾシン塩酸塩 and the total area of the peaks other than the peak of prazosin from the sample solution is not larger than 5 times the peak area of prazosin from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 254 nm). Column: A stainless steel column 4.6 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticlediameter).

C19H21N5O4.HCl: 419.86 Column temperature: A constant temperature of about 1-(4-Amino-6,7-dimethoxy-quinazolin-2-yl)- 259C. 4-(2-furoyl)piperazine monohydrochloride Mobile phase: Dissolve 3.484 g of sodium 1-pentane sul- [19237-84-4] fonate and 18 mL of tetramethylammonium hydroxide in 900 mL of water, adjust the pH to 5.0 with acetic acid (100), Prazosin Hydrochloride, when dried, contains not and add water to make 1000 mL. To this solution add 1000 less than 97.0z and not more than 103.0z of mL of methanol. C19H21N5O4.HCl. Flow rate: Adjust the flow rate so that the retention time of prazosin is about 9 minutes. Description Prazosin Hydrochloride occurs as a white crys- Time span of measurement: About 6 times as long as the talline powder. retention time of prazosin. It is slightly soluble in methanol, very slightly soluble in System suitability— ethanol (99.5) and practically insoluble in water. Test for required detectability: Pipet 5 mL of the standard It gradually turns pale yellowish white on exposure to solution, and add the mobile phase to make exactly 10 mL. light. Confirm that the peak area of prazosin obtained from 20 mL Melting point: about 2709C (with decomposition). of this solution is equivalent to 35 to 65z of that of prazosin Identification (1) Determine the absorption spectrum of a from 20 mL of the standard solution. solution of Prazosin Hydrochloride in 0.01 mol/L hydro- System performance: When the procedure is run with 20 chloric acid-methanol TS (1 in 200,000) as directed under mL of the standard solution under the above operating con- Ultraviolet-visible Spectrophotometry <2.24>,andcompare ditions, the number of theoretical plates and the symmetry the spectrum with the Reference Spectrum or the spectrum factor of the peak of prazosin are not less than 4000 and not of a solution of Prazosin Hydrochloride RS prepared in the more than 2.0, respectively. same manner as the sample solution: both spectra exhibit System repeatability: When the test is repeated 6 times similar intensities of absorption at the same wavelengths. with 20 mL of the standard solution under the above operat- (2) Determine the infrared absorption spectrum of ing conditions, the relative standard deviation of the peak Prazosin Hydrochloride as directed in the potassium chlo- area of prazosin is not more than 2.0z. ride disk method under Infrared Spectrophotometry <2.25>, (3) Residual solvent—Being specified separately. and compare the spectrum with the Reference Spectrum or Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, the spectrum of Prazosin Hydrochloride RS: both spectra 2 hours). exhibit similar intensities of absorption at the same wave numbers. Residue on ignition <2.44> Not more than 0.2z (1 g). (3) To 0.1 g of Prazosin Hydrochloride add 5 mL of Assay Weigh accurately about 25 mg each of Prazosin Hy- water and 1 mL of ammonia TS, shake, allow to stand for 5 drochloride and Prazosin Hydrochloride RS, previously minutes, and filter. Render the filtrate acid with acetic acid dried, and dissolve each in methanol to make exactly 50 mL. (100): the solution responds to the Qualitative Tests <1.09> Pipet 3 mL each of these solutions, and add a mixture of for chloride. methanol and water (7:3) to make exactly 100 mL, and use Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of these solutions as the sample solution and the standard solu- Prazosin Hydrochloride according to Method 4, and per- tion, respectively. Perform the test with exactly 10 mLeach form the test. Prepare the control solution with 1.0 mL of of the sample solution and standard solution as directed Standard Lead Solution (not more than 10 ppm). under Liquid Chromatography <2.01> according to the fol-

(2) Related substances—Dissolve 20 mg of Prazosin Hy- lowing conditions, and determine the peak areas, AT and AS, drochloride in 20 mL of the mobile phase, and use this solu- of prazosin in each solution. tion as the sample solution. Pipet 1 mL of the sample solu- Amount (mg) of prazosin hydrochloride (C H N O .HCl) tion, and add the mobile phase to make exactly 100 mL. 19 21 5 4 ＝ M × A /A Pipet 1 mL of this solution, add the mobile phase to make S T S exactly 10 mL, and use this solution as the standard solution. MS: Amount (mg) of Prazosin Hydrochloride RS Perform the test with exactly 20 mLeachofthesamplesolu- Operating conditions— tion and standard solution as directed under Liquid Chroma- Detector: An ultraviolet absorption photometer (wave- tography <2.01> according to the following conditions. De- length: 254 nm). termine each peak area of both solutions by the automatic Column: A stainless steel column 4.6 mm in inside diame- JP XVI Official Monographs / Prednisolone 1285 ter and 25 cm in length, packed with silica gel for liquid 0.2 g, ethanol (95), 20 mL, 100 mm). chromatography (5 mm in particle diameter). Purity (1) Selenium—To 0.10 g of Prednisolone add 0.5 Column temperature: A constant temperature of about mL of a mixture of perchloric acid and sulfuric acid (1:1) 259C. and 2 mL of nitric acid, and heat on a water bath until no Mobile phase: A mixture of methanol, water, acetic acid more brown gas evolves and the solution becomes to be a (100) and diethylamine (3500:1500:50:1). light yellow clear solution. After cooling, add 4 mL of nitric Flow rate: Adjust the flow rate so that the retention time acid to this solution, then add water to make exactly 50 mL, of prazosin is about 8 minutes. and use this solution as the sample solution. Separately, System suitability— pipet 3 mL of Standard Selenium Solution, add 0.5 mL of a System performance: When the procedure is run with 10 mixture of perchloric acid and sulfuric acid (1:1) and 6 mL mL of the standard solution under the above operating con- of nitric acid, then add water to make exactly 50 mL, and ditions, the number of theoretical plates and the symmetry use this solution as the standard solution. Perform the test factor of the peak of prazosin are not less than 5000 and not with the sample solution and standard solution as directed more than 2.0, respectively. under Atomic Absorption Spectrophotometry <2.23> accord- System repeatability: When the test is repeated 6 times ing to the following conditions, and determine constant ab- with 10 mL of the standard solution under the above operat- sorbances, A and A , obtained on a recorder after rapid ing conditions, the relative standard deviation of the peak T S increasing of the absorption: A is smaller than A (not more area of prazosin is not more than 1.0z. T S than 30 ppm). Containers and storage Containers—Well-closed contain- Perform the test by using a hydride generating system and ers. a thermal absorption cell. Storage—Light-resistant. Lamp: A selenium hollow cathode lamp. Wavelength: 196.0 nm. Temperature of sample atomizer: When an electric fur- Prednisolone nace is used, about 10009C. Carrier gas: Nitrogen or argon. プレドニゾロン (2) Related substances—Dissolve 20 mg of Prednisolone in exactly 2 mL of a mixture of methanol and chloroform (1:1), and use this solution as the sample solution. Sepa- rately, dissolve 20 mg of hydrocortisone and 10 mg of prednisolone acetate each in a mixture of methanol and chloroform (1:1) to make exactly 100 mL, and use these solutions as the standard solution (1) and standard solution (2). Per- form the test with these solutions as directed under Thin-

C21H28O5:360.44 layer Chromatography <2.03>. Spot 5 mLeachofthesample 11b,17,21-Trihydroxypregna-1,4-diene-3,20-dione solution and standard solutions (1) and (2) on a plate of [50-24-8] silica gel for thin-layer chromatography. Develop the plate with a mixture of acetone, toluene and diethylamine Prednisolone, when dried, contains not less than (55:45:2) to a distance of about 15 cm, and air-dry the plate 97.0z and not more than 102.0z of C21H28O5. (do not dip the filter paper in the developing vessel). Spray evenly alkaline blue tetrazolium TS on the plate: the spots Description Prednisolone occurs as a white, crystalline from the sample solution corresponding to those from the powder. standard solutions (1) and (2) are not more intense than the It is soluble in methanol and in ethanol (95), slightly solu- spots from the standard solutions (1) and (2), and no spots ble in ethyl acetate and in chloroform, and very slightly solu- other than the principal spot, hydrocortisone and predniso- ble in water. lone acetate appear from the sample solution. Melting point: about 2359C (with decomposition). Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C, Identification (1) To 2 mg of Prednisolone add 2 mL of 3 hours). sulfuric acid, and allow to stand for 2 to 3 minutes: a deep red color, without fluorescence, develops. To this solution Residue on ignition <2.44> Not more than 0.1z (0.5 g). add 10 mL of water cautiously: the color disappears and a Assay Dissolve about 25 mg each of Prednisolone and gray, flocculent precipitate is formed. Prednisolone RS, previously dried, and accurately weighed, (2) Determine the infrared absorption spectrum of Pred- in 50 mL of methanol, add exactly 25 mL of the internal nisolone, previously dried, as directed in the potassium bro- standard solution to each, and add methanol to make 100 mide disk method under Infrared Spectrophotometry <2.25>, mL. To 1 mL each of these solutions add the mobile phase to and compare the spectrum with the Reference Spectrum or make 10 mL, and use these solutions as the sample solution the spectrum of previously dried Prednisolone RS: both and standard solution. Perform the test with 20 mLeachof spectra exhibit similar intensities of absorption at the same these solutions as directed under Liquid Chromatography wave numbers. If any difference appears between the spec- <2.01> according to the following conditions, and calculate tra, dissolve Prednisolone and Prednisolone RS in ethyl the ratios, Q and Q , of the peak area of prednisolone to acetate, respectively, then evaporate the ethyl acetate to T S that of the internal standard. dryness, and repeat the test on the residues.

20 Amount (mg) of C21H28O5 ＝ MS × QT/QS Optical rotation <2.49> [a]D : ＋113 – ＋1199(after drying, 1286 Prednisolone Tablets / Official Monographs JP XVI

MS: Amount (mg) of Prednisolne RS add methanol to make exactly 100 mL. Centrifuge this solution, pipet V mL of the supernatant liquid, and add metha- Internal standard solution—A solution of methyl parahy- nol to make exactly V?mL to provide a solution that con- droxybenzoate in methanol (1 in 2000). tains about 10 mg of prednisolone (C H O ) per ml, and use Operating conditions— 21 28 5 this solution as the sample solution. Separately, weigh accu- Detector: An ultraviolet absorption photometer (wave- rately about 10 mg of Prednisolone RS, previously dried at length: 247 nm). 1059C for 3 hours, dissolve in 10 mL of water and 50 mL of Column: A stainless steel column 4.6 mm in inside diame- methanol, and add methanol to make exactly 100 mL. Pipet ter and 15 cm in length, packed with fluorosilanized silica gel 5 mL of this solution, add methanol to make exactly 50 mL, for liquid chromatography (5 mminparticlediameter). and use this solution as the standard solution. Determine the Column temperature: A constant temperature of about absorbances, A and A , of the sample solution and stand- 409C. T S ard solution at 242 nm as directed under Ultraviolet-visible Mobile phase: A mixture of water and methanol (13:7). Spectrophotometry <2.24>. Flow rate: Adjust the flow rate so that the retention time of prednisolone is about 15 minutes. Amount (mg) of prednisolone (C21H18O5) System suitability— ＝ MS × AT/AS × V?/V × 1/10 System performance: Dissolve 25 mg of Prednisolone and M : Amount (mg) of Prednisolone RS 25 mg of hydrocortisone in 100 mL of methanol. To 1 mL of S this solution add the mobile phase to make 10 mL. When the Dissolution <6.10> When the test is performed at 100 revo- procedure is run with 20 mL of this solution under the above lutions per minute according to the Paddle method, using operating conditions, hydrocortisone and prednisolone are 900 mL of water as the dissolution medium, the dissolution eluted in this order with the resolution between these peaks rate in 20 minutes of Prednisolone Tablets is not less than being not less than 1.5. 70z. System repeatability: When the test is repeated 6 times Start the test with 1 tablet of Prednisolone Tablets, with 20 mL of the standard solution under the above operat- withdraw not less than 20 mL of the medium at the specified ing conditions, the relative standard deviation of the ratios minute after starting the test, and filter through a membrane of the peak area of prednisolone to that of the internal filter with a pore size not exceeding 0.8 mm. Discard the first standard is not more than 1.0z. 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 10 mg Containers and storage Containers—Tight containers. of Prednisolone RS, previously dried at 1059C for 3 hours, and dissolve in ethanol (95) to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and Prednisolone Tablets use this solution as the standard solution. Determine the absorbances, A and A , of the sample solution and stand- プレドニゾロン錠 T S ard solution at the maximum wavelength at about 242 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, Prednisolone Tablets contain not less than 90.0z using water as the blank. and not more than 110.0z of the labeled amount of Dissolution rate (z) with respect to the labeled amount prednisolone (C21H28O5: 360.44). of prednisolone (C21H28O5) Method of preparation Prepare as directed under Tablets, ＝ MS × AT/AS × 1/C × 45 with Prednisolone. MS: Amount (mg) of Prednisolone RS Identification (1) Weigh a quantity of powdered Pred- C: Labeled amount (mg) of prednisolone (C21H28O5)in1 nisolone Tablets, equivalent to 0.05 g of Prednisolone ac- tablet cording to the labeled amount, add 10 mL of chloroform, Assay Weigh accurately and powder not less than 20 Pred- shake for 15 minutes, and filter. Evaporate the filtrate on a nisolone Tablets using an agate mortar. Weigh accurately a water bath to dryness. Dry the residue at 1059C for 1 hour, portion of the powder, equivalent to about 5 mg of predniso- and proceed as directed in the Identification (1) under Pred- lone (C H O ), add 1 mL of water, and shake gently. Add nisolone. 21 28 5 exactly 5 mL of the internal standard solution and 15 mL of (2) Determine the infrared absorption spectra of the methanol, and shake vigorously for 20 minutes. To 1 mL of residue obtained in (1) and Prednisolone RS, previously this solution add the mobile phase to make 10 mL, and filter dried, as directed in the potassium bromide disk method through a membrane filter with pore size of 0.45 mm. Dis- under Infrared Spectrophotometry <2.25>: both spectra ex- card the first 3 mL of the filtrate, and use the subsequent fil- hibit similar intensities of absorption at the same wave num- trate as the sample solution. Separately, weigh accurately bers. If any difference appears, dissolve the sample and the about 25 mg of Prednisolone RS, previously dried at 1059C RS in ethyl acetate, evaporate to dryness, and repeat the test for 3 hours, dissolve in 50 mL of methanol, add exactly 25 on the residues. mL of the internal standard solution, and add methanol to Uniformity of dosage units <6.02> Perform the test accord- make 100 mL. To 1 mL of this solution add the mobile phase ing to the following method: it meets the requirement of the to make 10 mL, and use this solution as the standard solu- Content uniformity test. tion. Proceed as directed in the Assay under Prednisolone Transfer 1 tablet of Prednisolone Tablets to a volumetric with these solutions. flask, and shake with 10 mL of water until the tablet is disintegrated. Add 50 mL of methanol, shake for 30 minutes, and JP XVI Official Monographs / Prednisolone Acetate 1287

Amount (mg) of prednisolone (C21H28O5) directed under Thin-layer Chromatography <2.03>. Spot 5 ＝ MS × QT/QS × 1/5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer M : Amount (mg) of Prednisolone RS S chromatography. Develop the plate with a mixture of Internal standard solution—A solution of methyl parahy- dichloromethane, diethyl ether, methanol and water droxybenzoate in methanol (1 in 2000). (385:75:40:6) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (wavelength: 254 mm): Containers and storage Containers—Tight containers. the spots from the sample solution corresponding to those from the standard solution are not more intense than the spots from the standard solution, and any spot from the Prednisolone Acetate sample solution other than the principal spot and the spots from prednisolone, cortisone acetate and hydrocortisone プレドニゾロン酢酸エステル acetate does not appear. Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g). Assay Dissolve about 10 mg each of Prednisolone Acetate and Prednisolone Acetate RS, previously dried and accurately weighed, in 60 mL each of methanol, add exactly 2

C23H30O6:402.48 mL each of the internal standard solution, then add metha- 11b,17,21-Trihydroxypregna-1,4-diene-3,20-dione nol to make 100 mL, and use these solutions as the sample 21-acetate solution and standard solution. Perform the text with 10 mL [52-21-1] each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-

Prednisolone Acetate, when dried, contains not less lowing conditions, and calculate the ratios, QT and QS,of than 96.0z and not more than 102.0z of C23H30O6. the peak height of prednisolone acetate to that of the internal standard. Description Prednisolone Acetate occurs as a white, crystalline powder. Amount (mg) of C23H30O6 ＝ MS × QT/QS It is slightly soluble in methanol, in ethanol (95), in M : Amount (mg) of Prednisolone Acetate RS ethanol (99.5), and in chloroform, and practically insoluble S in water. Internal standard solution—A solution of butyl parahydrox- Melting point: about 2359C (with decomposition). ybenzoate in methanol (3 in 1000). Operating conditions— Identification (1) To 2 mg of Prednisolone Acetate add 2 Detector: An ultraviolet absorption photometer (wave- mL of sulfuric acid, and allow to stand for 2 to 3 minutes: a length: 254 nm). deep red color, without fluorescence, develops. To this solu- Column: A stainless steel column 4.0 mm in inside diame- tion add 10 mL of water cautiously: the color disappears and ter and 15 cm in length, packed with octadecylsilanized silica a gray, flocculent precipitate is formed. gel for liquid chromatography (5 mminparticlediameter). (2) Determine the infrared absorption spectra of Pred- Column temperature: A constant temperature of about nisolone Acetate, previously dried, as directed in the potas- 259C. sium bromide disk method under Infrared Spectrophotome- Mobile phase: A mixture of water and acetonitrile (3:2). try <2.25>, and compare the spectrum in a range between Flow rate: Adjust the flow rate so that the retention time 4000 cm－1 and 650 cm－1 with the Infrared Reference Spec- of prednisolone acetate is about 10 minutes. trum or the spectrum of previously dried Prednisolone System suitability— Acetate RS: both spectra exhibit similar intensities of ab- System performance: When the procedure is run with 10 sorption at the same wave numbers. If any difference ap- mL of the standard solution under the above operating con- pears, dissolve the sample and the RS in ethanol (99.5), ditions, prednisolone acetate and the internal standard are respectively, evaporate to dryness, and repeat the test on the eluted in this order with the resolution between these peaks residues. being not less than 10. 20 Optical rotation <2.49> [a]D : ＋128 – ＋1379(after drying, System repeatability: When the test is repeated 6 times 70 mg, methanol, 20 mL, 100 mm). with 10 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios Purity Related substanes—Dissolve 0.20 g of Prednisolone of the peak height of prednisolone acetate to that of the in- Acetate in exactly 10 mL of a mixture of chloroform and ternal standard is not more than 1.0z. methanol (9:1), and use this solution as the sample solution. Separately, dissolve 20 mg each of prednisolone, cortisone Containers and storage Containers—Tight containers. acetate and hydrocortisone acetate in exactly 10 mL of a mixture of chloroform and methanol (9:1). Pipet 1 mL of this solution, add a mixture of chloroform and methanol (9:1) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as 1288 Prednisolone Sodium Phosphate / Official Monographs JP XVI

40) to make 10 mL. To 2.5 mL of this solution add diluted Prednisolone Sodium Phosphate hydrochloric acid (1 in 40) to make 100 mL. (2) Heavy metals <1.07>—Proceed with 0.5 g of Pred- プレドニゾロンリン酸エステルナトリウム nisolone Sodium Phosphate according to Method 3, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 40 ppm). (3) Free phosphoric acid—Weigh accurately about 0.25 g of Prednisolone Sodium Phosphate, dissolve in water to make exactly 100 mL, and use this solution as the sample solution. Pipet 5 mL each of the sample solution and Phos- phoric Acid Standard Solution, add 2.5 mL of hexaammo-

C21H27Na2O8P: 484.39 nium heptamolybdate-sulfuric acid TS and 1 mL of 1-amino- 11b,17,21-Trihydroxypregna-1,4-diene 3,20-dione 2-naphtol-4-sulfonic acid TS, shake, add water to make ex- 21-(disodium phosphate) actly 25 mL, and allow to stand at 20 ± 19C for 30 minutes. [125-02-0] Perform the test with these solutions as directed under Ultra- violet-visible Spectrophotometry <2.24>, using a solution Prednisolone Sodium Phosphate contains not prepared with 5 mL of water in the same manner as the less than 97.0z and not more than 103.0z of blank. Determine the absorbances, AT and AS, of each solu- C21H27Na2O8P, calculated on the anhydrous basis. tion from the sample solution and standard solution at 740 nm: the content of free phosphoric acid is not more than Description Prednisolone Sodium Phosphate occurs as a 1.0z. white to pale yellow powder.

It is freely soluble in water, soluble in methanol, and prac- Content (z) of free phosphoric acid (H3PO4) tically insoluble in ethanol (99.5). ＝ 1/M × AT/AS × 258.0 It is hygroscopic. M: Amount (mg) of Prednisolone Sodium Phosphate, cal- Identification (1) Moisten 1.0 g of Prednisolone Sodium culated on the anhydrous basis Phosphate with a small amount of sulfuric acid, and gradu- (4) Related substances—Dissolve 10 mg of Prednisolone ally heat to incinerate. After cooling, dissolve the residue in Sodium Phosphate in 100 mL of the mobile phase, and use 10 mL of dilute nitric acid, and heat in a water bath for 30 this solution as the sample solution. Pipet 2 mL of the sam- minutes. After cooling, filter if necessary. This solution re- ple solution, add the mobile phase to make exactly 100 mL, sponds to the Qualitative Tests <1.09> for phosphate. and use this solution as the standard solution. Perform the (2) Dissolve 2 mg of Prednisolone Sodium Phosphate in test with exactly 20 mL each of the sample solution and 2 mL of sulfuric acid, and allow to stand for 2 minutes: a standard solution as directed under Liquid Chromatography deep red color, without fluorescence, develops. <2.01> according to the following conditions. Determine each (3) Determine the absorption spectrum of a solution of peak area of both solutions by the automatic integration Prednisolone Sodium Phosphate (1 in 50,000) as directed method: the area of each peak other than the peak of pred- under Ultraviolet-visible Spectrophotometry <2.24>,and nisolone phosphate from the sample solution is not larger compare the spectrum with the Reference Spectrum: both than 1.5 times the peak area of prednisolone phosphate from spectra exhibit similar intensities of absorption at the same the standard solution, and the total area of the peaks other wavelengths. than the peak of prednisolone phosphate from the sample (4) Determine the infrared absorption spectrum of Pred- solution is not larger than 2.5 times the peak area of prednisolone Sodium Phosphate as directed in the potassium bro- nisolone phosphate from the standard solution. mide disk method under Infrared Spectrophotometry <2.25>, Operating conditions— and compare the spectrum with the Reference Spectrum: Detector: An ultraviolet absorption photometer (wave- both spectra exhibit similar intensities of absorption at the length: 245 nm). same wave numbers. Column: A stainless steel column 4.6 mm in inside diame- (5) The solution obtained in (1) responds to the Qualita- ter and 10 cm in length, packed with octadecylsilanized silica tive Tests <1.09> for sodium salt. gel for liquid chromatography (3 mminparticlediameter). 20 Optical rotation <2.49> [a]D : ＋96 – ＋1039(1 g, calculated Column temperature: A constant temperature of about on the anhydrous basis, phosphate buffer solution, pH 7.0, 409C. 100 mL, 100 mm). Mobile phase: Dissolve 6.80 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust the pH to pH <2.54> Dissolve 1.0 g of Prednisolone Sodium Phos- 2.5 with phosphoric acid. To 1000 mL of this solution add phate in 100 mL of water: the pH of the solution is between 250 mL of acetonitrile. 7.5 and 9.0. Flow rate: Adjust the flow rate so that the retention time Purity (1) Clarity and color of solution—Dissolve 1.0 g of prednisolone phosphate is about 7 minutes. of Prednisolone Sodium Phosphate in 10 mL of water: the Time span of measurement: About 4 times as long as the solution is clear and not more colored than the following retention time of prednisolone phosphate. control solution. System suitability— Control solution: To a mixture of 3.0 mL of Cobalt (II) Test for required detectability: Pipet 5 mL of the standard Chloride CS, 3.0 mL of Iron (III) Chloride CS and 2.4 mL solution, and add the mobile phase to make exactly 50 mL. of Copper (II) Sulfate CS add diluted hydrochloric acid (1 in Confirm that the peak area of prednisolone phosphate ob- JP XVI Official Monographs / Prednisolone Succinate 1289 tained from 20 mL of this solution is equivalent to 7 to 13z C25H32O8. of that of prednisolone phosphate from 20 mL of the stand- Description Prednisolone Succinate occurs as a white, fine, ard solution. crystalline powder. It is odorless. System performance: When the procedure is run with 20 It is freely soluble in methanol, soluble in ethanol (95), mL of the standard solution under the above operating con- and very slightly soluble in water and in diethyl ether. ditions, the number of theoretical plates and the symmetry Melting point: about 2059C (with decomposition). factor of the peak of prednisolone phosphate are not less than 3000 and not more than 2.0, respectively. Identification (1) To 2 mg of Prednisolone Succinate add System repeatability: When the test is repeated 6 times 2 mL of sulfuric acid, and allow to stand for 2 to 3 minutes: with 20 mL of the standard solution under the above operat- a deep red color, without fluorescence, develops. To this so- ing conditions, the relative standard deviation of the peak lution add 10 mL of water cautiously: the color disappears area of prednisolone phosphate is not more than 2.0z. and a gray, flocculent precipitate is formed. (5) Residual solvent—Being specified separately. (2) Determine the infrared absorption spectrum of Pred- nisolone Succinate as directed in the potassium bromide disk Water <2.48> Not more than 8.0z (0.1 g, volumetric titra- method under Infrared Spectrophotometry <2.25>,andcom- tion, direct titration). pare the spectrum with the Reference Spectrum or the spec- Assay Weigh accurately about 0.1 g of Prednisolone So- trum of Prednisolone Succinate RS: both spectra exhibit dium Phosphate, and dissolve in water to make exactly 100 similar intensities of absorption at the same wave numbers. mL. Pipet 2 mL of this solution, add 1 mL of alkaline phos- Optical rotation <2.49> [a]20: ＋114 – ＋1209(after drying, phatase TS, and allow to stand for 2 hours with occasional D 67 mg, methanol, 10 mL, 100 mm). shaking. To this solution add exactly 20 mL of 1-octanol, and shake vigorously. Centrifuge this solution, pipet 10 mL Purity Related substances—Dissolve 0.10 g of Predniso- of the 1-octanol layer, add 1-octanol to make exactly 50 mL, lone Succinate in methanol to make exactly 10 mL, and use and use this solution as the sample solution. Separately, this solution as the sample solution. Separately, dissolve 30 weigh accurately about 25 mg of Prednisolone RS, previ- mg of prednisolone in methanol to make exactly 10 mL. ously dried at 1059C for 3 hours, and dissolve in 1-octanol to Pipet 1 mL of the solution, add methanol to make exactly 10 make exactly 100 mL. Pipet 6 mL of this solution, add a so- mL, and use this solution as the standard solution. Perform lution prepared by adding 1 mL of alkaline phosphatase TS the test with these solutions as directed under Thin-layer to 2 mL water and being allowed to stand for 2 hours with Chromatography <2.03>. Spot 5 mL of the sample solution occasional gentle shaking, add exactly 14 mL of 1-octanol, and standard solution on a plate of silica gel with fluorescent and shake vigorously. Proceed in the same manner as the indicator for thin-layer chromatography. Develop the plate sample solution to make the standard solution. Perform the with a mixture of ethyl acetate and ethanol (95) (2:1) to a dis- test with the sample solution and standard solution as di- tance of about 10 cm, and air-dry the plate. Examine the rected under Ultraviolet-visible Spectrophotometry <2.24>, plate under ultraviolet light (main wavelength: 254 nm): the using 1-octanol as the blank, and determine the absorbances, spots other than the principal spot from the sample solution

AT and AS,at245nm. are not more intense than the spot from the standard solution. Amount (mg) of prednisolone sodium phosphate

(C21H27Na2O8P) Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- ＝ MS × AT/AS × 3 × 1.344 um, phosphorus (V) oxide, 609C, 6 hours).

