1050 Hydrate / O‹cial Monographs JP XV

Weigh accurately an amount of Pyrrolnitrin and Pyrrolnitrin less than 98.5z of quinidine sulfate [(C20H24N2O2)2.H2 Reference Standard, equivalent to about 50 mg (potency), SO4:746.91]. dissolve in diluted acetonitrile (3 in 5) to make exactly 50 mL. Description Quinidine Sulfate Hydrate occurs as white Pipet 10 mL each of these solutions, add exactly 10 mL of the crystals. It is odorless, and has a very bitter taste. internal standard solution, add diluted acetonitrile (3 in 5) to It is freely soluble in (95) and in boiling water, make 100 mL, and use these solutions as the sample solution sparingly soluble in water, and practically insoluble in diethyl and standard solution. Perform the test with 5 mLeachofthe ether. Quinidine Sulfate Hydrate, previously dried, is freely sample solution and standard solution as directed under Liq- soluble in . uid Chromatography <2.01> according to the following con- It darkens gradually by light. ditions, and determine the ratios, Q and Q , of the peak T S Optical rotation [a]20: ＋275 – ＋2879(after drying, 0.5 g, area of pyrrolnitrin to that of the internal standard. D 0.1 molWL hydrochloric acid VS, 25 mL, 100 mm). Amount [ mg (potency)] of C H Cl N O 10 6 2 2 2 Identiˆcation (1) Dissolve 0.01 g of Quinidine Sulfate Hy- ＝W ×(Q /Q )×1000 S T S drate in 10 mL of water and 2 to 3 drops of dilute sulfuric

WS: Amount [mg (potency)] of Pyrrolnitrin Reference acid: a blue ‰uorescence is produced. Standard (2) To 5 mL of an aqueous solution of Quinidine Sulfate Hydrate (1 in 1000) add 1 to 2 drops of bromine TS, then add Internal standard solution—A solution of benzyl benzoate in 1 mL of ammonia TS: a green color develops. diluted acetonitrile (3 in 5) (3 in 500). (3) To 5 mL of an aqueous solution of Quinidine Sulfate Operating conditions— Hydrate(1in100)add1mLofsilvernitrateTS,stirwithag- Detector: An ultraviolet absorption photometer lass rod, and allow to stand for a short interval: a white (wavelength: 254 nm). precipitate is produced, and it dissolves on addition of nitric Column: A stainless steel column 4 mm in inside diameter acid. and 15 cm in length, packed with octylsilanized silica gel for (4) Dissolve 0.4 g of Quinidine Sulfate Hydrate in 20 mL liquid chromatography (5 mm in particle diameter). of water and 1 mL of dilute hydrochloric acid: the solution Column temperature: A constant temperature of about responds to the Qualitative Tests <1.09> for sulfate. 259C. Mobile phase: A mixture of water and acetonitrile (11:9). pH <2.54> Dissolve 1.0 g of Quinidine Sulfate Hydrate in Flow rate: Adjust the ‰ow rate so that the retention time of 100 mL of freshly boiled and cooled water: the pH of this so- pyrrolnitrin is about 9 minutes. lution is between 6.0 and 7.0. System suitability— Purity (1) Chloroform-ethanol-insoluble substances— System performance: When the procedure is run with 5 mL Warm 2.0 g of Quinidine Sulfate Hydrate with 15 mL of a of the standard solution under the above operating condi- mixture of chloroform and ethanol (99.5) (2:1) at about 509C tions, pyrrolnitrin and the internal standard are eluted in this for 10 minutes. After cooling, ˆlter through a tared glass order with the resolution between these peaks being not less ˆlter (G4) by gentle suction. Wash the residue with ˆve than 3. 10-mL portions of a mixture of chloroform and ethanol System repeatability: When the test is repeated 6 times with (99.5) (2:1), and dry at 1059C for 1 hour: the mass of the 5 mL of the standard solution under the above operating con- residue is not more than 2.0 mg. ditions, the relative standard deviation of the ratios of the (2) Related substances—Dissolve 20 mg of Quinidine peak area of pyrrolnitrin to that of the internal standard is SulfateHydrateinthemobilephasetomakeexactly100mL, not more than 1.0z. and use this solution as the sample solution. Separately, dis- Containers and storage Containers—Tight containers. solve 25 mg of cinchonine in the mobile phase to make exact- Storage—Light-resistant. ly 100 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 50 mLeachofthesam- Quinidine Sulfate Hydrate ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi- キニジン硫酸塩水和物 tions. Determine each peak area of the sample solution by the automatic integration method, and calculate their amount by the area percentage method: the amount of dihydroquinidine sulfate is not more than 15.0z, and those of quinine sulfate and dihydroquinine sulfate are not more than 1.0z.The total area of the peaks other than the principal peak and the above peaks is not larger than the peak area of cinchonine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 235 nm). (C20H24N2O2)2.H2SO4.2H2O: 782.94 Column: A column about 4 mm in inside diameter and (9S)-6?-Methoxycinchonan-9-ol hemisulfate about 25 cm in length, packed with octadecylsilanized silica monohydrate [6591-63-5] gel (10 mm in particle diameter). Temperature: Room temperature. Quinidine Sulfate Hydrate, when dried, contains not Mobile phase: A mixture of water, acetonitrile, methane- JP XV O‹cial Monographs / Quinine Ethyl Carbonate 1051 sulfonic acid TS and a solution of diethylamine (1 in 10) It is very soluble in , freely soluble in ethanol (95) (43:5:1:1). and in ethanol (99.5), soluble in , and practically Flow rate: Adjust the ‰ow rate so that the retention time of insoluble in water. quinidine is about 10 minutes. It dissolves in dilute hydrochloric acid. Selection of column: Dissolve 0.01 g each of Quinidine Sul- Identiˆcation (1) Determine the absorption spectrum of a fate Hydrate and quinine sulfate in 5 mL of methanol, and solution of Quinine Ethyl Carbonate in methanol (1 in add the mobile phase to make 50 mL. Proceed with 50 mLof 20,000) as directed under Ultraviolet-visible Spectrophoto- this solution under the above operating conditions, and cal- metry <2.24>, and compare the spectrum with the Reference culate the resolution. Use a column giving elution of quini- Spectrum: both spectra exhibit similar intensities of absorp- dine, quinine, dihydroquinidine and dihydroquinine in this tion at the same wavelengths. order with a resolution between quinidine and quinine and (2) Determine the infrared absorption spectrum of Qui- that between quinine and dihydroquinidine being not less nine Ethyl Carbonate as directed in the potassium than 1.2, respectively. disk method under Infrared Spectrophotometry <2.25>,and Detection sensitivity: Adjust the detection sensitivity so compare the spectrum with the Reference Spectrum: both that the peak height of cinchonine obtained from 50 mLof spectra exhibit similar intensities of absorption at the same the standard solution is between 5 mm and 10 mm. wave numbers. Time span of measurement: About twice as long as the 20 retention time of quinidine beginning after the solvent peak. Optical rotation <2.49> [a]D : －42.2 – －44.09(0.5 g, calcu- (3) Readily carbonizable substances <1.15>—Take 0.20 g lated on the dehydrated basis, methanol, 50 mL, 100 mm). of Quinidine Sulfate Hydrate and perform the test: the solu- Melting point <2.60> 91–959C tion has no more color than Matching ‰uid M. Purity (1) Chloride <1.03>—Dissolve 0.30 g of Quinine Loss on drying <2.41> Not more than 5.0z (1 g, 130 9C, Ethyl Carbonate in 10 mL of dilute nitric acid and 20 mL of 3 hours). water. To 5 mL of the solution add 2 to 3 drops of silver ni- Residue on ignition <2.44> Not more than 0.1z (1 g). trate TS: no color develops. (2) Sulfate <1.14>—Dissolve 1.0 g of Quinine Ethyl Car- Assay Weigh accurately about 0.5 g of Quinidine Sulfate bonate in 5 mL of dilute hydrochloric acid and water to make Hydrate, previously dried, dissolve in 20 mL of acetic acid 50 mL, and perform the test using this solution as the test so- (100), and add 80 mL of acetic anhydride, and titrate <2.50> lution. Prepare the control solution with 1.0 mL of 0.005 mol with 0.1 molWL perchloric acid VS until the color of the solu- WL sulfuric acid VS, 5 mL of dilute hydrochloric acid and tion changes from purple through blue to blue-green (indica- water to make 50 mL (not more than 0.048z). tor: 3 drops of TS). Perform a blank determina- (3) Heavy metals <1.07>—Proceed with 2.0 g of Quinine tion, and make any necessary correction. Ethyl Carbonate according to Method 2, and perform the Each mL of 0.1 molWL perchloric acid VS test. Prepare the control solution with 2.0 mL of Standard

＝24.90 mg of (C20H24N2O2)2.H2SO4 Lead Solution (not more than 10 ppm). (4) Related substances—Dissolve 20 mg of Quinine Ethyl Containers and storage Containers—Well-closed contain- Carbonate in the mobile phase to make exactly 100 mL, and ers. use this solution as the sample solution. Separately, dissolve Storage—Light-resistant. 25 mg of quinine sulfate in the mobile phase to make exactly 100 mL. Pipet 2 mL of this solution, add mobile phase to make exactly 100 mL, and use this solution as the standard Quinine Ethyl Carbonate solution. Perform the test with exactly 10 mLeachofthesam- ple solution and standard solution as directed under Liquid キニーネエチル炭酸エステル Chromatography <2.01> according to the following condi- tions. Determine each peak area of these solutions by the au- tomatic integration method, and calculate the amount of a main impurity in the sample solution which appears at about 1.2 times of the retention time of quinine ethyl carbonate by the area percentage method: it is not more than 10.0z.The total peak area other than the principal peak and above men- tioned peak from the sample solution is not larger than the peak area of Quinine from the standard solution.

C23H28N2O4:396.48 Operating conditions— Ethyl (8S,9R)-6?-methoxycinchonan-9-yl carbonate Detector: An ultraviolet absorption photometer [83-75-0] (wavelength: 235 nm). Column: A stainless steel column about 4 mm in inside di- Quinine Ethyl Carbonate contains not less than ameter and about 15 cm in length, packed with octadecyl- 98.5z of C23H28N2O4, calculated on the dehydrated silanized silica gel for liquid chromatography (5 mminparti- basis. cle diameter). Column temperature: A constant temperature of about Description Quinine Ethyl Carbonate occurs as white crys- 409C. tals. It is odorless, and tasteless at ˆrst but slowly develops a Mobile phase: Dissolve 1.2 g of sodium 1-octanesulfonate bitter taste. in 1000 mL of a mixture of water and methanol (1:1), and ad- 1052 Quinine Hydrochloride Hydrate / O‹cial Monographs JP XV just to pH 3.5 with diluted phosphoric acid (1 in 20). tion add 100 mL of water and 1 drop of dilute sulfuric acid: a Flow rate: Adjust the ‰ow rate so that the retention time of blue ‰uorescence is produced. the peak of quinine ethyl carbonate is about 20 minutes. (2) To 5 mL of a solution of Quinine Hydrochloride Hy- Selection of column: Dissolve 5 mg each of Quinine Ethyl drate (1 in 1000) add 1 to 2 drops of bromine TS and 1 mL of Carbonate and quinine sulfate in the mobile phase to make ammonia TS: a green color develops. 50 mL. Proceed with 10 mL of this solution under the above (3) To 5 mL of a solution of Quinine Hydrochloride Hy- operating conditions, and calculate the resolution. Use a drate (1 in 50) add 1 mL of dilute nitric acid and 1 mL of sil- column giving elution of quinine, dihydroquinine, quinine ver nitrate TS: a white precipitate is produced. Collect the ethyl carbonate and the main impurity of quinine ethyl car- precipitate, and add an excess of ammonia TS: it dissolves. bonate in this order with the resolution between the peaks of Optical rotation <2.49> [a]20: －245 – －2559(after drying, quinine and dihydroquinine being not less than 2.7, and be- D 0.5 g, 0.1 molWL hydrochloric acid VS, 25 mL, 100 mm). tween the peaks of quinine and quinine ethyl carbonate being not less than 5. pH <2.54> Dissolve 1.0 g of Quinine Hydrochloride Hy- Detection sensitivity: Adjust the detection sensitivity so drate in 100 mL of freshly boiled and cooled water: the pH of that the peak height of quinine obtained from 10 mLofthe this solution is between 6.0 and 7.0. standard solution is between 5 mm and 10 mm. Purity (1) Sulfate <1.14>—Perform the test with 1.0 g of Time span of measurement: About 2 times as long as the Quinine Hydrochloride Hydrate. Prepare the control solu- retention time of quinine ethyl carbonate. tion with 1.0 mL of 0.005 molWL sulfuric acid VS (not more Water <2.48> Not more than 3.0z (0.5 g, direct titration). than 0.048z). (2) Barium—Dissolve 0.5 g of Quinine hydrochloride Residue on ignition <2.44> Not more than 0.1z (1 g). Hydrate in 10 mL of water by warming, and add 1 mL of di- Assay Weigh accurately about 0.3 g of Quinine Ethyl Car- lute sulfuric acid: no turbidity is produced. bonate, dissolve in 60 mL of acetic acid (100), add 2 mL of (3) Chloroform-ethanol-insoluble substances—Warm 2.0 acetic anhydride, and titrate <2.50> with 0.1 molWLperchloric g of Quinine Hydrochloride Hydrate with 15 mL of a mixture acid VS (potentiometric titration). Perform a blank determi- of chloroform and ethanol (99.5) (2:1) at 509Cfor10 nation, and make any necessary correction. minutes. After cooling, ˆlter through a tared glass ˆlter (G4) by gentle suction. Wash the residue with ˆve 10-mL portions Each mL of 0.1 molWL perchloric acid VS of a mixture of chloroform and ethanol (99.5) (2:1), dry at ＝ 19.82 mg of C H N O 23 28 2 4 1059C for 1 hour, and weigh: the mass of the residue so ob- Containers and storage Containers—Well-closed contain- tained is not more than 2.0 mg. ers. (4) Related substances—Dissolve 20 mg of Quinine Hydrochloride Hydrate in the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Quinine Hydrochloride Hydrate Separately, dissolve 25 mg of cinchonidine in the mobile phase to make exactly 100 mL. Pipet 2 mL of this solution, キニーネ塩酸塩水和物 add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex- actly 50 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 the sample solution by the automatic integration method, and calculate the amount of dihydroquinine hydrochloride by the area percentage method: it is not more than 10.0z.Thetotal area of the peaks other than the main peak and the above peaks is not larger than the peak area of cinchonidine from C H N O .HCl.2H O: 396.91 20 24 2 2 2 the standard solution. (8S,9R)-6?-Methoxycinchonan-9-ol monohydrochloride Operating conditions— dihydrate [6119-47-7] Detector: An ultraviolet absorption photometer (wavelength: 235 nm). Quinine Hydrochloride Hydrate, when dried, con- Column: A stainless steel column about 4 mm in inside di- tains not less than 98.5z of quinine hydrochloride ameter and about 25 cm in length, packed with octadecyl- (C H N O .HCl: 360.88). 20 24 2 2 silanized silica gel (10 mm in particle diameter). Description Quinine Hydrochloride Hydrate occurs as Column temperature: Room temperature. white crystals. It is odorless, and has a very bitter taste. Mobile phase: A mixture of water, acetonitrile, It is very soluble in ethanol (99.5), freely soluble in acetic methanesulfonic acid TS and a solution of diethylamine (1 in acid (100), in acetic anhydride and in ethanol (95), soluble in 10) (43:5:1:1). water, and practically insoluble in diethyl ether. Quinine Flow rate: Adjust the ‰ow rate so that the retention time of Hydrochloride Hydrate, previously dried, is freely soluble in quinine is about 10 minutes. chloroform. Selection of column: Dissolve 10 mg each of Quinine It gradually changes to brown by light. Hydrochloride and quinidine sulfate in 5 mL of methanol, and add the mobile phase to make 50 mL. Proceed with 50 Identiˆcation (1) A solution of Quinine Hydrochloride mL of this solution under the above operating conditions. Use Hydrate (1 in 50) shows no ‰uorescence. To 1 mL of the solu- JP XV O‹cial Monographs / Quinine Sulfate Hydrate 1053 a column giving elution of quinidine, quinine, dihydroquini- spectra exhibit similar intensities of absorption at the same dine and dihydroquinine in this order with the resolution be- wavelengths. tween quinidine and quinine, and that between quinine and (2) Determine the infrared absorption spectrum of Qui- dihydroquinidine being not less than 1.2, respectively. nine Sulfate Hydrate, previously dried, as directed in the Detection sensitivity: Adjust the detection sensitivity so disk method under Infrared Spec- that the peak height of cinchonidine from 50 mLofthestan- trophotometry <2.25>, and compare the spectrum with the dard solution is between 5 mm and 10 mm. Reference Spectrum: both spectra exhibit similar intensities Time span of measurement: About twice as long as the of absorption at the same wave numbers. retention time of quinine beginning after the solvent peak. (3) To 0.4 g of Quinine Sulfate Hydrate add 20 mL of (5) Readily carbonizable substances <1.15>—Perform the water and 1 mL of dilute hydrochloric acid: the solution test with 0.25 g of Quinine Hydrochloride Hydrate. The solu- responds to the Qualitative Tests <1.09> for sulfate. tion has no more color than Matching Fluid M. 20 Optical rotation <2.49> [a ]D : －235 – －2459(after drying, Loss on drying <2.41> Not more than 10.0z (1 g, 1059C, 0.5 g, 0.1 molWL hydrochloric acid VS, 25 mL, 100 mm). 5 hours). pH <2.54> Shake 2.0 g of Quinine Sulfate Hydrate in 20 mL Residue on ignition <2.44> Not more than 0.1z (1 g). of freshly boiled and cooled water, and ˆlter: the pH of this ˆltrate is between 5.5 and 7.0. Assay Weigh accurately about 0.4 g of Quinine Hydrochlo- ride Hydrate, previously dried, dissolve in 100 mL of a mix- Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of ture of acetic anhydride and acetic acid (100) (7:3) by warm- Quinine Sulfate Hydrate according to Method 2, and per- ing, cool, and titrate <2.50> with 0.1 molWL perchloric acid form the test. Prepare the control solution with 2.0 mL of VS (potentiometric titration). Perform a blank determina- Standard Lead Solution (not more than 10 ppm). tion, and make any necessary correction. (2) Chloroform-ethanol-insoluble substances—Warm 2.0 g of Quinine Sulfate Hydrate with 15 mL of a mixture of Each mL of 0.1 molWL perchloric acid VS chloroform and ethanol (99.5) (2:1) at 509C for 10 minutes. ＝ 18.04 mg of C H N O .HCl 20 24 2 2 After cooling, ˆlter through a tared glass ˆlter (G4) by gentle Containers and storage Containers—Well-closed contain- suction. Wash the residue with ˆve 10-mL portions of a mix- ers. ture of chloroform and ethanol (99.5) (2:1), dry at 1059Cfor Storage—Light-resistant. 1 hour, and weigh: the mass of the residue is not more than 2.0 mg. (3) Related substances—Dissolve 20 mg of Quinine Sul- Quinine Sulfate Hydrate fate Hydrate in the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Separately, dis- キニーネ硫酸塩水和物 solve 25 mg of cinchonidine in the mobile phase to make ex- actly 100 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 50 mLeach of the sample solution and standard solution as directed un- der Liquid Chromatography <2.01> according to the follow- ing conditions. Determine each peak area from the sample solution by the automatic integration method, and calculate the amount of dihydroquinine sulfate by the area percentage method: it is not more than 5z. The total area of the peaks other than the main peak and the above peak is not larger (C20H24N2O2)2.H2SO4.2H2O: 782.94 than the peak area of cinchonidine from the standard solu- (8S,9R)-6?-Methoxycinchonan-9-ol hemisulfate tion. monohydrate [6119-70-6] Operating conditions— Detector: An ultraviolet absorption photometer Quinine Sulfate Hydrate contains not less than (wavelength: 235 nm). 98.5z of quinine sulfate [(C20H24N2O2)2.H2SO4: Column: A column about 4 mm in inside diameter and 746.91], calculated on the dried basis. about 25 cm in length, packed with octadecylsilanized silica Description Quinine Sulfate Hydrate occurs as white crys- gel (10 mm in particle diameter). tals or crystalline powder. It is odorless, and has a very bitter Temperature: Room temperature taste. Mobile phase: A mixture of water, acetonitrile, methane It is freely soluble in acetic acid (100), slightly soluble in sulfonic acid TS and a solution of diethylamine (1 in 10) water, in ethanol (95), in ethanol (99.5) and in chloroform, (43:5:1:1). and practically insoluble in diethyl ether. Flow rate: Adjust the ‰ow rate so that the retention time of It gradually changes to brown by light. quinine is about 10 minutes. Selection of column: Dissolve 0.01 g each of Quinine Sul- Identiˆcation (1) Determine the absorption spectrum of a fate Hydrate and quinidine sulfate in 5 mL of methanol, and solution of Quinine Sulfate Hydrate (1 in 20,000) as directed add the mobile phase to make 50 mL. Proceed with 50 mLof under Ultraviolet-visible Spectrophotometry <2.24>,and this solution under the above operating conditions, and cal- compare the spectrum with the Reference Spectrum: both culate the resolution. Use a column giving elution of quini- 1054 Freeze-dried Inactivated Tissue Culture Rabies Vaccine / O‹cial Monographs JP XV dine, quinine, dihydroquinidine and dihydroquinine in this Description Ranitidine Hydrochloride occurs as a white to order with the resolution between quinidine and quinine and pale yellow, crystalline or ˆne granular powder. that between quinine and dihydroquinidine being not less It is very soluble in water, freely soluble in methanol, and than 1.2. slightly soluble in ethanol (99.5). Detection sensitivity: Adjust the detection sensitivity so It is hygroscopic. that the peak height of cinchonidine obtained from 50 mLof It is gradually colored by light. the standard solution is between 5 mm and 10 mm. Melting point: about 1409C (with decomposition). Time span of measurement: About twice as long as the retention time of quinine beginning after the solvent peak. Identiˆcation (1) Determine the absorption spectrum of a solution of Ranitidine Hydrochloride (1 in 100,000) as direct- Loss on drying <2.41> 3.0z –5.0z (1 g, 1059C, 3 hours). ed under Ultraviolet-visible Spectrophotometry <2.24>,and Residue on ignition <2.44> Not more than 0.1z (1 g). compare the spectrum with the Reference Spectrum or the spectrum of a solution of Ranitidine Hydrochloride Refer- Assay Weigh accurately about 0.5 g of Quinine Sulfate Hy- ence Standard prepared in the same manner as the sample so- drate, dissolve in 20 mL of acetic acid (100), add 80 mL of lution: both spectra exhibit similar intensities of absorption acetic anhydride, and titrate <2.50> with 0.1 molWL perchloric at the same wavelengths. acid VS until the color of the solution changes from purple (2) Determine the infrared absorption spectrum of through blue to blue-green (indicator: 2 drops of crystal vio- Ranitidine Hydrochloride as directed in the paste method un- let TS). Perform a blank determination, and make any neces- der Infrared Spectrophotometry <2.25>, and compare the sary correction. spectrum with the Reference Spectrum or the spectrum of Each mL of 0.1 molWL perchloric acid VS previously dried Ranitidine Hydrochloride Reference Stan-