MS: Amount (mg) of Prednisolone RS Residue on ignition <2.44> Not more than 0.1z (1 g). Containers and storage Containers—Tight containers. Assay Weigh accurately about 10 mg each of Prednisolone Succinate and Prednisolone Succinate RS, previously dried, and dissolve each in methanol to make exactly 100 mL. Pipet Prednisolone Succinate 5 mL each of these solutions, add methanol to make exactly 50 mL, and use these solutions as the sample solution and

プレドニゾロンコハク酸エステル standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 242 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>.

Amount (mg) of C25H32O8 ＝ MS × AT/AS

MS: Amount (mg) of Prednisolone Succinate RS Containers and storage Containers—Tight containers.

C25H32O8:460.52 11b,17,21-Trihydroxypregna-1,4-diene-3,20-dione 21-(hydrogen succinate) [2920-86-7]

Prednisolone Succinate, when dried, contains not less than 97.0z and not more than 103.0z of 1290 Prednisolone Sodium Succinate for Injection / Official Monographs JP XVI

water: the solution is clear and colorless. Prednisolone Sodium Succinate Loss on drying <2.41> Not more than 2.0z (0.15 g, in for Injection vacuum, phosphorus (V) oxide, 609C, 3 hours). Bacterial endotoxins <4.01> Less than 2.4 EU/mg of pred- 注射用プレドニゾロンコハク酸エステルナトリウム nisolone (C21H28O5). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the requirement. C H NaO :482.50 25 31 8 Sterility <4.06> Perform the test according to the Mem- Monosodium 11b,17,21-trihydroxypregna-1,4-diene-3,20- brane filtration method: it meets the requirement. dione 21-succinate [1715-33-9] Assay Take a quantity of sealed containers of Prednisolone Sodium Succinate for Injection, equivalent to about 0.1 g of

Prednisolone Sodium Succinate for Injection is a prednisolone (C21H28O5), and dissolve the contents in a suita- preparation for injection which is dissolved before ble amount of diluted methanol (1 in 2), and transfer to a used. 100-mL volumetric flask. Wash each container with diluted It contains not less than 72.4z and not more methanol (1 in 2), collect the washings in the volumetric than 83.2z of prednisolone sodium succinate flask, and add diluted methanol (1 in 2) to make volume. (C25H31NaO8), and the equivalent of not less than Pipet 4 mL of this solution, add diluted methanol (1 in 2) to 90.0z and not more than 110.0z of the labeled make exactly 50 mL. Pipet 5 mL of this solution, add exactly amount of prednisolone (C21H28O5: 360.44). 5 mL of the internal standard solution, mix, and use this so- The amount should be stated as the amount of pred- lution as the sample solution. Separately, weigh accurately nisolone (C21H28O5). about 25 mg of Prednisolone Succinate RS, previously dried in a desiccator for 6 hours (in vacuum, phosphorus (V) Method of preparation Prepare as directed under Injec- oxide, 609C), dissolve in methanol to make exactly 25 mL. tions, with Prednisolone Succinate and Dried Sodium Car- Pipet 5 mL of this solution, add diluted methanol (1 in 2) to bonate or Sodium Hydroxide. make exactly 50 mL. Pipet 5 mL of this solution, add exactly It contains a suitable buffer agent. 5 mL of the internal standard solution, mix, and use this so- Description Prednisolone Sodium Succinate for Injection lution as the standard solution. Perform the test with 10 mL occurs as a white powder or porous, friable mass. of the sample solution and standard solution as directed It is freely soluble in water. under Liquid Chromatography according <2.01> to the fol- It is hygroscopic. lowing conditions, and calculate the ratios, QT and QS,of the peak area of prednisolone succinate to that of the inter- Identification (1) To 2 mg of Prednisolone Sodium Suc- nal standard. cinate for Injection add 2 mL of sulfuric acid, and allow to stand for 2 to 3 minutes: a deep red color, without fluores- Amount (mg) of prednisolone sodium succinate cence, develops. To this solution add 10 mL of water cau- (C25H31NaO8) tiously: the color disappears and a gray, flocculent precipi- ＝ MS × QT/QS × 5 × 1.048 tate is formed. Amount (mg) of prednisolone (C H O ) (2) Dissolve 0.01 g of Prednisolone Sodium Succinate 21 28 5 ＝ M × Q /Q × 5 × 0.783 for Injection in 1 mL of methanol, add 1 mL of Fehling's S T S TS, and heat: an orange to red precipitate is formed. MS: Amount (mg) of Prednisolone Succinate RS (3) Dissolve 0.1 g of Prednisolone Sodium Succinate for Internal standard solution—A solution of propyl parahy- Injection in 2 mL of sodium hydroxide TS, allow to stand droxybenzoate in diluted methanol (1 in 2) (1 in 25,000). for 10 minutes, and filter. Add 1 mL of dilute hydrochloric Operating conditions— acid to the filtrate, shake, and filter if necessary. Adjust the Detector: An ultraviolet absorption photometer (wave- solution with diluted ammonia TS (1 in 10) to a pH of about length: 254 nm). 6, and add 2 to 3 drops of iron (III) chloride TS: a brown Column: A stainless steel column 4.6 mm in inside diame- precipitate is formed. ter and 25 cm in length, packed with octadecylsilanized silica (4) Prednisolone Sodium Succinate for Injection re- gel for liquid chromatography (5 mminparticlediameter). sponds to the Qualitative Tests <1.09> (1) for sodium salt. Column temperature: A constant temperature of about pH <2.54> Dissolve 1.0 g of Prednisolone Sodium Suc- 259C. cinate for Injection in 40 mL of water: the pH of the solu- Mobile phase: Dissolve 0.32 g of tetra n-butylammonium tion is between 6.5 and 7.2. bromide, 3.22 g of disodium hydrogen phosphate dodacahy- drate and 6.94 g of potassium dihydrogen phosphate in 1000 Purity Clarity and color of solution—Dissolve 0.25 g of mL of water. To 840 mL of this solution add 1160 mL of Prednisolone Sodium Succinate for Injection in 10 mL of methanol. JP XVI Official Monographs / Primidone 1291

Flow rate: Adjust the flow rate so that the retention time form the test with 2 mL of the sample solution and standard of prednisolone succinate is about 15 minutes. solution as directed under Gas Chromatography <2.02> System suitability— according to the following conditions, and calculate the

System performance: When the procedure is run with 10 ratios, QT and QS, of the peak area of 2-ethyl-2-phenyl- mL of the standard solution under the above operating con- malonediamide to that of the internal standard: QT is not ditions, prednisolone succinate and the internal standard are more than QS. eluted in this order with the resolution between these peaks Internal standard solution—A solution of stearylalcohol in being not less than 6. pyridine (1 in 2000). System repeatability: When the test is repeated 6 times Operating conditions— with 10 mL of the standard solution under the above operat- Detector: A hydrogen flame-ionization detector. ing conditions, the relative standard deviation of the ratios Column: A glass column 3 mm in inside diameter and 150 of the peak area of prednisolone succinate to that of the in- cm in length, packed with siliceous earth for gas chromatog- ternal standard is not more than 1.0z. raphy (125 to 150 mm in particle diameter) coated with 50z phenyl-methyl silicon polymer for gas chromatography at Containers and storage Containers—Hermetic containers. the ratio of 3z. Column temperature: A constant temperature of about 1959C. Primidone Carrier gas: Nitrogen. Flow rate: Adjust the flow rate so that the retention time プリミドン of stearylalcohol is about 10 minutes. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condition, 2-ethyl-2-phenylmalonediamide and the internal standard are eluted in this order with the resolution between these peaks being not less than 3.

C12H14N2O2: 218.25 System repeatability: When the test is repeated 5 times 5-Ethyl-5-phenyl-2,3-dihyropyrimidine-4,6(1H,5H)-dione with 2 mL of the standard solution under the above operating [125-33-7] conditions, the relative standard deviation of the ratios of the peak area of 2-ethyl-2-phenylmalonediamide to that of Primidone, when dried, contains not less than the internal standard is not more than 1.5z. 98.5z of C H N O . 12 14 2 2 Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, Description Primidone occurs as a white, crystalline pow- 2 hours). der or granules. It is odorless and has a slightly bitter taste. Residue on ignition <2.44> Not more than 0.2z (1 g). It is soluble in N,N-dimethylformamide, sparingly soluble in pyridine, slightly soluble in ethanol (95), very slightly Assay Weigh accurately about 20 mg each of Primidone soluble in water, and practically insoluble in diethyl ether. and Primidone RS, previously dried, dissolve each in 20 mL of ethanol (95) by warming, and after cooling, add ethanol Identification (1) Heat 0.5 g of Primidone with 5 mL of (95) to make exactly 25 mL, and use these solutions as the diluted sulfuric acid (1 in 2): the odor of formaldehyde is sample solution and the standard solution, respectively. perceptible. Determine the absorbance, A , of the sample solution and (2) Mix 0.2 g of Primidone with 0.2 g of anhydrous sodi- 1 standard solution at the wavelength of maximum absorption um carbonate, and heat: the gas evolved changes moistened at about 257 nm, and the absorbances, A and A ,atthe red litmus paper to blue. 2 3 wavelength of minimum absorption at about 254 nm and at Melting point <2.60> 279–2849C about 261 nm, as directed under Ultraviolet-visible Spectro- photometry <2.24>, using ethanol (95) as the blank. Purity (1) Clarity and color of solution—Dissolve 0.10 g of Primidone in 10 mL of N,N-dimethylformamide: the so- Amount (mg) of C12H14N2O2 lution is clear and colorless. ＝ MS × (2A1 － A2 － A3)T/(2A1 － A2 － A3)S (2) Heavy metals <1.07>—Proceed with 2.0 g of Primi- M : Amount (mg) of Primidone RS done according to Method 2, and perform the test. Prepare S the control solution with 2.0 mL of Standard Lead Solution where, (2A1 － A2 － A3)T is the value from the sample solu- (not more than 10 ppm). tion, and (2A1 － A2 － A3)S is from the standard solution. (3) 2-Ethyl-2-phenylmalonediamide—Dissolve 0.10 g of Containers and storage Containers—Tight containers. Primidone in 2 mL of pyridine, add exactly 2 mL of the internal standard solution, then add 1 mL of bis-trimethyl silyl acetamide, shake well, and heat at 1009C for 5 minutes. Cool, add pyridine to make 10 mL, and use this solution as the sample solution. Separately, dissolve 50 mg of 2-ethyl-2- phenylmalonediamide in pyridine to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 2 mL of the internal standard solution, proceed in thesamemannerasPrimi- done, and use this solution as the standard solution. Per- 1292 Probenecid / Official Monographs JP XVI

Assay Weigh accurately about 0.5 g of Probenecid, previ- Probenecid ously dried, and dissolve in 50 mL of neutralized ethanol. Titrate <2.50> with 0.1 mol/L sodium hydroxide VS (indica- プロベネシド tor: 3 drops of phenolphthalein TS). Each mL of 0.1 mol/L sodium hydroxide VS

＝ 28.54 mg of C13H19NO4S Containers and storage Containers—Well-closed containers.

C13H19NO4S: 285.36 4-(Dipropylaminosulfonyl)benzoic acid [57-66-9] Probenecid Tablets

Probenecid, when dried, contains not less than プロベネシド錠 98.0z of C13H19NO4S. Description Probenecid occurs as white crystals or crystal- Probenecid Tablets contain not less than 95.0z and line powder. It is odorless, and has a slightly bitter taste, fol- not more than 105.0z of the labeled amount of lowed by unpleasant bitter. probenecid (C13H19NO4S: 285.36). It is sparingly soluble in ethanol (99.5), and practically in- Method of preparation Prepare as directed under Tablets, soluble in water. with Probenecid. It dissolves in sodium hydroxide TS and in ammonia TS. Melting point: 198 – 2009C Identification (1) Weigh a quantity of powdered Probenecid Tablets, equivalent to 0.5 g of Probenecid ac- Identification (1) Heat Probenecid strongly: the odor of cording to the labeled amount, add 50 mL of ethanol (95) sulfur dioxide is perceptible. and 1 mL of 1 mol/L hydrochloric acid TS, shake, and (2) Determine the absorption spectrum of a solution of filter. Evaporate the filtrate on a water bath to about 20 mL. Probenecid in ethanol (99.5) (1 in 50,000) as directed under After cooling, collect produced crystals, recrystallize with 50 Ultraviolet-visible Spectrophotometry <2.24>,andcompare mL of dilute ethanol, and dry at 1059C for 4 hours: it melts the spectrum with the Reference Spectrum or the spectrum <2.60> between 1969Cand2009C. With the crystals so ob- of a solution of Probenecid RS prepared in the same manner tained, proceed as directed in the Identification (1) under as the sample solution: both spectra exhibit similar inten- Probenecid. sities of absorption at the same wavelengths. (2) Determine the absorption spectrum of a solution of Purity (1) Acidity—To 2.0 g of Probenecid add 100 mL the dried crystals obtained in (1) in ethanol (99.5) (1 in of water, heat on a water bath with occasional shaking for 50,000) as directed under Ultraviolet-visible Spectropho- 30 minutes, cool, and filter. To the filtrate add 1 drop of tometry <2.24>, and compare the spectrum with the Refer- phenolphthalein TS and 0.50 mL of 0.1 mol/L sodium hy- ence Spectrum or the spectrum of a solution of Probenecid droxide VS: a red color develops. RS prepared in the same manner as the sample solution: (2) Chloride <1.03>—To 1.0 g of Probenecid add 100 mL both spectra exhibit similar intensities of absorption at the of water and 1 mL of nitric acid, and heat on a water bath same wavelengths. with occasional shaking for 30 minutes. After cooling, add, Uniformity of dosage units <6.02> Perform the test accord- if necessary, water to make 100 mL, and filter. Perform the ing to the following method: it meets the requirement of the test using 50 mL of the filtrate as the test solution. Prepare Content uniformity test. the control solution with 0.30 mL of 0.01 mol/L hydrochlo- To 1 tablet of Probenecid Tablets add 30 mL of water and ric acid VS (not more than 0.021z). 2 mL of 1 mol/L hydrochloric acid TS, treat with ultrasonic (3) Sulfate <1.14>—To 1.0 g of Probenecid add 100 mL waves with occasional shaking to disintegrate the tablet com- of water and 1 mL of hydrochloric acid, and heat on a water pletely, and add ethanol (99.5) to make exactly 100 mL. Cen- bath with occasional shaking for 30 minutes. After cooling, trifuge this solution, pipet 3 mL of the supernatant liquid, add, if necessary, water to make 100 mL, and filter. Perform and add 1 mL of 1 mol/L hydrochloric acid TS and ethanol the test using 50 mL of the filtrate as the test solution. Pre- (99.5) to make exactly 50 mL. Pipet 5 mL of this solution, pare the control solution with 0.40 mL of 0.005 mol/L sulfu- and add ethanol (99.5) to make exactly V mL so that each ric acid VS (not more than 0.038z). mL contains about 15 mg of probenecid (C H NO S), and (4) Heavy metals <1.07>—Proceed with 2.0 g of 13 19 4 use this solution as the sample solution. Separately, weigh Probenecid according to Method 2, and perform the test. accurately about 0.125 g of Probenecid RS, previously dried Prepare the control solution with 2.0 mL of Standard Lead at 1059C for 4 hours, dissolve in 15 mL of water, 1 mL of 1 Solution (not more than 10 ppm). mol/L hydrochloric acid TS and ethanol (99.5) to make ex- (5) Arsenic <1.11>—Prepare the test solution with 1.0 g actly 50 mL. Pipet 3 mL of this solution, and add 1 mL of 1 of Probenecid according to Method 3, and perform the test mol/L hydrochloric acid TS and ethanol (99.5) to make ex- (not more than 2 ppm). actly 50 mL. Pipet 5 mL of this solution, add ethanol (99.5) Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, to make exactly 50 mL, and use this solution as the standard 4 hours). solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spec- Residue on ignition <2.44> Not more than 0.1z (1 g). trophotometry <2.24>, using a solution, prepared by adding JP XVI Official Monographs / Probucol 1293 ethanol (99.5) to 1 mL of 0.1 mol/L hydrochloric acid TS to MS: Amount (mg) of Probenecid RS make exactly 50 mL, as the blank, and determine the absor- Containers and storage Containers—Well-closed contain- bances, A and A ,at248nm. T S ers.

Amount (mg) of probenecid (C13H19NO4S) ＝ MS × AT/AS × V/25 Probucol MS: Amount (mg) of Probenecid RS Dissolution <6.10> When the test is performed at 50 revolu- プロブコール tions per minute according to the Paddle method, using 900 mL of the 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 30 minutes of Probenecid Tablets is not less than 80z. Start the test with 1 tablet of Probenecid Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard the first C31H48O2S2: 516.84 10 mL of the filtrate, pipet V mL of the subsequent filtrate, 4,4?-[Propan-2,2-diylbis(sulfandiyl)]bis[2,6-bis(1,1- add the dissolution medium to make exactly V?mL so that dimethylethyl)phenol] each mL contains about 14 mg of probenecid (C13H19NO4S) [23288-49-5] according to the labeled amount, and use this solution as the sample solution. Separately, weigh accurately about 70 mg Probucol, when dried, contains not less than 98.5z of Probenecid RS, previously dried at 1059C for 4 hours, and not more than 101.0z of C31H48O2S2. and dissolve in the dissolution medium to make exactly 100 Description Probucol occurs as a white crystalline powder. mL. Pipet 1 mL of this solution, add the dissolution medium It is very soluble in tetrahydrofuran, freely soluble in to make exactly 50 mL, and use this solution as the standard ethanol (99.5), soluble in methanol, and practically insoluble solution. Determine the absorbances, A and A ,at244nm T S in water. of the sample solution and standard solution as directed It gradually turns light yellow on exposure to light. under Ultraviolet-visible Spectrophotometry <2.24>. Identification (1) Determine the absorption spectrum of a Dissolution rate (z) with respect to the labeled amount solution of Probucol in methanol (1 in 100,000) as directed of probenecid (C H NO S) 13 19 4 under Ultraviolet-visible Spectrophotometry <2.24>,and ＝ M × A /A × V?/V × 1/C × 18 S T S compare the spectrum with the Reference Spectrum or the

MS: Amount (mg) of Probenecid RS spectrum of a solution of Probucol RS prepared in the same C: Labeled amount (mg) of probenecid (C13H19NO4S) in 1 manner as the sample solution: both spectra exhibit similar tablet intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Assay Weigh accurately, and powder not less than 20 Probucol as directed in the potassium bromide disk method Probenecid Tablets. Weigh accurately a portion of the pow- under Infrared Spectrophotometry <2.25>, and compare the der, equivalent to about 0.25 g of probenecid (C H NO S), 13 19 4 spectrum with the Reference Spectrum or the spectrum of add 30 mL of water and 2 mL of 1 mol/L hydrochloric acid Probucol RS: both spectra exhibit similar intensities of ab- TS, shake, add 30 mL of ethanol (99.5), disperse the parti- sorption at the same wave numbers. cles with the aid of ultrasonic waves, and add ethanol (99.5) to make exactly 100 mL. Centrifuge the solution, pipet 3 mL Melting point <2.60> 125 – 1289C of the supernatant liquid, add 1 mL of 1 mol/L hydrochloric Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of acid TS, and add ethanol (99.5) to make exactly 50 mL. Probucol according to Method 2, and perform the test. Pre- Pipet 5 mL of the solution, add ethanol (99.5) to make ex- pare the control solution with 2.0 mL of Standard Lead So- actly 50 mL, and use this solution as the sample solution. lution (not more than 10 ppm). Separately, weigh accurately about 0.125 g of Probenecid (2) Related substances—Conduct this procedure using RS, previously dried at 1059C for 4 hours, add 15 mL of light-resistant vessels. Dissolve 0.40 g of Probucol in 5 mL of water and 1 mL of 1 mol/L hydrochloric acid TS, then add ethanol (99.5), add the mobile phase to make 20 mL, and use ethanol (99.5) to make exactly 50 mL. Pipet 3 mL of this so- this solution as the sample solution. Pipet 1 mL of the sam- lution, add 1 mL of 1 mol/L hydrochloric acid TS, and add ple solution, and add the mobile phase to make exactly 50 ethanol (99.5) to make exactly 50 mL. Pipet 5 mL of the so- mL. Pipet 1 mL of this solution, add the mobile phase to lution, add ethanol (99.5) to make exactly 50 mL, and use make exactly 100 mL, and use this solution as the standard this solution as the standard solution. Determine the absor- solution. Perform the test with exactly 5 mLeachofthesam- bances, A and A , of the sample solution and standard so- T S ple solution and standard solution as directed under Liquid lution at 248 nm as directed under Ultraviolet-visible Spec- Chromatography <2.01> according to the following condi- trophotometry <2.24>, using a solution, prepared by mixing tions. Determine each peak area of both solutions by the au- 1 mL of 0.1 mol/L hydrochloric acid TS and sufficient tomatic integration method: the area of the peak having the ethanol (99.5) to make exactly 50 mL, as the blank. relative retention time of about 0.9 with respect to probucol

Amount (mg) of probenecid (C13H19NO4S) from the sample solution is not larger than the peak area of ＝ MS × AT/AS × 2 probucol from the standard solution; the area of peak hav- 1294 Probucol Fine Granules / Official Monographs JP XVI ing the relative retention time of about 1.9 with respect to and add the mobile phase to make 50 mL. probucol from the sample solution is not larger than 25 times Operating conditions— the peak area of probucol from the standard solution; and Detector: An ultraviolet absorption photometer (wave- the area of each peak other than the peak of probucol and length: 242 nm). other than the peaks mentioned above is not larger than 5 Column: A stainless steel column 4.6 mm in inside diame- times the peak area of probucol from the standard solution. ter and 25 cm in length, packed with octadecylsilanized silica Furthermore, the total area of the peaks other than probucol gel for liquid chromatography (5 mminparticlediameter). from the sample solution is not larger than 50 times the peak Column temperature: A constant temperature of about area of probucol from the standard solution. For this calcu- 409C. lation, use the areas of the peaks, having the relative reten- Mobile phase: A mixture of acetonitrile and water (93:7). tion times of about 0.9 and about 1.9 with respect to Flow rate: Adjust the flow rate so that the retention time probucol, after multiplying by their relative response fac- of probucol is about 13 minutes. tors, 1.2 and 1.4, respectively. System suitability— Operating conditions— System performance: When the procedure is run with 10 Detector, column, column temperature, mobile phase and mL of the standard solution under the above operating con- flow rate: Proceed as directed in the operating conditions in ditions, the internal standard and probucol are eluted in this the Assay. order with the resolution between these peaks being not less Time span of measurement: About 3 times as long as the than 6. retention time of probucol, beginning after the solvent peak, System repeatability: When the test is repeated 6 times excluding the peak having the relative retention time of with 10 mL of the standard solution under the above operat- about 0.5 with respect to probucol. ing conditions, the relative standard deviation of the ratio of System suitability— the peak area of probucol to that of the internal standard is Test for required detectability: Pipet 2 mL of the standard not more than 1.0z. solution, and add the mobile phase to make exactly 10 mL. Containers and storage Containers—Tight containers. Confirm that the peak area of probucol obtained from 5 mL Storage—Light-resistant. of this solution is equivalent to 14 to 26z of that of probucol from 5 mL of the standard solution. System performance: To 1 mL of the sample solution add the mobile phase to make 50 mL. To 1 mL of this solution Probucol Fine Granules add 1 mL of a solution of phthalic acid bis(cis-3,3,5- プロブコール細粒 trimethylcyclohexyl) in the mobile phase (1 in 1000), 5 mL of ethanol (99.5), and the mobile phase to make 20 mL. When the procedure is run with 5 mL of this solution under the Probucol Fine Granules contain not less than 95.0z above operating conditions, phthalic acid bis(cis-3,3,5- and not more than 105.0z of the labeled amount of trimethylcyclohexyl) and probucol are eluted in this order probucol (C31H48O2S2: 516.84). with the resolution between these peaks being not less than 6. Method of preparation Prepare as directed under Gran- System repeatability: When the test is repeated 6 times ules, with Probucol. with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area Identification To an amount of pulverized Probucol Fine of probucol is not more than 5z. Granules, equivalent to 50 mg of Probucol according to the (3) Residual solvent—Being specified separately. labeled amount, add 100 mL of methanol, shake, and filter. To 2 mL of the filtrate add methanol to make 100 mL. De- Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- termine the absorption spectrum of this solution as directed um, 809C, 1 hour). under Ultraviolet-visible Spectrophotometry <2.24>:itexhib- Residue on ignition <2.44> Not more than 0.1z (1 g). its a maximum between 240 nm and 244 nm. Assay Weigh accurately about 60 mg each of Probucol and Uniformity of dosage units <6.02> Perform the test accord- Probucol RS, previously dried, dissolve each in 5 mL of ing to the following method: the granules in single-unit con- tetrahydrofuran, and add the mobile phase to make exactly tainers meet the requirement of the Content uniformity test. 50 mL. Pipet 5 mL each of these solutions, add exactly 5 mL To the total amount of the content of 1 container of of the internal standard solution and the mobile phase to Probucol Fine Granules add 70 mL of methanol, shake thor- make 100 mL, and use these solutions as the sample solution oughly, and add methanol to make exactly 100 mL. Centri- and standard solution. Perform the test with 10 mLeachof fuge, pipet V mL of the supernatant liquid, equivalent to the sample solution and standard solution as directed under about 5 mg of probucol (C31H48O2S2), add exactly 5 mL of Liquid Chromatography <2.01> according to the following the internal standard solution, add methanol to make 100 conditions, and calculate the ratios, QT and QS, of the peak mL, and use this solution as the sample solution. Then, area of probucol to that of the internal standard. proceed as directed in the Assay.

Amount (mg) of probucol (C31H48O2S2) Amount (mg) of probucol (C31H48O2S2) ＝ MS × QT/QS ＝ MS × QT/QS × 10/V

MS: Amount (mg) of Probucol RS MS: Amount (mg) of Probucol RS Internal standard solution—Dissolve 0.2 g of bis(cis-3,3,5- Internal standard solution—A solution of bis(cis-3,3,5- trimethylcyclohexyl) phthalate in 1 mL of tetrahydrofuran, trimethylcyclohexyl) phthalate in methanol (1 in 250). JP XVI Official Monographs / Probucol Tablets 1295

Particle size <6.03> It meets the requirements of Fine gran- termine the absorption spectrum of this solution as directed ules. under Ultraviolet-visible Spectrophotometry <2.24>:itexhibits a maximum between 240 nm and 244 nm. Assay Weigh accurately an amount of pulverized Probucol Fine Granules, equivalent to about 0.25 g of probucol Uniformity of dosage units <6.02> Perform the test accord-

(C31H48O2S2), add 70 mL of methanol, shake thoroughly, ing to the following method: it meets the requirement of the and add methanol to make exactly 100 mL. Centrifuge, pipet Content uniformity test. 2 mL of the supernatant liquid, add exactly 5 mL of the in- Shake 1 tablet of Probucol Tablets with a suitable amount ternal standard solution, add methanol to make 100 mL, and of methanol until the tablet is disintegrated, and add metha- use this solution as the sample solution. Separately, weigh nol to make exactly V mL so that each mL of the solution accurately about 50 mg of Probucol RS, previously dried contains about 2.5 mg of probucol (C31H48O2S2). Centrifuge under reduced pressure at 809C for 1 hour, and dissolve in the solution, pipet 2 mL of the supernatant liquid, add ex- methanol to make exactly 20 mL. Pipet 2 mL of this solu- actly 5 mL of the internal standard solution, then add meth- tion, add exactly 5 mL of the internal standard solution, add anol to make 100 mL, and use this solution as the sample so- methanol to make 100 mL, and use this solution as the lution. Then, proceed as directed in the Assay. standard solution. Perform the test with 10 mLeachofthe Amount (mg) of probucol (C H O S ) sample solution and standard solution as directed under Liq- 31 48 2 2 ＝ M × Q /Q × V/20 uid Chromatography <2.01> according to the following con- S T S ditions, and calculate the ratios, QT and QS, of the peak area MS: Amount (mg) of Probucol RS of probucol to that of the internal standard. Internal standard solution—A solution of bis(cis-3,3,5-

Amount (mg) of probucol (C31H48O2S2) trimethylcyclohexyl) phthalate in methanol (1 in 250). ＝ M × Q /Q × 5 S T S Disintegration <6.09> It meets the requirement. M : Amount (mg) of Probucol RS S Assay Weigh accurately the mass of 20 Probucol Tablets, Internal standard solution—A solution of bis(cis-3,3,5- and powder the tablets. Weigh accurately a portion of trimethylcyclohexyl) phthalate in methanol (1 in 250). the powder, equivalent to about 0.25 g of probucol

Operating conditions— (C31H48O2S2), add 70 mL of methanol, shake thoroughly, Detector, column temperature, mobile phase, and flow and add methanol to make exactly 100 mL. Centrifuge, pipet rate: Proceed as directed in the operating conditions in the 2 mL of the supernatant liquid, add exactly 5 mL of the in- Assay under Probucol. ternal standard solution, add methanol to make 100 mL, and Column: A stainless steel column 4.6 mm in inside diame- use this solution as the sample solution. Separately, weigh ter and 15 cm in length, packed with octadecylsilanized silica accurately about 50 mg of Probucol RS, previously dried gel for liquid chromatography (5 mm in particle diameter). under reduced pressure at 809C for 1 hour, and dissolve in System suitability— methanol to make exactly 20 mL. Pipet 2 mL of this solu- System performance: When the procedure is run with 10 tion, add exactly 5 mL of the internal standard solution, add mL of the standard solution under the above operating con- methanol to make 100 mL, and use this solution as the ditions, the internal standard and probucol are eluted in this standard solution. Perform the test with 10 mLeachofthe order with the resolution between these peaks being not less sample solution and standard solution as directed under Liq- than 3. uid Chromatography <2.01> according to the following con-

System repeatability: When the test is repeated 6 times ditions, and calculate the ratios, QT and QS, of the peak area with 10 mL of the standard solution under the above operat- of probucol to that of the internal standard. ing conditions, the relative standard deviation of the ratio of Amount (mg) of probucol (C H O S ) the peak area of probucol to that of the internal standard is 31 48 2 2 ＝ M × Q /Q × 5 not more than 1.0z. S T S M : Amount (mg) of Probucol RS Containers and storage Containers—Well-closed contain- S ers. Internal standard solution—A solution of bis(cis-3,3,5- trimethylcyclohexyl) phthalate in methanol (1 in 250). Operating conditions— Probucol Tablets Detector, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the プロブコール錠 Assay under Probucol. Column: A stainless steel column 4.6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica Probucol Tablets contain not less than 95.0z and gel for liquid chromatography (5 mminparticlediameter). not more than 105.0z of probucol (C H O S : 31 48 2 2 System suitability— 516.84). System performance: When the procedure is run with 10 Method of preparation Prepare as directed under Tablets, mL of the standard solution under the above operating con- with Probucol. ditions, the internal standard and probucol are eluted in this order with the resolution between these peaks being not less Identification To an amount of pulverized Probucol than 3. Tablets, equivalent to 50 mg of Probucol according to the System repeatability: When the test is repeated 6 times labeled amount, add 100 mL of methanol, shake, and filter. with 10 mL of the standard solution under the above operat- To 2 mL of the filtrate add methanol to make 100 mL. De- 1296 Procainamide Hydrochloride / Official Monographs JP XVI ing conditions, the relative standard deviation of the ratio of integration method: the total area of the peaks other than the peak area of probucol to that of the internal standard is procainamide from the sample solution is not larger than the not more than 1.0z. peak area of procainamide from the standard solution. Operating conditions— Containers and storage Containers—Well-closed contain- Detector: An ultraviolet absorption photometer (wave- ers. length: 270 nm). Column: A stainless steel column 4.6 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica Procainamide Hydrochloride gel for liquid chromatography (5 mminparticlediameter). Column temperature: A constant temperature of about プロカインアミド塩酸塩 409C. Mobile phase: A mixture of 0.02 mol/L phosphate buffer solution, pH 3.0 and methanol (9:1). Flow rate: Adjust the flow rate so that the retention time of procainamide is about 9 minutes. Time span of measurement: About 2 times as long as the retention time of procainamide. C H N O.HCl: 271.79 13 21 3 System suitability— 4-Amino-N-(2-diethylaminoethyl)benzamide Test for required detectability: Pipet 10 mL of the stand- monohydrochloride ard solution, and add the mobile phase to make exactly 20 [614-39-1] mL. Confirm that the peak area of procainamide obtained from 10 mL of this solution is equivalent to 40 to 60z of that Procainamide Hydrochloride, when dried, contains of procainamide from 10 mL of the standard solution. not less than 98.0z and not more than 101.0z of System performance: When the procedure is run with 10 C H N O.HCl. 13 21 3 mL of the standard solution under the above operating con- Description Procainamide Hydrochloride occurs as a white ditions, the number of theoretical plates and the symmetry to light yellow crystalline powder. factor of the peak of procainamide are not less than 10,000 It is very soluble in water and soluble in ethanol (99.5). and not more than 1.5, respectively. It is hygroscopic. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- Identification (1) Determine the infrared absorption spec- ing conditions, the relative standard deviation of the peak trum of Procainamide Hydrochloride, previously dried, as area of procainamide is not more than 2.0z. directed in the potassium chloride disk method under In- frared Spectrophotometry <2.25>, and compare the spectrum Loss on drying <2.41> Not more than 0.3z (2 g, 1059C, with the Reference Spectrum: both spectra exhibit similar in- 4 hours). tensities of absorption at the same wave numbers. Residue on ignition <2.44> Not more than 0.1z (2 g). (2) A solution of Procainamide Hydrochloride (1 in 20) responds to the Qualitative Tests <1.09> for chloride. Assay Weigh accurately about 0.5 g of Procainamide Hy- drochloride, previously dried, dissolve in 50 mL of a mixture pH <2.54> Dissolve 1.0 g of Procainamide Hydrochloride of acetic anhydride and acetic acid (100) (7:3), and titrate in 10 mL of water: the pH of this solution is between 5.0 and <2.50> with 0.1 mol/L perchloric acid VS (potentiometric 6.5. titration). Perform a blank determination, and make any Melting point <2.60> 165–1699C necessary correction. Purity (1) Clarity and color of solution—Dissolve 1.0 g Each mL of 0.1 mol/L perchloric acid VS of Procainamide Hydrochloride in 10 mL of water: the solu- ＝ 27.18 mg of C13H21N3O.HCl tion is clear and colorless. Containers and storage Containers—Tight containers. (2) Heavy metals <1.07>—Proceed with 2.0 g of Procain- amide Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand- ard Lead Solution (not more than 10 ppm). Procainamide Hydrochloride (3) Arsenic <1.11>—Prepare the test solution with 1.0 g Injection of Procainamide Hydrochloride according to Method 1, and perform the test (not more than 2 ppm). プロカインアミド塩酸塩注射液 (4) Related substances—Dissolve 50 mg of Procainamide Hydrochloride in 100 mL of the mobile phase, and use this Procainamide Hydrochloride Injection is an solution as the sample solution. Pipet 1 mL of the sample so- aqueous solution for injection. lution, and add the mobile phase to make exactly 50 mL. It contains not less than 95.0 and not more than Pipet 2 mL of this solution, add the mobile phase to make z exactly 20 mL, and use this solution as the standard solution. 105.0z of the labeled amount of procainamide hydrochloride (C H N O.HCl: 271.79). Perform the test with exactly 10 mLeachofthesamplesolu- 13 21 3 tion and standard solution as directed under Liquid Chroma- Method of preparation Prepare as directed under Injec- tography <2.01> according to the following conditions. De- tions, with Procainamide Hydrochloride. termine each peak area of both solutions by the automatic JP XVI Official Monographs / Procainamide Hydrochloride Tablets 1297

Description Procainamide Hydrochloride Injection is a amines. clear, colorless or light yellow liquid. Uniformity of dosage units <6.02> Perform the test accord- pH:4.0–6.0 ing to the following method: it meets the requirement of the Identification (1) To a volume of Procainamide Hydro- Content uniformity test. chloride Injection, equivalent to 10 mg of Procainamide Hy- To 1 tablet of Procainamide Hydrochloride Tablets add drochloride according to the labeled amount, add 1 mL of 3V/5 mL of 0.02 mol/L phosphate buffer solution, pH 3.0, dilute hydrochloric acid and water to make 5 mL: the solu- treat with ultrasonic waves to disintegrate the tablet com- tion responds to the Qualitative Tests <1.09> (1) for primary pletely, add 0.02 mol/L phosphate buffer solution, pH 3.0, aromatic amines. to make exactly V mL so that each mL contains about 2.5

(2) To a volume of Procainamide Hydrochloride Injec- mg of procainamide hydrochloride (C13H21N3O.HCl), and tion, equivalent to 0.1 g of Procainamide Hydrochloride ac- shake for 5 minutes. Centrifuge this solution, pipet 1 mL of cording to the labeled amount, add water to make 100 mL. the supernatant liquid, add 0.02 mol/L phosphate buffer so- To 1 mL of this solution add water to make 100 mL. Deter- lution, pH 3.0, to make exactly 250 mL, and use this solu- mine the absorption spectrum of this solution as directed tion as the sample solution. Proceed as directed in the Assay. under Ultraviolet-visible Spectrophotometry <2.24>:itexhib- Amount (mg) of procainamide hydrochloride its a maximum between 277 nm and 281 nm. (C H N O.HCl) (3) Procainamide Hydrochloride Injection responds to 13 21 3 ＝ M × A /A × V/20 the Qualitative Tests <1.09> (2) for chloride. S T S M : Amount (mg) of procainamide hydrochloride for Bacterial endotoxins <4.01> Less than 0.30 EU/mg. S assay Extractable volume <6.05> It meets the requirement. Dissolution <6.10> When the test is performed at 50 revolu- Foreign insoluble matter <6.06> Perform the test according tions per minute according to the Paddle method, using 900 to Method 1: it meets the requirement. mL of water as the dissolution medium, the dissolution rate in 30 minutes of Procainamide Hydrochloride Tablets is not Insoluble particulate matter <6.07> It meets the require- less than 80z. ment. Start the test with 1 tablet of Procainamide Hydrochloride Sterility <4.06> Perform the test according to the Mem- Tablets, withdraw not less than 30 mL of the medium at the brane filtration method: it meets the requirement. specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Dis- Assay Dilute an accurately measured volume of Procain- card the first 10 mL of the filtrate, pipet V mL of the subse- amide Hydrochloride Injection, equivalent to about 0.5 g of quent filtrate, add 2nd fluid for dissolution test to make procainamide hydrochloride (C H N O.HCl), with 5 mL of 13 21 3 exactly V?mL so that each mL contains about 7 mgof hydrochloric acid and water to 50 mL, add 10 mL of potas- procainamide hydrochloride (C H N O.HCl) according to sium bromide solution (3 → 10), cool to 159C or lower, and 13 21 3 the labeled amount, and use this solution as the sample titrate <2.50> with 0.1 mol/L sodium nitrite VS (potentiome- solution. Separately, weigh accurately about 0.125 g of tric titration method or amperometric titration). procainamide hydrochloride for assay, previously dried at Each mL of 0.1 mol/L sodium nitrite VS 1059C for 4 hours, and dissolve in water to make exactly

＝ 27.18 mg of C13H21N3O.HCl 1000 mL. Pipet 5 mL of this solution, add 2nd fluid for dissolution test to make exactly 100 mL, and use this solution Containers and storage Containers—Hermetic containers. as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet- Procainamide Hydrochloride visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS,at278nm. Tablets Dissolution rate (z) with respect to the labeled amount of procainamide hydrochloride (C H N O.HCl) プロカインアミド塩酸塩錠 13 21 3 ＝ MS × AT/AS × V?/V × 1/C × 9/2 M : Amount (mg) of procainamide hydrochloride for Procainamide Hydrochloride Tablets contain not S assay less than 95.0 and not more than 105.0 of the z z C: Labeled amount (mg) of procainamide hydrochloride labeled amount of procainamide hydrochloride (C13H21N3O.HCl) in 1 tablet (C13H21N3O.HCl: 271.79). Assay To 10 Procainamide Hydrochloride Tablets add Method of preparation Prepare as directed under Tablets, about 300 mL of 0.02 mol/L phosphate buffer solution, pH with Procainamide Hydrochloride. 3.0, and treat with ultrasonic waves to disintegrate the Identification To a quantity of powdered Procainamide tablets completely. To this solution add 0.02 mol/L phos- Hydrochloride Tablets, equivalent to 1.5 g of Procainamide phate buffer solution, pH 3.0, to make exactly 500 mL, and Hydrochloride according to the labeled amount, add 30 mL stir for 5 minutes. Centrifuge this solution, pipet V mL of of water, shake well, filter, and use the filtrate as the sample the supernatant liquid, and add 0.02 mol/L phosphate solution. To 0.2 mL of the sample solution add 1 mL of buffer solution, pH 3.0, to make exactly V?mL so that each dilute hydrochloric acid and 4 mL of water: the solution re- mL contains about 10 mg of procainamide hydrochloride sponds to the Qualitative Tests <1.09> for primary aromatic (C13H21N3O.HCl). Filter this solution through a membrane 1298 Procaine Hydrochloride / Official Monographs JP XVI filter with a pore size not exceeding 0.45 mm, discard the first It is very soluble in water, soluble in ethanol (95), and 10 mL of the filtrate, and use the subsequent filtrate as the practically insolubleindiethylether. sample solution. Separately, weigh accurately about 50 mg Identification (1) Determine the absorption spectrum of a of procainamide hydrochloride for assay, previously dried at solution of Procaine Hydrochloride (1 in 100,000) as di- 1059C for 4 hours, dissolve in 0.02 mol/L phosphate buffer rected under Ultraviolet-visible Spectrophotometry <2.24>, solution, pH 3.0, to make exactly 100 mL. Pipet 2 mL of this and compare the spectrum with the Reference Spectrum: solution, add 0.02 mol/L phosphate buffer solution, pH 3.0, both spectra exhibit similar intensities of absorption at the to make exactly 100 mL, and use this solution as the stand- same wavelengths. ard solution. Perform the test with exactly 10 mLeachofthe (2) Determine the infrared absorption spectrum of sample solution and standard solution as directed under Liq- Procaine Hydrochloride, previously dried, as directed in the uid Chromatography <2.01> according to the following con- potassium chloride disk method under Infrared Spectropho- ditions, and determine the peak areas, A and A ,ofpro- T S tometry <2.25>, and compare the spectrum with the Refer- cainamide in each solution. ence Spectrum: both spectra exhibit similar intensities of ab- Amount (mg) of procainamide hydrochloride sorption at the same wave numbers.

(C13H21N3O.HCl) (3) A solution of Procaine Hydrochloride (1 in 10) re- ＝ MS × AT/AS × V?/V × 1/10 sponds to the Qualitative Tests <1.09> for chloride.

MS: Amount (mg) of procainamide hydrochloride for pH <2.54> The pH of a solution prepared by dissoluing assay 1.0 g of Procaine Hydrochloride in 20 mL of water is between 5.0 and 6.0. Operating conditions— Detector: An ultraviolet absorption photometer (wave- Melting point <2.60> 155 – 1589C length: 270 nm). Purity (1) Clarity and color of solution—Dissolve 1.0 g Column: A stainless steel column 4.6 mm in inside diame- of Procaine Hydrochloride in 10 mL of water: the solution is ter and 25 cm in length, packed with octadecylsilanized silica clear and colorless. gel for liquid chromatography (5 mm in particle diameter). (2) Heavy metals <1.07>—Proceed with 1.0 g of Procaine Column temperature: A constant temperature of about Hydrochloride according to Method 1, and perform the test. 409C. Prepare the control solution with 2.0 mL of Standard Lead Mobile phase: A mixture of 0.02 mol/L phosphate buffer Solution (not more than 20 ppm). solution, pH 3.0, and methanol (9:1). (3) Related substances—To 1.0 g of Procaine Hydro- Flow rate: Adjust the flow rate so that the retention time chloride add 5 mL of ethanol (95), dissolve by mixing well, of procainamide is about 9 minutes. add water to make exactly 10 mL, and use this solution as System suitability— the sample solution. Separately, dissolve 10 mg of 4- System performance: When the procedure is run with 10 aminobenzoic acid in ethanol (95) to make exactly 20 mL, mL of the standard solution under the above operating con- then pipet 1 mL of this solution, add 4 mL of ethanol (95) ditions, the number of theoretical plates and the symmetry and water to make exactly 10 mL, and use this solution as factor of the peak of procainamide are not less than 10,000 the standard solution. Perform the test with these solutions and not more than 1.5, respectively. as directed under Thin-layer Chromatography <2.03>. Spot 5 System repeatability: When the test is repeated 6 times mL each of the sample solution and standard solution on a with 10 mL of the standard solution under the above operat- plate of silica gel with fluorescent indicator for thin-layer ing conditions, the relative standard deviation of the peak chromatography. Develop the plate with a mixture of dibutyl area of procainamide is not more than 1.0z. ether, n-hexane and acetic acid (100) (20:4:1) to a distance of Containers and storage Containers—Tight containers. about 10 cm, and air-dry the plate. After drying the plate more at 1059C for 10 minutes, examine under ultraviolet light (main wavelength: 254 nm): the spots other than the Procaine Hydrochloride principal spot from the sample solution are not more intense than the spot from the standard solution. The principal spot プロカイン塩酸塩 from the sample solution stays at the origin. Loss on drying <2.41> Not more than 0.5z (1 g, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.4 g of Procaine Hydro- chloride, previously dried, dissolve in 5 mL of hydrochloric C H N O .HCl: 272.77 13 20 2 2 acid and 60 mL of water, add 10 mL of a solution of potas- 2-(Diethylamino)ethyl 4-aminobenzoate monohydrochloride sium bromide (3 in 10), cool to below 159C, and titrate [51-05-8] <2.50> with 0.1 mol/L sodium nitrite VS (potentiometric titration or amperometric titration). Procaine Hydrochloride, when dried, contains not less than 99.0z of C13H20N2O2.HCl. Each mL of 0.1 mol/L sodium nitrite VS ＝ 27.28 mg of C H N O .HCl Description Procaine Hydrochloride occurs as white crys- 13 20 2 2 tals or crystalline powder. Containers and storage Containers—Well-closed contain- JP XVI Official Monographs / Procarbazine Hydrochloride 1299 ers. Internal standard solution—A solution of caffeine in the mobile phase (1 in 1000). Operating conditions— Procaine Hydrochloride Injection Detector: An ultraviolet absorption photometer (wavelength: 254 nm). プロカイン塩酸塩注射液 Column: A stainless steel column about 6 mm in inside diameter and about 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparti- Procaine Hydrochloride Injection is an aqueous so- cle diameter). lution for injection. Column temperature: A constant temperature of about It contains not less than 95.0z and not more than 409C. 105.0z of the labeled amount of procaine hydrochlo- Mobile phase: Adjust the pH of 0.05 mol/L potassium ride (C H N O .HCl: 272.77). 13 20 2 2 dihydrogen phosphate TS to 3.0 with phosphoric acid, and Method of preparation Prepare as directed under Injec- add an amount of sodium 1-pentane sulfonate to make a so- tions, with Procaine Hydrochloride. lution so that containing 0.1z. To 800 mL of this solution add 200 mL of methanol. Description Procaine Hydrochloride Injection is a clear, Flow rate: Adjust the flow rate so that the retention time colorless liquid. of procaine is about 10 minutes. Identification (1) To a volume of Procaine Hydrochlo- System suitability— ride Injection, equivalent to 0.01 g of Procaine Hydrochlo- System performance: When the procedure is run with 5 mL ride according to the labeled amount, add water to make of the standard solution under the above operating condi- 1000 mL. Determine the absorption spectrum of this solutions, procaine and the internal standard are eluted in this tion as directed under Ultraviolet-visible Spectrophotometry order with the resolution between these peaks being not less <2.24>: it exhibits maxima between 219 nm and 223 nm, and than 8. between 289 nm and 293 nm. System repeatability: When the test is repeated 6 times (2) Procaine Hydrochloride Injection responds to the with 5 mL of the standard solution under the above operating Qualitative Tests <1.09> (2) for chloride. conditions, the relative standard deviation of the ratios of the peak area of procaine to that of the internal standard is pH <2.54> 3.3–6.0 not more than 1.0z. Bacterial endotoxins <4.01> Less than 0.02 EU/unit. Apply Containers and storage Containers—Hermetic containers. to the preparations intended for intraspinal administration. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according Procarbazine Hydrochloride to Method 1: it meets the requirement. プロカルバジン塩酸塩 Insoluble particulate matter <6.07> It meets the requirement. Sterility <4.06> Perform the test according to the Mem- brane filtration method: it meets the requirement. Assay To an exactly measured volume of Procaine Hydro- chloride Injection, equivalent to about 20 mg of procaine C12H19N3O.HCl: 257.76 hydrochloride (C13H20N2O2.HCl), add the mobile phase to N-(1-Methylethyl)- make exactly 20 mL. Pipet 5 mL of this solution, add exactly 4-[(2-methylhydrazino)methyl]benzamide 5 mL of the internal standard solution and the mobile phase monohydrochloride to make 20 mL, and use this solution as the sample solution. [366-70-1] Separately, weigh accurately about 50 mg of procaine hydrochloride for assay, previously dried in a desiccator (silica gel) Procarbazine Hydrochloride, when dried, contains for 4 hours, dissolve in the mobile phase to make exactly 50 not less than 98.5z and not more than 101.0z of mL. Pipet 5 mL of this solution, add exactly 5 mL of the in- C12H19N3O.HCl. ternal standard solution and the mobile phase to make 20 Description Procarbazine Hydrochloride occurs as white to mL, and use this solution as the standard solution. Perform light yellowish white crystals or crystalline powder. the test with 5 mL each of the sample solution and standard It is freely soluble in water, and slightly soluble in ethanol solution as directed under Liquid Chromatography <2.01> (99.5). according to the following conditions, and calculate the It dissolves in dilute hydrochloric acid. ratios, Q and Q , of the peak area of procaine hydrochlo- T S Melting point: about 2239C (with decomposition). ride to that of the internal standard. Identification (1) Dissolve 0.01 g of Procarbazine Hydro- Amount (mg) of procaine hydrochloride chloride in 1 mL of diluted copper (II) sulfate TS (1 in 10), (C H N O .HCl) 13 20 2 2 and add 4 drops of sodium hydroxide TS: a green precipitate ＝ M × Q /Q × 2/5 S T S is formed immediately, and the color changes from green

MS: Amount (mg) of procaine hydrochloride for assay through yellow to orange. 1300 Procaterol Hydrochloride Hydrate / Official Monographs JP XVI