＝ 24.90 mg of (C20H24N2O2)2.H2SO4 dard: both spectra exhibit similar intensities of absorption at thesamewavenumbers. Containers and storage Containers—Well-closed contain- (3) A solution of Ranitidine Hydrochloride (1 in 50) ers. responds to the Qualitative Tests <1.09> for chloride. Storage—Light-resistant. pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Ranitidine Hydrochloride in 100 mL of water is between Freeze-dried Inactivated Tissue 4.5 and 6.0. Culture Rabies Vaccine Purity (1) Clarity and color of solution—A solution of Ranitidine Hydrochloride (1 in 10) is clear and pale yellow to 乾燥組織培養不活化狂犬病ワクチン light yellow. (2) Heavy metals <1.07>—Proceed with 2.0 g of Raniti- dine Hydrochloride according to Method 2, and perform the Freeze-dried Inactivated Tissue Culture Rabies Vac- test. Prepare the control solution with 2.0 mL of Standard cine is a dried preparation containing inactivated rabies Lead Solution (not more than 10 ppm). virus. (3) Arsenic <1.11>—Prepare the test solution with 1.0 g It conforms to the requirements of Freeze-dried In- of Ranitidine Hydrochloride according to Method 4, and per- activated Tissue Culture Rabies Vaccine in the Mini- form the test (not more than 2 ppm). mum Requirements of Biologic Products. (4) Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 0.22 g Description Freeze-dried Inactivated Tissue Culture Rabies of Ranitidine Hydrochloride in methanol to make exactly 10 Vaccine becomes a colorless or light yellow-red clear liquid mL, and use this solution as the sample solution. Pipet 0.5 on addition of solvent. mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution (1). Pipet 6 mL, 4 mL, 2 mL and 1 mL of the standard solution (1), add Ranitidine Hydrochloride to each methanol to make exactly 10 mL, and use these solu- tions as the standard solution (2), the standard solution (3), ラニチジン塩酸塩 the standard solution (4) and the standard solution (5), re- spectively. Separately, dissolve 12.7 mg of ranitidinediamine in methanol to make exactly 10 mL, and use this solution as the standard solution (6). Perform the test with these solu- tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu- tions (1), (2), (3), (4) and (5) on a plate of silica gel for thin- C H N O S.HCl: 350.86 13 22 4 3 layer chromatography. Separately, spot 10 mLofthesample (1EZ)-N｛- 2-[(｛5-[(Dimethylamino)methyl]furan- solution on the plate, then spot 10 mL of the standard solu- 2-yl｝methyl)sulfanyl]ethyl｝-N?-methyl-2-nitroethene- tion (6) on the spotted position of the sample solution. Imme- 1,1-diamine monohydrochloride [66357-59-3 ] diately develop the plate with a mixture of ethyl acetate, 2- propanol, ammonia solution (28) and water (25:15:5:1) to a Ranitidine Hydrochloride, when dried, contains not distance of about 15 cm, and air-dry the plate. Allow the less than 97.5z and not more than 102.0z of C H N O S.HCl. plate to stand in iodine vapor until the spot from the standard 13 22 4 3 solution (5) appears: the spot obtained from the standard so- JP XV O‹cial Monographs / 1055 lution (6) is completely separated from the principal spot of Brassica campestris Linnáe subsp. napus Hooker ˆl. from the sample solution. The spot having Rf value of about et Anderson var. nippo-oleifera Makino (Cruciferae). 0.7 from the sample solution is not more intense than the spot Description Rape Seed Oil is a clear, pale yellow, slightly from the standard solution (1), the spots other than the prin- viscous oil. It is odorless or has a slight odor and a mild taste. cipal spot and the spot of Rf 0.7 from the sample solution are It is miscible with diethyl ether and with petroleum diethyl not more intense than the spot from the standard solution ether. It is slightly soluble in ethanol (95). (2), and the total amount of these related substances, calcu- Speciˆc gravity d25: 0.906 – 0.920 lated by comparison with the spots from the standard solu- 25 tions (1), (2), (3), (4) and (5), is not more than 1.0z. Acid value <1.13> Not more than 0.2. Loss on drying <2.41> Not more than 0.75z (1 g, in vacu- Saponiˆcation value <1.13> 169 – 195 um, 609C, 3 hours). Unsaponiˆable matters <1.13> Not more than 1.5z. Residue on ignition <2.44> Not more than 0.1z (1 g). Iodine value <1.13> 95–127 Assay Weigh accurately about 20 mg of Ranitidine Containers and storage Containers—Tight containers. Hydrochloride and Ranitidine Hydrochloride Reference Standard, previously dried, dissolve each in the mobile phase to make exactly 200 mL, and use these solutions as the sam- ple solution and the standard solution. Perform the test with Reserpine exactly 10 mL each of the sample solution and the standard レセルピン solution as directed under Liquid Chromatography <2.01> ac- cording to the following conditions, and determine the peak areas, AT and AS, of ranitidine.

Amount (mg) of C13H22N4O3S.HCl ＝WS×(AT/AS)

WS: Amount (mg) of Ranitidine Hydrochloride Reference Standard Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 322 nm). Column: A stainless steel column 4.6 mm in inside di- ameter and 20 cm in length, packed with octadecylsilanized C33H40N2O9:608.68 silica gel for liquid chromatography (10 mminparticledi- Methyl (3S,16S,17R,18R,20R)-11,17-dimethoxy-18- ameter). (3,4,5-trimethoxybenzoyloxy)yohimban-16-carboxylate Column temperature: A constant temperature of about [50-55-5 ] 259C. Mobile phase: A mixture of methanol and diluted 0.5 Reserpine, when dried, contains not less than 96.0z mol/L ammonium acetate TS (1 in 5) (17:3). of C33H40N2O9. Flow rate: Adjust the ‰ow rate so that the retention time of ranitidine is about 5 minutes. Description Reserpine occurs as white to pale yellow crys- System suitability— tals or crystalline powder. System performance: Dissolve 20 mg of Ranitidine It is freely soluble in acetic acid (100) and in chloroform, Hydrochloride and 5 mg of benzalphthalide in 200 mL of the slightly soluble in acetonitrile, very slightly soluble in ethanol mobile phase. When the procedure is run with 10 mLofthis (95), and practically insoluble in water and in diethyl ether. solution under the above operating conditions, ben- It is aŠected by light. zalphthalide and ranitidine are eluted in this order with the Identiˆcation (1) To 1 mg of Reserpine add 1 mL of vanil- resolution between these peaks being not less than 2.0. lin-hydrochloric acid TS, and warm: a vivid red-purple color System repeatability: When the test is repeated 6 times with develops. 10 mL of the standard solution under the above operating (2) Determine the absorption spectrum of a solution of conditions, the relative standard deviation of the peak area of Reserpine in acetonitrile (1 in 50,000) as directed under ranitidine is not more than 1.0z. Ultraviolet-visible Spectrophotometry <2.24>,andcompare Containers and storage Containers—Tight containers. the spectrum with the Reference Spectrum or the spectrum of Storage—Light-resistant. a solution of Reserpine Reference Standard prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Rape Seed Oil (3) Determine the infrared absorption spectrum of Reser- pine, previously dried, as directed in the potassium bromide Oleum Rapae disk method under Infrared Spectrophotometry <2.25>,and compare the spectrum with the Reference Spectrum or the ナタネ油 spectrum of previously dried Reserpine Reference Standard: both spectra exhibit similar intensities of absorption at the same wave numbers. Rape Seed Oil is the ˆxed oil obtained from the seed 1056 Reserpine / O‹cial Monographs JP XV

20 Optical rotation <2.49> [a]D : －114 – －1279(after drying, Internal standard solution—A solution of butyl parahydrox- 0.25 g, chloroform, 25 mL, 100 mm). ybenzoate in acetonitrile (1 in 50,000). Operating conditions— Purity Related substances—Conduct this procedure Detector: An ultraviolet absorption photometer without exposure to daylight, using light-resistant vessels. (wavelength: 268 nm). Dissolve 50 mg of Reserpine in 50 mL of acetonitrile, and use Column: A stainless steel column 4 mm in inside diameter this solution as the sample solution. Pipet 3 mL of the sample and 25 cm in length, packed with octadecylsilanized silica gel solution, add acetonitrile to make exactly 100 mL, and use for liquid chromatography (5 mminparticlediameter). this solution as the standard solution. Perform the test with Column temperature: A constant temperature of about exactly 10 mL each of the sample solution and standard solu- 409C. tion as directed under Liquid Chromatography <2.01> ac- Mobile phase: A mixture of 0.05 mol/L potassium di- cording to the following conditions. Determine each peak , pH 3.0 and acetonitrile (11:9). area from these solutions by the automatic integration Flow rate: Adjust the ‰ow rate so that the retention time of method: the total area of all peaks other than reserpine peak reserpine is about 10 minutes. from the sample solution is not larger than the peak area of System suitability— reserpine from the standard solution. System performance: When the procedure is run with 20 Operating conditions— mL of the standard solution under the above operating condi- Detector, column, and column temperature: Proceed as tions, reserpine and the internal standard are eluted in this directed in the operating conditions in the Assay. order with the resolution between these peaks being not less Mobile phase: A mixture of 0.05 mol/L potassium di- than 2.0. hydrogen phosphate, pH 3.0 and acetonitrile (13:7). System repeatability: When the test is repeated 6 times with Flow rate: Adjust the ‰ow rate so that the retention time of 20 mL of the standard solution under the above operating reserpine is about 20 minutes. conditions, the relative standard deviation of the ratios of the Time span of measurement: About twice as long as the peak area of reserpine to that of the internal standard is not retention time of reserpine. more than 2.0z. System suitability— Test for required detection: To exactly 2 mL of the stan- Containers and Storage Containers—Well-closed contain- dard solution add acetonitorile to make exactly 50 mL. Con- ers. ˆrm that the peak area of reserpine obtained from 10 mLof Storage—Light-resistant. this solution is equivalent to 3 to 5z of that of reserpine ob- tained from 10 mL of the standard solution. System performance: Dissolve 0.01 g of Reserpine and 4 Reserpine Injection mg of butyl parahydroxybenzoate in 100 mL of acetonitrile. To 5 mL of this solution add acetonitrile to make 50 mL. レセルピン注射液 When the procedure is run with 20 mL of this solution accord- ing to the operating conditions in the Assay, reserpine and butyl parahydroxybenzoate are eluted in this order with the Reserpine Injection is an aqueous solution for injec- resolution between these peaks being not less than 2.0. tion. It contains not less than 90z and not more than System repeatability: When the test is repeated 6 times with 110z of the labeled amount of reserpine (C33H40N2O9: 10 mL of the standard solution under the above operating 608.68). conditions, the relative standard deviation of the peak area of Method of preparation Prepare as directed under Injec- reserpine is not more than 2.0z. tions, with Reserpine. Loss on drying <2.41> Not more than 0.5z (0.2 g, in vacu- Description Reserpine Injection is a clear, colorless or pale um, 609C, 3 hours). yellow liquid. Residue on ignition <2.44> Not more than 0.2z (0.2 g). pH:2.5–4.0 Assay Conduct this procedure without exposure to day- Identiˆcation Measure a volume of Reserpine Injection, light, using light-resistant vessels. Weigh accurately about 10 equivalent to 1.5 mg of Reserpine according to the labeled mg each of Reserpine and Reserpine Reference Standard, amount, add 10 mL of diethyl ether, shake for 10 minutes, previously dried, and dissolve each in acetonitrile to make ex- and take the aqueous layer. If necessary, add 10 mL of actly 100 mL. Pipet 5 mL each of these solutions, add exactly diethyl ether to the aqueous layer, and shake for 10 minutes 10 mL of the internal standard solution, 5 mL of acetonitrile to repeat the process. To the aqueous layer add water to and water to make 50 mL, and use these solutions as the sam- make 50 mL, and determine the absorption spectrum of this ple solution and the standard solution, respectively. Perform solution as directed under Ultraviolet-visible Spectrophoto- the test with 20 mL each of the sample solution and standard metry <2.24>: it exhibits a maximum between 265 nm and 269 solution as directed under Liquid Chromatography <2.01> ac- nm. cording to the following conditions, and calculate the ratios, Extractable volume <6.05> It meets the requirement. Q T and QS, of the peak area of reserpine to that of the inter- nal standard. Assay Measure exactly a volume of Reserpine Injection, equivalent to about 4 mg of reserpine (C H N O ). Amount (mg) of C H N O ＝W ×(Q /Q ) 33 40 2 9 33 40 2 9 S T S Separately, weigh accurately about 4 mg of Reserpine Refer- WS: Amount (mg) of Reserpine Reference Standard ence Standard, previously dried in vacuum at 609Cfor3 JP XV O‹cial Monographs / Reserpine Tablets 1057 hours. Transfer them to separate separator, add 10 mL each vacuum for 3 hours, dissolve in acetonitrile to make exactly of water and 5 mL each of ammonia TS, and extract with one 100 mL. Pipet 5 mL of this solution, add exactly 10 mL of 20-mL portion of chloroform, then with three 10-mL por- the internal standard solution, 5 mL of acetonitrile and water tions of chloroform with shaking vigorously. Combine the to make 50 mL, and use this solution as the standard solu- chloroform extracts, wash with two 50-mL portions of dilut- tion. Proceed with the sample solution and standard solution ed hydrochloric acid (1 in 1000), and combine the washings. as directed in the Assay under Reserpine. Then wash the chloroform extract with two 50-mL portions Amount (mg) of reserpine (C H N O ) of a solution of sodium hydrogen carbonate (1 in 100), and 33 40 2 9 ＝W ×(Q /Q )×(1/20) combine the all washings. Extract the combined washing with S T S two 10-mL portions of chloroform, and combine the wash- WS: Amount (mg) of Reserpine Reference Standard ings with the former chloroform extract. Transfer the chlo- Internal standard solution—A solution of butyl parahydrox- roform solution to a 100-mL volumetric ‰ask through a ybenzoate in acetonitrile (1 in 50,000). pledget of absorbent cotton previously wetted with chlo- roform, wash with a small amount of chloroform, dilute with Containers and storage Containers—Well-closed contain- chloroform to make 100 mL, and use these solutions as the ers. sample solution and the standard solution, respectively. De- Storage—Light-resistant. termine the absorbances, AT and AS, of the sample solution and the standard solution, respectively, at 295 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Reserpine Tablets Amount (mg) of reserpine (C H N O ) 33 40 2 9 レセルピン錠 ＝WS×(AT/AS)

WS: Amount (mg) of Reserpine Reference Standard Containers and storage Containers—Hermetic containers, Reserpine Tablets contain not less than 90z and not and colored containers may be used. more than 110z of the labeled amount of reserpine Storage—Light-resistant. (C33H40N2O9:608.68). Method of preparation Prepare as directed under Tablets, with Reserpine. 0.1 Reserpine Powder Identiˆcation Take a portion of powdered Reserpine Reserpine Powder Tablets, equivalent to 0.4 mg of Reserpine according to the labeled amount, add 20 mL of acetonitrile, shake for 30 レセルピン散 0.1 minute, and centrifuge. Determine the absorption spectrum of the supernatant liquid as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 265 0.1z Reserpine Powder contains not less than nm and 269 nm, and between 294 nm and 298 nm. 0.09z and not more than 0.11z of reserpine Uniformity of dosage units <6.02> Perform the test accord- (C33H40N2O9: 608.68). ing to the following method: it meets the requirement of the Method of preparation Content uniformity test. Conduct this procedure without exposure to daylight, us- Reserpine 1 g ing light-resistant vessels. To one tablet of Reserpine Tablets Lactose Hydrate a su‹cient quantity add 2 mL of water, disintegrate by warming at 509Cfor15 To make 1000 g minutes while shaking. After cooling, add exactly 2 mL of the internal standard solution per 0.1 mg of reserpine Prepare as directed under Powders, with the above in- (C33H40N2O9) according to the labeled amount, add 2 mL of gredients. acetonitrile,warmat509C for 15 minutes while shaking, and Identiˆcation To 0.4 g of 0.1z Reserpine Powder add 20 after cooling, add water to make 10 mL. Centrifuge the solu- mL of acetonitrile, shake for 30 minutes, and centrifuge. De- tion, and use the supernatant liquid as the sample solution. termine the absorption spectrum of the supernatant liquid as Separately, weigh accurately about 10 mg of Reserpine Refer- directed under Ultraviolet-visible Spectrophotometry <2.24>: ence Standard, previously dried at 609C in vacuum for 3 it exhibits maxima between 265 nm and 269 nm, and between hours, dissolve in acetonitrile to make exactly 100 mL. Pipet 294 nm and 298 nm. 5 mL of this solution add exactly 10 mL of the internal stan- dard solution, 5 mL of acetonitrile and water to make 50 mL, Assay Conduct this procedure without exposure to day- and use this solution as the standard solution. Proceed with light, using light-resistant vessels. Weigh accurately a quanti- the sample solution and standard solution as directed in the ty of 0.1z Reserpine Powder, equivalent to about 0.5 mg of Assay under Reserpine. reserpine (C33H40N2O9), disperse in 12 mL of water, add ex- actly 10 mL of the internal standard solution and 10 mL of Amount (mg) of reserpine (C33H40N2O9) acetonitrile, and dissolve by warming at 509C for 15 minutes, ＝WS×(QT/QS)×(C/10) then add water to make 50 mL, and use this solution as the WS: Amount (mg) of Reserpine Reference Standard sample solution. Separately, weigh accurately about 10 mg of C: Labeled amount (mg) of reserpine in each tablet. Reserpine Reference Standard, previously dried at 609Cin Internal standard solution—A solution of butyl parahydrox- 1058 Retinol Acetate / O‹cial Monographs JP XV ybenzoate in acetonitrile (1 in 50,000). Containers and storage Containers—Well-closed contain- Dissolution <6.10> Perform the test according to the follow- ers. ing method: it meets the requirement. Storage—Light-resistant. Take 1 tablet of Reserpine Tablets, and perform the test at 100 revolutions per minute with 500 mL of a solution of poly- sorbate 80 (1 in 20,000) in diluted dilute acetic acid (1 in 200) Retinol Acetate as the test solution according to the Paddle method. Take 20 mL or more of the dissolved solution 30 minutes after start- Vitamin A Acetate ing the dissolution test, ˆlter through a ˆlter laminated with polyester ˆbers, discard the ˆrst 10 mL of the ˆltrate, and use レチノール酢酸エステル the subsequent ˆltrate as the sample solution. Separately, dry Reserpine Reference Standard at 609C in vacuum for 3 hours, weigh accurately an amount 100 times the labeled amount, dissolve in 1 mL of chloroform and 80 mL of ethanol (95), and add a solution of polysorbate 80 in diluted dilute acetic acid (1 in 200) (1 in 20,000) to make exactly 200 C22H32O2: 328.49 mL. Pipet 1 mL of this solution, add a solution of polysor- (2E,4E,6E,8E )-3,7-Dimethyl-9-(2,6,6-trimethylcyclohex- bate 80 in diluted dilute acetic acid (1 in 200) (1 in 20,000) to 1-en-1-yl)nona-2,4,6,8-tetraen-1-yl acetate [127-47-9] make exactly 250 mL, and use this solution as the standard solution. Pipet 5 mL each of the sample solution and stan- Retinol Acetate is synthetic retinol acetate or syn- dard solution, transfer to glass-stoppered brown test tubes T thetic retinol acetate diluted with ˆxed oil. and S, respectively, add exactly 5 mL each of ethanol (99.5), It contains not less than 2,500,000 Vitamin A Units shake well, add exactly 1 mL each of diluted vanadium (V) per gram. oxide (1 in 2), shake vigorously, and allow to stand for 30 A suitable antioxidant may be added. minutes. Perform the test with these solutions as directed un- Retinol Acetate contains not less than 95.0z and not der Fluorometry <2.22>, and determine the intensity of more than 105.0z of the labeled Units. ‰uorescence, FT and FS, at the wavelength of excitation at 400 nm and at the wavelength of ‰uorescence at 500 nm. Dissolu- Description Retinol Acetate occurs as pale yellow to yellow- tion rate of Reserpine Tablets after 30 minutes should be not red crystals or an ointment-like substance, and has a faint, less than 70z. characteristic odor, but has no rancid odor. It is freely soluble in petroleum ether, soluble in ethanol Dissolution rate (z) to the labeled (95), and practically insoluble in water. amount of reserpine (C33H40N2O9) It is decomposed by air and by light. ＝WS×(FT/FS)×(1/ C ) Identiˆcation Dissolve Retinol Acetate and Retinol Acetate WS: Amount (mg) of Reserpine Reference Standard. Reference Standard, equivalent to 15,000 Units, in 5 mL of C: Labeled amount (mg) of reserpine (C33H40N2O9)ineach petroleum ether, and use these solutions as the sample solu- tablet. tion and standard solution. Perform the test with these solu- Assay Conduct this procedure without exposure to day- tions as directed under Thin-layer Chromatography <2.03>. light, using light-resistant vessels. Weigh accurately and pow- Spot 5 mL each of the sample solution and standard solution der not less than 20 Reserpine Tablets. Weigh accurately a on a plate of silica gel for thin-layer chromatography. De- quantity of the powder, equivalent to about 0.5 mg of reser- velop with a mixture of cyclohexane and diethyl ether (12:1) to a distance of about 10 cm, and air-dry the plate. Spray pine (C33H40N2O9), add 3 mL of water, and warm at 509Cfor 15 minutes while shaking. After cooling, add exactly 10 mL evenly antimony (III) chloride TS: the principal spot ob- of the internal standard solution, 10 mL of acetonitrile and tained from the sample solution is the same in color tone and warm at 509C for 15 minutes while shaking. After cooling, Rf value with the blue spot from the standard solution. add water to make 50 mL, centrifuge, and use the super- Purity (1) Acid value <1.13>—Take exactly 5.0 g of natant liquid as the sample solution. Separately, weigh ac- Retinol Acetate, and perform the test: not more than 2.0. curately about 10 mg of Reserpine Reference Standard, (2) —Weigh accurately about 5 g of Retinol previously dried at 609C in vacuum for 3 hours, and dissolve Acetate, transfer in a 250-mL glass-stoppered conical ‰ask, in acetonitrile to make exactly 100 mL. Pipet 5 mL of this so- add 50 mL of a mixture of acetic acid (100) and isooctane lution, add exactly 10 mL of the internal standard solution, 5 (3:2), and gently mix to dissolve completely. Replace the air mL of acetonitrile and water to make 50 mL, and use this so- of the inside gradually with about 600 mL of Nitrogen, then lution as the standard solution. Proceed with the sample solu- add 0.1 mL of saturated TS under a current tion and standard solution as directed in Assay under Reser- of Nitrogen. Immediately stopper tightly, and mix with a pine. swirling motion for 1 minute. Add 30 mL of water, stopper

Amount (mg) of reserpine (C33H40NO9) tightly, and shake vigorously for 5 to 10 seconds. Titrate <2.50> this solution with 0.01 mol/L sodium VS ＝WS×(QT/QS)×(1/20) until the blue color of the solution disappears after addition WS: Amount (mg) of Reserpine Reference Standard of 0.5 mL of starch TS near the end point where the solution Internal standard solution—A solution of butyl parahydrox- is a pale yellow color. Calculate the amount of peroxide by ybenzoate in acetonitrile (1 in 50,000). the following formula: not more than 10 meq/kg. JP XV O‹cial Monographs / Ribo‰avin 1059

of Nitrogen. Immediately stopper tightly, and mix with a Amount (meq/kg) of peroxide＝(V/W)×10 swirling motion for 1 minute. Add 30 mL of water, stopper V: Volume (mL) of 0.01 mol/L VS tightly, and shake vigorously for 5 to 10 seconds. Titrate consumed <2.50> this solution with 0.01 mol/L sodium thiosulfate VS W: Amount (g) of the sample until the blue color of the solution disappears after addition of 0.5 mL of starch TS near the end point where the solution Assay Proceed as directed in Method 1-1 under Vitamin A is a pale yellow color. Calculate the amount of peroxide by Assay <2.55>. the following formula: not more than 10 meq/kg. Containers and storage Containers—Tight containers. Amount (meq/kg) of peroxide ＝(V/W)×10 Storage—Light-resistant, and almost well-ˆlled, or under Nitrogen atmosphere, and in a cold place. V: Volume (mL) of 0.01 mol/L sodium thiosulfate VS W: Amount (g) of the sample Assay Proceed as directed in Method 1-1 under the Vitamin Retinol Palmitate AAssay<2.55>. Vitamin A Palmitate Containers and storage Containers—Tight containers. Storage—Light-resistant, and almost well-ˆlled, or under レチノールパルミチン酸エステル Nitrogen atmosphere, and in a cold place.

Ribo‰avin

Vitamin B2 C H O :524.86 36 60 2 リボフラビン (2E,4E,6E,8E )-3,7-Dimethyl-9-(2,6,6-trimethylcyclohex- 1-en-1-yl)nona-2,4,6,8-tetraen-1-yl palmitate [79-81-2]

Retinol Palmitate is a synthetic retinol palmitate or a synthetic retinol palmitate diluted with ˆxed oil, and contains not less than 1,500,000 Vitamin A Units in each gram. It may contain a suitable antioxidant. Retinol Palmitate contains not less than 95.0z and not more than 105.0z of the labeled Units.

Description Retinol Palmitate occurs as a light yellow to C17H20N4O6:376.36 yellow-red, ointment-like or an oily substance. It has a faint, 7,8-Dimethyl-10-[(2S,3S,4R)-2,3,4,5- characteristic odor, but has no rancid odor. tetrahydroxypentyl]benzo[g]pteridine-2,4(3H,10H )-dione It is very soluble in petroleum ether, slightly soluble in [83-88-5] ethanol (95), and practically insoluble in water. It is decomposed by air and by light. Ribo‰avin, when dried, contains not less than 98.0z of C H N O . Identiˆcation Dissolve Retinol Palmitate and Retinol 17 20 4 6 Palmitate Reference Standard, equivalent to 15,000 Units, in Description Ribo‰avin occurs as yellow to orange-yellow 5 mL of petroleum ether, and use these solutions as the sam- crystals. It has a slight odor. ple solution and standard solution. Perform the test with It is very slightly soluble in water, practically insoluble in these solutions as directed under Thin-layer Chromatography ethanol (95), in acetic acid (100), and in diethyl ether. <2.03>.Spot5mL each of the sample solution and standard It dissolves in TS. solution on a plate of silica gel for thin-layer chro- A saturated solution of Ribo‰avin is neutral. matography. Develop with a mixture of cyclohexane and It is decomposed by light. diethyl ether (12:1) to a distance of about 10 cm, and air-dry Melting point: about 2909C (with decomposition). the plate. Spray evenly antimony (III) chloride TS: the prin- Identiˆcation (1) A solution of Ribo‰avin (1 in 100,000) is cipal spot obtained from the sample solution is the same in light yellow-green in color and has an intense yellow-green color tone and Rf value with the blue spot from the standard ‰uorescence. The color and ‰uorescence of the solution dis- solution. appear upon the addition of 0.02 g of sodium hydrosulˆte to Purity (1) Acid value <1.13>—Take exactly 5.0 g of 5 mL of the solution, and reappear on shaking the mixture in Retinol Palmitate, and perform the test: not more than 2.0. air. This ‰uorescence disappears upon the addition of dilute (2) Peroxide—Weigh accurately about 5 g of Retinol hydrochloric acid or sodium hydroxide TS. Palmitate, transfer in a 250-mL glass-stoppered conical ‰ask, (2) To 10 mL of a solution of Ribo‰avin (1 in 100,000) add 50 mL of a mixture of acetic acid (100) and isooctane placed in a glass-stoppered test tube add 1 mL of sodium (3:2), and gently mix to dissolve completely. Replace the air hydroxide TS, and after illumination with a ‰uorescence of the inside gradually with about 600 mL of Nitrogen, then lamp of 10 to 30 watts at 20-cm distance for 30 minutes be- add 0.1 mL of saturated potassium iodide TS under a current tween 209Cand409C, acidify with 0.5 mL of acetic acid (31), 1060 Ribo‰avin Powder / O‹cial Monographs JP XV and shake with 5 mL of chloroform: the chloroform layer with Ribo‰avin. shows a yellow-green ‰uorescence. Identiˆcation Shake a portion of Ribo‰avin Powder, (3) Determine the absorption spectrum of a solution of equivalent to 1 mg of Ribo‰avin according to the labeled Ribo‰avin in phosphate buŠer solution, pH 7.0 (1 in amount, with 100 mL of water, ˆlter, and proceed with the 100,000) as directed under Ultraviolet-visible Spectrophoto- ˆltrate as directed in the Identiˆcation (1) and (2) under metry <2.24>, and compare the spectrum with the Reference Ribo‰avin. Spectrum or the spectrum of a solution of Rivo‰avin Refer- ence Standard prepared in the same manner as the sample so- Purity Rancidity—Ribo‰avin Powder is free from any un- lution: both spectra exhibit similar intensities of absorption pleasant or rancid odor or taste. at the same wavelengths. Assay The procedure should be performed under protec- 20 Optical rotation <2.49> [a]D : －128 – －1429 Weigh ac- tion from direct sunlight and in light-resistant vessels. Weigh curately about 0.1 g of dried Ribo‰avin, dissolve in exactly 4 accurately Ribo‰avin Powder equivalent to about 15 mg of mL of dilute sodium hydroxide TS, add 10 mL of freshly ribo‰avin (C17H20N4O6), add 800 mL of diluted acetic acid boiled and cooled water, add exactly 4 mL of aldehyde-free (100) (1 in 400), and extract by warming for 30 minutes with while shaking, add freshly boiled and cooled water to occasional shaking. Cool, dilute with water to make exactly make exactly 20 mL, and determine the rotation in a 100-mm 1000 mL, and ˆlter through a glass ˆlter (G4). Use this ˆltrate cell within 30 minutes after preparing the solution. as the sample solution, and proceed as directed in the Assay under Ribo‰avin. Purity Lumi‰avin—Shake 25 mg of Ribo‰avin with 10 mL of ethanol-free chloroform for 5 minutes, and ˆlter: the Amount (mg) of ribo‰avin (C17H20N4O6) ˆltrate has no more color than the following control solution. ＝WS×｛(AT－AT?)/(AS－AS?)｝ Control solution: To 2.0 mL of 1/60 molWL potassium W : Amount (mg) of Ribo‰avin Reference Standard dichromate VS add water to make 1000 mL. S Containers and storage Containers—Tight containers. Loss on drying <2.41> Not more than 1.5z (0.5 g, 1059C, Storage—Light-resistant. 2 hours). Residue on ignition <2.44> Not more than 0.2z (1 g). Assay Conduct this procedure without exposure to day- Ribo‰avin Butyrate light, using light-resistant vessels. Weigh accurately about リボフラビン酪酸エステル 15 mg of Ribo‰avin, previously dried, dissolve in 800 mL of diluted acetic acid (100) (1 in 400) by warming, cool, add water to make exactly 1000 mL, and use this solution as the sample solution. Dry Ribo‰avin Reference Standard at 1059C for 2 hours, weigh accurately about 15 mg, dissolve in 800 mL of diluted acetic acid (100) (1 in 400) by warming, cool, add water to make exactly 1000 mL, and use this solu- tion as the standard solution. Perform the test with the sam- ple solution and standard solution as directed under Ultrav- iolet-visible Spectrophotometry <2.24>,usingwaterasthe blank, and determine the absorbances, AT and AS,at445nm. Add 0.02 g of sodium hydrosulˆte to 5 mL of each solution, shake until decolorized, and immediately measure the absor- C33H44N4O10:656.72 bances, AT? and AS?, of the solutions. (2R,3S,4S )-5-(7,8-Dimethyl-2,4-dioxo-3,4- dihydrobenzo[g]pteridin-10(2H )-yl)pentan-1,2,3,4-tetrayl Amount (mg) of C17H20N4O6 tetrabutanoate [752-56-7 ] ＝WS×｛(AT－AT?)/(AS－AS?)｝

WS: Amount (mg) of Ribo‰avin Reference Standard Ribo‰avin Butyrate, when dried, contains not less than 98.5z of C H N O . Containers and storage Containers—Tight containers. 33 44 4 10 Storage—Light-resistant. Description Ribo‰avin Butyrate occurs as orange-yellow crystals or crystalline powder. It has a slight, characteristic odor and a slightly bitter taste. Ribo‰avin Powder It is freely soluble in methanol, in ethanol (95) and in chlo- roform, slightly soluble in diethyl ether, and practically in- soluble in water. Vitamin B2 Powder It is decomposed by light. リボフラビン散 Identiˆcation (1) A solution of Ribo‰avin Butyrate in ethanol (95) (1 in 100,000) shows a light yellow-green color Ribo‰avin Powder contains not less than 95z and with a strong yellowish green ‰uorescence. To the solution not more than 115z of the labeled amount of add dilute hydrochloric acid or sodium hydroxide TS: the ribo‰avin (C H N O : 376.36). ‰uorescence disappears. 17 20 4 6 (2) Dissolve 0.01 g of Ribo‰avin Butyrate in 5 mL of Method of preparation Prepare as directed under Powders, JP XV O‹cial Monographs / Ribo‰avin Sodium Phosphate 1061 ethanol (95), add 2 mL of a mixture of a solution of hydrox- this solution as the sample solution. Separately, weigh ac- ylammonium chloride (3 in 20) and a solution of sodium curately about 50 mg of Ribo‰avin Reference Standard, hydroxide (3 in 20) (1:1), and shake well. To this solution add previously dried at 1059C for 2 hours, dissolve in 150 mL of 0.8 mL of hydrochloric acid and 0.5 mL of (III) chloride dilutedaceticacid(100)(2in75)bywarming,andaftercool- TS, and add 8 mL of ethanol (95): a deep red-brown color de- ing, add water to make exactly 500 mL. Pipet 5 mL of this so- velops. lution, add ethanol (95) to make exactly 50 mL, and use this (3) Determine the absorption spectrum of the sample so- solution as the standard solution. Determine the absor- lution obtained in the Assay as directed under Ultraviolet- bances, AT and AS, of the sample solution and standard solu- visible Spectrophotometry <2.24>, and compare the spectrum tion at 445 nm as directed under Ultraviolet-visible Spec- with the Reference Spectrum: both spectra exhibit similar in- trophotometry <2.24>. tensities of absorption at the same wavelengths. Amount (mg) of C33H44N4O10 Melting point <2.60> 146 – 1509C ＝WS×(AT/AS)×1.7449×(1/2)