(2) Determine the absorption spectrum of a solution of Procarbazine Hydrochloride in 0.1 mol/L hydrochloric acid Procaterol Hydrochloride Hydrate TS (1 in 100,000) as directed under Ultraviolet-visible Spec- trophotometry <2.24>, and compare the spectrum with the プロカテロール塩酸塩水和物 Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (3) Determine the infrared absorption spectrum of Procarbazine Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro- photometry <2.25>, and compare the spectrum with the Ref- erence Spectrum: both spectra exhibit similar intensities of 1 C H N O .HCl. H O: 335.83 absorption at the same wave numbers. 16 22 2 3 2 2 8-Hydroxy-5-{(1RS,2SR)-1-hydroxy- (4) A solution of Procarbazine Hydrochloride (1 in 20) 2-[(1-methylethyl)amino]butyl]}quinolin-2(1H)-one responds to the Qualitative Tests <1.09> for chloride. monohydrochloride hemihydrate pH <2.54> Dissolve 0.10 g of Procarbazine Hydrochloride [62929-91-3, anhydride] in 10 mL of water: the pH of this solution is between 3.0 and 5.0. Procaterol Hydrochloride Hydrate contains not less than 98.5z of procaterol hydrochloride Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of (C H N O .HCl: 326.82), calculated on the anhy- Procarbazine Hydrochloride according to Method 4, and 16 22 2 3 drous basis. perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Description Procaterol Hydrochloride Hydrate occurs as (2) Related substances—Dissolve 50 mg of Procarbazine white to pale yellowish white crystals or crystalline powder. Hydrochloride in 5.0 mL of a solution of L-cysteine hydro- It is soluble in water, in formic acid and in methanol, chloride monohydrate in diluted methanol (7 in 10) (1 in slightly soluble in ethanol (95), and practically insoluble in 200), and use this solution as the sample solution. Pipet 1 diethyl ether. mL of the sample solution, add a solution of L-cysteine The pH of a solution of Procaterol Hydrochloride Hy- hydrochloride monohydrate in diluted methanol (7 in 10) (1 drate (1 in 100) is between 4.0 and 5.0. in 200) to make exactly 50 mL, and use this solution as the It is gradually colored by light. standard solution. Perform the test with these solutions as The solution of Procaterol Hydrochloride Hydrate (1 in directed under Thin-layer Chromatography <2.03>.Immerse 20) shows no optical rotation. slowly, by inclining, a plate of silica gel with fluorescent Melting point: about 1959C (with decomposition). indicator for thin-layer chromatography in a solution of Identification (1) Determine the absorption spectrum of a L-cysteine hydrochloride monohydrate in diluted methanol solution of Procaterol Hydrochloride Hydrate (7 in (7 in 10) (1 in 200), allow to stand for 1 minute, lift the plate 1,000,000) as directed under Ultraviolet-visible Spectropho- from the solution, dry it in cold wind for 10 minutes, then tometry <2.24>, and compare the spectrum with the Refer- dry in warm wind for 5 minutes, and then dry at 609Cfor5 ence Spectrum: both spectra exhibit similar intensities of ab- minutes. After cooling, spot 5 mLeachofthesamplesolu- sorption at the same wavelengths. tion and standard solution on the plate. Develop the plate (2) Determine the infrared absorption spectrum of with a mixture of methanol and ethyl acetate (1:1) to a dis- Procaterol Hydrochloride Hydrate as directed in the potas- tance of about 12 cm, and air-dry the plate. Examine under sium bromide disk method under Infrared Spectrophotome- ultraviolet light (main wavelength: 254 nm): not more than 1 try <2.25>, and compare the spectrum with the Reference spot other than the principal spot and the spot of the starting Spectrum: both spectra exhibit similar intensities of absorp- point from the sample solution appears, and is not more tion at the same wave numbers. intense than the spot from the standard solution. (3) A solution of Procaterol Hydrochloride Hydrate (1 Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, in 50) responds to the Qualitative Tests <1.09> for chloride. 2 hours). Purity (1) Clarity and color of solution—Dissolve 1.0 g Residue on ignition <2.44> Not more than 0.1z (1 g). of Procaterol Hydrochloride Hydrate in 30 mL of water: the solution is clear, and has no more color than the following Assay Weigh accurately about 0.15 g of Procarbazine Hy- control solution. drochloride, previously dried, place in a glass-stoppered Control solution: To 3.0 mL of Iron (III) Chloride CS add flask, dissolve in 25 mL of water, add 25 mL of hydrochloric water to make 50 mL. acid, and cool to room temperature. To this solution add 5 (2) Heavy metals <1.07>—Proceedwith2.0gof mL of chloroform, and titrate <2.50>, while shaking, with Procaterol Hydrochloride Hydrate according to Method 2, 0.05 mol/L potassium iodate VS until the purple color of the and perform the test. Prepare the control solution with 2.0 chloroform layer disappears. The end point is reached when mL of Standard Lead Solution (not more than 10 ppm). the red-purple color of the chloroform layer no more reap- (3) Related substances—Dissolve 0.10 g of Procaterol pears within 5 minutes after the purple color disappeared. Hydrochloride Hydrate in 100 mL of diluted methanol (1 in Each mL of 0.05 mol/L potassium iodate VS 2), and use this solution as the sample solution. Pipet 1 mL

＝ 8.592 mg of C12H19N3O.HCl of the sample solution, add diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard Containers and storage Containers—Tight containers. solution. Perform the test with exactly 2 mLeachofthesam- JP XVI Official Monographs / Prochlorperazine Maleate 1301 ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi- Prochlorperazine Maleate tions. Determine each peak area of these solutions by the automatic integration method: the total area of the peaks other プロクロルペラジンマレイン酸塩 than procaterol from the sample solution is not larger than the peak area of procaterol from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 254 nm). Column: A stainless steel column about 4 mm in inside diameter and about 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparti- C H ClN S.2C H O : 606.09 cle diameter). 20 24 3 4 4 4 2-Chloro-10-[3-(4-methylpiperazin-1-yl)propyl]- Column temperature: A constant temperature of about 10H-phenothiazine dimaleate 409C. [84-02-6] Mobile phase: Dissolve 0.87 g of sodium 1-pentanesulfonate in 1000 mL of water. To 760 mL of this solution add Prochlorperazine Maleate, when dried, contains not 230 mL of methanol and 10 mL of acetic acid (100). less than 98.0z of C H ClN S.2C H O . Flow rate: Adjust the flow rate so that the retention time 20 24 3 4 4 4 of procaterol is about 15 minutes. Description Prochlorperazine Maleate occurs as a white to Selection of column: Dissolve 20 mg each of Procaterol light yellow powder. It is odorless, and has a slightly bitter Hydrochloride Hydrate and threoprocaterol hydrochloride taste. in 100 mL of diluted methanol (1 in 2). To 15 mL of this so- It is slightly soluble in acetic acid (100), very slightly solu- lution add diluted methanol (1 in 2) to make 100 mL. Pro- ble in water and in ethanol (95), and practically insoluble in ceed with 2 mL of this solution under the above operating diethyl ether. conditions, and calculate the resolution. Use a column giving It gradually acquires a red tint by light. elution of procaterol and threoprocaterol in this order with Melting point: 195 – 2039C (with decomposition). the resolution of these peaks being not less than 3. Identification (1) Dissolve 5 mg of Prochlorperazine Detection sensitivity: Adjust the detection sensitivity so Maleate in 5 mL of sulfuric acid: a red color develops, which that the peak height of procaterol obtained from 2 mLofthe darkens slowly on standing. Warm a half of the solution: the standard solution is not less than 10 mm. color changes to red-purple. To the remainder add 1 drop of Time span of measurement: 2.5 times as long as the reten- potassium dichromate TS: a green-brown color develops, tion time of procaterol beginning after the solvent peak. which changes to brown on standing. Water <2.48> 2.5 – 3.3z (0.5 g, volumetric titration, direct (2) Boil 0.5 g of Prochlorperazine Maleate with 10 mL of titration). hydrobromic acid under a reflux condenser for 10 minutes. After cooling, add 100 mL of water, and filter through glass Residue on ignition <2.44> Not more than 0.1z (1 g). filter (G4). Wash the residue with three 10-mL portions of Assay Weigh accurately about 0.25 g of Procaterol Hydro- water, and dry at 1059C for 1 hour: it melts <2.60> between chloride Hydrate, add 2 mL of formic acid, dissolve by 1959Cand1989C (with decomposition). warming, and add exactly 15 mL of 0.1 mol/L perchloric (3) Dissolve 0.2 g of Prochlorperazine Maleate in 5 mL acid VS. Add 1 mL of acetic anhydride, heat on a water bath of a solution of sodium hydroxide (1 in 10), and extract with for 30 minutes, cool, add 60 mL of acetic anhydride, and three 3-mL portions of diethyl ether [reserve the aqueous titrate <2.50> the excess perchloric acid with 0.1 mol/L sodi- layer, and use for test (4)]. Evaporate the combined diethyl um acetate VS (potentiometric titration). Perform a blank etherextractsonawaterbathto dryness, dissolve the residue determination. in 10 mL of methanol by warming, and pour into 30 mL of a solution of 2,4,6-trinitrophenol in methanol (1 in 75), previ- Each mL of 0.1 mol/L perchloric acid VS ously warmed to 509C. Allow to stand for 1 hour, collect the ＝ 32.68 mg of C H N O .HCl 16 22 2 3 crystals, wash with a small amount of methanol, and dry at Containers and storage Containers—Well-closed contain- 1059C for 1 hour: the crystals melt <2.60> between 2529C ers. and 2589C (with decomposition). Storage—Light-resistant. (4) To the aqueous layer reserved in (3) add boiling chips, and heat on a water bath for 10 minutes. Cool, add 2 mL of bromine TS, heat on a water bath for 10 minutes, and heat the solution to boil. After cooling, add 2 drops of this solution to 3 mL of a solution of resorcinol in sulfuric acid (1 in 300), and heat on a water bath for 15 minutes: a red- purple color is produced. Purity Heavy metals <1.07>—Proceed with 1.0 g of Pro- chlorperazine Maleate according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Stand- ard Lead Solution (not more than 10 ppm). 1302 Prochlorperazine Maleate Tablets / Official Monographs JP XVI

Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, of potassium permanganate TS: the red color of the test so- 3 hours). lution is discharged immediately. Residue on ignition <2.44> Not more than 0.1z (1 g). Uniformity of dosage units <6.02> Perform the test according to the following method: it meets the requirement of the Assay Weigh accurately about 0.3 g of Prochlorperazine Content uniformity test. Maleate, previously dried, dissolve in 60 mL of acetic acid Conduct this procedure using light-resistant vessels. To 1 (100) while stirring and warming. Cool, and titrate <2.50> tablet of Prochlorperazine Maleate Tablets add 3V/5 mL of with 0.05 mol/L perchloric acid VS until the color of the so- a mixture of dilute phosphoric acid (1 in 500) and ethanol lution changes from orange to green (indicator: 0.5 mL of p- (99.5) (1:1), treat with ultrasonic waves until the tablet is dis- naphtholbenzein TS). Perform a blank determination, and integrated, and shake vigorously for 10 minutes. Add exactly make any necessary correction. V/20 mL of the internal standard solution, and a mixture of Each mL of 0.05 mol/L perchloric acid VS dilute phosphoric acid (1 in 500) and ethanol (99.5) (1:1) to

＝ 15.15 mg of C20H24ClN3S.2C4H4O4 make V mL so that each mL contains about 80 mgof prochlorperazine maleate (C H ClN S.2C H O ). Centri- Containers and storage Containers—Tight containers. 20 24 3 4 4 4 fuge this solution, and use the supernatant liquid as the sam- Storage—Light-resistant. ple solution. Proceed as directed in the Assay. Amount (mg) of prochlorperazine maleate Prochlorperazine Maleate Tablets (C20H24ClN3S.2C4H4O4) ＝ MS × QT/QS × V/250 プロクロルペラジンマレイン酸塩錠 MS: Amount (mg) of Prochlorperazine Maleate RS Internal standard solution—A solution of butyl parahy- Prochlorperazine Maleate Tablets contain not droxybenzoate in a mixture of diluted phosphoric acid (1 in less than 95.0z and not more than 105.0z of 500) and ethanol (99.5) (1:1) (1 in 1000). the labeled amount of prochlorperazine maleate (C20H24ClN3S.2C4H4O4: 606.09). Dissolution <6.10> When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 Method of preparation Prepare as directed under Tablets, mL of 2nd fluid for dissolution test as the dissolution me- with Prochlorperazine Maleate. dium, the dissolution rate in 45 minutes of Prochlorperazine Identification (1) Weigh a quantity of powdered Pro- Maleate Tablets is not less than 75z. chlorperazine Maleate Tablets, equivalent to 5 mg of Pro- Start the test with 1 tablet of Prochlorperazine Maleate chlorperazine Maleate according to the labeled amount, add Tablets, withdraw not less than 20 mL of the medium at the 15 mL of acetic acid (100), shake, and filter. To 5 mL of the specified minute after starting the test, and filter through a filtrate add 3 mL of sulfuric acid, and shake: a light red membrane filter with a pore size not exceeding 0.45 mm. Dis- color develops. To this solution add 1 drop of potassium card the first 10 mL of the filtrate, pipet V mL of the subse- dichromate TS: a green-brown color is produced and quent filtrate, add the dissolution medium to make exactly changes to brown on standing. V?mL so that each mL contains about 9 mg of prochlorpera-

(2) Weigh a quantity of powdered Prochlorperazine zine maleate (C20H24ClN3S.2C4H4O4) according to the Maleate Tablets, equivalent to 0.08 g of Prochlorperazine labeled amount, and use this solution as the sample solution. Maleate according to the labeled amount, add 15 mL of Separately, weigh accurately about 18 mg of Prochlorpera- methanol and 1 mL of dimethylamine, shake, centrifuge, zine Maleate RS, previously dried at 1059C for 3 hours, and and use the supernatant liquid as the sample solution. Sepa- dissolve in methanol to make exactly 100 mL. Pipet 5 mL of rately, dissolve 0.08 g of Prochlorperazine Maleate RS in 15 this solution, add the dissolution medium to make exactly mL of methanol and 1 mL of dimethylamine, and use this 100 mL, and use this solution as the standard solution. Per- solution as the standard solution. Perform the test with these form the test with the sample solution and standard solution solutions as directed under Thin-layer Chromatography as directed under Ultraviolet-visible Spectrophotometry <2.03>. Spot 10 mL each of the sample solution and standard <2.24>, using the dissolution medium as the blank, and deter- solution on a plate of silica gel for thin-layer chromatogra- mine the absorbances, AT and AS,at255nm. phy. Develop the plate with a mixture of 1-butanol and am- Dissolution rate (z) with respect to the labeled amount monia TS (15:2) to a distance of about 10 cm, and air-dry of prochlorperazine maleate (C H ClN S.2C H O ) the plate. Spray evenly palladium (II) chloride TS on the 20 24 3 4 4 4 ＝ M × A /A × V?/V × 1/C × 45 plate: the spots obtained from the sample solution and S T S standard solution show a red-purple color, and has the same MS: Amount (mg) of Prochlorperazine Maleate RS Rf value. C: Labeled amount (mg) of prochlorperazine maleate

(3) To a quantity of powdered Prochlorperazine Maleate (C20H24ClN3S.2C4H4O4)in1tablet Tablets, equivalent to 0.04 g of Prochlorperazine Maleate Assay Conduct this procedure using light-resistant vessels. according to the labeled amount, add 10 mL of 1 mol/L hy- Weigh accurately the mass of not less than 20 Prochlorpera- drochloric acid TS and 20 mL of diethyl ether, shake, and zine Maleate Tablets, and powder in an agate mortar. Weigh centrifuge. Transfer the diethyl ether layer to a separator, accurately a portion of the powder, equivalent to about 8 mg wash with 5 mL of 0.05 mol/L sulfuric acid TS, and evapo- of prochlorperazine maleate (C H ClN S.2C H O ), add 60 rate on a water bath to dryness. Dissolve the residue in 5 mL 20 24 3 4 4 4 mL of a mixture of diluted phosphoric acid (1 in 500) and of sulfuric acid TS, filter, if necessary, and add 1 to 2 drops ethanol (99.5) (1:1), and shake vigorously for 10 minutes. JP XVI Official Monographs / Progesterone 1303

Add exactly 5 mL of the internal standard solution, and add a mixture of diluted phosphoric acid (1 in 500) and ethanol Progesterone (99.5) (1:1) to make 100 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, プロゲステロン weigh accurately about 20 mg of Prochlorperazine Maleate RS, previously dried at 1059C for 3 hours, and dissolve in a mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) to make exactly 25 mL. Pipet 10 mL of this solution, add exactly 5 mL of the internal standard solution and a mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) to make 100 mL, and use this solution as the standard solution. Perform the test with 5 mLeachofthe C21H30O2:314.46 sample solution and standard solution as directed under Liq- Pregn-4-ene-3,20-dione uid Chromatography <2.01> according to the following con- [57-83-0] ditions, and calculate the ratios, QT and QS, of the peak area of prochlorperazine to that of the internal standard. Progesterone, when dried, contains not less than 97.0z and not more than 103.0z of C H O . Amount (mg) of prochlorperazine maleate 21 30 2

(C20H24ClN3S.2C4H4O4) Description Progesterone occurs as white crystals or crys- ＝ MS × QT/QS × 2/5 talline powder. It is soluble in methanol and in ethanol (99.5), and practi- M : Amount (mg) of Prochlorperazine Maleate RS S cally insoluble in water. Internal standard solution—A solution of butyl parahy- Identification (1) Determine the absorption spectrum of a droxybenzoate in a mixture of diluted phosphoric acid (1 in solution of Progesterone in ethanol (99.5) (1 in 100,000) as 500) and ethanol (99.5) (1:1) (1 in 1000). directed under Ultraviolet-visible Spectrophotometry <2.24>, Operating conditions— and compare the spectrum with the Reference Spectrum or Detector: An ultraviolet absorption photometer (wave- the spectrum of a solution of Progesterone RS prepared in length: 257 nm). the same manner as the sample solution: both spectra exhibit Column: A stainless steel column 4.6 mm in inside diame- similar intensities of absorption at the same wavelengths. ter and 15 cm in length, packed with octadecylsilanized silica (2) Determine the infrared absorption spectrum of gel for liquid chromatography (5 mm in particle diameter). Progesterone, as directed in the potassium bromide disk Column temperature: A constant temperature of about method under Infrared Spectrophotometry <2.25>,and 259C. compare the spectrum with the Reference Spectrum or the Mobile phase: A mixture of diluted 0.05 mol/L sodium di- spectrum of Progesterone RS: both spectra exhibit similar hydrogen phosphate TS (1 in 2) and acetonitrile (11:9). intensities of absorption at the same wave numbers. If any Flow rate: Adjust the flow rate so that the retention time difference appears between the spectra, dissolve Progester- of prochlorperazine is about 5 minutes. one and Progesterone RS in ethanol (95), respectively, then System suitability— evaporate the ethanol to dryness, and repeat the test on the System performance: When the procedure is run with 5 mL residues. of the standard solution under the above operating condi- 20 tions, prochlorperazine and the internal standard are eluted Optical rotation <2.49> [a]D : ＋184 – ＋1949(after drying, in this order with the resolution between these peaks being 0.2 g, ethanol (99.5), 10 mL, 100 mm). not less than 10. Melting point <2.60> 128 – 1339C or 120 – 1229C System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating Purity Related substances—Dissolve 80 mg of Progester- conditions, the relative standard deviation of the ratio of the one in 2 mL of methanol, and use this solution as the sample peak area of prochlorperazine to that of the internal stand- solution. Pipet 1 mL of the sample solution, add methanol ard is not more than 1.0z. to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as di- Containers and storage Containers—Tight containers. rected under Thin-layer Chromatography <2.03>. Spot 5 mL Storage—Light-resistant. each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of diethyl ether and diethylamine (19:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacuum, phosphorus (V) oxide, 4 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g). Assay Weigh accurately about 10 mg each of Progesterone 1304 Progesterone Injection / Official Monographs JP XVI and Progesterone RS, previously dried, and dissolve each in accurately about 10 mg of Progesterone RS, previously dried ethanol (99.5) to make exactly 100 mL. Pipet 5 mL each of in vacuum for 4 hours using phosphorus (V) oxide as the these solutions, add ethanol (99.5) to make exactly 50 mL, desiccant, dissolve in 2 mL of tetrahydrofuran, and add and use these solution as the sample solution and the stand- ethanol (99.5) to make exactly 20 mL. Pipet 2 mL of this so- ard solution, respectively. Determine the absorbances, AT lution, add exactly 10 mL of the internal standard solution and AS, of the sample solution and standard solution at the and ethanol (99.5) to make 20 mL, and use this solution as wavelength of maximum absorption at about 241 nm as di- the standard solution. Perform the test with 5 mLeachofthe rected under Ultraviolet-visible Spectrophotometry <2.24>. sample solution and standard solution as directed under Liq- uid Chromatography <2.01> according to the following con- Amount (mg) of progesterone (C21H30O2) ＝ MS × AT/AS ditions, and calculate the ratios, QT and QS, of the peak area MS: Amount (mg) of Progesterone RS of progesterone to that of the internal standard.

Containers and storage Containers—Tight containers. Amount (mg) of progesterone (C21H30O2) Storage—Light-resistant. ＝ MS × QT/QS × V/20

MS: Amount (mg) of Progesterone RS Progesterone Injection Internal standard solution—A solution of testosterone propionate in ethanol (99.5) (1 in 4000). プロゲステロン注射液 Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 241 nm). Progesterone Injection is an oily solution for injec- Column: A stainless steel column 4.6 mm in inside diame- tion. ter and 15 cm in length, packed with octadecylsilanized silica It contains not less than 95.0z and not more than gel for liquid chromatography (5 mminparticlediameter). 105.0z of the labeled amount of progesterone Column temperature: A constant temperature of about (C H O : 314.46). 21 30 2 359C. Method of preparation Prepare as directed under Injec- Mobile phase: A mixture of acetonitrile and water (7:3). tions, with Progesterone. Flow rate: Adjust the flow rate so that the retention time of progesterone is about 6 minutes. Description Progesterone Injection is a clear, colorless to System suitability— pale yellow, oily liquid. System performance: When the procedure is run with 5 mL Identification To 1 mL of Progesterone Injection add 1 mL of the standard solution under the above operating condi- of diluted ethanol (9 in 10), shake well, take the ethanol tions, progesterone and the internal standard are eluted in layer, shake well with 1 mL of petroleum benzin, and use the this order with the resolution between these peaks being not ethanol layer as the sample solution. Separately, dissolve less than 9. about 5 mg of Progesterone RS in 1 mL of ethanol (99.5), System repeatability: When the test is repeated 6 times and use this solution as the standard solution. Perform the with 5 mL of the standard solution under the above operating test with these solutions as directed under Thin-layer Chro- conditions, the relative standard deviation of the ratio of the matography <2.03>. Spot 2 mL each of the sample solution peak area of progesterone to that of the internal standard is and standard solution on a plate of silica gel for thin-layer not more than 1.0z. chromatography. Develop the plate with a mixture of diethyl Containers and storage Containers—Hermetic containers. ether and diethylamine (19:1) to a distance of about 10 cm, Storage—Light-resistant. and air-dry the plate. Spray evenly sulfuric acid on the plate, and heat the plate at 1059C for 10 minutes: the principal spot obtained from the sample solution has the same Rf value as the spot from the standard solution. Proglumide Extractable volume <6.05> It meets the requirement. プログルミド Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the requirement. Sterility <4.06> Perform the test according to the Direct in- oculation method: it meets the requirement. C H N O : 334.41 Assay Measure the specific gravity of Progesterone Injec- 18 26 2 4 (4RS)-4-Benzoylamino-N,N-dipropylglutaramic acid tion. Weigh accurately the mass of Progesterone Injection, [6620-60-6] equivalent to about 1 mL, mix with 2 mL of tetrahydrofuran, and add ethanol (99.5) to make exactly V mL so that Proglumide, when dried, contains not less than each mL contains about 0.5 mg of progesterone (C H O ). 21 30 2 98.5z of C H N O . Pipet 2 mL of this solution, add exactly 10 mL of the inter- 18 26 2 4 nal standard solution and ethanol (99.5) to make 20 mL, and Description Proglumide occurs as white crystals or crystal- use this solution as the sample solution. Separately, weigh line powder. JP XVI Official Monographs / L-Proline 1305

It is freely soluble in methanol, soluble in ethanol (95), sparingly soluble in diethyl ether, and very slightly soluble in L-Proline water. A solution of Proglumide in methanol (1 in 10) shows no L-プロリン optical rotation. Identification (1) Put 0.5 g of Proglumide in a round bottom tube, add 5 mL of hydrochloric acid, seal the tube, and heat the tube carefully at 1209C for 3 hours. After cooling, C H NO :115.13 open the tube, filter the content to collect crystals separated 5 9 2 (2S)-Pyrrolidine-2-carboxylic acid out, wash the crystals with 50 mL of cold water, and dry at [147-85-3] 1009C for 1 hour: the melting point <2.60> of the crystals is between 1219Cand1249C. L-Proline contains not less than 99.0z and not (2) Determine the infrared absorption spectrum of more than 101.0z of C H NO ,calculatedonthe Proglumide, previously dried, as directed in the potassium 5 9 2 dried basis. bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- Description L-Proline occurs as white crystals or crystalline trum: both spectra exhibit similar intensities of absorption at powder. It has a slightly sweet taste. the same wave numbers. Itisverysolubleinwaterandinformicacid,andslightly soluble in ethanol (99.5). Absorbance <2.24> E1z (225 nm): 384 – 414 (after drying, 1cm It is deliquescent. 4 mg, methanol, 250 mL). Identification Determine the infrared absorption spectrum Melting point <2.60> 148–1509C of L-Proline as directed in the potassium bromide disk Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of method under Infrared Spectrophotometry <2.25>,andcom- Proglumide according to Method 2, and perform the test. pare the spectrum with the Reference Spectrum: both spectra Prepare the control solution with 2.0 mL of Standard Lead exhibit similar intensities of absorption at the same wave Solution (not more than 20 ppm). numbers. (2) Arsenic <1.11>—To 1.0 g of Proglumide add 10 mL Optical rotation <2.49> [a]20: －84.0 – －86.09(1 g, calcu- of a solution of magnesium nitrate hexahydrate in ethanol D lated on the dried basis, water, 25 mL, 100 mm). (95) (1 in 10) and 1.5 mL of hydrogen peroxide (30), burn the ethanol, and prepare the test solution according to Method pH <2.54> The pH of a solution of 1.0 g of L-Proline in 10 3, and perform the test (not more than 2 ppm). mL of water is 5.9 to 6.9. (3) Related substances—Dissolve 0.10 g of Proglumide Purity (1) Clarity and color of solution—Dissolve 1.0 g in 5 mL of methanol, and use this solution as the sample so- of L-Proline in 10 mL of water: the solution is clear and col- lution. Pipet 1 mL of the sample solution, add methanol to orless. make exactly 200 mL, and use this solution as the standard (2) Chloride <1.03>—Perform the test with 0.5 g of solution. Perform the test with these solutions as directed L-Proline. Prepare the control solution with 0.30 mL of 0.01 under Thin-layer Chromatography <2.03>. Spot 10 mLeach mol/L hydrochloric acid VS (not more than 0.021z). of the sample solution and standard solution on a plate of (3) Sulfate <1.14>—Perform the test with 0.6 g of L-Pro- silica gel with fluorescent indicator for thin-layer chromatog- line. Prepare the control solution with 0.35 mL of 0.005 raphy. Develop the plate with a mixture of cyclohexane, mol/L sulfuric acid VS (not more than 0.028z). ethyl acetate, acetic acid (100) and methanol (50:18:5:4) to a (4) Ammonium <1.02>—Perform the test with 0.25 g of distance of about 10 cm, and air-dry the plate. Examine L-Proline. Prepare the control solution with 5.0 mL of under ultraviolet light (main wavelength: 254 nm): the spots Standard Ammonium Solution (not more than 0.02z). other than the principal spot from the sample solution are (5) Heavy metal <1.07>—Proceed with 1.0 g of L-Proline not more intense than the spot from the standard solution. according to Method 1, and perform the test. Prepare the Loss on drying <2.41> Not more than 0.10z (1 g, reduced control solution with 1.0 mL of Standard Lead Solution (not pressure, phosphorus (V) oxide, 609C, 3 hours). more than 10 ppm). (6) Iron <1.10>—Prepare the test solution with 1.0 g of Residue on ignition <2.44> Not more than 0.1z (1 g). L-Proline according to Method 1, and perform the test ac- Assay Weigh accurately about 0.16 g of Proglumide, previ- cording to Method A. Prepare the control solution with 1.0 ously dried, dissolve in 40 mL of methanol, add 10 mL of mL of Standard Iron Solution (not more than 10 ppm). water, and titrate <2.50> with 0.1 mol/L sodium hydroxide (7) Related substances—Weigh accurately about 0.5 g of VS (potentiometric titration). Perform a blank determina- L-Proline, and dissolve in 0.5 mL of hydrochloric acid and tion, and make any necessary correction. water to make exactly 100 mL. Pipet 10 mL of this solution, add 0.02 mol/L hydrochloric acid TS to make exactly 50 Each mL of 0.1 mol/L sodium hydroxide VS mL, and use this solution as the sample solution. Separately, ＝ 33.44 mg of C H N O 18 26 2 4 weigh accurately an amount, equivalent to 2.5 mmol, of Containers and storage Containers—Well-closed contain- L-aspartic acid, L-threonine, L-serine, L-glutamic acid, L-pro- ers. line, glycine, L-alanine, L-cystine, L-valine, L-methionine, L-isoleucine, L-leucine, L-tyrosine, L-phenylalanine, L-lysine hydrochloride, ammonium chloride, L-histidine and L-argi- 1306 Promethazine Hydrochloride / Official Monographs JP XVI nine, dissolve them in 0.1 mol/L hydrochloric acid TS to 5 minutes, add 81 mg of sodium borohydride, pass nitrogen make exactly 1000 mL, and use this solution as the standard for 30 minutes, and use this solution as Solution (II). Pre- stock solution. Pipet 5 mL of the standard stock solution, pare a mixture with an equal volume of the Solution (I) and andadd0.02mol/LhydrochloricacidTStomakeexactly (II). (Prepare before use). 100 mL. Pipet 4 mL of this solution, add 0.02 mol/L hydro- Flow rate of mobile phase: 0.20 mL per minute. chloric acid TS to make exactly 50 mL, and use this solution Flow rate of reaction regent: 0.24 mL per minute. as the standard solution. Perform the test with exactly 20 mL System suitability— each of the sample solution and standard solution as directed System performance: When the test is run with 20 mLof under Liquid Chromatography <2.01> according to the fol- the standard solution under the above operating conditions, lowing conditions, and calculate the mass percentage of each the resolution between the peaks of glycine and alanine is not amino acid, using the mass of amino acid other than proline less than 1.2. in 1 mL of the sample solution obtained from the height of System repeatability: When the test is repeated 6 times the peaks obtained from the sample and standard solution: with 20 mL of the standard solution under the above operat- the amount of each amino acid other than proline is not ing conditions, the relative standard deviations of the peak more than 0.1z. height of each amino acid other than proline in the standard Operating conditions— solution is not more than 5.0z, and the relative standard Detector: A visible absorption photometer (wavelength: deviation of the retention time is not more than 1.0z. 570 nm). (8) Residual solvent—Being specified separately. Column: A stainless steel column 4.6 mm in inside diame- Loss on drying <2.41> Not more than 0.3z (1 g, 1059C, ter and 8 cm in length, packed with strongly acidic ion- 3 hours). exchange resin for liquid chromatography composed with a sulfonated polystyrene (3 mminparticlediameter)(Natype). Residue on ignition <2.44> Not more than 0.1z (1 g). Column temperature: A constant temperature of about Assay Weigh accurately about 0.12 g of L-Proline, dissolve 579C. in 3 mL of formic acid, add 50 mL of acetic acid (100), and Chemical reaction vessel temperature: A constant temper- titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio- ature of about 1309C. metric titration). Perform a blank determination in the same Reaction time: About 1 minute. manner, and make any necessary correction. Mobile phase: Prepare the mobile phases A, B, C, D and E according to the following table, and add 0.1 mL each of Each mL of 0.1 mol/L perchloric acid VS caprylic acid. ＝ 11.51 mg of C5H9NO2 Containers and storage Containers—Tight containers. Mobile phase A B C D E

Citric acid 19.80 g 22.00 g 12.80 g 6.10 g — monohydrate Promethazine Hydrochloride Trisodium citrate 6.19 g 7.74 g 13.31 g 26.67 g — dihydrate プロメタジン塩酸塩 Sodium chloride 5.66 g 7.07 g 3.74 g 54.35 g — Sodium hydroxide ————8.00g Ethanol (99.5) 130 mL 20 mL 4 mL — 100 mL Thiodiglycol 5 mL 5 mL 5 mL — — Benzyl alcohol — — — 5 mL — Lauromacrogol solution (1 in 4) 4mL 4mL 4mL 4mL 4mL asufficientasufficienta sufficient asufficientasufficient C17H20N2S.HCl: 320.88 Water amount amount amount amount amount (2RS)-N,N-Dimethyl-1-(10H-phenothiazin-10-yl)propan- 2-ylamine monohydrochloride Total amount 1000 mL 1000 mL 1000 mL 1000 mL 1000 mL [58-33-3]

Switching of mobile phase: Switch the mobile phases A, Promethazine Hydrochloride, when dried, contains B, C, D and E sequentially so that when proceed with 20 mL not less than 98.0z of C17H20N2S.HCl. of the standard solution under the above conditions, aspartic acid, threonine, serine, glutamic acid, proline, glycine, ala- Description Promethazine Hydrochloride occurs as a white nine, cystine, valine, methionine, isoleucine, leucine, tyro- to light yellow powder. sine, phenylalanine, lysine, ammonia, histidine and arginine It is very soluble in water, freely soluble in ethanol (95) are eluted in this order with the resolution between the peaks and in acetic acid (100), sparingly soluble in acetic anhy- of isoleucine and leucine being not less than 1.2. dride, and practically insoluble in diethyl ether. Reaction reagent: Dissolve 204 g of lithium acetate dihy- It is gradually colored by light. drate in an appropriate amount of water, add 123 mL of A solution of Promethazine Hydrochloride (1 in 25) shows acetic acid (100), 401 mL of 1-methoxy-2-propanol and on optical rotation. water to make 1000 mL, pass nitrogen for 10 minutes, and Melting point: about 2239C (with decomposition). use this solution as Solution (I). Separately, to 979 mL of 1- Identification (1) Determine the absorption spectrum of a methoxy-2-propanol add 39 g of ninhydrin, pass nitrogen for solution of Promethazine Hydrochloride (1 in 100,000) as JP XVI Official Monographs / Propafenone Hydrochloride 1307 directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: Propafenone Hydrochloride both spectra exhibit similar intensities of absorption at the same wavelengths. プロパフェノン塩酸塩 (2) Determine the infrared absorption spectrum of Promethazine Hydrochloride, previously dried, as directed in the potassium bromide disk method under Infrared Spec- trophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. C H NO .HCl: 377.90 (3) Dissolve 0.5 g of Promethazine Hydrochloride in 5 21 27 3 1-{2-[(2RS)-2-Hydroxy- mL of water, add 2 mL of ammonia TS, and filter. To 5 mL 3-(propylamino)propyloxy]phenyl}-3-phenylpropan-1-one of the filtrate add dilute nitric acid to make acidic: the solu- monohydrochloride tion responds to the Qualitative Tests <1.09> (2) for chloride. [34183-22-7] pH <2.54> The pH of a solution of Promethazine Hydro- chloride (1 in 10) is between 4.0 and 5.5. Propafenone Hydrochloride, when dried, contains not less than 98.5z and not more than 101.0z of Purity (1) Clarity and color of solution—Dissolve 1.0 g C H NO .HCl. of Promethazine Hydrochloride in 10 mL of water, protect- 21 27 3 ing from direct sunlight: the solution is clear and colorless. Description Propafenone Hydrochloride occurs as white (2) Heavy metals <1.07>—Proceed with 1.0 g of crystals or a white crystalline powder. Promethazine Hydrochloride according to Method 2, and It is freely soluble in formic acid, sparingly soluble in perform the test. Prepare the control solution with 2.0 mL of methanol, and slightly soluble in water and in ethanol (99.5). Standard Lead Solution (not more than 20 ppm). A solution of Propafenone Hydrochloride in methanol (3) Related substances—Perform the test under the pro- (1 in 100) shows no optical rotation. tection from sunlight. Dissolve 0.10 g of Promethazine Identification (1) Dissolve 0.1 g of Propafenone Hydro- Hydrochloride in exactly 5 mL of ethanol (95), and use this chloride in 20 mL of water by warming. After cooling, to 3 solution as the sample solution. Pipet 1 mL of the sample so- mL of this solution add water to make 500 mL. Determine lution, add ethanol (95) to make exactly 200 mL, and use the absorption spectrum of this solution as directed under this solution as the standard solution (1). Separately, dis- Ultraviolet-visible Spectrophotometry <2.24>,andcompare solve 20 mg of isopromethazine hydrochloride for thin-layer the spectrum with the Reference Spectrum: both spectra chromatography in ethanol (95) to make exactly 100 mL, exhibit similar intensities of absorption at the same wave- and use this solution as the standard solution (2). Perform lengths. the test with these solutions as directed under Thin-layer (2) Determine the infrared absorption spectrum of Chromatography <2.03>. Spot 10 mLeachofthesamplesolu- Propafenone Hydrochloride as directed in the potassium tion and standard solutions (1) and (2) on a plate of silica gel chloride disk method under Infrared Spectrophotometry with fluorescent indicator for thin-layer chromatography. <2.25>, and compare the spectrum with the Reference Spec- Develop the plate with a mixture of methanol and diethyla- trum: both spectra exhibit similar intensities of absorption at mine (19:1) to a distance of about 12 cm, and air-dry the the same wave numbers. plate. Examine under ultraviolet light (main wavelength: 254 (3) Dissolve 0.1 g of Propafenone Hydrochloride in 20 nm): the spots from the sample solution corresponding to mL of water by warming. After cooling, to 10 mL of this so- the spots from the standard solution (2) are not more intense lution add 1 mL of dilute nitric acid, and filter to separate than the spot from the standard solution (2), and any spot formed precipitate: the filtrate responds to the Qualitative other than the principal spot from the sample solution is not Tests <1.09> (2) for chloride. more intense than the spot from the standard solution (1). Melting point <2.60> 172 – 1759C Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Propafenone Hydrochloride according to Method 4, and Residue on ignition <2.44> Not more than 0.1z (1 g). perform the test. Prepare the control solution with 2.0 mL of Assay Weigh accurately about 0.5 g of Promethazine Hy- Standard Lead Solution (not more than 20 ppm). drochloride, previously dried, dissolve in 50 mL of a mixture (2) Related substances—Dissolve 0.10 g of Propafenone of acetic anhydride and acetic acid (100) (7:3), and titrate Hydrochloride in 20 mL of the mobile phase in the operating <2.50> with 0.1 mol/L perchloric acid VS (potentiometric conditions 1, and use this solution as the sample solution. titration). Perform a blank determination, and make any Pipet 2 mL of the sample solution, and add the mobile phase necessary correction. in the operating conditions 1 to make exactly 50 mL. Pipet 2.5 mL of this solution, add 2.5 mL of a solution of diphenyl Each mL of 0.1 mol/L perchloric acid VS phthalate in methanol (1 in 2000), add the mobile phase in ＝ 32.09 mg of C H N S.HCl 17 20 2 the operating conditions 1 to make exactly 100 mL, and use Containers and storage Containers—Tight containers. this solution as the standard solution. Perform the test with Storage—Light-resistant. exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions 1 and 2. Determine each 1308 Propafenone Hydrochloride Tablets / Official Monographs JP XVI peak area of both solutions by the automatic integration acid, add 50 mL of acetic anhydride, and titrate <2.50> with method: the area of each peak other than the peak of 0.05 mol/L perchloric acid VS (potentiometric titration). propafenone from the sample solution is not larger than the Perform a blank determination in the same manner, and peak area of propafenone from the standard solution. make any necessary correction. Operating conditions 1— Each mL of 0.05 mol/L perchloric acid VS Detector: An ultraviolet absorption photometer (wave- ＝ 18.90 mg of C H NO .HCl length: 254 nm). 21 27 3 Column: A stainless steel column 4.6 mm in inside diame- Containers and storage Containers—Well-closed contain- ter and 15 cm in length, packed with octadecylsilanized silica ers. gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Propafenone Hydrochloride Tablets Mobile phase: Dissolve 4.6 g of sodium 1-nonanesulfonate and 2.3 g of phosphoric acid in water to make 1000 mL, and プロパフェノン塩酸塩錠 filter through a membrane filter with a pore size not exceeding 0.45 mm. To 900 mL of the filtrate add 600 mL of Propafenone Hydrochloride Tablets contain not acetonitrile. less than 96.0z and not more than 104.0z of Flow rate: Adjust the flow rate so that the retention time the labeled amount of propafenone hydrochloride of diphenyl phthalate is about 39 minutes. (C H NO .HCl: 377.90). Time span of measurement: Beginning after the solvent 21 27 3 peak to the retention time of diphenyl phthalate. Method of preparation Prepare as directed under Tablets, System suitability 1— with Propafenone Hydrochloride. System performance: Dissolve 12 mg of Propafenone Hy- Identification To a quantity of Propafenone Hydrochlo- drochloride and 50 mg of isopropyl benzoate in 100 mL of ride Tablets, equivalent to 0.3 g of Propafenone Hydrochlo- methanol. When the procedure is run with 10 mLofthisso- ride according to the labeled amount, add 60 mL of water, lution under the above operating conditions 1, propafenone and disintegrate by warming. After cooling, centrifuge, and and isopropyl benzoate are eluted in this order with the reso- to 3 mL of the supernatant liquid add water to make 500 lution between these peaks being not less than 5. mL. Determine the absorption spectrum of this solution as System repeatability: When the test is repeated 6 times directed under Ultraviolet-visible Spectrophotometry <2.24>: with 10 mL of the standard solution under the above operat- it exhibits maxima between 247 nm and 251 nm, and between ing conditions 1, the relative standard deviation of the peak 302 nm and 306 nm. Separately, determine the both maximal area of propafenone is not more than 2.0z. absorbances, A and A , of the solution, the ratio of A /A Operating conditions 2— 1 2 1 2 is between 2.30 and 2.55. Detector, column and column temperature: Proceed as directed in the operation conditions 1. Uniformity of dosage units <6.02> Perform the test accord- Mobile phase: Dissolve 7.33 g of sodium 1-decanesul- ing to the following method: it meets the requirement of the fonate and 2.3 g of phosphoric acid in water to make 1000 Content uniformity test. mL, and filter through a membrane filter with a pore size To 1 tablet of Propafenone Hydrochloride Tablets add 30 not exceeding 0.45 mm. To 700 mL of the filtrate add 700 mL mL of a mixture of water and acetonitrile (1:1), shake well to of acetonitrile. disintegrate, add a mixture of water and acetonitrile (1:1) to Flow rate: Adjust the flow rate so that the retention time make exactly 50 mL, and centrifuge. Pipet V mL of the su- of diphenyl phthalate is about 11 minutes. pernatant liquid, equivalent to about 6 mg of propafenone

Time span of measurement: About 2.5 times as long as the hydrochloride (C21H27NO3.HCl), add exactly 5 mL of the in- retention time of diphenyl phthalate, beginning after the ternal standard solution, add methanol to make 50 mL, and retention time of diphenyl phthalate. use this solution as the sample solution. Proceed as directed System suitability 2— in the Assay. System performance: When the procedure is run with 10 Amount (mg) of propafenone hydrochloride mL of the standard solution under the above operating con- (C H NO .HCl) ditions 2, propafenone and diphenyl phthalate are eluted in 21 27 3 ＝ M × Q /Q × 10/V this order with the resolution between these peaks being not S T S less than 21. MS: Amount (mg) of propafenone hydrochloride for assay System repeatability: When the test is repeated 6 times Internal standard solution—A solution of isopropyl benzo- with 10 mL of the standard solution under the above operat- ate in methanol (1 in 200). ing conditions 2, the relative standard deviation of the peak area of propafenone is not more than 2.0z. Dissolution <6.10> When the test is performed at 50 revolu- (3) Residual solvent—Being specified separately. tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, in 30 minutes of Propafenone Hydrochloride Tables is not 2 hours). less than 75z. Residue on ignition <2.44> Not more than 0.1z (1 g). Start the test with 1 tablet of Propafenone Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the Assay Weigh accurately about 0.3 g of Propafenone Hy- specified minute after starting the test, and filter through a drochloride, previously dried, dissolve in 2 mL of formic membrane filter with a pore size not exceeding 0.5 mm. Dis- JP XVI Official Monographs / Propantheline Bromide 1309 card the first 10 mL of the filtrate, pipet V mL of the subse- mL of the standard solution under the above operating con- quent filtrate, add water to make exactly V?mL so that each ditions, propafenone and the internal standard are eluted in mL contains about 67 mg of propafenone hydrochloride this order with the resolution between these peaks being not

(C21H27NO3.HCl) according to the labeled amount, and use less than 5. this solution as the sample solution. Separately, weigh accu- System repeatability: When the test is repeated 6 times rately about 13 mg of propafenone hydrochloride for assay, with 10 mL of the standard solution under the above operat- previously dried at 1059C for 2 hours, dissolve in water to ing conditions, the relative standard deviation of the ratio of make exactly 200 mL, and use this solution as the standard the peak area of propafenone to that of the internal standard solution. Determine the absorbances, AT and AS,ofthesam- is not more than 1.0z. ple solution and standard solution at 305 nm as directed Containers and storage Containers—Tight containers. under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of propafenone hydrochloride (C21H27NO3.HCl) Propantheline Bromide ＝ MS × AT/AS × V?/V × 1/C × 450 プロパンテリン臭化物 MS: Amount (mg) of propafenone hydrochloride for assay C: Labeled amount (mg) of propafenone hydrochloride

(C21H27NO3.HCl) in 1 tablet Assay To a quantity of Propafenone Hydrochloride Tablets, equivalent to 1.5 g of propafenone hydrochloride

(C21H27NO3.HCl), add 70 mL of a mixture of water and acetonitrile (1:1), shake well to disintegrate, shake well for another 5 minutes, add a mixture of water and acetonitrile C23H30BrNO3: 448.39 (1:1) to make exactly 100 mL, and centrifuge. Pipet 4 mL of N-Methyl-N,N-bis(1-methylethyl)-2-[(9H-xanthen- the supernatant liquid, and add methanol to make exactly 50 9-ylcarbonyl)oxy]ethylaminium bromide mL. Pipet 5 mL of the solution, add exactly 5 mL of the in- [50-34-0] ternal standard solution, add methanol to make 50 mL, and use this solution as the sample solution. Separately, weigh Propantheline Bromide, when dried, contains not accurately about 30 mg of propafenone hydrochloride for less than 98.0z and not more than 102.0z of assay, previously dried at 1059C for 2 hours, and dissolve in C23H30BrNO3. methanol to make exactly 50 mL. Pipet 10 mL of this solu- Description Propantheline Bromide occurs as a white to tion, add exactly 5 mL of the internal standard solution, add yellowish white, crystalline powder. It is odorless and has a methanol to make 50 mL, and use this solution as the stand- very bitter taste. ard solution. Perform the test with 10 mLeachofthesample It is very soluble in water, in ethanol (95), in acetic acid solution and standard solution as directed under Liquid (100) and in chloroform, soluble in acetic anhydride, and Chromatography <2.01> according to the following condi- practically insolubleindiethylether. tions, and calculate the ratios, Q and Q , of the peak area T S The pH of a solution of Propantheline Bromide (1 in 50) is of propafenone to that of the internal standard. between 5.0 and 6.0. Amount (mg) of propafenone hydrochloride Melting point: about 1619C (with decomposition, after

(C21H27NO3.HCl) drying). ＝ M × Q /Q × 50 S T S Identification (1) To 5 mL of a solution of Propantheline

MS: Amount (mg) of propafenone hydrochloride for assay Bromide (1 in 20) add 10 mL of sodium hydroxide TS, heat to boil for 2 minutes. Cool to 609C, and add 5 mL of dilute Internal standard solution—A solution of isopropyl benzo- hydrochloric acid. After cooling, collect the precipitates, and ate in methanol (1 in 200). wash with water. Recrystallize from dilute ethanol, and dry Operating conditions— at 1059C for 1 hour: the crystals melt <2.60> between 2179C Detector: An ultraviolet absorption photometer (wave- and 2229C. length: 254 nm). (2) Dissolve 0.01 g of the crystals obtained in (1) in 5 mL Column: A stainless steel column 4.6 mm in inside diame- of sulfuric acid: a vivid yellow to yellow-red color develops. ter and 15 cm in length, packed with octadecylsilanized silica (3) To 5 mL of a solution of Propantheline Bromide (1 gel for liquid chromatography (5 mm in particle diameter). in 10) add 2 mL of dilute nitric acid: this solution responds Column temperature: A constant temperature of about to the Qualitative Tests <1.09> (1) for bromide. 409C. Mobile phase: Dissolve 4.6 g of sodium 1-nonanesulfonate Purity Xanthene-9-carboxylic acid and xanthone—Dis- and 2.3 g of phosphoric acid in water to make 1000 mL, and solve 10 mg of Propantheline Bromide in exactly 2 mL of filter through a membrane filter with a pore size not chloroform, and use this solution as the sample solution. exceeding 0.45 mm. To 900 mL of the filtrate add 600 mL of Separately, dissolve 1.0 mg of xanthene-9-carboxylic acid acetonitrile. and 1.0 mg of xanthone in exactly 40 mL of chloroform, and Flow rate: Adjust the flow rate so that the retention time use this solution as the standard solution. Perform the test of propafenone is about 8 minutes. immediately with these solutions as directed under Thin- System suitability— layer Chromatography <2.03>. Spot 25 mLeachofthesample System performance: When the procedure is run with 10 solution and standard solution on a plate of silica gel with 1310 Propiverine Hydrochloride / Official Monographs JP XVI fluorescent indicator for thin-layer chromatography, and (3) To 5 mL of a solution of Propiverine Hydrochloride air-dry the plate for 10 minutes. Develop the plate with a (1 in 100) add 6 mL of ethyl acetate, and add 3 drops of sil- mixture of 1,2-dichloroethane, methanol, water and formic ver nitrate TS: a white precipitate is formed, which does not acid (56:24:1:1) to a distance of about 12 cm, and air-dry the dissolve on the addition of 0.5 mL of dilute nitric acid and plate. Examine under ultraviolet light: the spots from the shaking. The precipitate dissolves on the addition of 2 mL of sample solution corresponding to the spots from the stand- ammonia TS and shaking. ard solution are not more intense than those from the stand- Melting point <2.60> 213 – 2189C ard solution. Purity (1) Sulfate <1.14>—Perform the test with 0.40 g of Loss on drying <2.41> Not more than 0.5z (2 g, 1059C, Propiverine Hydrochloride. Prepare the control solution 4 hours). with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more Residue on ignition <2.44> Not more than 0.1z (1 g). than 0.048z). (2) Heavy metals <1.07>—Proceedwith1.0gof Assay Weigh accurately about 1 g of Propantheline Propiverine Hydrochloride according to Method 2, and per- Bromide, previously dried, dissolve in 50 mL of a mixture of form the test. Prepare the control solution with 2.0 mL of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> Standard Lead Solution (not more than 20 ppm). with 0.1 mol/L perchloric acid VS (potentiometric titration). (3) Related substances—Dissolve 50 mg of Propiverine Perform a blank determination, and make any necessary Hydrochloride in 100 mL of the mobile phase, and use this correction. solution as the sample solution. Pipet 1 mL of the sample so- Each mL of 0.1 mol/L perchloric acid VS lution, add the mobile phase to make exactly 100 mL, and

＝ 44.84 g of C23H30BrNO3 use this solution as the standard solution. Perform the test with exactly 15 mL each of the sample solution and standard Containers and storage Containers—Well-closed contain- solution as directed under Liquid Chromatography <2.01> ers. according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak, having the relative retention time about 0.28 to Propiverine Hydrochloride propiverine, obtained from the sample solution is not larger than 3/10 times the peak area of propiverine from the stand- プロピベリン塩酸塩 ard solution, the area of the peak other than propiverine and above mentioned peak from the sample solution is not larger than 1/10 times the peak area of propiverine from the standard solution, and the total area of the peaks other than propiverine from the sample solution is not larger than 1/2 times the peak area of propiverine from the standard solution.