Purity (1) Chloride—Dissolve 2.0 g of Ribo‰avin WS: Amount (mg) of Ribo‰avin Reference Standard Butyrate in 10 mL of methanol, and add 24 mL of dilute Containers and storage Containers—Tight containers. nitric acid and water to make 100 mL. After shaking well, al- Storage—Light-resistant. low to stand for 10 minutes, ˆlter, discard the ˆrst 10 mL of the ˆltrate, and use the subsequent ˆltrate as the sample solu- tion. To 25 mL of the sample solution add water to make 50 mL, then add 1 mL of silver nitrate TS, and allow to stand Ribo‰avin Sodium Phosphate for 5 minutes: the turbidity of the solution is not thicker than that of the following control solution. Ribo‰avin Phosphate Control solution: To 25 mL of the sample solution add 1 Vitamin B2 Phosphate Ester mL of silver nitrate TS, allow to stand for 10 minutes, and ˆlter. Wash the precipitate with four 5-mL portions of water, リボフラビンリン酸エステルナトリウム and combine the washings with the ˆltrate. To this solution add 0.30 mL of 0.01 molWL hydrochloric acid VS and water to make 50 mL, add 1 mL of water, and mix (not more than 0.021z). (2) Heavy metals <1.07>—Proceed with 2.0 g of Ribo‰avin Butyrate 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) Free acid—To 1.0 g of Ribo‰avin Butyrate add 50 mL of freshly boiled and cooled water, shake, and ˆlter. To 25 mL of the ˆltrate add 0.50 mL of 0.01 molWLsodium C17H20N4NaO9P: 478.33 hydroxide VS and 2 drops of phenolphthalein TS: the solu- Monosodium (2R,3S,4S)-5-(7,8-dimethyl-2,4-dioxo-3,4- tion shows a red color. dihydrobenzo[g]pteridin-10(2H )-yl)-2,3,4-trihydroxypentyl (4) Related substances—Dissolve 0.10 g of Ribo‰avin monohydrogenphosphate [130-40-5] Butyrate in 10 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add Ribo‰avin Sodium Phosphate contains not less than chloroform to make exactly 50 mL. Pipet 5 mL of this solu- 92z of C17H20N4NaO9P, calculated on the anhydrous tion, add chloroform to make exactly 20 mL, and use this so- basis. lution as the standard solution. Perform the test with these Description Ribo‰avin Sodium Phosphate is a yellow to solutions as directed under Thin-layer Chromatography orange-yellow, crystalline powder. It is odorless, and has a <2.03>.Spot10mL each of the sample solution and standard slightly bitter taste. solution on a plate of silica gel with ‰uorescent indicator for It is soluble in water, and practically insoluble in ethanol thin-layer chromatography. Develop the plate with a mixture (95), in chloroform and in diethyl ether. of chloroform and 2-propanol (9:1) to a distance of about 10 It is decomposed on exposure to light. cm, and air-dry the plate. Examine under ultraviolet light It is very hygroscopic. (main wavelength: 254 nm): the spots other than the principal spot from the sample solution are not more intense than the Identiˆcation (1) A solution of Ribo‰avin Sodium Phos- spot from the standard solution. phate (1 in 100,000) is light yellow-green in color and has an intense yellow-green ‰uorescence. The color and ‰uorescence Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum, of the solution disappear upon the addition of 0.02 g of sodi- silica gel, 4 hours). um hydrosulˆte to 5 mL of the solution, and reappear on Residue on ignition <2.44> Not more than 0.1z (1 g). shaking the mixture in air. This ‰uorescence disappears upon the addition of dilute hydrochloric acid or sodium hydroxide Assay Conduct this procedure without exposure to day- TS. light, using light-resistant vessels. Weigh accurately about 40 (2) To 10 mL of a solution of Ribo‰avin Sodium Phos- mg of Ribo‰avin Butyrate, previously dried, dissolve in phate (1 in 100,000) placed in a glass-stoppered test tube add ethanol (95) to make exactly 500 mL, and pipet 10 mL of this 1 mL of sodium hydroxide TS, and after illumination with a solution, add ethanol (95) to make exactly 50 mL, and use ‰uorescence lamp of 10 to 30 watts at 20-cm distance for 30 1062 Ribo‰avin Sodium Phosphate Injection / O‹cial Monographs JP XV minutes between 209Cand409C, acidify with 0.5 mL of acet- light, using light-resistant vessels. To about 0.1 g of ic acid (31), and shake with 5 mL of chloroform: the chlo- Ribo‰avin Sodium Phosphate, accurately weighed, dissolve roform layer shows a yellow-green ‰uorescence. in diluted acetic acid (100) (1 in 500) to make exactly 1000 (3) Determine the absorption spectrum of a solution of mL, then pipet 10 mL of this solution, and add diluted acetic Ribo‰avin Sodium Phosphate in phosphate buŠer solution, acid (100) (1 in 500) to make exactly 50 mL. Use this solution pH 7.0, (1 in 100,000) as directed under Ultraviolet-visible as the sample solution. Separately, dry Ribo‰avin Reference Spectrophotometry <2.24>, and compare the spectrum with Standard at 1059C for 2 hours, weigh accurately about 15 the Reference Spectrum: both spectra exhibit similar intensi- mg, dissolve in 800 mL of diluted acetic acid (100) (1 in 400) ties of absorption at the same wavelengths. by warming, cool, add water to make exactly 1000 mL, and (4) To 0.05 g of Ribo‰avin Sodium Phosphate add 10 mL use this solution as the standard solution. Perform the test of nitric acid, evaporate on a water bath to dryness, and ig- with the sample solution and standard solution as directed nite. Boil the residue with 10 mL of nitric acid (1 in 50) for 5 under Ultraviolet-visible Spectrophotometry <2.24>,using minutes, after cooling, neutralize this solution with ammonia water as the blank, and determine the absorbances, AT and A TS, and ˆlter, if necessary: the solution responds to the S,at445nm.Add0.02gofsodiumhydrosulˆteto5mLof Qualitative Tests <1.09> for sodium salt and phosphate. each solution, shake until decolorized, and immediately

20 measure the absorbances, AT′ and AS′, of the solutions. Optical rotation <2.49> [a]D : ＋38 – ＋439(0.3 g, calculat- ed on the anhydrous basis, 5 molWL hydrochloric acid TS, 20 Amount (mg) of C17H20N4NaO9P mL, 100 mm). ＝WS×｛(AT－AT?)/(AS－AS?)｝×1.2709×5 pH <2.54> Dissolve 0.20 g of Ribo‰avin Sodium Phosphate WS: Amount (mg) of Ribo‰avin Reference Standard in 20 mL of water: the pH of the solution is between 5.0 and Containers and storage Containers—Tight containers. 6.5. Storage—Light-resistant. Purity (1) Clarity and color of solution—Dissolve 0.20 g of Ribo‰avin Sodium Phosphate in 10 mL of water: the solu- tion is clear and yellow to orange-yellow in color. Ribo‰avin Sodium Phosphate (2) Lumi‰avin—To 35 mg of Ribo‰avin Sodium Phos- phate add 10 mL of ethanol-free chloroform, and shake for 5 Injection minutes, then ˆlter: the ˆltrate has no more color than the control solution. Ribo‰avin Phosphate Injection Control solution: To 3.0 mL of 1/60 molWL potassium Vitamin B2 Phosphate Ester Injection dichromate VS add water to make 1000 mL. (3) Free phosphoric acid—Weigh accurately about 0.4 g リボフラビンリン酸エステルナトリウム注射液 of Ribo‰avin Sodium Phosphate, dissolve in water to make exactly 100 mL, and use this solution as the sample solution. Measure exactly 5 mL each of the sample solution and Phos- Ribo‰avin Sodium Phosphate Injection is an aque- phoric Acid Standard Solution, transfer to separate 25-mL ous solution for injection. volumetric ‰asks, add 2.5 mL of hexaammonium hep- It contains not less than 95z and not more than tamolybdate-sulfuric acid TS and 1 mL of 1-amino-2- 120z of the labeled amount of ribo‰avin (C17H20N4O6: naphthol-4-sulfonicacidTStoeachofthese‰asks,mix,and 376.36). add water to make 25 mL. Allow to stand for 30 minutes at The concentration of Ribo‰avin Sodium Phosphate 20 ± 19C, and perform the test with these solutions as direct- Injection should be stated as the amount of ribo‰avin ed under Ultraviolet-visible Spectrophotometry <2.24>,using (C17H20N4O6). asolutionpreparedwith5mLofwaterinthesamemanneras Method of preparation Prepare as directed under Injec- a blank. Determine the absorbances, AT and AS, of the subse- tions, with Ribo‰avin Sodium Phosphate. quent solutions of the sample solution and standard phos- phoric acid solution at 740 nm: the free phosphoric acid con- Description Ribo‰avin Sodium Phosphate Injection is a tent is not more than 1.5z. clear, yellow to orange-yellow liquid. pH:5.0–7.0 Content (z) of free phosphoric acid (H3PO4) Identiˆcation (1) To a measured volume of Ribo‰avin So- ＝(AT/AS)×(1/W)×257.8 dium Phosphate Injection, equivalent to 1 mg of Ribo‰avin according to the labeled amount, add water to make 100 mL, W: Amount (mg) of Ribo‰avin Sodium Phosphate calcu- and proceed with this solution as directed in the Identiˆcation lated on the anhydrous basis. (1) and (2) under Ribo‰avin Sodium Phosphate. Water <2.48> Place 25 mL of a mixture of methanol for (2) To a measured volume of Ribo‰avin Sodium Phos- Karl Fischer method and ethylene glycol for Karl Fischer phate Injection, equivalent to 0.05 g of Ribo‰avin according method (1:1) in a dry ‰ask for titration, and titrate with water to the labeled amount, and evaporate on a water bath to dry- determinationTStotheendpoint.Weighaccuratelyabout ness. Proceed with this residue as directed in the Identiˆca- 0.1 g of Ribo‰avin Sodium Phosphate, place quickly into the tion (4) under Ribo‰avin Sodium Phosphate. ‰ask, add a known excess volume of Karl Fischer TS, mix for Extractable volume <6.05> It meets the requirement. 10 minutes, and perform the test: the water content is not more than 10.0z. Assay Conduct this procedure without exposure to day- light, using light-resistant vessels. To an accurately measured Assay Conduct this procedure without exposure to day- JP XV O‹cial Monographs / Ribostamycin Sulfate 1063 volume of Ribo‰avin Sodium Phosphate Injection, equiva- 0.2z ninhydrin-water saturated 1-butanol TS, and heat at lent to about 15 mg of ribo‰avin (C17H20N4O6), add diluted a- 1009C for 10 minutes: the principal spots obtained from the cetic acid (100) (1 in 500) to make exactly 1000 mL, and use sample solution and standard solution show a purple-brown this solution as the sample solution. Proceed as directed in color and the same Rfvalue. the Assay under Ribo‰avin Sodium Phosphate. (3) To 2 mL of a solution of Ribostamycin Sulfate (1 in 5) add 1 drop of barium chloride TS: a white turbidity is pro- Amount (mg) of Ribo‰avin (C H N O ) 17 20 4 6 duced. ＝WS×｛(AT－AT?)/(AS－AS?)｝ Optical rotation <2.49> [a]20: ＋42 – ＋499(after drying, W : Amount (mg) of Ribo‰avin Reference Standard D S 0.25 g, water, 25 mL, 100 mm). Containers and storage Containers—Hermetic containers. pH <2.54> The pH of a solution obtained by dissolving 1.0 g Storage—Light-resistant. of Ribostamycin Sulfate in 20 mL of water is between 6.0 and 8.0. Ribostamycin Sulfate Purity (1) Clarity and color of solution—Dissolve 1.0 g of Ribostamycin Sulfate in 5 mL of water: the solution is clear, リボスタマイシン硫酸塩 and colorless or pale yellow. (2) Heavy metals <1.07>—Proceed with 1.0 g of Ribostamycin Sulfate according to Method 1, and perform the test. Prepare the control solution with 3.0 mL of Stan- dard Lead Solution (not more than 30 ppm). (3) Arsenic <1.11>—Prepare the test solution with 1.0 g of Ribostamycin Sulfate according to Method 1, and perform the test (not more than 2 ppm). (4) Related substances—Dissolve 0.12 g of Ribostamycin Sulfate in water to make exactly 20 mL, and use this solution as the sample solution. Pipet 5 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>.Spot5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a solution of potassium dihydrogen phosphate (3 in 40) to a distance of about 10 cm, and air-dry the plate. C17H34N4O10.xH2SO4 Sprayevenly0.2z ninhydrin-water saturated 1-butanol TS 2,6-Diamino-2,6-dideoxy-a-D-glucopyranosyl-(1ª4)- on the plate, and heat at 1009C for 10 minutes: the spot other [b-D-ribofuranosyl-(1ª5)]-2-deoxy-D-streptamine sulfate than the principal spot obtained from the sample solution is [53797-35-6] not more intense than the spot from the standard solution. Ribostamycin Sulfate is the sulfate of an Loss on drying <2.41> Not more than 5.0z (0.5 g, reduced aminoglycoside substance having antibacterial activity pressure not exceeding 0.67 kPa, 609C, 3 hours). produced by the growth of Streptomyces ribosidiˆcus. Residue on ignition <2.44> Not more than 1.0z (1 g). It contains not less than 680 mg (potency) and not more than 780 mg (potency) per mg, calculated on the Assay Perform the test according to the Cylinder-plate dried basis. The potency of Ribostamycin Sulfate is method as directed under <4.02> Microbial Assay for An- expressed as mass (potency) of ribostamycin tibiotics according to the following conditions. (C17H34N4O10: 454.47). (i) Test organism—Bacillus subtilis ATCC 6633 (ii) Culture medium—Use the medium i in 1) Medium for Description Ribostamycin Sulfate occurs as a white to yel- test organism [5] under (1) Agar media for seed and base lay- lowish white powder. er. It is freely soluble in water, and practically insoluble in (iii) Standard solutions—Weigh accurately an amount of ethanol (95). Ribostamycin Sulfate Reference Standard, previously dried, Identiˆcation (1) Dissolve 20 mg of Ribostamycin Sulfate equivalent to about 20 mg (potency), dissolve in diluted phos- in 2 mL of phosphate buŠer solution, pH 6.0, add 1 mL of phate buŠer solution, pH 6.0 (1 in 2) to make exactly 50 mL, ninhydrin TS, and boil: a blue-purple color develops. and use this solution as the standard stock solution. Keep the (2) Dissolve 0.12 g each of Ribostamycin Sulfate and standard stock solution at 5 to 159C and use within 20 days. Ribostamycin Sulfate Reference Standard in 20 mL of water, Take exactly a suitable amount of the standard stock solution and use these solutions as the sample solution and standard before use, add 0.1 mol/L phosphate buŠer solution, pH 8.0 solution. Perform the test with these solutions as directed un- to make solutions so that each mL contains 20 mg (potency) der Thin-layer Chromatography <2.03>.Spot5mLeachof and 5 mg (potency), and use these solutions as the high con- the sample solution and standard solution on a plate of silica centrationstandardsolutionandlowconcentrationstandard gel for thin-layer chromatography. Develop the plate with a solution, respectively. solution of potassium dihydrogen phosphate (3 in 40) to a (iv) Sample solutions—Weigh accurately an amount of distance of about 10 cm, and air-dry the plate. Spray evenly Ribostamycin Sulfate, equivalent to about 20 mg (potency), 1064 / O‹cial Monographs JP XV and dissolve in water to make exactly 50 mL. Take exactly a Solution (not more than 20 ppm). suitable amount of this solution, add 0.1 mol/L phosphate (2) Arsenic <1.11>—Prepare the test solution with 1.0 g buŠer solution, pH 8.0 to make solutions so that each mL of Rifampicin according to Method 3, and perform the test contains 20 mg (potency) and 5 mg (potency), and use these so- (not more than 2 ppm). lutions as the high concentration sample solution and low (3) Related substances—Perform the test immediately af- concentration sample solution, respectively. ter preparing of the sample and standard solutions. Dissolve 0.10 g of Rifampicin in 50 mL of acetonitrile, and use this so- Containers and storage Containers—Tight containers. lution as the sample stock solution. Pipet 5 mL of the sample stock solution, add citric acid-phosphate-acetonitrile TS to make exactly 50 mL, and use this solution as the sample solu- Rifampicin tion. Separately, pipet 1 mL of the sample stock solution, and add acetonitrile to make exactly 100 mL. Pipet 5 mL of リファンピシン this solution, add citric acid-phosphate-acetonitrile TS to make exactly 50 mL, and use this solution as the standard so- lution. Perform the test with exactly 50 mLeachofthesample solutionandstandardsolutionasdirectedunderLiquid Chromatography <2.01> according to the following condi- tions, and determine each peak area by the automatic integra- tion method: the area of the peak appeared at the relative retention time of about 0.7 with respect to rifampicin from the sample solution is not more than 1.5 times the peak area of rifampicin from the standard solution, the area of the peak other than rifampicin and the peak mentioned above from the sample solution is not more than the peak area of rifampicin from the standard solution, and the total area of C H N O :822.94 43 58 4 12 the peaks other than rifampicin and the peak mentioned (2S,12Z,14E,16S,17S,18R,19R,20R,21S,22R,23S,24E)- above from the sample solution is not more than 3.5 times the 5,6,9,17,19-Pentahydroxy-23-methoxy- peak area of rifampicin from the standard solution. 2,4,12,16,18,20,22-heptamethyl-8-(4-methylpiperazin-1- Operating conditions— yliminomethyl)-1,11-dioxo-1,2-dihydro-2,7- Detector, column, column temperature, mobile phase, and (epoxypentadeca[1,11,13]trienimino)naphtho[2,1-b]furan- ‰ow rate: Proceed as directed in the operating conditions in 21-yl acetate [13292-46-1] the Assay. Time span of measurement: About 3 times as long as the Rifampicin is a derivative of a substance having an- retention time of rifampicin beginning after the peak of the tibacterial activity produced by the growth of Strep- solvent. tomyces mediterranei. System suitability— It contains not less than 970 mg (potency) and not Test for required detectability: Measure exactly 2 mL of more than 1020 mg (potency) per mg, calculated on the the standard solution, and add citric acid-phosphate-acetoni- dried basis. The potency of Rifampicin is expressed as trile TS to make exactly 20 mL. Conˆrm that the peak area of mass (potency) of rifampicin (C H N O ). 43 58 4 12 rifampicin obtained from 50 mL of this solution is equivalent Description Rifampicin occurs as orange-red to red-brown, to 7 to 13z of that from 50 mL of the standard solution. crystals or crystalline powder. System performance: Proceed as directed in the system It is slightly soluble in water, in acetonitrile, in methanol suitability in the Assay. and in ethanol (95). System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operating Identiˆcation (1) To 5 mL of a solution of Rifampicin in conditions, the relative standard deviation of the peak area of methanol (1 in 5000) add 0.05 mol/L phosphate buŠer solu- rifampicin is not more than 2.0z. tion, pH 7.0 to make 100 mL. Determine the absorption spec- trum of this solution as directed under Ultraviolet-visible Loss on drying <2.41> Not more than 2.0z (1 g, reduced Spectrophotometry <2.24>, and compare the spectrum with pressure not exceeding 0.67 kPa, 609C, 3 hours). the Reference Spectrum or the spectrum of a solution of Residue on ignition <2.44> Not more than 0.1z (1 g). Rifampicin Reference Standard prepared in the same manner as the sample solution: both spectra exhibit similar intensities Assay WeighaccuratelyanamountofRifampicinand of absorption at the same wavelengths. Rifampicin Reference Standard, equivalent to about 40 mg (2) Determine the infrared absorption spectrum of (potency), and dissolve each in acetonitrile to make exactly Rifampicin as directed in the potassium bromide disk method 200 mL. Pipet 10 mL each of these solutions, add citric acid- under the Infrared Spectrophotometry <2.25>,andcompare phosphate-acetonitrile TS to make exactly 100 mL, and use the spectrum with the Reference Spectrum or the spectrum of these solutions as the sample solution and standard solution. Rifampicin Reference Standard: both spectra exhibit similar Perform the test with exactly 50 mL each of the sample solu- intensities of absorption at the same wave numbers. tion and standard solution as directed under Liquid Chro- matography <2.01> according to the following conditions, Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of and determine the peak areas, A and A , of rifampicin. Rifampicin according to Method 2, and perform the test. T S

Prepare the control solution with 2.0 mL of Standard Lead Amount [ mg (potency)] of C43H58N4O12 JP XV O‹cial Monographs / Rifampicin Capsules 1065

＝WS×(AT/AS)×1000 Pipet 2 mL of this solution, add a mixture of acetonitrile and methanol (1:1) to make exactly 20 mL, and use this solution W : Amount [mg (potency)] of Rifampicin Reference S as the sample solution. Separately, weigh accurately about 20 Standard mg (potency) of Rifampicin Reference Standard, and dis- Operating conditions— solve in acetonitrile to make exactly 10 mL. Pipet 2 mL of Detector: An ultraviolet absorption photometer this solution, and add the mixture of acetonitrile and (wavelength: 254 nm). methanol (1:1) to make exactly 20 mL. Pipet 1 mL of this so- Column: A stainless steel column 4.6 mm in inside di- lution, add the mixture of acetonitrile and methanol (1:1) to ameter and 10 cm in length, packed with octylsilanized silica make exactly 50 mL, and use this solution as the standard so- gel for liquid chromatography (5 mminparticlediameter). lution. Perform the test with exactly 20 mLeachofthesample Column temperature: A constant temperature of about solutionandstandardsolutionasdirectedunderLiquid 259C. Chromatography <2.01> according to the following condi- Mobile phase: Dissolve 4.2 g of citric acid monohydrate tions, and determine each peak area by the automatic integra- and 1.4 g of sodium perchlorate in 1000 mL of a mixture of tion method: the amount of the peaks of quinone substance water, acetonitrile and phosphate buŠer solution, pH 3.1 and N-oxide substance, having the relative retention time of (11:7:2). about 0.5 and about 1.2 with respect to rifampicin, respec- Flow rate: Adjust the ‰ow rate so that the retention time of tively,arenotmorethan4.0z and not more than 1.5z,re- rifampicin is about 8 minutes. spectively. The amount of the peak other than the peaks men- System suitability— tioned above is not more than 1.0z, and the total amount of System performance: To 5 mL of a solution of Rifampicin these related substances is not more than 2.0z.Forthesecal- in acetonitrile (1 in 5000) add 1 mL of a solution of butyl culations, use the areas of the peaks of the quinone substance parahydroxybenzoate in acetonitrile (1 in 5000) and citric and N-oxide substance after multiplying by their relative acid-phosphate-acetonitrile TS to make 50 mL. When the response factors, 1.24 and 1.16, respectively. procedure is run with 50 mL of this solution under the above Amount (mg) of quinone substance operating conditions, butyl parahydroxybenzoate and rifam- ＝(W /W )×(A /A )×2.48 picin are eluted in this order with the resolution between these S T Ta S peaks being not less than 1.5. Amount (mg) of N-oxide substance