C23H29NO3.HCl: 403.94 Operating conditions— 1-Methylpiperidin-4-yl 2,2-diphenyl-2-propoxyacetate Detector, column, column temperature, mobile phase, and monohydrochloride flow rate: Proceed as directed in the operating conditions in [54556-98-8] the Assay. Time span of measurement: About 2.5 times as long as the Propiverine Hydrochloride, when dried, contains retention time of propiverine, beginning after the solvent not less than 98.5z and not more than 101.5z of peak. C23H29NO3.HCl. System suitability— Test for required detectability: Pipet 1 mL of the standard Description Propiverine Hydrochloride occurs as white solution, and add the mobile phase to make exactly 20 mL. crystals or a white crystalline powder. Confirm that the peak area of propiverine obtained with 15 It is soluble in water and in ethanol (99.5). mL of this solution is equivalent to 3.5 to 6.5z of that with Identification (1) Dissolve 50 mg of Propiverine Hydro- 15 mL of the standard solution. chloride in 20 mL of water, and add acetonitrile to make 100 System performance: When the procedure is run with 15 mL. Determine the absorption spectrum of this solution as mL of the standard solution under the above operating con- directed under Ultraviolet-visible Spectrophotometry <2.24>, ditions, the number of theoretical plates and the symmetry and compare the spectrum with the Reference Spectrum or factor of the peak of propiverine are not less than 7000 and the spectrum of a solution of Propiverine Hydrochrolide RS not more than 1.5, respectively. prepared in the same manner as the sample solution: both System repeatability: When the test is repeated 6 times spectra exhibit similar intensities of absorption at the same with 15 mL of the standard solution under the above operat- wavelengths. ing conditions, the relative standard deviation of the peak (2) Determine the infrared absorption spectrum of area of propiverine is not more than 2.0z. Propiverine Hydrochloride, previously dried, as directed in (4) Residual solvent—Being specified separately. the potassium chloride disk method under Infrared Spectro- Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, photometry <2.25>, and compare the spectrum with the Ref- 1 hour). erence Spectrum or the spectrum of dried Propiverine Hy- drochloride RS: both spectra exhibit similar intensities of Residue on ignition <2.44> Not more than 0.1z (1 g). absorption at the same wave numbers. JP XVI Official Monographs / Propiverine Hydrochloride Tablets 1311

Assay Weigh accurately about 50 mg each of Propiverine Purity Related substances—Shake vigorously a quantity of Hydrochloride and Propiverine Hydrachloride RS, both pre- powdered Propiverine Hydrochloride Tablets, equivalent to viously dried, and dissolve each in the mobile phase to make 50 mg of Propiverine Hydrochloride according to the labeled exactly 100 mL. Pipet 10 mL each of these solutions, add the amount, with the mobile phase, add the mobile phase to mobile phase to make exactly 50 mL, and use these solutions make 100 mL, centrifuge, and use the supernatant liquid as as the sample solution and the standard solution, respec- the sample solution. Pipet 1 mL of the sample solution, add tively. Perform the test with exactly 15 mLeachofthesam- the mobile phase to make exactly 100 mL, and use this solu- ple solution and standard solution as directed under Liquid tion as the standard solution. Perform the test with exactly Chromatography <2.01> according to the following condi- 15 mL each of the sample solution and standard solution as tions, and determine the peak areas, AT and AS, of propiver- directed under Liquid Chromatography <2.01> according to ine from each solution. the following conditions. Determine each peak area by the automatic integration method: the area of the peak, having Amount (mg) of propiverine hydrochloride the relative retention time about 0.28 to propiverine, ob- (C H NO .HCl) 23 29 3 tained from the sample solution is not larger than 3/10 times ＝ M × A /A S T S the peak area of propiverine from the standard solution, the

MS: Amount (mg) of Propiverine Hydrochloride RS area of the peak other than propiverine and the peak mentioned above from the sample solution is not larger than 1/5 System suitability— times the peak area of propiverine from the standard solu- Detector: An ultraviolet absorption photometer (wave- tion, and the total area of the peaks other than propiverine length: 210 nm). from the sample solution is not larger than 7/10 times the Column: A stainless steel column 4.6 mm in inside diame- peak area of propiverine from the standard solution. ter and 15 cm in length, packed with phenylated silica gel for Operating conditions— liquid chromatography (5 mminparticlediameter). Detector, column, column temperature, mobile phase, and Column temperature: A constant temperature of about flow rate: Proceed as directed in the operating conditions in 409C. the Assay under Propiverine Hydrochloride. Mobile phase: Dissolve 2.21 g of potassium dihydrogen Time span of measurement: About 2.5 times as long as the phosphate and 1.51 g of sodium 1-octane sulfonate in 650 retention time of propiverine, beginning after the solvent mL of water, adjust to pH 3.2 with phosphoric acid, and peak. add 350 mL of acetonitrile. System suitability— Flow rate: Adjust the flow rate so that the retention time Test for required detectability: Pipet 1 mL of the standard of propiverine is about 17 minutes. solution, and add the mobile phase to make exactly 20 mL. System suitability— Confirm that the peak area of propiverine obtained with 15 System performance: When the procedure is run with 15 mL of this solution is equivalent to 3.5 to 6.5z of that with mL of the standard solution under the above operating con- 15 mL of the standard solution. ditions, the number of theoretical plates and the symmetry System performance: When the procedure is run with 15 factor of the peak of propiverine are not less than 6000 and mL of the standard solution under the above operating con- not more than 2.0, respectively. ditions, the number of theoretical plates and the symmetry System repeatability: When the test is repeated 6 times factor of the peak of propiverine are not less than 7000 and with 15 mL of the standard solution under the above operat- not more than 1.5, respectively. ing conditions, the relative standard deviation of the peak System repeatability: When the test is repeated 6 times area of propiverine is not more than 1.0z. with 15 mL of the standard solution under the above operat- Containers and storage Containers—Tight containers. ing conditions, the relative standard deviation of the peak area of propiverine is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord- Propiverine Hydrochloride Tablets ing to the following method: it meets the requirement of the Content uniformity test. プロピベリン塩酸塩錠 To 1 tablet of Propiverine Hydrochloride Tablets add the mobile phase, shake vigorously, add the mobile phase to Propiverine Hydrochloride Tablets contain not less make exactly V mL so that each mL contains about 0.1 mg than 95.0z and not more than 105.0z of propiverine of propiverine hydrochloride (C23H29NO3.HCl), centrifuge, hydrochloride (C23H29NO3.HCl: 403.94). and use the supernatant liquid as the sample solution. Sepa- rately, weigh accurately about 50 mg of Propirevine Hydro- Method of preparation Prepare as directed under Tablets, chloride RS, previously dried at 1059C for 1 hour, and dis- with Propiverine Hydrochloride. solve in the mobile phase to make exactly 100 mL. Pipet 10 Identification Shake vigorously a quantity of powdered mL of this solution, add the mobile phase to make exactly 50 Propiverine Hydrochloride Tablets, equivalent to 50 mg of mL, and use this solution as the standard solution. Then, Propiverine Hydrochloride according to the labeled amount, proceed as directed in the Assay under Propiverine Hydro- with 20 mL of water. Add acetonitrile to make 100 mL, cen- chloride. trifuge, and filter the supernatant liquid, if necessary. Deter- Amount (mg) of propiverine hydrochloride mine the absorption spectrum of the supernatant liquid or (C H NO .HCl) the filtrate under Ultraviolet-visible Spectrophotometry 23 29 3 ＝ M × A /A × V/500 <2.24>: it exhibits a maximum between 257 nm and 261 nm. S T S 1312 Propranolol Hydrochloride / Official Monographs JP XVI

MS: Amount (mg) of Propiverine Hydrochloride RS tion of the powder, equivalent to about 50 mg of propiverine hydrochloride (C H NO .HCl), add the mobile phase, Dissolution <6.10> When the test is performed at 50 revolu- 23 29 3 shake vigorously, and add the mobile phase to make exactly tions per minute according to Paddle method, using 900 mL 100 mL. Centrifuge this solution, pipet 10 mL of the super- of 2nd fluid for dissolution test as the dissolution medium, natant liquid, add the mobile phase to make exactly 50 mL, the dissolution rate in 20 minutes of Propiverine Hydrochlo- and use this solution as the sample solution. Separately, ride Tablets is not less than 85z. weigh accurately about 50 mg of Propiverine Hydrochloride Start the test with 1 tablet of Propiverine Hydrochloride RS, previously dried at 1059C for 1 hour, and dissolve in the Tablets, withdraw not less than 25 mL of the dissolved solu- mobile phase to make exactly 100 mL. Pipet 10 mL of this tion at the specified minute after starting the test, and filter solution, add the mobile phase to make exactly 50 mL, and through a membrane filter with a pore size not exceeding use this solution as the standard solution. Then, proceed as 0.45 mm. Discard the first 10 mL of the filtrate, pipet V mL directed in the Assay under Propiverine Hydrochloride. of the subsequent filtrate, add the dissolution medium to make exactly V?mL so that each mL contains about 11 mgof Amount (mg) of propiverine hydrochloride propiverine hydrochloride (C23H29NO3.HCl) according to (C23H29NO3.HCl) the labeled amount. Pipet 15 mL of this solution, add ex- ＝ MS × AT/AS actly 2 mL of 0.1 mol/L hydrochloric acid TS, and use this M : Amount (mg) of Propiverine Hydrochloride RS solution as the sample solution. Separately, weigh accurately S about 28 mg of Propiverine Hydrochloride RS, previously Containers and storage Container—Tight containers. dried at 1059C for 1 hour, and dissolve in the dissolution medium to make exactly 100 mL. Pipet 4 mL of this solution, and add the dissolution medium to make exactly 100 Propranolol Hydrochloride mL. Further, pipet 15 mL of this solution, add exactly 2 mL of 0.1 mol/L hydrochloric acid TS, and use this solution as プロプラノロール塩酸塩 the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of propiverine of both solutions. Dissolution rate (z) with respect to the labeled amount C H NO .HCl: 295.80 of propiverine hydrochloride (C H NO .HCl) 16 21 2 23 29 3 (2RS)-1-(1-Methylethyl)amino-3-(naphthalen- ＝ M × A /A × V?/V × 1/C × 36 S T S 1-yloxy)propan-2-ol monohydrochloride

MS: Amount (mg) of Propiverine Hydrochloride RS [318-98-9] C: Labeled amount (mg) of propiverine hydrochloride

(C23H29NO3.HCl) in 1 tablet Propranolol Hydrochloride, when dried, contains not less than 99.0z and not more than 101.0z of Operating conditions— C H NO .HCl. Detector: An ultraviolet absorption photometer (wave- 16 21 2 length: 220 nm). Description Propranolol Hydrochloride occurs as a white, Column: A stainless steel column 4.6 mm in inside diame- crystalline powder. ter and 15 cm in length, packed with octadecylsilanized silica It is freely soluble in methanol, soluble in water and in gel for liquid chromatography (5 mm in particle diameter). acetic acid (100), and sparingly soluble in ethanol (99.5). Column temperature: A constant temperature of about A solution of Propranolol Hydrochloride in methanol 259C. (1 in 40) shows no optical rotation. Mobile phase: To diluted 0.02 mol/L potassium dihydro- It is gradualy colored to yellowish white to light brown by gen phosphate TS (1 → 2) add phosphoric acid, and adjust light. to pH 2.0. To 560 mL of this solution add 440 mL of aceto- Identification (1) Determine the absorption spectrum of a nitrile. solution of Propranolol Hydrochloride in methanol (1 in Flow rate: Adjust the flow rate so that the retention time 50,000) as directed under Ultraviolet-visible Spectropho- of propirevine is about 6 minutes. tometry <2.24>, and compare the spectrum with the Refer- System suitability— ence Spectrum: both spectra exhibit similar intensities of ab- System performance: When the procedure is run with 20 sorption at the same wavelengths. mL of the standard solution under the above operating con- (2) Determine the infrared absorption spectrum of ditions, the number of theoretical plates and the symmetry Propranolol Hydrochloride, previously dried, as directed in factor of the peak of propiverine are not less than 4000 and the potassium chloride disk method under Infrared Spectro- not more than 2.0, respectively. photometry <2.25>, and compare the spectrum with the Ref- System repeatability: When the test is repeated 6 times erence Spectrum: both spectra exhibit similar intensities of with 20 mL of the standard solution under the above opera- absorption at the same wave numbers. tions conditions, the relative standard deviation of the peak (3) A solution of Propranolol Hydrochloride (1 in 50) area of propiverine is not more than 2.0z. responds to the Qualitative Tests <1.09> (2) for chloride. Assay Weigh accurately and powder not less than 20 pH <2.54> The pH of a solution prepared by dissolving Propiverine Hydrochloride Tablets. Weigh accurately a por- 0.5 g of Propranolol Hydrochloride in 50 mL of water is JP XVI Official Monographs / Propranolol Hydrochloride Tablets 1313

5.0–6.0. Assay Weigh accurately about 0.5 g of Propranolol Hydro- chloride, previously dried, dissolove in 50 mL of a mixture Melting point <2.60> 163–1669C of acetic anhydride and acetic acid (100) (7:3), and titrate Purity (1) Clarity and color of solution—Dissolve 1.0 g <2.50> with 0.1 mol/L perchloric acid VS (potentiometric of Propranolol Hydrochloride in 20 mL of water: the solu- titration). Perform a blank determination in the same man- tion is clear and colorless. ner, and make any necessary correction. (2) Heavy metals <1.07>—Proceed with 1.0 g of Each mL of 0.1 mol/L perchloric acid VS Propranolol Hydrochloride according to Method 4, and per- ＝ 29.58 mg of C H NO .HCl form the test. Prepare the control solution with 2.0 mL of 16 21 2 Standard Lead Solution (not more than 20 ppm). Containers and storage Containers—Well-closed contain- (3) Related substances—Dissolve 20 mg of Propranolol ers. Hydrochloride in 10 mL of the mobile phase, and use this Storage—Light-resistant. solution as the sample solution. Pipet 2 mL of the sample solution, and add the mobile phase to make exactly 100 mL. Pipet 1 mL of this solution, add the mobile phase to make Propranolol Hydrochloride Tablets exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 20 mLeachofthesamplesolu- プロプラノロール塩酸塩錠 tion and standard solution as directed under Liquid Chroma- tography <2.01> according to the following conditions, and Propranolol Hydrochloride Tablets contain not determine each peak area by the automatic integration less than 95.0z and not more than 105.0z of method: the area of the peak other than propranolol from the labeled amount of propranolol hydrochloride the sample solution is not larger than 1/2 times the peak area (C H NO .HCl: 295.80). of propranolol from the standard solution, and the total 16 21 2 area of the peaks other than the peak of propranolol is not Method of preparation Prepare as directed under Tablets, larger than 2 times the peak area of propranolol from the with Propranolol Hydrochloride. standard solution. Identification Determine the absorption spectrum of the Operating conditions— sample solution obtained in the Assay as directed under Detector: An ultraviolet absorption photometer (wave- Ultraviolet-visible Spectrophotometry <2.24>:itexhibits length: 292 nm). maxima between 288 nm and 292 nm, and between 317 nm Column: A stainless steel column 4.6 mm in inside diame- and 321 nm. ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Uniformity of dosage units <6.02> Perform the test accord- Column temperature: A constant temperature of about ing to the following method: it meets the requirement of the 259C. Content uniformity test. Mobile phase: Dissolve 1.6 g of sodium lauryl sulfate and To 1 tablet of Propranolol Hydrochloride Tablets add 20 0.31 g of tetrabutylammonium phosphate in 450 mL of mL of water, and shake until the tablet is completely disinte- water, add 1 mL of sulfuric acid and 550 mL of acetonitrile grated. Add 50 mL of methanol, shake vigorously for 10 for liquid chromatography, and adjust to pH 3.3 with 2 minutes, then add methanol to make exactly 100 mL, and mol/L sodium hydroxide TS. filter. Discard the first 20 mL of the filtrate, pipet V mL of Flow rate: Adjust the flow rate so that the retention time the subsequent filtrate, add methanol to make exactly V?mL of propranolol is about 4 minutes. so that each mL contains about 20 mg of propranolol hydro-

Time span of measurement: About 5 times as long as the chloride (C16H21NO2.HCl), and use this solution as the sam- retention time of propranolol. ple solution. Separately, weigh accurately about 50 mg of System suitability— propranolol hydrochloride for assay, previously dried at Test for required detectability: Measure exactly 5 mL of 1059C for 4 hours, and dissolve in methanol to make exactly the standard solution, and add the mobile phase to make ex- 50 mL. Pipet 2 mL of this solution, add methanol to make actly 20 mL. Confirm that the peak area of propranolol ob- exactly 100 mL, and use this solution as the standard solu- tained with 20 mL of this solution is equivalent to 17 to 33z tion. Determine the absorbances, AT and AS,ofthesample of that with 20 mL of the standard solution. solution and standard solution at 290 nm as directed under System performance: When the procedure is run with 20 Ultraviolet-visible Spectrophotometry <2.24>. mL of the standard solution under the above operating con- Amount (mg) of propranolol hydrochloride ditions, the number of theoretical plates and the symmetry (C H NO .HCl) factor of the peak of propranolol is not less than 3000 and 16 21 2 ＝ M × A /A × V?/V × 1/25 not more than 2.0, respectively. S T S

System repeatability: When the test is repeated 6 times MS: Amount (mg) of propranolol hydrochloride for assay with 20 mL of the standard solution under the above operat- Dissolution <6.10> When the test is performed at 50 revolu- ing conditions, the relative standard deviation of the peak tions per minute according to the Paddle method, using 900 area of propranolol is not more than 2.0z. mL of water as the dissolution medium, the dissolution rate Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, in 15 minutes of Propranolol Hydrochloride Tablets is not 4 hours). less than 80z. Start the test with 1 tablet of Propranolol Hydrochloride Residue on ignition <2.44> Not more than 0.1z (1 g). Tablets, withdraw not less than 20 mL of the medium at the 1314 Propyl Parahydroxybenzoate / Official Monographs JP XVI

 specified minute after starting the test, and filter through a that are not harmonized are marked with symbols ( ). membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet V mL of the Propyl Parahydroxybenzoate contains not less than subsequent filtrate, add water to make exactly V?mL so that 98.0z and not more than 102.0z of C10H12O3. each mL contains about 10 mg of propranolol hydrochloride Description Propyl Parahydroxybenzoate occurs as col- (C H NO .HCl) according to the labeled amount, and use 16 21 2 orless crystals or a white, crystalline powder. this solution as the sample solution. Separately, weigh accu- It is freely soluble in ethanol (95) and in acetone, and very rately about 50 mg of propranolol hydrochloride for assay, slightly soluble in water. previously dried at 1059C for 4 hours, and dissolve in water  to make exactly 50 mL. Pipet 1 mL of this solution, add Identification (1) The melting point <2.60> of Propyl Par- water to make exactly 100 mL, and use this solution as the ahydroxybenzoate is between 969Cand999C.  standard solution. Determine the absorbances, AT and AS, (2) Determine the infrared absorption spectrum of of the sample solution and standard solution at 290 nm as Propyl Parahydroxybenzoate as directed in the potassium directed under Ultraviolet-visible Spectrophotometry <2.24>. bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- Dissolution rate (z) with respect to the labeled amount trum: both spectra exhibit similar intensities of absorption at of propranolol hydrochloride (C16H21NO2.HCl) the same wave numbers. ＝ MS × AT/AS × V?/V × 1/C × 18 Purity (1) Clarity and color of solution—Dissolve 1.0 g M : Amount (mg) of propranolol hydrochloride for assay S of Propyl Parahydroxybenzoate in 10 mL of ethanol (95): C: Labeled amount (mg) of propranolol hydrochloride the solution is clear and not more intensely colored than the (C H NO .HCl) in 1 tablet 16 21 2 following control solution. Assay Weigh accurately the mass of not less than 20 Control solution: To 5.0 mL of Cobalt (II) Chloride CS, Propranolol Hydrochloride Tablets, and powder. Weigh 12.0 mL of Iron (III) Chloride CS and 2.0 mL of Copper (II) accurately a portion of the powder, equivalent to about 20 Sulfate CS add water to make 1000 mL. mg of propranolol hydrochloride (C16H21NO2.HCl), add 60 (2) Acidity—Dissolve 0.20 g of Propyl Parahydrox- mL of methanol, shake for 10 minutes, and add methanol to ybenzoate in 5 mL of ethanol (95), add 5 mL of freshly make exactly 100 mL. Filter, discard the first 20 mL of the boiled and cooled water and 0.1 mL of bromocresol green- filtrate, pipet 10 mL of the subsequent filtrate, add methanol sodium hydroxide-ethanol TS, then add 0.1 mL of 0.1 to make exactly 100 mL, and use this solution as the sample mol/L sodium hydroxide VS: the solution shows a blue solution. Separately, weigh accurately about 50 mg of color. propranolol hydrochloride for assay, previously dried at (3) Heavy metals <1.07>—Dissolve 1.0 g of Propyl Par- 1059C for 4 hours, and dissolve in methanol to make exactly ahydroxybenzoate in 25 mL of acetone, add 2 mL of dilute 50 mL. Pipet 2 mL of this solution, add methanol to make acetic acid and water to make 50 mL, and perform the test exactly 100 mL, and use this solution as the standard solu- using this solution as the test solution. Prepare the control tion. Determine the absorbances, AT and AS,ofthesample solution as follows: to 2.0 mL of Standard Lead Solution solution and standard solution at 290 nm as directed under add 25 mL of acetone, 2 mL of dilute acetic acid, and water

Ultraviolet-visible Spectrophotometry <2.24>. to make 50 mL (not more than 20 ppm). (4) Related substances—Dissolve 0.10 g of Propyl Par- Amount (mg) of propranolol hydrochloride ahydroxybenzoate in 10 mL of acetone, and use this solution (C H NO .HCl) 16 21 2 as the sample solution. Pipet 0.5 mL of the sample solution, ＝ M × A /A × 2/5 S T S add acetone to make exactly 100 mL, and use this solution as

MS: Amount (mg) of propranolol hydrochloride for assay the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 Containers and storage Containers—Well-closed contain- mL each of the sample solution and standard solution on a ers. plate of silica gel with fluorescent indicator for thin-layer Storage—Light-resistant. chromatography. Develop the plate with a mixture of methanol, water and acetic acid (100) (70:30:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultravio- Propyl Parahydroxybenzoate let light (main wavelength: 254 nm): the spot other than the principal spot obtained with is not more intense than the パラオキシ安息香酸プロピル spot with the standard solution. Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 1.0 g of Propyl Parahydrox- ybenzoate add exactly 20 mL of 1 mol/L sodium hydroxide VS, heat at about 709C for 1 hour, and immediately cool in

C10H12O3:180.20 ice. Titrate <2.50> the excess sodium hydroxide with 0.5 Propyl 4-hydroxybenzoate mol/LsulfuricacidVSuptothe second equivalent point [94-13-3] (potentiometric titration). Perform a blank determination. Each mL of 1 mol/L sodium hydroxide VS This monograph is harmonized with the European Phar- ＝ 180.2 mg of C H O macopoeia and the U.S. Pharmacopeia. The parts of the text 10 12 3 JP XVI Official Monographs / Propylthiouracil 1315

Containers and storage Containers—Well-closed contain- more than 0.005z. ers.  Distilling range <2.57> 184 – 1899C, not less than 95 volz. Containers and storage Containers—Tight containers. Propylene Glycol プロピレングリコール Propylthiouracil

プロピルチオウラシル

C3H8O2: 76.09 (2RS)-Propane-1,2-diol [57-55-6]