System repeatability: When the test is repeated 5 times with ＝(WS/WT)×(ATb/AS)×2.32 50 mL of the standard solution under the above operating Each amount (mg) of related substances other than quinone conditions, the relative standard deviation of the peak area of and N-oxide substances＝(W /W )×(A /A )×2 rifampicin is not more than 1.0z. S T Ti S W : Amount [mg (potency)] of Rifampicin Reference Containers and storage Containers—Tight containers. S Standard

WT: Amount [mg (potency)] of sample Rifampicin Capsules AS: Peak area of the standard solution ATa: Peak area of quinone substance A :PeakareaofN-oxide substance リファンピシンカプセル Tb ATi: Each peak area of related substances other than quinone and N-oxide substances Rifampicin Capsules contain not less than 93.0z Operating conditions— and not more than 105.0 z of rifampicin Detector: An ultraviolet absorption photometer (C43H58N4O12: 822.94). (wavelength: 254 nm). Method of preparation Prepare as directed under Capsules, Column: A stainless steel column 4.6 mm in inside di- with Rifampicin. ameter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Identiˆcation Dissolve an amount of the content of Rifam- Column temperature: A constant temperature of about picin Capsules, equivalent to 20 mg (potency) of Rifampicin 259C. according to the labeled amount, in methanol to make 100 Mobile phase: Dissolve 2.1 g of sodium perchlorate, 6.5 g mL, and ˆlter. To 5 mL of the ˆltrate add 0.05 mol/L phos- of citric acid monohydrate and 2.3 g of potassium dihydro- phate buŠer solution, pH 7.0 to make 100 mL. Determine the gen phosphate in 1100 mL of water, and add 900 mL of absorption spectrum of this solution as directed under acetonitrile. Ultraviolet-visible Spectrophotometry <2.24>: it exhibits max- Flow rate: Adjust the ‰ow rate so that the retention time of ima between 234 nm and 238 nm, between 252 nm and 256 rifampicin is about 12 minutes. nm, between 331 nm and 335 nm, and between 472 nm and Time span of measurement: About 2.5 times as long as the 476 nm. retention time of rifampicin. Purity Related substances—Perform the test quickly after System suitability— the sample solution and the standard solution are prepared. Test for required detectability: To exactly 1 mL of the stan- Open the capsules of not less than 20 Rifampicin Capsules, dard solution add a mixture of acetonitrile and methanol carefully take out the content, weigh accurately, and powder. (1:1) to make exactly 20 mL. Conˆrm that the peak area of Weigh accurately a portion of the powder, equivalent to rifampicin obtained with 20 mLofthissolutionisequivalent about 20 mg (potency) of Rifampicin according to the labeled to 3.5 to 6.5z of that with 20 mL of the standard solution. amount, and dissolve in acetonitrile to make exactly 10 mL. System performance: When the procedure is run with 20 1066 Ringer's Solution / O‹cial Monographs JP XV mL of the standard solution under the above operating condi- rifampicin is not more than 1.0z. tions, the number of theoretical plates and the symmetry fac- Containers and storage Containers—Tight containers. tor of the peak of rifampicin is not less than 2500 and not more than 4.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operating Ringer's Solution conditions, the relative standard deviation of the peak area of リンゲル液 rifampicin is not more than 2.0z. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Ringer's Solution is an aqueous solution for injec- Assay Open the capsules of not less than 20 Rifampicin tion. Capsules, take out the content, weigh accurately the mass of It contains not less than 0.53 wWvz and not more the content, and powder. Weigh accurately a portion of the than 0.58 wWvz of chlorine [as (Cl: 35.45)], and not powder, equivalent to about 75 mg (potency) of Rifampicin, less than 0.030 wWvz and not more than 0.036 wWvz dissolve in a mixture of acetonitrile and methanol (1:1) to of chloride hydrate (CaCl2.2H2O: 147.01). make exactly 50 mL. Pipet 10 mL of this solution, and add Method of preparation acetonitrile to make exactly 50 mL. Pipet 5 mL of this solu- Sodium Chloride 8.6 g tion prepared by dissolving 2.1 g of citric acid monohydrate, Potassium Chloride 0.3 g 27.6 g of disodium hydrogen phosphate dodecahydrate and Calcium Chloride Hydrate 0.33 g 3.1 g of potassium dihydrogen phosphate in 1000 mL of a Water for Injection a su‹cient quantity mixture of water and acetonitrile (3:1) to make exactly 50 mL, and use this solution as the sample solution. Separately, To make 1000 mL weigh accurately about 30 mg (potency) of Rifampicin Refer- Prepare as directed under Injections, with the above in- ence Standard, dissolve in 20 mL of a mixture of acetonitrile gredients. and methanol (1:1), and add acetonitrile to make exactly 100 No preservative may be added. mL. Pipet 5 mL of this solution, add the solution prepared by dissolving 2.1 g of citric acid monohydrate, 27.6 g of diso- Description Ringer's Solution is a clear and colorless liquid. dium hydrogen phosphate dodecahydrate and 3.1 g of potas- It has a slightly taste. sium dihydrogen phosphate in 1000 mL of the mixture of Identiˆcation (1) Evaporate 10 mL of Ringer's Solution water and acetonitrile (3:1) to make exactly 50 mL, and use to 5 mL: the solution responds to the Qualitative Tests <1.09> this solution as the standard solution. Perform the test with for potassium salt and calcium salt. exactly 50 mL each of the sample solution and standard solu- (2) Ringer's Solution responds to the Qualitative Tests tion as directed under Liquid Chromatography <2.01> ac- <1.09> for sodium salt and chloride. cording to the following conditions, and determine the peak areas, AT and AS, of rifampicin. pH <2.54> 5.0–7.5

Amount [mg (potency)] of rifampicin (C43H58N4O12) Purity (1) Heavy metals <1.07>—Evaporate 100 mL of ＝WS×(AT/AS)×(5/2) Ringer's Solution to about 40 mL on a water bath. Add 2 mL of dilute acetic acid and water to make 50 mL, and perform W : Amount [mg (potency)] of Rifampicin Reference S the test using this solution as the test solution. Control solu- Standard tion: to 3.0 mL of Standard Lead Solution add 2 mL of dilute Operating conditions— acetic acid and water to make 50 mL (not more than 0.3 Proceed as directed in the operating conditions in the As- ppm). say under Rifampicin. (2) Arsenic <1.11>—Perform the test with 20 mL of Rin- System suitability— ger's Solution as the test solution (not more than 0.1 ppm). System performance: Dissolve 30 mg (potency) of Rifam- Bacterial endotoxins <4.01> Less than 0.50 EUWmL. picin Reference Standard in 20 mL of the mixture of acetoni- trile and methanol (1:1), and add acetonitrile to make 100 Extractable volume <6.05> It meets the requirement. mL. To 5 mL of this solution add 2 mL of a solution of butyl Assay (1) Chlorine—To 20 mL of Ringer's Solution, ac- parahydroxybenzoate in the mixture of acetonitrile and curately measured, add 30 mL of water. Titrate <2.50> with methanol (1:1) (1 in 5000), then add the solution prepared by 0.1 molWL silver nitrate VS while shaking vigorously (indica- dissolving 2.1 g of citric acid monohydrate, 27.6 g of disodi- tor: 3 drops of sodium ‰uorescein TS). um hydrogen phosphate dodecahydrate and 3.1 g of potassi- um dihydrogen phosphate in 1000 mL of a mixture of water Each mL of 0.1 molWL silver nitrate VS and acetonitrile (3:1) to make exactly 50 mL. When the ＝3.545 mg of Cl procedure is run with 50 mL of this solution under the above (2) Calcium chloride Hydrate—To 50 mL of Ringer's So- operating conditions, butyl parahydroxybenzoate and rifam- lution, exactly measured, add 2 mL of 8 molWL potassium picin are eluted in this order with the resolution between these hydroxide TS and 0.05 g of NN indicator, and titrate <2.50> peaks being not less than 1.5. immediately with 0.01 mol WL disodium dihydrogen System repeatability: When the test is repeated 5 times with ethylenediamine tetraacetate VS, until the color of the solu- 50 mL of the standard solution under the above operating tion changes from red-purple to blue. conditions, the relative standard deviation of the peak area of Each mL of 0.01 molWL disodium dihydrogen JP XV O‹cial Monographs / Ritodrine Hydrochloride 1067

ethylenediamine tetraacetate VS Hydrochloride in 20 mL of the mobile phase, and use this so-

＝1.470 mg of CaCl2.2H2O lution as the sample solution. Pipet 1 mL of the sample solu- tion, add the mobile phase to make exactly 200 mL, and use Containers and storage Containers—Hermetic containers. this solution as the standard solution. Perform the test with Plastic containers for aqueous infusions may be used. exactly 10 mL each of the sample solution and standard solu- tion as directed under Liquid Chromatography <2.01> ac- cording to the following conditions, and determine each peak Ritodrine Hydrochloride area by the automatic integration method: the area of the peak of ritodrine threo-isomer, having the relative retention リトドリン塩酸塩 time of about 1.2 with respect to ritodrine, is not larger than 4/5 times the peak area of ritodrine from the standard solu- tion, the area of the peak other than ritodrine and ritodrine threo-isomer is not larger than 3/10 times the peak area of ritodrine from the standard solution, and the total area of the peaks other than ritodrine and ritodrine threo-isomer is not C17H21NO3･HCl: 323.81 larger than 4 times the peak area of ritodrine from the stan- (1RS,2SR)-1-(4-Hydroxyphenyl)-2- dard solution. ｛[2-(4-hydroxyphenyl)ethyl]amino｝propan-1-ol Operating conditions— monohydrochloride Column, column temperature, and mobile phase: Proceed [23239-51-2] as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer Ritodrine Hydrochloride, when dried, contains not (wavelength: 220 nm). less than 98.0z and not more than 102.0z of Flow rate: Adjust the ‰ow rate so that the retention time of C17H21NO3.HCl. ritodrine is about 10 minutes. Time span of measurement: About 3 times as long as the Description Ritodrine Hydrochloride occurs as a white retention time of ritodrine beginning after the solvent peak. crystalline powder. System suitability— It is freely soluble in water, in methanol and in ethanol Test for required detectability: To exactly 5 mL of the stan- (99.5). dard solution add exactly 50 mL of the mobile phase. Con- It dissolves in 0.01 mol/L hydrochloric acid TS. ˆrm that the peak area of ritodrine obtained with 10 mLof A solution of Ritodrine Hydrochloride (1 in 10) shows no this solution is equivalent to 7 to 13z of that with 10 mLof optical rotation. the standard solution. Melting point: about 1969C (with decomposition). System performance: To about 20 mg of ritodrine It is gradually colored to a light yellow by light. hydrochloride add 50 mL of the mobile phase and 5.6 mL of Identiˆcation (1) Determine the absorption spectrum of a sulfuric acid, and add the mobile phase to make 100 mL. solution of Ritodrine Hydrochloride (1 in 20,000) as directed Heat a portion of this solution at about 859C for about 2 under Ultraviolet-visible Spectrophotometry <2.24>,and hours, and allow to cool. Pipet 10 mL of this solution, and compare the spectrum with the Reference Spectrum or the add exactly 10 mL of 2 mol/L sodium hydroxide TS. When spectrum of a solution of Ritodrine Hydrochloride Reference the procedure is run with 10 mL of this solution under the Standard prepared in the same manner as the sample solu- above operating conditions, ritodrine and the threo-isomer tion: both spectra exhibit similar intensities of absorption at are eluted in this order with the resolution between these the same wavelengths. peaks being not less than 3. (2) Determine the infrared absorption spectrum of Rito- System repeatability: When the test is repeated 6 times with drine Hydrochloride as directed in the potassium chloride 10 mL of the standard solution under the above operating disk method under Infrared Spectrophotometry <2.25>,and conditions, the relative standard deviation of the peak area of compare the spectrum with the Reference Spectrum or the ritodrine is not more than 2.0z. spectrum of Ritodrine Hydrochloride Reference Standard: Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 both spectra exhibit similar intensities of absorption at the hours). same wave numbers. (3) A solution of Ritodrine Hydrochloride (1 in 50) Residue on ignition <2.44> Not more than 0.2z (1 g). responds to the Qualitative Tests <1.09> (2) for chloride. Assay Weigh accurately about 30 mg each of Ritodrine pH <2.54> The pH of a solution obtained by dissolving 1.0 g Hydrochloride and Ritodrine Hydrochloride Reference Stan- of Ritodrine Hydrochloride in 50 mL of water is between 4.5 dard, previously dried, and dissolve in methanol to make ex- and 5.5. actly 50 mL. Pipet 25 mL of these solutions, add exactly 5 mL of the internal standard solution, then add water to make Purity (1) Clarity and color of solution—A solution ob- 50 mL, and use these solutions as the sample solution and tained by dissolving 1.0 g of Ritodrine Hydrochloride in 10 standard solution. Perform the test with 10 mLofthesample mL of water is clear and colorless. solutionandstandardsolutionasdirectedunderLiquid (2) Heavy metals <1.07>—Proceed with 2.0 g of Ritodrine Chromatography <2.01> according to the following condi- Hydrochloride according to Method 4, and perform the test. tions, and determine the ratios, Q and Q ,ofthepeakarea Prepare the control solution with 2.0 mL of Standard Lead T S of ritodrine to that of the internal standard. Solution (not more than 10 ppm).

(3) Related substances—Dissolve 20 mg of Ritodrine Amount (mg) of C17H21NO3.HCl 1068 Ritodrine Hydrochloride Tablets / O‹cial Monographs JP XV

＝WS×(QT/QS) this solution as the sample solution. Separately, weigh ac- curately about 25 mg of Ritodrine Hydrochloride Reference W : Amount (mg) of Ritodrine Hydrochloride Reference S Standard, previously dried at 1059C for 2 hours, and dissolve Standard in 0.01 mol/L hydrochloric acid TS to make exactly 50 mL. Internal standard solution—A solution of methyl para- Pipet 3 mL of this solution, add exactly 1 mL of the internal hydroxybenzoate in methanol (3 in 5000） standard solution, and use this solution as the standard solu- Operating conditions— tion. Perform the test with 10 mL each of the sample solution Detector: An ultraviolet absorption photometer and standard solution as directed under Liquid Chro- (wavelength: 274 nm). matography <2.01> according to the following conditions,

Column: A stainless steel column 4.6 mm in inside di- and determine the ratios, QT and QS, of the peak area of rito- ameter and 15 cm in length, packed with octadecylsilanized drine to that of the internal standard. silica gel for liquid chromatography (5 mminparticledi- Amount (mg) of ritodrine hydrochloride (C H NO .HCl) ameter). 17 21 3 ＝W ×(Q /Q )×(1/5) Column temperature: A constant temperature of about S T S 259C. W : Amount (mg) of Ritodrine Hydrochloride Reference Mobile phase: Dissolve 6.6 g of diammonium hydrogen S Standard phosphate and 1.1 g of sodium 1-heptanesulfonate in 700 mL of water, and add 300 mL of methanol. Adjust to pH 3.0 Internal standard solution—A solution of methyl para- with phosphoric acid. hydroxybenzoate in methanol (3 in 10,000) Flow rate: Adjust the ‰ow rate so that the retention time of Operating conditions— ritodrine is about 6 minutes. Proceed as directed in the Assay. System suitability— System suitability— System performance: When the procedure is run with 10 System performance: When the procedure is run with 10 mL of the standard solution under the above operating condi- mL of the standard solution under the above operating condi- tions, ritodrine and the internal standard are eluted in this tions, ritodrine and the internal standard are eluted in this order with the resolution between these peaks being not less order with the resolution between these peaks being not less then 3. than 3. System repeatability: When the test is repeated 6 times with System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operating 10 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the conditions, the relative standard deviation of the ratio of the peak area of ritodrine to that of the internal standard is not peak area of ritodrine to that of the internal standard is not more than 1.0z. more than 1.0z. Containers and storage Containers—Tight containers. Dissolution <6.10> Perform the test according to the follow- Storage—Light-resistant. ing method: it meets the requirement. Perform the test with 1 tablet of Ritodrine Hydrochloride Tablets at 50 revolutions per minute according to the Paddle Ritodrine Hydrochloride Tablets method using 900 mL of water as the dissolution medium. Withdraw 20 mL or more of the dissolution medium 15 リトドリン塩酸塩錠 minutes after starting the test, and ˆlter through a membrane ˆlterwithporesizeofnotmorethan0.45mm. Discard the ˆrst 10 mL of the ˆltrate, pipet the subsequent V mL, add Ritodrine Hydrochloride Tablets contain not less water to make exactly V? mL so that each mL contains about than 93.0z and not more than 107.0z of the labeled 5.6 mg of ritodrine hydrochloride (C17H21NO3.HCl) accord- amount of ritodrine hydrochloride (C17H21NO3.HCl: ing to the labeled amount, and use this solution as the sample 323.81). solution. Separately, weigh accurately about 28 mg of Rito- Method of preparation Prepare as directed under Tablets, drine Hydrochloride Reference Standard, previously dried at with Ritodrine Hydrochloride. 1059C for 2 hours, and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly Identiˆcation To 10 mL of the ˆltrate obtained in the Assay 100 mL, and use this solution as the standard solution. Per- add 0.01 mol/L hydrochloric acid TS to make 100 mL. De- form the test with exactly 80 mL each of the sample solution termine the absorption spectrum of this solution as directed and standard solution as directed under Liquid Chro- under Ultraviolet-visible Spectrophotometry <2.24>:itex- matography <2.01> according to the following conditions, hibits a maximum between 272 nm and 276 nm. and determine the peak areas, AT and AS, of ritodrine. The Uniformity of dosage units <6.02> Perform the test accord- dissolution rate in 15 minutes is not less than 80z. ing to the following method: it meets the requirement of the Dissolution rate (z) with respect to the labeled amount of Content uniformity test. ritodrine hydrochloride (C H NO .HCl) To 1 tablet of Ritodrine Hydrochloride Tablets add 9 mL 17 21 3 ＝W ×(A /A )×(V?/V)×(1/C)×18 of 0.01 mol/L hydrochloric acid TS, shake until the tablet is S T S completely disintegrated, add 0.01 mol/L hydrochloric acid WS: Amount (mg) of Ritodrine Hydrochloride Reference TS to make exactly 10 mL, and ˆlter through a membrane Standard ˆlterhavingporesizeof0.45mm. Pipet 3 mL of the ˆltrate, C: Labeled amount (mg) of ritodrine hydrochloride add exactly 1 mL of the internal standard solution, and use (C17H21NO3.HCl) in 1 tablet JP XV O‹cial Monographs / Rokitamycin 1069