Description Propylene Glycol is a clear, colorless, viscous C7H10N2OS: 170.23 liquid. It is odorless, and has a slightly bitter taste. 6-Propyl-2-thiouracil It is miscible with water, with methanol, with ethanol (95) [51-52-5] and with pyridine. It is freely soluble in diethyl ether. Propylthiouracil, when dried, contains not less than It is hygroscopic. 98.0z of C7H10N2OS. Identification (1) Mix 2 to 3 drops of Propylene Glycol Description Propylthiouracil occurs as a white powder. It with 0.7 g of triphenylchloromethane, add 1 mL of pyridine, is odorless, and has a bitter taste. and heat under a reflux condenser on a water bath for 1 It is sparingly soluble in ethanol (95), and very slightly hour. After cooling, dissolve the mixture in 20 mL of ace- soluble in water and in diethyl ether. tone by warming, shake with 0.02 g of activated charcoal, It dissolves in sodium hydroxide TS and in ammonia TS. and filter. Concentrate the filtrate to about 10 mL, and cool. Identification (1) Shake well 0.02 g of Propylthiouracil Collect the separated crystals, and dry in a desiccator (silica with 7 mL of bromine TS for 1 minute, and heat until the gel) for 4 hours: the crystals melt <2.60> between 1749Cand color of bromine TS disappears. Cool, filter, and add 10 mL 1789C. of barium hydroxide TS to the filtrate: a white precipitate is (2) Heat gently 1 mL of Propylene Glycol with 0.5 g of produced. The color of the precipitate does not turn purple potassium hydrogen sulfate: a characteristic odor is evolved. within 1 minute. 20 Specific gravity <2.56> d 20: 1.035 – 1.040 (2) To 5 mL of a hot saturated solution of Propylthio- uracil add 2 mL of a solution of sodium pentacyanoammine Purity (1) Acidity—Mix 10.0 mL of Propylene Glycol ferroate (II) n-hydrate (1 in 100): a green color develops. with 50 mL of freshly boiled and cooled water, and add 5 drops of phenolphthalein TS and 0.30 mL of 0.1 mol/L so- Melting point <2.60> 218 – 2219C dium hydroxide VS: the solution has a red color. Purity (1) Sulfate <1.14>—Triturate Propylthiouracil (2) Chloride <1.03>—Perform the test with 2.0 g of finely in a mortar. To 0.75 g of the powder add 25 mL of Propylene Glycol. Prepare the control solution with 0.40 mL water, heat for 10 minutes on a water bath, cool, filter, and of 0.01 mol/L hydrochloric acid VS (not more than wash the residue with water until the volume of the filtrate 0.007z). becomes 30 mL. To 10 mL of the filtrate add 1 mL of dilute (3) Sulfate <1.14>—Perform the test with 10.0 g of hydrochloric acid and water to make 50 mL, and perform Propylene Glycol. Prepare the control solution with 0.40 mL the test using this solution as the test solution. Prepare the of 0.005 mol/L sulfuric acid VS (not more than 0.002z). control solution with 0.40 mL of 0.005 mol/L sulfuric acid (4) Heavy metals <1.07>—Perform the test with 5.0 g of VS (not more than 0.077z). Propylene Glycol according to Method 1. Prepare the con- (2) Thiourea—Dissolve 0.30 g of Propylthiouracil in 50 trol solution with 2.5 mL of Standard Lead Solution (not mL of water by heating under a reflux condenser for 5 more than 5 ppm). minutes, cool, and filter. To 10 mL of the filtrate add 3 mL (5) Arsenic <1.11>—Prepare the test solution with 1.0 g of ammonia TS, shake well, and add 2 mL of silver nitrate of Propylene Glycol according to Method 1, and perform TS: the solution has no more color than the following con- the test (not more than 2 ppm). trol solution. (6) Glycerin—Heat 1.0 g of Propylene Glycol with 0.5 g Control solution: Weigh exactly 60 mg of thiourea, and of potassium hydrogen sulfate and evaporate to dryness: no dissolve in water to make exactly 100 mL. Pipet 1 mL of this odor of acrolein is perceptible. solution, add water to make exactly 100 mL, and proceed Water <2.48> Not more than 0.5z (2 g, volumetric titra- with 10 mL of this solution in the same manner. tion, direct titration). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, Residue on ignition <2.44> Weigh accurately about 20 g of 2 hours). Propylene Glycol in a tared crucible, and heat to boiling. Residue on ignition <2.44> Not more than 0.1z (1 g). Stop heating, and immediately ignite to burn. Cool, moisten the residue with 0.2 mL of sulfuric acid, and heat strongly Assay Weigh accurately about 0.3 g of Propylthiouracil, with care to constant mass: the mass of the residue is not previously dried, and add 30 mL of water. Add 30 mL of 0.1 1316 Propylthiouracil Tablets / Official Monographs JP XVI mol/L sodium hydroxide VS from a burette, heat to boil, 10 mL of the filtrate, pipet V mL of the subsequent filtrate, and dissolve by stirring. Wash down the solid adhering to the add the dissolution medium to make exactly V?mL so that wall of the flask with a small amount of water, and add 50 each mL contains about 5.6 mg of propylthiouracil mL of 0.1 mol/L silver nitrate VSwithstirring.Boilgently (C7H10N2OS) according to the labeled amount, and use this for 5 minutes, add 1 to 2 mL of bromothymol blue TS, and solution as the sample solution. Separately, weigh about 50 titrate <2.50> with 0.1 mol/L sodium hydroxide VS until a mg of propylthiouracil for assay, previously dried at 1059C persistent blue-green color develops. Determine the total for 3 hours, and dissolve in the dissolution medium to make volume of 0.1 mol/L sodium hydroxide VS consumed. exactly 1000 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 50 mL, and use this solution Each mL of 0.1 mol/L sodium hydroxide VS as the standard solution. Proceed as directed in the Assay. ＝ 8.512 mg of C7H10N2OS Dissolution rate (z) with respect to the labeled amount Containers and storage Containers—Well-closed contain- of propylthiouracil (C H N OS) ers. 7 10 2 ＝ M × A /A × V?/V × 1/C × 9 Storage—Light-resistant. S T S

MS: Amount (mg) of propylthiouracil for assay C: Labeled amount (mg) of propylthiouracil (C7H10N2OS) Propylthiouracil Tablets in 1 tablet Assay Weigh accurately the mass of not less than 20 プロピルチオウラシル錠 Propylthiouracil Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg of

Propylthiouracil Tablets contain not less than propylthiouracil (C7H10N2OS), add 150 mL of 2nd fluid for 93.0z and not more than 107.0z of the labeled dissolution test, disperse finely the particles with the aid of amount of propylthiouracil (C7H10N2OS: 170.23). ultrasonic waves, and add 2nd fluid for dissolution test to make exactly 200 mL. Filter this solution through a mem- Method of preparation Prepare as directed under Tablets, brane filter with a pore size not exceeding 0.45 mm, discard with Propylthiouracil. the first 5 mL of the filtrate, pipet 2 mL of the subsequent Identification To a quantity of powdered Propylthiouracil filtrate, add 2nd fluid for dissolution test to make exactly Tablets, equivalent to 0.3 g of Propylthiouracil according to 100 mL, and use this solution as the sample solution. Sepa- the labeled amount, add 5 mL of ammonia TS, allow to rately, weigh accurately about 50 mg of propylthiouracil for stand for 5 minutes with occasional shaking, add 10 mL of assay, previously dried at 1059C for 2 hours, and dissolve in water, and centrifuge. To the supernatant liquid add acetic 2nd fluid for dissolution test to make exactly 200 mL. Pipet acid (31), collect the precipitate produced, recrystallize from 2 mL of this solution, add 2nd fluid for dissolution test to water, and dry at 1059C for 1 hour: it melts <2.60> between make exactly 100 mL, and use this solution as the standard

2189C and 2219C. Proceed with the residue as directed in the solution. Determine the absorbance at 274 nm, AT and AS, Identification under Propylthiouracil. of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Uniformity of dosage units <6.02> Perform the test according to the following method: it meets the requirement of the Amount (mg) of propylthiouracil (C7H10N2OS) Content uniformity test. ＝ MS × AT/AS To 1 tablet of Propylthiouracil Tablets add 3V/4 mL of M : Amount (mg) of propylthiouracil for assay 2nd fluid for dissolution test, treat with ultrasonic waves S until the tablet is disintegrated, and add 2nd fluid for disso- Containers and storage Containers—Well-closed contain- lution test to make exactly V mL so that each mL contains ers. about 0.25 mg of propylthiouracil (C7H10N2OS). Filter this Storage—Light-resistant. solution through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 5 mL of the filtrate, pipet 2 mL of the subsequent filtrate, add 2nd fluid for dis- Protamine Sulfate solution test to make exactly 100 mL, and use this solution as the sample solution. Proceed as directed in the Assay. プロタミン硫酸塩

Amount (mg) of propylthiouracil (C7H10N2OS) ＝ MS × AT/AS × V/200 Protamine Sulfate is the sulfate of protamine prepared from the mature spermary of fish belonging to M : Amount (mg) of propylthiouracil for assay S the family Salmonidae. Dissolution <6.10> When the test is performed at 75 revolu- It has a property to bind with heparin. tions per minute according to the Paddle method, using 900 It binds with not less than 100 Units of heparin per mL of 2nd fluid for dissolution test as the dissolution me- mg,calculatedonthedriedbasis. dium, the dissolution rate in 30 minutes of Propylthiouracil Description Protamine Sulfate occurs as a white powder. Tablets is not less than 80z. It is sparingly soluble in water. Start the test with 1 tablet of Propylthiouracil Tablets, withdraw not less than 20 mL of the medium at the specified Identification (1) Dissolve 1 mg of Protamine Sulfate in 2 minute after starting the test, and filter through a membrane mL of water, add 5 drops of a solution prepared by dissolv- filter with a pore size not exceeding 0.8 mm. Discard the first ing 0.1 g of 1-naphthol in 100 mL of diluted ethanol (7 in 10) JP XVI Official Monographs / Protamine Sulfate Injection 1317 and 5 drops of sodium hypochlorite TS, then add sodium hy- Amount (heparin Unit) of heparin bound to 1 mg droxide TS until the solution becomes alkaline: a vivid red of Protamine Sulfate color develops. ＝ S × V × 50/MT × d (2) Dissolve 5 mg of Protamine Sulfate in 1 mL of water S: Amount (heparin Unit) of heparin sodium in 1 mL of by warming, add 1 drop of a solution of sodium hydroxide the standard solution (1 in 10) and 2 drops of copper (II) sulfate TS: a red-purple M : Amount (mg) of the sample, calculated on the dried color develops. T basis (3) An aqueous solution of Protamine Sulfate (1 in 20) d: Dilution factor for each sample solution from the sam- responds to the Qualitative Tests <1.09> for sulfate. ple solution (a) pH <2.54> Dissolve 1.0 g of Protamine Sulfate in 100 mL Sulfate content Weigh accurately about 0.15 g of Prota- of water: the pH of this solution is between 6.5 and 7.5. mine Sulfate, dissolve in 75 mL of water, add 5 mL of 3 Purity (1) Clarity and color of solution—Dissolve 0.10 g mol/L hydrochloric acid TS, and heat to boil. Add gradually of Protamine Sulfate in 10 mL of water: the solution is clear 10 mL of barium chloride TS while boiling, and allow to and colorless. stand for 1 hour while heating. Filter the precipitate formed, (2) Absorbance—Dissolve 0.10 g of Protamine Sulfate in wash the precipitate with warm water several times, and 10 mL of water, and determine the absorption spectrum as transfer the precipitate into a tared crucible. Dry the precipi- directed under Ultraviolet-visible Spectrophotometry <2.24>: tate, and incinerate by ignition to constant mass: the amount the absorbance between 260 nm and 280 nm is not more than of sulfate (SO4) is16–22z, calculated on the dried basis, 0.1. where 1 g of the residue is equivalent to 0.4117 g of SO4. Loss on drying <2.41> Not more than 5.0z (1 g, 1059C, Containers and storage Containers—Tight containers. 3 hours). Nitrogen content Weigh accurately about 10 mg of Prota- mine Sulfate, and perform the test as directed under Nitro- Protamine Sulfate Injection gen Determination <1.08>: the amount of nitrogen (N:14.01) プロタミン硫酸塩注射液 is 22.5 – 25.5z, calculated on the dried basis. Heparin-binding capacity Protamine Sulfate Injection is an aqueous solution (i) Sample solution (a)—Weigh accurately about 15 mg for injection. of Protamine Sulfate, and dissolve in water to make exactly It contains not less than 92.0z and not more than 100 mL. Repeat this procedure 3 times, and use the solutions 108.0z of the labeled amount of Protamine Sulfate. so obtained as the sample solutions (a ), (a )and(a). 1 2 3 It binds with not less than 100 Units of heparin per mg (ii) Sample solution (b)—Pipet 10 mL each of the sample of the labeled amount. solutions (a1), (a2)and(a3), add exactly 5 mL of water to them, and use these solutions as the sample solutions (b1), Method of preparation Prepare as directed under Injec- (b2)and(b3). tions, with Protamine Sulfate. (iii) Sample solution (c)—Pipet 10 mL each of the sam- Description Protamine Sulfate Injection is a colorless liq- ple solutions (a ), (a )and(a), add exactly 20 mL of water 1 2 3 uid. It is odorless or has the odor of preservatives. to them, and use these solutions as the sample solutions (c1), (c2)and(c3). Identification (1) Dilute a volume of Protamine Sulfate (iv) Standard solution—Dissolve Heparin Sodium RS in Injection, equivalent to 1 mg of Protamine Sulfate according water to make a solution containing exactly about 20 Units to the labeled amount, with water to make 2 mL, and per mL. proceed as directed in the Identification (1) under Protamine (v) Procedure—Transfer exactly 2 mL of the sample Sulfate. solution to a cell for spectrophotometer, add the standard (2) Dilute a volume of Protamine Sulfate Injection, solution dropwise while mixing, and determine the transmit- equivalent to 5 mg of Protamine Sulfate according to the tance at 500 nm as directed under Ultraviolet-visible Spectro- labeled amount, with water to make 1 mL, and proceed as photometry <2.24>. Continue the addition until a sharp directed in the Identification (2) under Protamine Sulfate. change in the transmittance is observed, and note the pH <2.54> 5.0–7.0 volume, V mL, of the standard solution added. Repeat this procedure 2 times for each sample solution. Bacterial endotoxins <4.01> Less than 6.0 EU/mg. (vi) Calculation—Calculate the amount of heparin Extractable volume <6.05> It meets the requirement. bound with 1 mg of the sample by the following formula from the volume of titrant on each sample solution, and cal- Foreign insoluble matter <6.06> Perform the test according culate the average of 18 results obtained. The assay is not to Method 1: it meets the requirement. valid unless each relative standard deviation of 6 results ob- Insoluble particulate matter <6.07> It meets the require- tained from the sample solution (a), sample solution (b) and ment. sample solution (c) is not more than 5z, respectively, and also unless each relative standard deviation of 6 results ob- Sterility <4.06> Perform the test according to the Mem- tained from 3 sets, (a1,b1,c1), (a2,b2,c2)and(a3,b3,c3)is brane filtration method: it meets the requirement. not more than 5z, respectively. Assay (1) Protein—Pipet a volume of Protamine Sulfate Injection, equivalent to about 10 mg of Protamine Sulfate, 1318 Prothionamide / Official Monographs JP XVI transfer to a Kjeldahl flask, evaporate on a water bath to Purity (1) Clarity and color of solution—Dissolve 0.5 g dryness with the aid of a current of air, determine the nitro- of Prothionamide in 20 mL of ethanol (95): the solution is gen as directed under Nitrogen Determination <1.08>,and clear, and shows a yellow color. calculate the amount of protein by converting 0.24 mg of (2) Acidity—Dissolve 3.0 g of Prothionamide in 20 mL nitrogen (N: 14.01) to 1 mg of protein. of methanol with warming. Add 100 mL of water to the so- (2) Heparin-binding activity—Proceed the test as dilution, cool in an ice water bath with agitation, and remove rected in the Heparin-binding capacity under Protamine any precipitate by filtration. Allow 80 mL of the filtrate to Sulfate, changing the sample solution (a) as below, and de- cool to room temperature, and add 0.8 mL of cresol red TS termine the amount of heparin bound to 1 mg of protein by and 0.20 mL of 0.1 mol/L sodium hydroxide VS: a red color dividing by the amount of protein. develops. (i) Sample solution (a)—Pipet a volume of Protamine (3) Heavy metals <1.07>—Proceed with 1.0 g of Pro- Sulfate Injection, equivalent to 15.0 mg of Protamine Sul- thionamide according to Method 2, and perform the test. fate, and add water to make exactly 100 mL. Repeat this Prepare the control solution with 2.0 mL of Standard Lead procedure two more times, and designate the solutions so Solution (not more than 20 ppm). obtained as the sample solutions (a1), (a2)and(a3). (4) Arsenic <1.11>—Prepare the test solution with 0.6 g of Prothionamide according to Method 3, and perform the Containers and storage Containers—Hermetic containers. test. To the test solution add 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 50), then add 1.5 mL of hydrogen peroxide (30), and ignite to burn (not Prothionamide more than 3.3 ppm). プロチオナミド Loss on drying <2.41> Not more than 0.5z (1 g, 809C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.3 g of Prothionamide, previously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the

C9H12N2S: 180.27 color of the solution changes from orange-red to dark 2-Propylpyridine-4-carbothioamide orange-brown (indicator: 2 mL of p-naphtholbenzein TS). [14222-60-7] Perform a blank determination. Each mL of 0.1 mol/L perchloric acid VS Prothionamide, when dried, contains not less than ＝ 18.03 mg of C9H12N2S 98.0z of C9H12N2S. Containers and storage Containers—Well-closed contain- Description Prothionamide occurs as yellow crystals or ers. crystalline powder. It has a slight, characteristic odor. Storage—Light-resistant. It is freely soluble in methanol and in acetic acid (100), soluble in ethanol (95), slightly soluble in diethyl ether, and practically insoluble in water. It dissolves in dilute hydrochloric acid and in dilute sulfu- Protirelin ric acid. プロチレリン Identification (1) Mix 0.05 g of Prothionamide with 0.1 g of 1-chloro-2,4-dinitrobenzene, transfer about 10 mg of this mixture to a test tube, and heat for several seconds over a small flame until the mixture is fused. Cool, and add 3 mL of potassium hydroxide-ethanol TS: a red to orange-red color develops. (2) Place 0.5 g of Prothionamide in a 100-mL beaker, and dissolve in 20 mL of sodium hydroxide TS by heating C16H22N6O4: 362.38 while shaking occasionally: the gas evolved turns a 5-Oxo-L-prolyl-L-histidyl-L-prolinamide moistened red litmus paper to blue. Boil gently, and evapo- [24305-27-9] rate the solution to 3 to 5 mL. After cooling, add gradually 20 mL of acetic acid (100), and heat on a water bath: the gas Protirelin contains not less than 98.5z of evolved darkens moistened lead (II) acetate paper. C16H22N6O4, calculated on the dehydrated basis. Evaporate the solution on a water bath to 3 to 5 mL with the Description Protirelin occurs as a white powder. aid of a current of air, cool, add 10 mL of water, and mix It is freely soluble in water, in methanol, in ethanol (95) well. Filter the crystals by suction, recrystallize from water and in acetic acid (100). immediately, and dry in a desiccator (in vacuum, silica gel) It is hygroscopic. for 6 hours: the crystals melt <2.60> between 1989Cand 2039C (with decomposition). Identification (1) Take 0.01 g of Protirelin in a test tube made of hard glass, add 0.5 mL of 6 mol/L hydrochloric Melting point <2.60> 142–1459C acid TS, seal the upper part of the tube, and heat carefully at JP XVI Official Monographs / Protirelin Tartrate Hydrate 1319

1109C for 5 hours. After cooling, open the seal, transfer the mol/L perchloric acid VS (potentiometric titration). Per- contents into a beaker, and evaporate on a water bath to form a blank determination, and make any necessary correc- dryness. Dissolve the residue in 1 mL of water, and use this tion. solution as the sample solution. Separately, dissolve 0.08 g Each mL of 0.02 mol/L perchloric acid VS of L-glutamic acid, 0.12 g of L-histidine hydrochloride ＝ 7.248 mg of C16H22N6O4 monohydrate and 0.06 g of L-proline in 20 mL of water, and use this solution as the standard solution. Perform the test Containers and storage Containers—Tight containers. with these solutions as directed under Thin-layer Chroma- tography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer Protirelin Tartrate Hydrate chromatography. Develop the plate with a mixture of 1- butanol, water, acetic acid (100) and pyridine (4:1:1:1) to a プロチレリン酒石酸塩水和物 distance of about 12 cm, and dry the plate at 1009Cfor30 minutes. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat at 809C for 5 minutes: the three spots obtained from the sample solution show the same color and the same Rf value as each corresponding spots obtained from the standard solution. (2) Determine the infrared absorption spectrum of Pro- tirelin, as directed in the potassium bromide disk method C H N O .C H O .H O: 530.49 under Infrared Spectrophotometry <2.25>, and compare the 16 22 6 4 4 6 6 2 5-Oxo-L-prolyl-L-histidyl-L-prolinamide monotartrate spectrum with the Reference Spectrum: both spectra exhibit monohydrate similar intensities of absorption at the same wave numbers. [24305-27-9, Protirelin] 20 Optical rotation <2.49> [a]D : －66.0 – －69.09(0.1 g calculated on the dehydrated basis, water, 20 mL, 100 mm). Protirelin Tartrate Hydrate, calculated on the anhydrous basis, contains not less than 98.5z of protirelin pH <2.54> Dissolve 0.20 g of Protirelin in 10 mL of water: tartrate (C H N O .C H O : 512.48). the pH of this solution is between 7.5 and 8.5. 16 22 6 4 4 6 6 Description Protirelin Tartrate Hydrate occurs as white to Purity (1) Clarity and color of solution—Dissolve 0.10 g pale yellowish white crystals or crystalline powder. of Protirelin in 10 mL of water: the solution is clear and col- It is freely soluble in water, sparingly soluble in acetic acid orless. (100), and practically insoluble in ethanol (95) and in diethyl (2) Heavy metals <1.07>—Proceed with 1.0 g of Protire- ether. lin according to Method 2, and perform the test. Prepare the Melting point: about 1879C (with decomposition). control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Identification (1) To 1 mL of a solution of Protirelin (3) Related substances—Dissolve 0.20 g of Protirelin in Tartrate Hydrate (1 in 1000) add 2 mL of a solution of 4- 10 mL of water, and use this solution as the sample solution. nitrobenzene diazonium fluoroborate (1 in 2000) and 2 mL Pipet 1 mL of the sample solution, add water to make ex- of boric acid-potassium chloride-sodium hydroxide buffer actly 200 mL, and use this solution as the standard solution. solution, pH 9.0: a red color develops. Perform the test with these solutions as directed under Thin- (2) Dissolve 0.03 g of Protirelin Tartrate Hydrate in 5 layer Chromatography <2.03>. Spot 5 mLeachofthesample mL of sodium hydroxide TS, add 1 drop of copper (II) sul- solution and standard solution on a plate (1) of silica gel for fate TS: a purple color develops. thin-layer chromatography, and spot 5 mL of the sample so- (3) To 0.20 g of Protirelin Tartrate Hydrate add 5.0 mL lution on a plate (2) of silica gel for thin-layer chromatogra- of 6 mol/L hydrochloric acid TS, and boil for 7 hours under phy. Develop the plates with a mixture of 1-butanol, water, a reflux condenser. After cooling, evaporate 2.0 mL of this pyridine and acetic acid (100) (4:2:1:1) to a distance of about solution on a water bath to dryness, dissolve the residue in 12 cm, and dry the plates at 1009C for 30 minutes. Spray 2.0 mL of water and use this solution as the sample solution. evenly a mixture of a solution of sulfanilic acid in 1 mol/L Separately, dissolve 22 mg of L-glutamic acid, 32 mg of L- hydrochloric acid TS (1 in 200) and a solution of sodium histidine hydrochloride monohydrate and 17 mg of L-proline nitrite (1 in 20) (1:1) on the plate (1), and air-dry the plates. in2.0mLof0.1mol/LhydrochloricacidTSbyheating,and Successively spray evenly a solution of sodium carbonate use this solution as the standard solution. Perform the test decahydrate (1 in 10) on it: the spots other than the principal with these solutions as directed under Thin-layer Chroma- spot from the sample solution are not more intense than the tography <2.03>. Spot 2 mL each of the sample solution and spot from the standard solution. Spray evenly a solution of standard solution on a plate of silica gel for thin-layer chro- ninhydrin in acetone (1 in 50) on the plate (2), and heat at matography. Develop the plate with a mixture of 1-butanol, 809C for 5 minutes: no colored spot appears. water, acetic acid (100) and pyridine (4:1:1:1) to a distance of about 12 cm, and dry at 1009C for 30 minutes. Spray Water <2.48> Not more than 5.0z (0.1 g, volumetric titra- evenly a solution of ninhydrin in acetone (1 in 50) on the tion, direct titration). plate, and dry at 809C for 5 minutes: the three spots ob- Residue on ignition <2.44> Not more than 0.3z (0.2 g). tained from the sample solution show, respectively, the same color and the same Rf value as the corresponding spot from Assay Weigh accurately about 70 mg of Protirelin dissolve the standard solution. in 50 mL of acetic acid (100), and titrate <2.50> with 0.02 1320 Pullulan / Official Monographs JP XVI

(4) A solution of Protirelin Tartrate Hydrate (1 in 40) responds to the Qualitative Tests <1.09> for tartrate. Pullulan Optical rotation <2.49> [a]20: －50.0 – －53.09(0.5 g calcu- D プルラン lated on the anhydrous basis, water, 25 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Protirelin Tartrate Hydrate in 100 mL of water: the pH of this solution is between 3.0 and 4.0. Purity (1) Clarity and color of solution—Dissolve 0.10 g of Protirelin Tartrate Hydrate in 10 mL of water: the solution is clear and colorless. (2) Heavy metals <1.07>—Proceed with 1.0 g of Protire- lin Tartrate Hydrate according to Method 2, and perform (C18H30O15)n the test. Prepare the control solution with 2.0 mL of Stand- Poly[6)-a-D-glucopyranosyl-(1→4)-a-D- ard Lead Solution (not more than 20 ppm). glucopyranosyl-(1→4)-a-D-glucopyranosyl-(1→] (3) Arsenic <1.11>—Take 1.0 g of Protirelin Tartrate Hy- [9057-02-7] drate in a porcelain crucible. Add 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10), Pullulan is a neutral simple polysaccharide pro- ignite the ethanol, and heat gradually to incinerate. If a car- duced by the growth of Aureobasidium pullulans.It bonized material still remains in this method, moisten with a has a chain structure of repeated a-1,6 binding of mal- small quantity of nitric acid, and ignite to incinerate. After totriose composed of three glucoses in a-1,4 binding. cooling, add 10 mL of dilute hydrochloric acid, heat on a Description Pullulan occurs as a white powder. water bath to dissolve the residue, use this solution as the It is freely soluble in water, and practically insoluble in test solution, and perform the test (not more than 2 ppm). ethanol (99.5). (4) Related substances—Dissolve 0.60 g of Protirelin Tartrate Hydrate in 10 mL of water, and use this solution as Identification (1) Dissolve 10 g of Pullulan in 100 mL of the sample solution. Pipet 1 mL of the sample solution, add water with stirring by adding in small portions: a viscous so- water to make exactly 200 mL, and use this solution as the lution is produced. standard solution. Perform the test with these solutions as (2) Mix 10 mL of the viscous solution obtained in (1) directed under Thin-layer Chromatography <2.03>. Spot 5 with 0.1 mL of pullulanase TS, and allow to stand: the solu- mL each of the sample solution and standard solution on a tion loses its viscosity. plate (1) of silica gel for thin-layer chromatography. Spot 5 (3) To 10 mL of a solution of Pullulan (1 in 50) add 2 mL of the sample solution on a plate (2) of silica gel for thin- mL of macrogol 600: a white precipitate is formed immedi- layer chromatography. Develop the plates with a mixture of ately. chloroform, methanol and ammonia solution (28) (6:4:1) to Viscosity <2.53> Take exactly 10.0 g of Pullulan, previously a distance of about 10 cm, and dry at 1009C for 30 minutes. dried, dissolve in water to make exactly 100 g, and perform Spray evenly a mixture of a solution of sulfanilic acid in 1 the test at 30 ± 0.19C as directed in Method 1: the kinematic mol/L hydrochloric acid TS (1 in 200) and a solution of so- viscosity is between 100 and 180 mm2/s. dium nitrite (1 in 20) (1:1) on the plate (1), and air-dry the plate. Then, spray evenly a solution of sodium carbonate pH <2.54> Dissolve 1.0 g of Pullulan in 10 mL of freshly decahydrate (1 in 10) on the plate: the spots other than the boiled and cooled water: the pH is between 4.5 and 6.5. principal spot from the sample solution are not more intense Purity (1) Heavy metals <1.07>—Proceed with 4.0 g of than those from the standard solution in color. On the other Pullulan according to Method 2, and perform the test. Pre- hand, spray evenly a solution of ninhydrin in acetone (1 in pare the control solution with 2.0 mL of Standard Lead So- 50) on the plate (2), and dry at 809C for 5 minutes: no lution (not more than 5 ppm). colored spot is obtained. (2) Nitrogen—Weigh accurately about 3 g of Pullulan, Water <2.48> Not more than 4.5z (0.2 g, volumetric titra- previously dried, and perform the test as directed under tion, direct titration). Nitrogen Determination <1.08>: the amount of nitrogen (N: 14.01) is not more than 0.05z. Use 12 mL of sulfuric acid Residue on ignition <2.44> Not more than 0.2z (0.5 g). for the decomposition, and add 40 mL of a solution of sodi- Assay Weigh accurately about 0.5 g of Protirelin Tartrate um hydroxide (2 in 5). Hydrate, dissolve in 80 mL of acetic acid (100) by warming, (3) Monosaccharide and oligosaccharides—Dissolve cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS 0.8 g of Pullulan, previously dried, in 100 mL of water, and (potentiometric titration). Perform a blank determination, designate this solution as the sample stock solution. To 1 mL and make any necessary correction. of the sample stock solution add 0.1 mL of potassium chloride saturated solution, and shake vigorously with 3 mL of Each mL of 0.1 mol/L perchloric acid VS methanol. Centrifuge, and use the supernatant liquid as the ＝ 51.25 mg of C H N O .C H O 16 22 6 4 4 6 6 sample solution. Separately, pipet 1 mL of the sample stock Containers and storage Containers—Well-closed contain- solution, add water to make exactly 50 mL, and use this so- ers. lution as the standard solution. Pipet 0.2 mL each of the sample solution, the standard solution and water, transfer them gently to each test tube containing 5 mL of a solution JP XVI Official Monographs / Pyrantel Pamoate 1321 of anthrone in diluted sulfuric acid (3 in 4) (1 in 500) and tion spectrum of the solution as directed under Ultraviolet- cooling in ice water, stir immediately, then heat at 909Cfor visible Spectrophotometry <2.24>, and compare the spectrum 10 minutes, and cool immediately. Perform the test with with the Reference Spectrum: both spectra exhibit similar in- these solutions so obtained as directed under Ultraviolet- tensities of absorption at the same wavelengths. visible Spectrophtometry <2.24> using water as a blank, and (4) Determine the infrared absorption spectrum of determine the absorbances at 620 nm, AT, AS and AB:the Pyrantel Pamoate, previously dried, as directed in the potas- amount of monosaccharide and oligosaccharides is not more sium bromide disk method under Infrared Spectrophotome- than 10.0z. try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp- Amount (z) of monosaccharide and oligosaccharides tion at the same wave numbers. ＝ (AT － AB)/(AS － AB) × 8.2 Purity (1) Chloride <1.03>—To 1.0 g of Pyrantel Pamo- Loss on drying <2.41> Not more than 6.0z (1 g, in vacu- ate add 10 mL of dilute nitric acid and 40 mL of water, and um, 909C, 6 hours). heat on a water bath with shaking for 5 minutes. After cool- Residue on ignition <2.44> Not more than 0.3z (2 g). ing, add water to make 50 mL, and filter. To 20 mL of the filtrate add 2 mL of dilute nitric acid and water to make 50 Containers and storage Containers—Well-closed contain- mL. Proceed the test using this solution as the test solution. ers. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.036z). (2) Sulfate <1.14>—To 0.75 g of Pyrantel Pamoate add 5 Pyrantel Pamoate mL of dilute hydrochloric acid and water to make 100 mL, andheatonawaterbathfor5minuteswithshaking.After ピランテルパモ酸塩 cooling, add water to make 100 mL, and filter. To 20 mL of the filtrate add water to make 50 mL. Proceed the test using this solution as the test solution. Prepare the control solution with 0.45 mL of 0.005 mol/L sulfuric acid VS (not more than 0.144z). (3) Heavy metals <1.07>—Proceed with 1.0 g of Pyrantel Pamoate according to Method 2, and perform the test. Pre- pare the control solution with 3.0 mL of Standard Lead So- lution (not more than 30 ppm).

C11H14N2S.C23H16O6: 594.68 (4) Arsenic <1.11>—Prepare the test solution with 1.0 g 1-Methyl-2-[(1E)-2-(thien-2-yl)vinyl]-1,4,5,6- of Pyrantel Pamoate according to Method 3, and perform tetrahydropyrimidine mono[4,4?-methylenebis(3- the test (not more than 2 ppm). hydroxy-2-naphthoate)] (1/1) (5) Related substances—The procedure should be per- [22204-24-6] formed under protection from direct sunlight in light- resistant vessels. Dissolve 0.10 g of Pyrantel Pamoate in 10 Pyrantel Pamoate, when dried, contains not less mL of N,N-dimethylformamide, and use this solution as the than 98.0z of C11H14N2S.C23H16O6. sample solution. Pipet 1 mL of the sample solution, add N,N-dimethylformamide to make exactly 100 mL, and use Description Pyrantel Pamoate occurs as a light yellow to this solution as the standard solution. Perform the test with yellow, crystalline powder. It is odorless and tasteless. these solutions as directed under Thin-layer Chromatogra- It is sparingly soluble in N,N-dimethylformamide, very phy <2.03>. Spot 5 mL each of the sample solution and stand- slightly soluble in methanol and in ethanol (95), and practi- ard solution on a plate of silica gel with fluorescent indicator cally insoluble in water, in ethyl acetate and in diethyl ether. for thin-layer chromatography. Develop the plate with a Melting point: 256 – 2649C (with decomposition). mixture of ethyl acetate, water and acetic acid (100) (3:1:1) Identification (1) To 0.05 g of Pyrantel Pamoate add 10 to a distance of about 10 cm, and air-dry the plate. Examine mL of methanol and 1 mL of a mixture of hydrochloric acid under ultraviolet light (main wavelength: 254 nm): the spots and methanol (1:1), and shake vigorously: a yellow precipi- other than the spot of pyrantel and the spot of pamoic acid tate is produced. Filter the solution, and use the filtrate as from the sample solution are not more intense than the spot the sample solution. Use the precipitate for the test (2). To of pyrantel (Rf value: about 0.3) from the standard solution. 0.5 mL of the sample solution add 1 mL of a solution of 2,3- Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, indolinedione in sulfuric acid (1 in 1000): a red color devel- 2 hours). ops. (2) Collect the precipitate obtained in the test (1), wash Residue on ignition <2.44> Not more than 0.3z (1 g). with methanol, and dry at 1059C for 1 hour. To 0.01 g of the Assay Weigh accurately about 0.5 g of Pyrantel Pamoate, dried precipitate add 10 mL of methanol, shake well, and previously dried, add 25 mL of chloroform and 25 mL of so- filter. To 5 mL of the filtrate add 1 drop of iron (III) chlo- dium hydroxide TS, shake for 15 minutes, and extract. Ex- ride TS: a green color develops. tract further with two 25-mL portions of chloroform. Filter (3) Dissolve 0.1 g of Pyrantel Pamoate in 50 mL of N,N- each extract through 5 g of anhydrous sodium sulfate on a dimethylformamide, and add methanol to make 200 mL. To pledget of absorbent cotton. Combine the chloroform ex- 2 mL of the solution add a solution of hydrochloric acid in tracts, add 30 mL of acetic acid (100), and titrate <2.50> with methanol (9 in 1000) to make 100 mL. Determine the absorp- 0.1 mol/L perchloric acid VS (indicator: 2 drops of crystal 1322 Pyrazinamide / Official Monographs JP XVI violet TS). Perform a blank determination, and make any Residue on ignition <2.44> Not more than 0.1z (1 g). necessary correction. Assay Weigh accurately about 0.1 g of Pyrazinamide, pre- Each mL of 0.1 mol/L perchloric acid VS viously dried, dissolve in 50 mL of acetic anhydride, and

＝ 59.47 mg of C11H14N2S.C23H16O6 titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the Containers and storage Containers—Tight containers. same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS Pyrazinamide ＝ 12.31 mg of C5H5N3O Containers and storage Containers—Well-closed contain- ピラジナミド ers.

Pyridostigmine Bromide

ピリドスチグミン臭化物

C5H5N3O: 123.11 Pyrazine-2-carboxamide [98-96-4]

Pyrazinamide, when dried, contains not less than 99.0z and not more than 101.0z of C5H5N3O.

Description Pyrazinamide occurs as white crystals or crys- C9H13BrN2O2: 261.12 talline powder. 3-Dimethylcarbamoyloxy-1-methylpyridinium bromide It is sparingly soluble in water and in methanol, and [101-26-8] slightly soluble in ethanol (99.5) and in acetic anhydride. Pyridostigmine Bromide, when dried, contains not Identification (1) Determine the absorption spectrum of a less than 98.5z of C H BrN O . solution of Pyrazinamide in 0.1 mol/L hydrochloric acid TS 9 13 2 2 (1 in 100,000) as directed under Ultraviolet-visible Spectro- Description Pyridostigmine Bromide occurs as a white, photometry <2.24>, and compare the spectrum with the Ref- crystalline powder. It is odorless or has a slightly characteris- erence Spectrum: both spectra exhibit similar intensities of tic odor. absorption at the same wavelengths. It is very soluble in water, freely soluble in ethanol (95) (2) Determine the infrared absorption spectrum of and in acetic acid (100), and practically insoluble in diethyl Pyrazinamide, previously dried, as directed in the potassium ether. bromide disk method under Infrared Spectrophotometry The pH of a solution of Pyridostigmine Bromide (1 in 10) <2.25>, and compare the spectrum with the Reference Spec- is between 4.0 and 6.0. trum: both spectra exhibit similar intensities of absorption at It is deliquescent. the same wave numbers. Identification (1) Dissolve 0.02 g of Pyridostigmine Melting point <2.60> 188–1939C Bromide in 10 mL of water, add 5 mL of Reinecke salt TS: a light red precipitate is produced. Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of (2) To 0.1 g of Pyridostigmine Bromide add 0.6 mL of Pyrazinamide according to Method 2, and perform the test. sodium hydroxide TS: the unpleasant odor of dimethylamine Prepare the control solution with 2.0 mL of Standard Lead is perceptible. Solution (not more than 20 ppm). (3) Determine the absorption spectrum of a solution of (2) Related substances—Dissolve 0.10 g of Pyrazinamide Pyridostigmine Bromide in 0.1 mol/L hydrochloric acid TS in 10 mL of methanol, and use this solution as the sample so- (1 in 30,000) as directed under Ultraviolet-visible Spectro- lution. Pipet 1 mL of the sample solution, add methanol to photometry <2.24>, and compare the spectrum with the Ref- make exactly 200 mL, and use this solution as the standard erence Spectrum: both spectra exhibit similar intensities of solution. Perform the test with these solutions as directed absorption at the same wavelengths. under Thin-layer Chromatography <2.03>. Spot 20 mLeach (4) A solution of Pyridostigmine Bromide (1 in 50) re- of the sample solution and standard solution on a plate of sponds to the Qualitative Tests <1.09> for Bromide. silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water Melting point <2.60> 153 – 1579C and acetic acid (100) (3:1:1) to a distance of about 10 cm, Purity (1) Clarity and color of solution—Dissolve 1.0 g and air-dry the plate. Examine under ultraviolet light (main of Pyridostigmine Bromide in 10 mL of water: the solution is wavelength: 254 nm): the spot other than the principal spot clear and colorless. obtained from the sample solution is not more intense than (2) Heavy metals <1.07>—Proceedwith1.0gof the spot from the standard solution. Pyridostigmine Bromide according to Method 1, and per- Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- form the test. Prepare the control solution with 2.0 mL of um, silica gel, 4 hours). Standard Lead Solution (not more than 20 ppm). (3) Arsenic <1.11>—Prepare the test solution with 1.0 g JP XVI Official Monographs / Pyridoxine Hydrochloride 1323 of Pyridostigmine Bromide according to Method 1, and perchloric acid TS (1 in 100,000) as directed under Ultraviolet- form the test (not more than 2 ppm). visible Spectrophotometry <2.24>, and compare the spectrum (4) Related substances—Dissolve 0.10 g of Pyridostig- with the Reference Spectrum or the spectrum of a solution of mine Bromide in 10 mL of ethanol (95), and use this solution Pyridoxine Hydrochloride RS prepared in the same manner as the sample solution. Pipet 2 mL of the sample solution, as the sample solution: both spectra exhibit similar inten- and add ethanol (95) to make exactly 10 mL. Pipet 1 mL of sities of absorption at the same wavelengths. this solution, add ethanol (95) to make exactly 25 mL, and (2) Determine the infrared absorption spectrum of use this solution as the standard solution. Perform the test Pyridoxine Hydrochloride as directed in the potassium chlo- with these solutions as directed under Thin-layer Chroma- ride disk method under Infrared Spectrophotometry <2.25>, tography <2.03>. Spot 10 mL each of the sample solution and and compare the spectrum with the Reference Spectrum or standard solution on a plate of silica gel with fluorescent in- the spectrum of Pyridoxine Hydrochloride RS: both spectra dicator for thin-layer chromatography. Develop the plate exhibit similar intensities of absorption at the same wave with a mixture of methanol, chloroform and ammonium numbers. chloride TS (5:4:1) to a distance of about 12 cm, and air-dry (3) A solution of Pyridoxine Hydrochloride (1 in 10) the plate. Examine under ultraviolet light (main wavelength: responds to the Qualitative Tests <1.09> for chloride. 254 nm): the spots other than the principal spot from the pH <2.54> The pH of a solution prepared by dissolving sample solution are not more intense than the spot from the 1.0 g of Pyridoxine Hydrochloride in 50 mL of water is be- standard solution in color. tween 2.5 and 3.5. Loss on drying <2.41> Not more than 2.0z (1 g, in vacu- Purity (1) Clarity and color of solution—Dissolve 1.0 g um, phosphorus (V) oxide, 1009C, 5 hours). of Pyridoxine hydrochloride in 20 mL of water: the solution Residue on ignition <2.44> Not more than 0.1z (1 g). is clear and colorless. (2) Heavy metals <1.07>—Proceedwith1.0gof Assay Weigh accurately about 0.3 g of Pyridostigmine Pyridoxine Hydrochloride according to Method 1, and per- Bromide, previously dried, dissolve in 10 mL of acetic acid form the test. Prepare the control solution with 3.0 mL of (100), add 40 mL of acetic anhydride, and titrate <2.50> with Standard Lead Solution (not more than 30 ppm). 0.1 mol/L perchloric acid VS (potentiometric titration). Per- (3) Related substances—Dissolve 1.0 g of Pyridoxine form a blank determination, and make any necessary correc- Hydrochloride in 10 mL of water, and use this solution as tion. the sample solution. Pipet 2.5 mL of the sample solution, Each mL of 0.1 mol/L perchloric acid VS and add water to make exactly 100 mL. Pipet 1 mL of this

＝ 26.11 mg of C9H13BrN2O2 solution, add water to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these Containers and storage Containers—Hermetic containers. solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra- Pyridoxine Hydrochloride phy, and air-dry the plate. Develop the plate with a mixture of acetone, tetrahydrofuran, hexane and ammonia solution Vitamin B 6 (28) (65:13:13:9) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of sodium carbonate in ピリドキシン塩酸塩 diluted ethanol (3 in 10) (1 in 20) on the plate. After air-dry ing, spray evenly a solution of 2,6-dibromo-N-chloro-1,4- benzoquinone monoimine in ethanol (99.5) (1 in 1000) on the plate, and air-dry: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. C H NO .HCl: 205.64 8 11 3 Loss on drying <2.41> Not more than 0.30z (1 g, in vacu- 4,5-Bis(hydroxymethyl)-2-methylpyridin-3-ol um, silica gel, 4 hours). monohydrochloride [58-56-0] Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.2 g of Pyridoxine Hydro- Pyridoxine Hydrochloride, when dried, contains chloride, previously dried, add 5 mL of acetic acid (100) and not less than 98.0z and not more than 101.0z of 5 mL of acetic anhydride, dissolve by gentle boiling, cool, C H NO .HCl. 8 11 3 add 30 mL of acetic anhydride, and titrate <2.50> with 0.1 Description Pyridoxine Hydrochloride occurs as a white to mol/L perchloric acid VS (potentiometric titration). Per- pale yellow, crystalline powder. form a blank determination, and make any necessary correc- It is freely soluble in water, slightly soluble in ethanol tion. (99.5), and practically insoluble in acetic anhydride and in Each mL of 0.1 mol/L perchloric acid VS acetic acid (100). ＝ 20.56 mg of C H NO .HCl It is gradually affected by light. 8 11 3 Melting point: about 2069C (with decomposition). Containers and storage Containers—Tight containers. Storage—Light-resistant. Identification (1) Determine the absorption spectrum of a solution of Pyridoxine Hydrochloride in 0.1 mol/L hydro- 1324 Pyridoxine Hydrochloride Injection / Official Monographs JP XVI

lution as the sample solution. Separately, weigh accurately Pyridoxine Hydrochloride Injection about 0.1 g of Pyridoxine Hydrochloride RS, previously dried in a desiccator (in vacuum, silica gel) for 4 hours, and Vitamin B6 Injection dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 200 mL, and use this so- ピリドキシン塩酸塩注射液 lution as the standard solution. Pipet 1 mL each of the sample solution and standard solution, add 2.0 mL of barbital buffer solution, 9.0 mL of 2-propanol and 2.0 mL of a Pyridoxine Hydrochloride Injection is an aqueous freshly prepared solution of 2,6-dibromo-N-chloro-1,4-ben- solution for injection. zoquinone monoimine in ethanol (95) (1 in 4000), shake well, It contains not less than 95.0z and not more than add 2-propanol to make exactly 25 mL, and allow to stand 105.0z of the labeled amount of pyridoxine hydro- for 90 minutes. Determine the absorbances, A and A ,of chloride (C H NO .HCl: 205.64). T S 8 11 3 the subsequent sample solution and subsequent standard so- Method of preparation Prepare as directed under Injec- lution, respectively, at 650 nm as directed under Ultraviolet- tions, with Pyridoxine Hydrochloride. visible Spectrophotometry <2.24>, using a solution, prepared in the same manner with 1 mL of water, as the blank. Description Pyridoxine Hydrochloride Injection is a colorless or pale yellow, clear liquid. Amount (mg) of pyridoxine hydrochloride

It is gradually affected by light. (C8N11NO3.HCl) pH:3.0–6.0 ＝ MS × AT/AS × 1/5

Identification (1) To a volume of Pyridoxine Hydrochlo- MS: Amount (mg) of Pyridoxine Hydrochloride RS ride Injection, equivalent to 0.05 g of Pyridoxine Hydrochlo- Containers and storage Containers—Hermetic containers, ride according to the labeled amount, add 0.1 mol/L hydro- and colored containers may be used. chloricacidTStomake100mL.To2mLofthissolution Storage—Light-resistant. add 0.1 mol/L hydrochloric acid TS to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible spectrophotometry <2.24>:it exhibits a maximum between 288 nm and 292 nm. Pyroxylin (2) To a volume of Pyridoxine Hydrochloride Injection, ピロキシリン equivalent to 0.01 g of Pyridoxine Hydrochloride according to the labeled amount, add water to make 10 mL, and use this solution as the sample solution. Separately, dissolve Pyroxylin is a nitric acid ester of cellulose. It is usu- 0.01 g of Pyridoxine Hydrochloride RS in 10 mL of water, ally moistened with 2-propanol or some other solvent. and use this solution as the standard solution. Perform the Description Pyroxylin occurs as a white cotton-like sub- test with these solutions as directed under Thin-layer Chro- stance or white flakes. matography <2.03>. Spot 2 mL each of the sample solution It is freely soluble in acetone, and very slightly soluble in and standard solution on a plate of silica gel for thin-layer diethyl ether. chromatography, and air-dry the plate. Develop the plate Upon heating or exposure to light, it is decomposed with with a mixture of acetone, tetrahydrofuran, hexane and am- the evolution of nitrous acid vapors. monia solution (28) (65:13:13:9) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of sodium Identification Ignite Pyroxylin: it burns very rapidly with a carbonate in diluted ethanol (3 in 10) (1 in 20) on the plate. luminous flame. After air-drying, spray evenly a solution of 2,6-dibromo-N- Purity (1) Clarity of solution—Dissolve 1.0 g of Pyroxy- chloro-1,4-benzoquinone monoimine in ethanol (99.5) (1 in lin, previously dried at 809C for 2 hours, in 25 mL of a mix- 1000) on the plate: the spots obtained from the sample solu- ture of diethyl ether and ethanol (95) (3:1): the solution is tion and the standard solution are blue in color and have the clear. same Rf value. (2) Acidity—Shake 1.0 g of Pyroxylin, previously dried Bacterial endotoxins <4.01> Less than 3.0 EU/mg. at 809C for 2 hours, with 20 mL of water for 10 minutes: the filtrate is neutral. Extractable volume <6.05> It meets the requirement. (3) Water-soluble substances—Evaporate 10 mL of the Foreign insoluble matter <6.06> Perform the test according filtrate obtained in (2) on a water bath to dryness, and dry at to Method 1: it meets the requirement. 1059C for 1 hour: the mass of the residue is not more than 1.5 mg. Insoluble particulate matter <6.07> It meets the require- (4) Residue on ignition—Weigh accurately about 2 g of ment. Pyroxylin, previously dried at 809C for 2 hours, and moisten Sterility <4.06> Perform the test according to the Mem- with 10 mL of a solution of castor oil in acetone (1 in 20) to brane filtration method: it meets the requirement. gelatinize the sample. Ignite the contents to carbonize the sample, heat strongly at about 5009C for 2 hours, and allow Assay Measure exactly a volume of Pyridoxine Hydrochlo- to cool in a desicator (silica gel): the amount of the residue is ride Injection, equivalent to about 20 mg of pyridoxine hy- not more than 0.30z. drochloride (C8H11NO3.HCl), dilute with water, if necessary, and add water to make exactly 100 mL. Pipet 25 mL of this Containers and storage Containers—Tight containers. solution, add water to make exactly 200 mL, and use this so- Storage—Light-resistant, packed loosely, remote from JP XVI Official Monographs / Quinapril Hydrochloride 1325 fire, and preferably in a cold place. Assay Conduct this procedure using light-resistant vessels. Weigh accurately an amount of Pyrrolnitrin and Pyrrolnitrin RS, equivalent to about 50 mg (potency) each, and dissolve Pyrrolnitrin separetely in diluted acetonitrile (3 in 5) to make exactly 50 mL. Pipet 10 mL each of these solutions, add exactly 10 mL ピロールニトリン of the internal standard solution, add diluted acetonitrile (3 in 5) to make 100 mL, and use these solutions as the sample solution and standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-

lowing conditions, and calculate the ratios, QT and QS,of the peak area of pyrrolnitrin to that of the internal standard.

Amount [mg (potency)] of C10H6Cl2N2O2 C10H6Cl2N2O2:257.07 ＝ MS × QT/QS × 1000 3-Chloro-4-(3-chloro-2-nitrophenyl)pyrrole M : Amount [mg (potency)] of Pyrrolnitrin RS [1018-71-9] S Internal standard solution—A solution of benzyl benzoate in Pyrrolnitrin contains not less than 970 mg (potency) diluted acetonitrile (3 in 5) (3 in 500). and not more than 1020 mg (potency) per mg, calcu- Operating conditions— lated on the dried basis. The potency of Pyrrolnitrin Detector: An ultraviolet absorption photometer (wave- is expressed as mass (potency) of pyrrolnitrin length: 254 nm). (C10H6Cl2N2O2). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octylsilanized silica gel for Description Pyrrolnitrin occurs as yellow to yellow-brown, liquid chromatography (5 mm in particle diameter). crystals or crystalline powder. Column temperature: A constant temperature of about It is freely soluble in methanol and in ethanol (95), and 259C. practically insoluble in water. Mobile phase: A mixture of water and acetonitrile (11:9). Identification (1) Determine the absorption spectrum of a Flow rate: Adjust the flow rate so that the retention time solution of Pyrrolnitrin in ethanol (95) (1 in 100,000) as di- of pyrrolnitrin is about 9 minutes. rected under Ultraviolet-visible Spectrophotometry <2.24>, System suitability— and compare the spectrum with the Reference Spectrum or System performance: When the procedure is run with 5 mL the spectrum of a solution of Pyrrolnitrin RS prepared in the of the standard solution under the above operating condi- same manner as the sample solution: both spectra exhibit tions, pyrrolnitrin and the internal standard are eluted in this similar intensities of absorption at the same wavelengths. order with the resolution between these peaks being not less (2) Determine the infrared absorption spectrum of Pyr- than 3. rolnitrin as directed in the potassium bromide disk method System repeatability: When the test is repeated 6 times under Infrared Spectrophotometry <2.25>, and compare the with 5 mL of the standard solution under the above operating spectrum with the Reference Spectrum or the spectrum of conditions, the relative standard deviation of the ratios of Pyrrolnitrin RS: both spectra exhibit similar intensities of the peak area of pyrrolnitrin to that of the internal standard absorption at the same wave numbers. is not more than 1.0z. Melting point <2.60> 124–1289C Containers and storage Containers—Tight containers. Storage—Light-resistant. Purity Related substances—Dissolve 0.10 g of Pyrrolnitrin in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add methanol to make exactly 100 mL. Pipet 3 mL of this solution, add Quinapril Hydrochloride methanol to make exactly 10 mL, and use this solution as the キナプリル塩酸塩 standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of xylene, ethyl acetate and formic acid (18:2:1) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly diluted sulfuric acid (1 in 3) on the plate, and heat at 1009C for 30 minutes: the spot C H N O .HCl: 474.98 other than the principal spot obtained from the sample solu- 25 30 2 5 (3S)-2-((2S)-2-{[(1S)-1-Ethoxycarbonyl- tion is not more intense than the spot from the standard so- 3-phenylpropyl]amino}propanoyl)-1,2,3,4-tetrahydroisoquinoline- lution. 3-carboxylic acid monohydrochloride Loss on drying <2.41> Not more than 0.5z (1 g, reduced [82586-55-8] pressure not exceeding 0.67 kPa, 609C, 3 hours). Quinapril Hydrochloride contains not less than Residue on ignition <2.44> Not more than 0.1z (1 g). 99.0z and not more than 101.0z of C25H30N2O5.