conditions, the relative standard deviation of the ratio of the Operating conditions— peak area of ritodrine to that of the internal standard is not Proceed as directed in the Assay. more than 1.0z. System suitability— System performance: When the procedure is run with 80 Containers and storage Containers—Tight containers. mL of the standard solution under the above operating condi- Storage—Light-resistant. tions, the number of theoretical plates and the symmetry fac- tor of the peak of ritodrine are not less than 3000 and not more than 1.5, respectively. Rokitamycin System repeatability: When the test is repeated 6 times with 80 mL of the standard solution under the above operating ロキタマイシン conditions, the relative standard deviation of the peak area of ritodrine is not more than 1.5z. Assay To 20 Ritodrine Hydrochloride Tablets add 150 mL of 0.01 mol/L hydrochloric acid TS, shake for 20 minutes, andadd0.01mol/LhydrochloricacidTStomakeexactly 200 mL. Filter through a glass ˆlter (G4), and discard the ˆrst 20 mL of the ˆltrate. Pipet 30 mL of the subsequent ˆltrate, add exactly 5 mL of the internal standard solution and 0.01 mol/L hydrochloric acid TS to make 50 mL, and use this so- lution as the sample solution. Separately, weigh accurately about 25 mg of Ritodrine Hydrochloride Reference Stan- dard, previously dried at 1059C for 2 hours, and dissolve in 0.01 mol/L hydrochloric acid TS to make exactly 50 mL. Pipet 30 mL of this solution, add exactly 5 mL of the internal standard solution and 0.01 mol/L hydrochloric acid TS to make 50 mL, and use this solution as the standard solution. C42H69NO15:827.99 Perform the test with 10 mL each of the sample solution and (3R,4S,5S,6R,8R,9R,10E,12E,15R)-5-[4-O-Butanoyl- standard solution as directed under Liquid Chromatography 2,6-dideoxy-3-C-methyl-3-O-propanoyl-a-L-ribo- <2.01> according to the following conditions, and determine hexopyranosyl-(1ª4)-3,6-dideoxy-3-dimethylamino-b-D- glucopyranosyloxy]-6-formylmethyl-3,9-dihydroxy-4- the ratios, QT and QS, of the peak area of ritodrine to that of the internal standard. methoxy-8-methylhexadeca-10,12-dien-15-olide [74014-51-0] Amount (mg) of ritodrine hydrochloride (C17H21NO3.HCl) ＝WS×(QT/QS)×4 Rokitamycin is a derivative of leucomycin A5,which is a macrolide antibiotic produced by the growth of the WS: Amount (mg) of Ritodrine Hydrochloride Reference mutants of Streptomyces kitasatoensis. Standard Rokitamycin contains not less than 900 mg(potency) Internal standard solution—A solution of methyl para- and not more than 1050 mg (potency) per mg, calculat- hydroxybenzoate in methanol (3 in 5000) ed on the anhydrous basis. The potency of Rokitamy- Operating conditions— cin is expressed as mass (potency) of rokitamycin Detector: An ultraviolet absorption photometer (C42H69NO15). (wavelength: 274 nm). Description Rokitamycin occurs as a white to yellowish Column: A stainless steel column 4.6 mm in inside di- white powder. ameter and 15 cm in length, packed with octadecylsilanized It is very soluble in methanol and in chloroform, freely silica gel for liquid chromatography (5 mminparticledi- soluble in ethanol (99.5) and in acetonitrile, and practically ameter). insoluble in water. Column temperature: A constant temperature of about 259C. Identiˆcation (1) Determine the absorption spectrum of a Mobile phase: Dissolve 6.6 g of diammonium hydrogen solution of Rokitamycin in methanol (1 in 50,000), as direct- phosphate and 1.1 g of sodium 1-heptanesulfonate in 700 mL ed under Ultraviolet-visible Spectrophotometry <2.24>,and of water, and add 300 mL of methanol. Adjust to pH 3.0 compare the spectrum with the Reference Spectrum or the with phosphoric acid. spectrum of a solution of Rokitamycin Reference Standard Flow rate: Adjust the ‰ow rate so that the retention time of prepared in the same manner as the sample solution: both ritodrine is about 6 minutes. spectra exhibit similar intensities of absorption at the same System suitability— wavelengths. System performance: When the procedure is run with 10 (2) Determine the infrared absorption spectrum of mL of the standard solution under the above operating condi- Rokitamycin as directed in the potassium bromide disk tions, ritodrine and the internal standard are eluted in this method under Infrared Spectrophotometry <2.25>,andcom- order with the resolution between these peaks being not less pare the spectrum with the Reference Spectrum or the spec- than 3. trum of Rokitamycin Reference Standard: both spectra ex- System repeatability: When the test is repeated 6 times with hibit similar intensities of absorption at the same wave num- 10 mL of the standard solution under the above operating bers. 1070 Roxatidine Acetate Hydrochloride / O‹cial Monographs JP XV

(3) Determine the spectrum of a solution of Rokitamycin the peak of rokitamycin are not less than 3000 and not more in deuterated chloroform for nuclear magnetic resonance than 1.5, respectively. spectroscopy (1 in 20), using tetramethylsilane for nuclear System repeatability: When the test is repeated 6 times with magnetic resonance spectroscopy as an internal reference 5 mL of the standard solution under the above operating con- compound, as directed under Nuclear Magnetic Resonance ditions, the relative standard deviation of the peak area of Spectroscopy <2.21> (1H): it exhibits single signals A, B, C rokitamycin is not more than 2.0z. and D at around d 1.4ppm,ataroundd 2.5 ppm, at around d Water <2.48> Not more than 3.0z (0.2 g, volumetric titra- 3.5 ppm and at around d 9.8 ppm, respectively. The ratio of tion, direct titration). integrated intensity of these signals, A:B:C:D, is about 3:6:3:1. Residue on ignition <2.44> Not more than 0.2z (1 g). Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Assay Perform the test according to the Cylinder-plate Rokitamycin according to Method 2, and perform the test. method as directed under Microbial Assay for Antibiotics Prepare the control solution with 2.0 mL of Standard Lead <4.02> according to the following conditions. Solution (not more than 10 ppm). (i) Test organism—Micrococcus luteus ATCC 9341 (2) Related substances—Dissolve 50 mg of Rokitamycin (ii) Culture medium—Use the medium i in 3) Medium for in 50 mL of acetonitrile, and use this solution as the sample other organisms under (1) Agar media for seed and base lay- solution. Pipet 3 mL of the sample solution, add acetonitrile er.AdjustthepH<2.54> of the medium so that it will be 7.8 to make exactly 100 mL, and use this solution as the standard to 8.0 after sterilization. solution. Perform the test with exactly 5 mLeachofthesam- (iii) Standard solutions—Weigh accurately an amount of ple solution and standard solution as directed under Liquid Rokitamycin Reference Standard equivalent to about 40 mg Chromatography <2.01> according to the following condi- (potency), dissolve in 50 mL of methanol, add 0.1 molWL tions, and determine each peak area by the automatic integra- phosphate buŠer solution, pH 4.5 to make exactly 100 mL, tion method: the peak areas of 3!-O-propionylleucomycin A7 and use this solution as the standard stock solution. Keep the having the relative retention time of about 0.72, 3!-O- standard stock solution at 59Corbelowandusewithin10 propionylisoleucomycin A5 having the relative retention time days. Take exactly a suitable amount of the standard stock of about 0.86 and 3!-O-propionylleucomycin A1 having the solution before use, add 0.1 molWL phosphate buŠer solu- relative retention time of about 1.36 with respect to tion, pH 8.0 containing 0.01z of polysorbate 80 to make so- rokitamycin obtained with the sample solution are not larger lutions so that each mL contains 2 mg (potency) and 0.5 mg than the peak area of rokitamycin with the standard solution, (potency), and use these solutions as the high concentration the area of the peak other than rokitamycin, 3!-O- standard solution and low concentration standard solution, propionylleucomycin A7,3!-O-propionylisoleucomycin A5 respectively. and 3!-O-propionylleucomycin A1 obtained with the sample (iv) Sample solutions—Weigh accurately an amount of solution is not larger than 23/100 times the peak area of Rokitamycin equivalent to about 40 mg (potency), dissolve in rokitamycin with the standard solution, and the total area of 50 mL of methanol, and add 0.1 molWL phosphate buŠer so- the peaks other than rokitamycin obtained with the sample lution, pH 4.5 to make exactly 100 mL. Take exactly a suita- solution is not larger than 3 times the peak area of rokitamy- ble amount of the solution, add 0.1 molWL phosphate buŠer cinwiththestandardsolution. solution, pH 8.0 containing 0.01z of polysorbate 80 to make Operating conditions— solutions so that each mL contains 2 mg (potency) and 0.5 mg Detector: An ultraviolet absorption photometer (potency), and use these solutions as the high concentration (wavelength: 232 nm). sample solution and low concentration sample solution, Column: A stainless steel column 4.0 mm in inside di- respectively. ameter and 15 cm in length, packed with octadecylsilanized Containers and storage Containers—Tight containers. silica gel for liquid chromatography (5 mminparticledi- ameter). Column temperature: A constant temperature of about 559C. Roxatidine Acetate Hydrochloride Mobile phase: A mixture of methanol, diluted 0.5 mol/L ロキサチジン酢酸エステル塩酸塩 ammonium acetate TS (2 in 5) and acetonitrile (124:63:13). Flow rate: Adjust the ‰ow rate so that the retention time of rokitamycin is about 11 minutes. Time span of measurement: About 2.5 times as long as the retention time of rokitamycin beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the sam- ple solution add acetonitrile to make exactly 10 mL. Conˆrm that the peak area of rokitamycin obtained with 5 mLofthis C19H28N2O4･HCl: 384.90 solution is equivalent to 7 to 13z of that with 5 mLofthe (3-｛3-[(Piperidin- standard solution. 1-yl)methyl]phenoxy｝propylcarbamoyl)methyl System performance: When the procedure is run with 5 mL acetate monohydrochloride [93793-83-0] of the sample solution under the above operating conditions, the number of theoretical plates and the symmetry factor of Roxatidine Acetate Hydrochloride, when dried, con- JP XV O‹cial Monographs / Roxatidine Acetate Hydrochloride Extended-release Capsules 1071 tains not less than 99.0z and not more than 101.0z of Time span of measurement: About 1.5 times as long as the C19H28N2O4.HCl. retention time of roxatidine acetate beginning after the sol- vent peak. Description Roxatidine Acetate Hydrochloride occurs as System suitability— white, crystals or crystalline powder. Test for required detectability: To exactly 5 mL of the stan- It is very soluble in water, freely soluble in acetic acid dard solution add ethanol (99.5) to make exactly 10 mL, and (100), and sparingly soluble in ethanol (99.5). use this solution as the solution for system suitability test. Identiˆcation (1) Determine the absorption spectrum of a Pipet 1 mL of the solution for system suitability test, and add solution of Roxatidine Acetate Hydrochloride in ethanol ethanol (99.5) to make exactly 10 mL. Conˆrm that the peak (99.5) (1 in 10,000) as directed under Ultraviolet-visible Spec- area of roxatidine acetate obtained with 10 mL of this solu- trophotometry <2.24>, and compare the spectrum with the tion is equivalent to 7 to 13z of that with 10 mLofthesolu- Reference Spectrum or the spectrum of a solution of Roxati- tion for system suitability test. dine Acetate Hydrochloride Reference Standard prepared in System performance: Dissolve 50 mg of Roxatidine the same manner as the sample solution: both spectra exhibit Acetate Hydrochloride and 10 mg of benzoic acid in 25 mL similar intensities of absorption at the same wavelengths. of ethanol (99.5). When the procedure is run with 10 mLof (2) Determine the infrared absorption spectrum of Rox- this solution under the above operating conditions, benzoic atidine Acetate Hydrochloride as directed in the potassium acid and roxatidine acetate are eluted in this order with the chloride disk method under Infrared Spectrophotometry resolution between these peaks being not less than 10. <2.25>, and compare the spectrum with the Reference Spec- System repeatability: When the test is repeated 6 times with trum or the spectrum of Roxatidine Acetate Hydrochloride 10 mL of the standard solution under the above operating Reference Standard: both spectra exhibit similar intensities conditions, the relative standard deviation of the peak area of of absorption at the same wave numbers. roxatidine acetate is not more than 1.0z. (3) A solution of Roxatidine Acetate Hydrochloride (1 in Loss on drying <2.41> Not more than 0.3z (1 g, in vacuum, 50) responds to the Qualitative Tests <1.09> (2) for chloride. phosphorus (V) oxide, 4 hours). pH <2.54> Dissolve 1.0 g of Roxatidine Acetate Hydrochlo- Residue on ignition <2.44> Not more than 0.1z (1 g). ride in 20 mL of water: the pH of this solution is between 4.0 and 6.0. Assay Weigh accurately about 0.3 g of Roxatidine Acetate Hydrochloride, previously dried, dissolve in 5 mL of acetic Melting point <2.60> 147 – 1519C (after drying). acid (100), add 50 mL of acetic anhydride, and titrate <2.50> Purity (1) Clarity and color of solution—Dissolve 1.0 g of with 0.1 mol/L perchloric acid VS (potentiometric titration). Roxatidine Acetate Hydrochloride in 10 mL of water: the so- Perform a blank determination in the same manner, and lution is clear and colorless. make any necessary correction. (2) Heavy metals <1.07>—Proceed with 2.0 g of Roxati- Each mL of 0.1 mol/L perchloric acid VS dine Acetate Hydrochloride according to Method 1, and per- ＝38.49 mg of C H N O .HCl form the test. Prepare the control solution with 2.0 mL of 19 28 2 4 Standard Lead Solution (not more than 10 ppm). Containers and storage Containers—Tight containers. (3) Related substances—Dissolve 50 mg of Roxatidine Acetate Hydrochloride in 10 mL of ethanol (99.5), and use this solution as the sample solution. Pipet 1 mL of the sample Roxatidine Acetate Hydrochloride solution, add ethanol (99.5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with Extended-release Capsules exactly 10 mL each of the sample solution and standard solu- ロキサチジン酢酸エステル塩酸塩徐放カプセル tion as directed under Liquid Chromatography <2.01> ac- cording to the following conditions, and determine each peak area by the automatic integration method: the area of the Roxatidine Acetate Hydrochloride Extended-release peak other than roxatidine acetate is not larger than 1/5 times Capsules contains not less than 93.0z and not more the peak area of roxatidine acetate obtained from the stan- than 107.0z of the labeled amount of roxatidine dard solution, and the total area of the peaks other than rox- acetate hydrochloride (C19H28N2O4.HCl: 384.90). atidine acetate is not larger than 1/2 times the peak area of roxatidine acetate from the standard solution. Method of preparation Prepare as directed under Capsules, Operating conditions— with Roxatidine Acetate Hydrochloride. Detector: An ultraviolet absorption photometer Identiˆcation To 1 mL of the ˆltrate obtained in the Assay (wavelength: 274 nm). add ethanol (99.5) to make 20 mL, and determine the absorp- Column: A stainless steel column 4 mm in inside diameter tion spectrum as directed under Ultraviolet-visible Spec- and 25 cm in length, packed with cyanopropylsilanized silica trophotometry <2.24>: it exhibits maxima between 275 nm gel for liquid chromatography (5 mminparticlediameter). and 278 nm, and between 282 nm and 285 nm. Column temperature: A constant temperature of about 359C. Uniformity of dosage units <6.02> Perform the test accord- Mobile phase: A mixture of hexane, ethanol (99.5), ing to the following method: it meets the requirement of the triethylamine and acetic acid (100) (384:16:2:1). Content uniformity test. Flow rate: Adjust the ‰ow rate so that the retention time of Take out the contents of 1 capsule of Roxatidine Acetate roxatidine acetate is about 10 minutes. Hydrochloride Extended-release Capsules, add exactly V mL of ethanol (99.5) so that each mL contains about 2.5 mg of 1072 Roxatidine Acetate Hydrochloride Extended-release Capsules / O‹cial Monographs JP XV

roxatidine acetate hydrochloride (C19H28N2O4.HCl) accord- ameter and 15 cm in length, packed with octadecylsilanized ing to the labeled amount, disperse the particles with the aid silica gel for liquid chromatography (5 mminparticledi- of ultrasonic wave, and ˆlter through a membrane ˆlter with ameter). pore size of not more than 1.0 mm. To exactly 8 mL of the Column temperature: A constant temperature of about ˆltrate add exactly 2 mL of the internal standard solution, 409C. mix, and use this solution as the sample solution. Proceed as Mobile phase: A mixture of water, acetonitrile, triethyla- directed in the Assay. mine and acetic acid (100) (340:60:2:1). Flow rate: Adjust the ‰ow rate so that the retention time of Amount (mg) of roxatidine acetate hydrochloride roxatidine acetate is about 5 minutes. (C H N O .HCl)＝W ×(Q /Q )×(V/20) 19 28 2 4 S T S System suitability—

WS: Amount (mg) of Roxatidine Acetate Hydrochloride System performance: When the procedure is run with 100 Reference Standard mL of the standard solution under the above operating condi- tions, the number of theoretical plates and the symmetry fac- Internal standard solution—A solution of benzoic acid in tor of the peak of roxatidine acetate are not less than 3000 ethanol (99.5) (1 in 500). and not more than 2.0, respectively. Dissolution <6.10> Perform the test according to the follow- System repeatability: When the test is repeated 6 times with ing method: it meets the requirement. 100 mL of the standard solution under the above operating Perform the test with 1 capsule of Roxatidine Acetate conditions, the relative standard deviation of the peak area of Hydrochloride Extended-release Capsules at 50 revolutions roxatidine acetate is not more than 1.0z. per minute according to the Paddle method, using the sinker, Assay Take out the contents of not less than 20 Roxatidine using 900 mL of water as the dissolution medium. Withdraw Acetate Hydrochloride Extended-release Capsules, weigh ac- exactly 20 mL each of the dissolution medium 45 minutes, 90 curately the mass of the contents, and powder. Weigh ac- minutes and 8 hours after starting the test for a 37.5-mg cap- curately a portion of the powder, equivalent to about 75 mg sule and 60 minutes, 90 minutes and 8 hours after starting the of roxatidine acetate hydrochloride (C H N O .HCl), add test for a 75-mg capsule. Supply exactly 20 mL of water, 19 28 2 4 exactly 30 mL of ethanol (99.5), shake, and ˆlter through a warmed at 37±0.59C, immediately after withdrawing of the membrane ˆlter with pore size of not more than 1.0 mm. To dissolution medium every time. Filter the dissolution media exactly 8 mL of the ˆltrate add exactly 2 mL of the internal withdrawn through a membrane ˆlter with pore size of not standard solution, mix, and use this solution as the sample more than 0.45 mm. Discard the ˆrst 10 mL of the ˆltrate, solution. Separately, weigh accurately about 50 mg of Rox- pipet V mL of the subsequent ˆltrate, add water to make ex- atidine Acetate Hydrochloride Reference Standard, previous- actly V? mL so that each mL contains about 42 mgofroxati- ly dried in a desiccator (in vacuum, phosphorus (V) oxide) for dine acetate hydrochloride (C H N O .HCl) according to 19 28 2 4 4 hours, and dissolve in ethanol (99.5) to make exactly 20 the labeled amount, and use these solutions as the sample so- mL. To exactly 8 mL of this solution add exactly 2 mL of the lutions. Separately, weigh accurately about 21 mg of Roxati- internal standard solution, mix, and use this solution as the dine Acetate Hydrochloride Reference Standard, previously standard solution. Perform the test with 10 mLeachofthe dried in a desiccator (in vacuum, phosphorus (V) oxide) for 4 sample solution and standard solution as directed under Liq- hours, and dissolve in water to make exactly 50 mL. Pipet 2 uid Chromatography <2.01> according to the following con- mL of this solution, add water to make exactly 20 mL, and ditions, and determine the ratios, Q and Q , of the peak area use this solution as the standard solution. Perform the test T S of roxatidine acetate to that of the internal standard. with exactly 100 mL each of the sample solutions and stan- dard solution as directed under Liquid Chromatography Amount (mg) of roxatidine acetate hydrochloride

<2.01> according to the following conditions, and determine (C19H28N2O4.HCl) the peak areas, AT(n) and AS, of roxatidine acetate of each so- ＝WS×(QT/QS)×(3/2) lution. The dissolution rates for a 37.5-mg capsule in 45 W : Amount (mg) of Roxatidine Acetate Hydrochloride minutes, in 90 minutes and in 8 hours are 10 – 40z,35–65 S Reference Standard z, and not less than 70z, respectively, and for a 75-mg cap- sule in 60 minutes, in 90 minutes and in 8 hours are 20 – 50z, Internal standard solution—A solution of benzoic acid in 35–65z, and not less than 70z, respectively. ethanol (99.5) (1 in 500). Operating conditions— Dissolution rate (z) with respect to the labeled amount of Detector: An ultraviolet absorption photometer roxatidine acetate hydrochloride (C H N O .HCl) on the 19 28 2 4 (wavelength: 274 nm). nth dissolution medium withdrawing (n＝1,2,3) Column: A stainless steel column 4.6 mm in inside di- A n－1 A 1 V? 1 ameter and 25 cm in length, packed with cyanopropyl- ＝W × T(n)＋ T(i)× × × ×180 S « A SØ A 45»$ V C silanized silica gel for liquid chromatography (5 mminparti- S i＝1 S cle diameter). W : Amount (mg) of Roxatidine Acetate Hydrochloride S Column temperature: A constant temperature of about Reference Standard 359C. C: Labeled amount (mg) of roxatidine acetate hydrochlo- Mobile phase: A mixture of hexane, ethanol (99.5), ride (C H N O .HCl) in 1 capsule 19 28 2 4 triethylamine and acetic acid (100) (384:16:2:1). Operating conditions— Flow rate: Adjust the ‰ow rate so that the retention time of Detector: An ultraviolet absorption photometer roxatidine acetate is about 10 minutes. (wavelength: 274 nm). System suitability— Column: A stainless steel column 4.6 mm in inside di- System performance: When the procedure is run with 10 JP XV O‹cial Monographs / Roxithromycin 1073 mL of the standard solution under the above operating condi- cin in the mobile phase A to make 10 mL, and use this solu- tions, the internal standard and roxatidine acetate are elute in tion as the sample solution. Separately, dissolve 20 mg of this order with the resolution between these peaks being not Roxithromycin Reference Standard in the mobile phase A to less than 10. make exactly 10 mL. Pipet 1 mL of this solution, add the mo- System repeatability: When the test is repeated 6 times with bile phase A to make exactly 100 mL, and use this solution as 10 mL of the standard solution under the above operating the standard solution. Perform the test with exactly 20 mL conditions, the relative standard deviation of the ratio of the each of the sample solution and standard solution as directed peak area of roxatidine acetate to that of the internal stan- under Liquid Chromatography <2.01> according to the fol- dard is not more than 1.0z. lowing conditions, and determine the peak areas by the auto- matic integration method: the area of a peak having the rela- Containers and storage Containers—Tight containers. tive retention time of about 1.05 to the retention time of rox- ithromycin from the sample solution is not larger than 2 times of the peak area of roxithromycin from the standard Roxithromycin solution. The areas of other than the peak of roxithromycin and the peak having the relative retention time of about 1.05 ロキシスロマイシン to the retention time of roxithromycin are not larger than the peak area of roxithromycin from the standard solution, and the total area of the peaks other than roxithromycin from the sample solution is not larger than 6 times of the peak area of roxithromycin from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 205 nm). Column: A stainless steel column 4.6 mm in inside di- ameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticledi- ameter). Column temperature: A constant temperature of about 259C. C41H76N2O15: 837.05 (2R,3S,4S,5R,6R,8R,10R,11R,12S,13R)- Mobile phase A: To 200 mL of a solution of ammonium 5-(3,4,6-Trideoxy-3-dimethylamino-b-D-xylo- dihydrogenphosphate (17 in 100) add 510 mL of water, and hexopyranosyloxy)-3-(2,6-dideoxy-3-C-methyl-3-O-methyl- adjust to pH 5.3 with 2 molWL sodium hydroxide TS. To this a-L-ribo-hexopyranosyloxy)-6,11,12-trihydroxy-9- solution add 315 mL of acetonitrile. (2-methoxyethoxy)methoxyimino-2,4,6,8,10,12- Mobile phase B: A mixture of acetonitrole and water (7:3). hexamethylpentadecan-13-olide [80214-83-1] Flowing of the mobile phase: Control the gradient by mix- ing the mobile phases A and B as directed in the following ta- Roxithromycin is a derivative of . ble. It contains not less than 970 mg (potency) per mg, calculated on the anhydrous basis. The potency of Time after injection Mobile phase Mobile phase Roxithromycinisexpressedasmass(potency)ofrox- of sample (min) A(volz) B(volz) ithromycin (C41H76N2O15). Description Roxithromycin occurs as a white crystalline 0–38 100 0 powder. 38–39 100ª90 0ª10 It is freely soluble in ethanol (95) and in acetone, soluble in 39–80 90 10 methanol, sparingly soluble in acetonitrile, and practically in- soluble in water. Flow rate: Adjust the ‰ow rate so that the retention time of Identiˆcation Determine the infrared absorption spectrum roxithromycin is about 21 minutes. of Roxithromycin as directed in the potassium bromide disk System suitability— method under Infrared Spectrophotometry <2.25>,andcom- Test for required detection: To exactly 2 mL of the stan- pare the spectrum with the Reference Spectrum or the spec- dard solution add the mobile phase A to make exactly 10 mL. trum of Roxithromycin Reference Standard: both spectra ex- Conˆrm that the peak area of roxithromycin obtained from hibit similar intensities of absorption at the same wave num- 20 mL of this solution is equivalent to 15 to 25z of that of bers. roxithromycin obtained from 20 mL of the standard solution. System performance: Dissolve 5 mg each of Roxithromy- 20 Optical rotation <2.49> [a]D : －93 – －969(0.5 g calculated cin Reference Standard and N-demethylroxithromycin in the on the anhydrous basis, acetone, 50 mL, 100 mm). mobile phase A to make 100 mL. When the procedure is run Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of with 20 mL of this solution under the above operating condi- Roxithromycin according to Method 2, and perform the test. tions, N-demethylroxithromycin and roxithromycin are elut- Prepare the control solution with 2.0 mL of Standard Lead ed in this order with the resolution between these peaks being Solution (not more than 10 ppm). not less than 6. (2) Related substances—Dissolve 40 mg of Roxithromy- System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operating 1074 Freeze-dried Live Attenuated Rubella Vaccine / O‹cial Monographs JP XV conditions, the relative standard deviation of the peak areas It conforms to the requirements of Freeze-dried Live of roxithromycin is not more than 2.0z. Attenuated Rubella Vaccine in the Minimum Require- ments for Biological Products. Water <2.48> Not more than 3.0z (0.3 g, volumetric titra- tion, direct titration). Description Freeze-dried Live Attenuated Rubella Vaccine becomes a colorless, yellowish or reddish clear liquid on addi- Residue on ignition <2.44> Not more than 0.1z (1 g). tion of solvent. Assay Weigh accurately an amount of Roxithromycin and Roxithromycin Reference Standard, equivalent to about 20 mg (potency), and dissolve separately in the mobile phase to Saccharated Pepsin make exactly 10 mL, and use these solutions as the sample so- lution and standard solution. Perform the test with exactly 20 含糖ペプシン mLeachofthesamplesolutionandstandardsolutionas directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the peak areas, A T Saccharated Pepsin is a mixture of pepsin obtained and A S, of roxithromycin. from the gastric mucosa of hog or cattle and Lactose Hydrate, and it is an enzyme drug having a proteolytic Amount [mg (potency)] of C41H76N2O15 ＝W ×(A /A )×1000 activity. S T S Saccharated Pepsin contains not less than 3800 units WS: Amount [mg (potency)] of Roxithromycin Reference and not more than 6000 units per g. Standard Description Saccharated Pepsin occurs as a white powder. It has a characteristic odor, and has a slightly sweet taste. Operating conditions— It dissolves in water to give a slightly turbid liquid, and Detector: An ultraviolet absorption photometer does not dissolve in ethanol (95) and in diethyl ether. (wavelength: 205 nm). It is slightly hygroscopic. Column: A stainless steel column 4.6 mm in inside di- ameter and 25 cm in length, packed with octadecylsilanized Purity (1) Rancidity—Saccharated Pepsin has no un- silica gel for liquid chromatography (5 mminparticledi- pleasant or rancid odor. ameter). (2) Acidity—Dissolve 0.5 g of Saccharated Pepsin in 50 Column temperature: A constant temperature of about mL of water, and add 0.50 mL of 0.1 molWLsodium 259C. hydroxide VS and 2 drops of phenolphthalein TS: the solu- Mobile phase: To 200 mL of a solution of ammonium di- tion is red in color. hydrogenphosphate (17 in 100) add 510 mL of water, and ad- Loss on drying <2.41> Not more than 1.0z (1 g, 809C, just to pH 5.3 with 2 molWL sodium hydroxide TS. To this 4 hours). solution add 315 mL of acetonitrile. Flow rate: Adjust the ‰ow rate so that the retention time of Residue on ignition <2.44> Not more than 0.5z (1 g). roxithromycin is about 11 minutes. Assay (i) Substrate solution—Use the substrate solution 1 System suitability— described in (2) Assay for protein digestive activity under the System performance: Dissolve 5 mg each of Roxithromy- Digestion Test <4.03> after adjusting the pH to 2.0. cin Reference Standard and N-demethylroxithromycin in the (ii) Sample solution—Weigh accurately an amount of mobile phase to make 100 mL. When the procedure is run Saccharated Pepsin equivalent to about 1250 units, dissolve with 20 mL of this solution under the above operating condi- in ice-cold 0.01 mol LhydrochloricacidTStomakeexactly tions, N-demethylroxithromycin and roxithromycin are elut- W 50 ml. ed in this order with the resolution between these peaks being (iii) Standard solution—Weigh accurately a suitable not less than 6 and the symmetry factor of the peak of rox- amount of Saccharated Pepsin Reference Standard, and dis- ithromycin is not more than 1.5. solve in ice-cold 0.01 mol LhydrochloricacidTStomakea System repeatability: When, the test is repeated 6 times W solution containing about 25 units per ml. with 20 mL of the standard solution under the above operat- (iv) Procedure—Proceed as directed in (2) Assay for pro- ing conditions, the relative standard deviation of the peak tein digestive activity under Digestion Test <4.03>,anddeter- areas of roxithromycin is not more than 1.0z. mine the absorbances, AT and ATB, of the sample solution, Containers and storage Containers—Tight containers. using trichloroacetic acid TS A as the precipitation reagent.

Separately, determine the absorbances, AS and ASB,ofthe standard solution in the same manner as the sample solution. Freeze-dried Live Attenuated Units in 1 g of Saccharated Pepsin Rubella Vaccine ＝US×｛(AT－ATB)/(AS－ASB)｝×(1/ W) U : Units per ml of the standard solution 乾燥弱毒生風しんワクチン S W: Amount (g) of Saccharated Pepsin per ml of the sam- ple solution Freeze-dried Live Attenuated Rubella Vaccine is a Containers and storage Containers—Tight containers. preparation for injection which is dissolved before use. Storage—Not exceeding 309C. It contains live attenuated rubella virus. JP XV O‹cial Monographs / Saccharin 1075

not larger than QS. Saccharin Internal standard solution—A solution of caŠeine in ethyl acetate (1 in 500). サッカリン Operating conditions— Detector: A hydrogen ‰ame-ionization detector. Column: A glass column 3 mm in inside diameter and 1 m in length, packed with siliceous earth for gas chro- matography coated 3z with diethylene glycol succinate poly- ester for gas chromatography (180 – 250 mminparticledi- ameter).

C7H5NO3S: 183.18 Column temperature: A constant temperature of about 1,2-Benzo[d]isothiazol-3(2H)-one 1,1-dioxide 2009C. [81-07-2] Temperature of injection port: A constant temperature of about 2259C. This monograph is harmonized with the European Temperature of detector: A constant temperature of about Pharmacopoeia and the U.S. Pharmacopeia. The parts 2509C. of the text that are not harmonized are marked with Carrier gas: Nitrogen  symbols ( ). Flow rate: Adjust the ‰ow rate so that the retention time of Saccharin contains not less than 99.0z and not caŠeine is about 6 minutes. more than 101.0z of C7H5NO3S, calculated on the System suitability— dried basis. System performance: When the procedure is run with 1 mL of the standard solution under the above operating condi- Description Saccharin occurs as colorless or white crystals tions, the internal standard and o- sulfonamide are or a white crystalline powder. It has a very sweet taste. eluted in this order with the resolution between these peaks It is sparingly soluble in ethanol (95), and slightly soluble being not less than 2.0. in water. System repeatability: When the test is repeated 6 times with It dissolves in sodium hydroxide TS.  1 mL of the standard solution under the above operating con- Identiˆcation Determine the infrared absorption spec- ditions, the relative standard deviation of the ratio of the trum of Saccharin as directed in the potassium bromide disk peak height of o-toluene sulfonamide to that of the internal method under Infrared Spectrophotometry <2.25>,andcom- standard is not more than 2.0z. pare the spectrum with the Reference Spectrum: both spectra (5) Readily carbonizable substances <1.15>—Perform the exhibit similar intensities of absorption at the same wave test with 0.20 g of Saccharin, by warming at 48 to 509Cfor10 numbers. minutes: the color of the solution is not more intense than the matching ‰uid A. Melting point <2.60> 226 – 2309C Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 Purity (1) Clarity and color of solution—Dissolve 1.0 g hours). of Saccharin in 30 mL of hot water or in 50 mL of ethanol

(95): the solution is clear and colorless in each case. Residue on ignition <2.44> Not more than 0.2z (1 g). (2) Heavy metals <1.07>—Proceed with 2.0 g of Saccha- Assay Weigh accurately about 0.5 g of Saccharin, dissolve rin according to Method 2, and perform the test. Prepare the in 40 mL of ethanol (95), add 40 mL of water, and titrate control solution with 2.0 mL of Standard Lead Solution (not <2.50> with 0.1 mol/L sodium hydroxide VS (indicator: 3 more than 10 ppm).  drops of phenolphthalein TS). Perform a blank determina- (3) Benzoate and salicylate—To 10 mL of a saturated so- tion in the same manner, and make any necessary correction. lution of Saccharin in hot water add 3 drops of iron (III) chloride TS: no precipitate is formed, and no red-purple to Each mL of 0.1 mol/L sodium hydroxide VS purple color develops. ＝18.32 mg of C7H5NO3S (4) o-Toluene sulfonamide—Dissolve 10 g of Saccharin Containers and storage Containers—Well-closed contain- in 70 mL of sodium hydroxide TS, and extract with three ers. 30-mL portions of ethyl acetate. Combine all the ethyl acetate extracts, wash with 30 mL of a solution of sodium chloride (1 in 4), dehydrate with 5 g of anhydrous sodium sul- fate, then evaporate the solvent. To the residue add exactly 5 mL of the internal standard solution to dissolve, and use this solution as the sample solution. Separately, dissolve 0.10 g of o-toluene sulfonamide in ethyl acetate to make exactly 100 mL. Pipet 1 mL of this solution, evaporate to dryness on a water bath, dissolve the residue in exactly 5 mL of the inter- nal standard solution, and use this solution as the standard solution. Perform the test with 1 mLeachofthesamplesolu- tion and standard solution as directed under Gas Chro- matography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak height of o- toluene sulfonamide to that of the internal standard: QT is 1076 Saccharin Sodium Hydrate / O‹cial Monographs JP XV

10 ppm). (4) Benzoate and salicylate—Dissolve 0.5 g of Saccharin Saccharin Sodium Hydrate Sodium Hydrate in 10 mL of water, add 5 drops of acetic acid (31) and 3 drops of iron (III) chloride TS: no turbidity is Saccharin Sodium produced, and no red-purple to purple color develops. (5) o-Toluene sulfonamide—Dissolve 10 g of Saccharin サッカリンナトリウム水和物 Sodium Hydrate in 50 mL of water, and extract with three 30-mL portions of ethyl acetate. Combine all the ethyl acetate extracts, wash with 30 mL of a solution of sodium chloride (1 in 4), dehydrate with 5 g of anhydrous sodium sul- fate, and evaporate ethyl acetate. To the residue add exactly 5 mL of the internal standard solution to dissolve, and use this solution as the sample solution. Separately, dissolve 0.10 g of C7H4NNaO3S.2H2O: 241.20 o-toluene sulfonamide in ethyl acetate to make exactly 100 mL. Pipet 1 mL of this solution, evaporate on a water bath to 2-Sodio-1,2-benzo[d]isothiazol-3(2H)-one 1,1-dioxide dryness, dissolve the residue in exactly 5 mL of the internal dihydrate [6155-57-3] standard solution, and use this solution as the standard solu- tion. Perform the test with 1 mL each of the sample solution This monograph is harmonized with the European and standard solution as directed under Gas Chro- Pharmacopoeia and the U.S. Pharmacopeia. The parts matography <2.02> according to the following conditions, of the text that are not harmonized are marked with and calculate the ratios, QT and QS, of the peak height of o-  toluene sulfonamide to that of the internal standard: QT is symbols ( ). Saccharin Sodium Hydrate, contains not less than not more than QS. 99.0z and not more than 101.0z of saccharin sodium Internal standard solution—A solution of caŠeine in ethyl acetate (1 in 500). (C6H4NNaO3S: 205.17), calculated on the anhydrous basis. Operating conditions— Detector: A hydrogen ‰ame-ionization detector. Description Saccharin Sodium Hydrate occurs as color- Column: A column 3 mm in inside diameter and 1 m in less crystals or a white, crystalline powder. It has an intensely length, packed with siliceous earth for gas chromatography sweet taste, even in 10,000 dilutions. (180 to 250 mm in diameter), coated with diethyleneglycol It is freely soluble in water and in methanol, and sparingly succinate polyester for gas chromatography at the ratio of 3 soluble in ethanol (95) and in acetic acid (100). z. It eŒoresces slowly and loses about half the amount of Column temperature: A constant temperature of about water of crystallization in air. 2009C. Identiˆcation (1) Determine the infrared absorption Injection port temperature: A constant temperature of spectrum of Saccharin Sodium Hydrate as directed in the about 2259C. potassium bromide disk method under Infrared Spec- Detector temperature: A constant temperature of about trophotometry <2.25>, and compare the spectrum with the 2509C. Reference Spectrum: both spectra exhibit similar intensities Carrier gas: Nitrogen Flow rate: Adjust the ‰ow rate so that the retention time of of absorption at the same wave numbers. (2) A solution of Saccharin Sodium Hydrate (1 in 10) caŠeine is about 6 minutes. responds to the Qualitative Tests <1.09> for sodium salt. System suitability— System performance: When the procedure is run with 1 mL Purity (1) Clarity and color of solution—Dissolve 1.0 g of the standard solution under the above operating condi- of Saccharin Sodium Hydrate in 1.5 mL of water or in 50 mL tions, the internal standard and o-toluene sulfonamide are of ethanol (95): the solution is clear and colorless. eluted in this order with the resolution between these peaks (2) Acidity or alkalinity—Dissolve 1.0 g of Saccharin So- being not less than 2.0. dium Hydrate in 10 mL of water, and add 1 drop of System repeatability: When the test is repeated 6 times with phenolphthalein TS: the solution is colorless. Add 1 drop of 1 mL of the standard solution under the above operating con- 0.1 mol/L sodium hydroxide VS to the solution: the color ditions, the relative standard deviation of the ratio of the changes to red. peak height of o-toluene sulfonamide to that of the internal (3) Heavy metals <1.07>—Dissolve 2.0 g of Saccharin standard is not more than 2.0z. Sodium Hydrate in 40 mL of water, add 0.7 mL of dilute (6) Readily carbonizable substances <1.15>—Perform the hydrochloric acid, dilute with water to make 50 mL, and rub test with 0.20 g of Saccharin Sodium Hydrate. Allow the so- the inner wall of the vessel with a glass rod until crystalliza- lution to stand between 489Cand509C for 10 minutes: the tion begins. Allow the solution to stand for 1 hour after the solution has no more color than Matching Fluid A. beginning of crystallization, and then ˆlter through dry ˆlter paper. Reject the ˆrst 10 mL of the ˆltrate, and take 25 mL Water <2.48> Not more than 15.0z (0.1 g, volumetric titra- of the subsequent ˆltrate. Add 2 mL of dilute acetic acid and tion, direct titration). water to make 50 mL, and perform the test, using this solu- Assay Weigh accurately about 0.15 g of Saccharin Sodium tion as the test solution. To 1.0 mL of Standard Lead Solu- Hydrate, dissolve in 50 mL of acetic acid (100), heat slightly tion add 2 mL of dilute acetic acid and water to make 50 mL, if necessary, and titrate <2.50> with 0.1 mol/L perchloric acid and use this solution as the control solution (not more than VS (potentiometric titration). Perform a blank determina- JP XV O‹cial Monographs / Salazosulfapyridine 1077 tion, and make any necessary correction. lution. Prepare the control solution with 0.40 mL of 0.01 molWL hydrochloric acid VS (not more than 0.014z). Each mL of 0.1 mol/L perchloric acid VS (2) Sulfate <1.14>—Dissolve 2.0 g of Salazosulfapyridine ＝20.52 mg of C H NNaO S 7 4 3 in 12 mL of sodium hydroxide TS and 36 mL of water, add 2 Containers and storage Containers—Well-closed contain- mL of hydrochloric acid, shake, and ˆlter. To 25 mL of the ers. ˆltrate add 1 mL of dilute hydrochloric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 1.0 mL of Salazosulfapyridine 0.005 molWL sulfuric acid VS (not more than 0.048z). (3) Heavy metals <1.07>—Proceed with 1.0 g of Sulfasalazine Salazosulfapyridine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard サラゾスルファピリジン Lead Solution (not more than 20 ppm). (4) Arsenic <1.11>—Take 1.0 g of Salazosulfapyridine in a decomposition ‰ask, add 20 mL of nitric acid, and heat gently until it becomes ‰uid. After cooling, add 5 mL of sul- furic acid, and heat until white fumes are evolved. Add, if necessary, 5 mL of nitric acid after cooling, and heat again. Repeat this operation until the solution becomes colorless to slightly yellow. After cooling, add 15 mL of a saturated solu-

C18H14N4O5S: 398.39 tion of ammonium oxalate monohydrate, and heat until 2-Hydroxy-5-[4-(pyridin-2-ylsulfamoyl)phenylazo]benzoic white fumes are evolved again. After cooling, add water to acid [599-79-1] make 25 mL. Perform the test with 5 mL of this solution as the test solution: the color of the test solution is not deeper Salazosulfapyridine, when dried, contains not less than that of the following standard stain. than 96.0z of C18H14N4O5S. Standard stain: Proceed in the same manner without Salazosulfapyridine, transfer 5 mL of the obtained solution Description Salazosulfapyridine occurs as a yellow to yel- to a generator bottle, add exactly 2 mL of Standard Arsenic low-brown, ˆne powder. It is odorless and tasteless. Solution, and proceed in the same manner as the test with the It is sparingly soluble in pyridine, slightly soluble in test solution (not more than 10 ppm). ethanol (95), practically insoluble in water, in chloroform (5) Related substances—Dissolve 0.20 g of Salazosul- andindiethylether. fapyridine in 20 mL of pyridine, and use this solution as the It dissolves in sodium hydroxide TS. sample solution. Pipet 1 mL of this solution, add pyridine to Melting point: 240 – 2499C (with decomposition). make exactly 100 mL, and use this solution as the standard Identiˆcation (1) Dissolve 0.1 g of Salazosulfapyridine in solution. Perform the test with these solutions as directed un- 20 mL of dilute sodium hydroxide TS: a red-brown color de- der Thin-layer Chromatography <2.03>.Spot10mLeachof velops. This color gradually fades upon gradual addition of the sample solution and standard solution on a plate of silica 0.5 g of sodium hydrosulˆte with shaking. Use this solution gel with ‰uorescent indicator for thin-layer chromatography. in the following tests (2) to (4). Develop the plate with diluted methanol (9 in 10) to a dis- (2) To 1 mL of the solution obtained in (1) add 40 mL of tance of about 10 cm, and air-dry the plate. Examine the water, neutralize with 0.1 molWL hydrochloric acid TS, and plate under ultraviolet light (main wavelength: 254 nm): the add water to make 50 mL. To 5 mL of this solution add 2 to 3 spots other than the principal spot from the sample solution drops of dilute iron (III) chloride TS: a red color develops are not more intense than the spot from the standard solu- and changes to purple, then fades when dilute hydrochloric tion. acid is added dropwise. (6) Salicylic acid—To 0.10 g of Salazosulfapyridine add (3) The solution obtained in (1) responds to the Qualita- 15 mL of diethyl ether, and shake vigorously. Add 5 mL of tive Tests <1.09> for primary aromatic amines. dilute hydrochloric acid, shake vigorously for 3 minutes, col- (4) To 1 mL of the solution obtained in (1) add 1 mL of lect the diethyl ether layer, and ˆlter. To the water layer add pyridine and 2 drops of copper (II) sulfate TS, and shake. 15 mL of diethyl ether, shake vigorously for 3 minutes, col- Add 3 mL of water and 5 mL of chloroform, shake, and al- lect the diethyl ether layer, ˆlter, and combine the ˆltrates. low to stand: a green color develops in the chloroform layer. Wash the residue on the ˆlter paper with a small quantity of (5) Determine the absorption spectrum of a solution of diethyl ether, and combine the washings and the ˆltrate. Salazosulfapyridine in dilute sodium hydroxide TS (1 in Evaporate the diethyl ether with the aid of air-stream at room 100,000) as directed under Ultraviolet-visible Spectrophoto- temperature. To the residue add dilute ammonium iron (III) metry <2.24>, and compare the spectrum with the Reference sulfate TS, shake, and ˆlter, if necessary. Wash the residue Spectrum: both spectra exhibit similar intensities of absorp- on the ˆlter paper with a small quantity of dilute ammonium tion at the same wavelengths. iron (III) sulfate TS, combine the washings and the ˆltrate, add dilute ammonium iron (III) sulfate TS to make exactly 20 Purity (1) Chloride <1.03>—Dissolve 2.0 g of Salazosul- mL, and use this solution as the sample solution. Separately, fapyridine in 12 mL of sodium hydroxide TS and 36 mL of weigh accurately about 10 mg of salicylic acid for assay, water, add 2 mL of nitric acid, shake, and ˆlter. To 25 mL of previously dried in a desiccator (silica gel) for 3 hours, dis- the ˆltrate add 6 mL of dilute nitric acid and water to make solve in dilute ammonium iron (III) sulfate TS to make exact- 50 mL, and perform the test using this solution as the test so- ly 400 mL, and use this solution as the standard solution. De- 1078 Salbutamol Sulfate / O‹cial Monographs JP XV

termine the absorbances, AT and AS,at535nmofthesample Salbutamol Sulfate in 20 mL of water: the solution is clear solution and the standard solution as directed under Ultrav- and colorless. iolet-visible Spectrophotometry <2.24>: salicylic acid content (2) Heavy metals <1.07>—Proceed with 1.0 g of Sal- is not more than 0.5z. butamol Sulfate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Content (z) of salicylic acid (C H O ) 7 6 3 Lead Solution (not more than 20 ppm). ＝ W ×(A /A )×0.05 S T S (3) Related substances—Dissolve 20 mg of Salbutamol

WS: Amount (mg) of salicylic acid for assay Sulfate in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to Loss on drying <2.41> Not more than 2.0z (1 g, 1059C, make exactly 100 mL, and use this solution as the standard 4 hours). solution. Perform the test with these solutions as directed un- Residue on ignition <2.44> Not more than 0.2z (1 g). der Thin-layer Chromatography <2.03>.Spot5mLeachof the sample solution and standard solution on a plate of silica Assay Weigh accurately about 20 mg of Salazosulfapyri- gel for thin-layer chromatography. Develop the plate with a dine, previously dried, and perform the test as directed in the mixture of ethyl acetate, 2-propanol, water and ammonia so- procedure of determination for sulfur under the lution (28) (25:15:8:2) to a distance of about 15 cm, and air- Flask Combustion Method <1.06>, using 10 mL of diluted dry the plate. Leave the plate in a well-closed vessel saturated hydrogen peroxide (30) (1 in 40) as an absorbing liquid. with diethylamine vapor for 5 minutes, and spray evenly 4- Each mL of 0.005 molWL barium perchlorate VS nitrobenzenediazonium chloride TS: the spots other than the

＝ 1.992 mg of C18H14N4O5S principal spot from the sample solution are not more intense than the spot from the standard solution in color. Containers and storage Containers—Tight containers. (4) Boron—Take 50 mg of Salbutamol Sulfate and 5.0 Storage—Light-resistant. mL of the boron standard solution, and transfer to a plati- num crucible. Add 5 mL of potassium carbonate-sodium car- bonate TS, evaporate on a water bath to dryness, and dry at Salbutamol Sulfate 1209C for 1 hour. Ignite the residue immediately. After cool- ing,add0.5mLofwaterand3mLofcurcuminTStothe サルブタモール硫酸塩 residue, warm gently in a water bath for 5 minutes. After cooling, add 3 mL of acetic acid-sulfuric acid TS, mix, and allow to stand for 30 minutes. Add ethanol (95) to make ex- actly 100 mL, and ˆlter. Discard the ˆrst 10 mL of the ˆltrate, and use the subsequent ˆltrate as the sample solution and standard solution. Perform the test with these solutions as directed under Ultraviolet-visible Spectrophotometry <2.24>, using ethanol (95) as the blank: the absorbance of the (C13H21NO3)2.H2SO4:576.70 sample solution at 555 nm is not larger than that of the stan- (1RS)-2-(1,1-Dimethylethyl)amino-1-(4-hydroxy-3- dard solution. hydroxymethylphenyl)ethanol hemisulfate [51022-70-9] Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum Salbutamol Sulfate, when dried, contains not less at a pressure not exceeding 0.67 kPa, 1009C, 3 hours). than 98.0z of (C13H21NO3)2.H2SO4. Residue on ignition <2.44> Not more than 0.1z (1 g). Description Salbutamol Sulfate occurs as a white powder. Assay Weigh accurately about 0.9 g of Salbutamol Sulfate, It is freely soluble in water, slightly soluble in ethanol (95), previously dried, and dissolve in 50 mL of acetic acid (100) by and in acetic acid (100) and practically insoluble in diethyl warming. After cooling, titrate <2.50> with 0.1 molWLper- ether. chloric acid VS until the color of the solution changes from A solution of Salbutamol Sulfate (1 in 20) shows no optical purple through blue to blue-green (indicator: 3 drops of crys- rotation. tal violet TS). Perform a blank determination, and make any Identiˆcation (1) Determine the absorption spectrum of a necessary correction. solution of Salbutamol Sulfate in 0.1 mol/L hydrochloric Each mL of 0.1 molWL perchloric acid VS acid TS (1 in 12,500) as directed under Ultraviolet-visible ＝ 57.67 mg of (C13H21NO3)2.H2SO4 Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi- Containers and storage Containers—Tight containers. ties of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Sal- butamol Sulfate, previously dried, as directed in the potassi- Salicylated Alum Powder um bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- サリチル・ミョウバン散 trum: both spectra exhibit similar intensities of absorption at thesamewavenumbers. (3) A solution of Salbutamol Sulfate (1 in 20) responds to Salicylated Alum Powder contains not less than the Qualitative Tests <1.09> for sulfate. 2.7z and not more than 3.3z of salicylic acid (C H O :138.12). Purity (1) Clarity and color of solution—Dissolve 1.0 g of 7 6 3 JP XV O‹cial Monographs / Salicylic Acid 1079

Method of preparation Salicylic Acid, ˆnely powdered 30 g Salicylic Acid Dried Aluminum Potassium Sulfate, very ˆnely powdered 640 g サリチル酸 Talc, very ˆnely powdered a su‹cient quantity To make 1000 g Prepare as directed under Powders, with the above in- gredients. C7H6O3: 138.12 Description Salicylated Alum Powder occurs as a white 2-Hydroxybenzoic acid [69-72-7 ] powder. Identiˆcation (1) The colored solution obtained in the As- Salicylic Acid, when dried, contains not less than say has a red-purple color and exhibits an absorption maxi- 99.5z of C7H6O3. mum <2.24> between 520 nm and 535 nm (salicylic acid). Description Salicylic Acid occurs as white crystals or crys- (2) Shake 0.3 g of Salicylated Alum Powder with 5 mL of talline powder. It is odorless and has a slightly acid, followed methanol, ˆlter, and use the ˆltrate as the sample solution. by an acrid taste. Separately, dissolve 0.01 g of salicylic acid in 5 mL of It is freely soluble in ethanol (95), in acetone and in diethyl methanol, and use this solution as the standard solution. Per- ether, soluble in hot water, and slightly soluble in water. form the test with these solutions as directed under Thin-lay- er Chromatography <2.03>.Spot5mLeachofthesampleso- Identiˆcation A solution of Salicylic Acid (1 in 500) lution and standard solution on the plate of silica gel with responds to the Qualitative Tests <1.09> (1)and(3)forsalicy- ‰uorescent indicator for thin-layer chromatography. Develop late. the plate with a mixture of chloroform, acetone and acetic Melting point <2.60> 158 – 1619C acid (100) (45:5:1) to a distance of about 10 cm, and air-dry the plate. Examine the plate under ultraviolet light (main Purity (1) Chloride <1.03>—Dissolve 5.0 g of Salicylic wavelength: 254 nm): the spot from the sample solution and Acid in 90 mL of water by heating, cool, dilute with water to that from the standard solution show the same Rfvalue. 100 mL, and ˆlter. Discard the ˆrst 20 mL of the ˆltrate, take Spray evenly iron (III) chloride TS upon the plate: the spot subsequent 30 mL of the ˆltrate, add 6 mL of dilute nitric from the standard solution and the corresponding spot from acid and water to make 50 mL, and perform the test using the sample solution reveal a purple color. this solution as the test solution. Prepare the control solution with 0.35 mL of 0.01 molWL hydrochloric acid VS (not more Assay Weigh accurately about 0.33 g of Salicylated Alum than 0.008z). Powder, add 80 mL of ethanol (95), and shake vigorously. (2) Sulfate <1.14>—To 30 mL of the ˆltrate obtained in Dilute with ethanol (95) to make exactly 100 mL, ˆlter, and (1) add 1 mL of dilute hydrochloric acid and water to make discard the ˆrst 10 mL of the ˆltrate. Use the subsequent 50 mL, and perform the test using this solution as the test so- ˆltrate as the sample solution. Dissolve about 0.1 g of salicyl- lution. Prepare the control solution with 0.35 mL of 0.005 ic acid for assay, previously dried in a desiccator (silica gel) molWL sulfuric acid VS (not more than 0.011z). for 3 hours and accurately weighed, in su‹cient ethanol (95) (3) Heavy metals <1.07>—Dissolve 2.0 g of Salicylic Acid to make exactly 100 mL. Pipet 10 mL of this solution, dilute in 25 mL of acetone, add 4 mL of sodium hydroxide TS, 2 with ethanol (95) to make exactly 100 mL, and use the solu- mL of dilute acetic acid and water to make 50 mL, and per- tion as the standard solution. Pipet 10 mL each of the sample form the test using this solution as the test solution. Prepare solution and standard solution into stoppered test tubes the control solution as follows: to 2.0 mL of Standard Lead respectively,toeachaddexactly5mLofasolutionofiron Solution add 25 mL of acetone, 2 mL of dilute acetic acid (III) nitrate enneahydrate (1 in 200), and dilute with and water to make 50 mL (not more than 10 ppm). hydrochloric acid-potassium chloride buŠer solution, pH (4) Readily carbonizable substances <1.15>—Perform the 2.0, to make exactly 25 mL. Determine the absorbances, AT test with 0.5 g of Salicylic Acid: the solution has no more and AS, of both solutions at 530 nm as directed under Ultrav- color than Matching Fluid C. iolet-visible Spectrophotometry <2.24>,usingasolutionpre- pared in the same manner with ethanol (95), instead of the Loss on drying <2.41> Not more than 0.5z (2 g, silica gel, 3 sample solution, as the blank. hours). Residue on ignition <2.44> Not more than 0.05z (1 g). Amount (mg) of salicylic acid (C7H6O3) ＝WS×(AT/AS)×(1/10) Assay Weigh accurately about 0.5 g of Salicylic Acid, previously dried, dissolve in 25 mL of neutralized ethanol, WS: Amount (mg) of salicylic acid for assay and titrate <2.50> with 0.1 molWL sodium hydroxide VS (indi- Containers and storage Containers—Well-closed contain- cator: 3 drops of phenolphthalein TS). ers. Each mL of 0.1 molWL sodium hydroxide VS

＝ 13.81 mg of C7H6O3 Containers and storage Containers—Well-closed contain- ers. 1080 Salicylic Acid Adhesive Plaster / O‹cial Monographs JP XV

200), dilute with hydrochloric acid-potassium chloride buŠer Salicylic Acid Adhesive Plaster solution, pH 2.0, to exactly 25 mL. Determine the absor- bances <2.24>, AT and AS, of both solutions at 530 nm, using サリチル酸絆創膏 a blank solution prepared in the same manner with water in- stead of the sample solution.

Method of preparation Amount (mg) of salicylic acid (C7H6O3) Adhesive Plaster consists of a mixture of the below in- ＝WS×(AT/AS) gredients with carefully selected rubber, resins, oxide W : Amount (mg) of salicylic acid for assay andothersubstances.Ithasadhesiveproperties.Itspreads S evenly on a fabric. Containers and storage Containers—Tight containers. Salicylic Acid, ˆnely powdered 500 g Adhesive plaster base a su‹cient quantity Compound Salicylic Acid Spirit To make 1000 g 複方サリチル酸精 Description The surface of Salicylic Acid Adhesive Plaster is whitish in color and adheres well to the skin. Containers and storage Containers—Well-closed contain- Compound Salicylic Acid Spirit contains not less ers. than 1.8 wWvz and not more than 2.2 wWvz of salicyl- Storage—Light-resistant. ic acid (C7H6O3: 138.12), and not less than 0.43 wWvz and not more than 0.53 wWvz of (C6H6O: 94.11). Salicylic Acid Spirit Method of preparation サリチル酸精 Salicylic Acid 20 g Liqueˆed Phenol 5 mL Glycerin 40 mL Salicylic Acid Spirit contains not less than 2.7 w Wvz Ethanol 800 mL and not more than 3.3 wWvz of salicylic acid (C7H6O3: Water or Puriˆed Water a su‹cient quantity 138.12). To make 1000 mL Method of preparation Prepare as directed under Medicated Spirits, with the Salicylic Acid 30 g above ingredients. Glycerin 50 mL Description Compound Salicylic Acid Spirit is a clear, Ethanol a su‹cient quantity colorless to light red liquid. 20 To make 1000 mL Speciˆc gravity d20: about 0.88 Prepare as directed under Medicated Spirits, with the Identiˆcation (1) To 1 mL of Compound Salicylic Acid above ingredients. Spirit add hydrochloric acid-potassium chloride buŠer solu- tion, pH 2.0, to make 200 mL, and to 5 mL of this solution Description Salicylic Acid Spirit is a clear, colorless liquid. add 5 mL of a solution of iron (III) nitrate enneahydrate (1 in Speciˆc gravity d20: about 0.86 20 200): a red-purple color is produced (salicylic acid). Identiˆcation The solution obtained in the Assay has a red- (2) To 1 mL of Compound Salicylic Acid Spirit add 20 purple color. Determine the absorption spectrum of the solu- mL of water and 5 mL of dilute hydrochloric acid, and ex- tion as directed under Ultraviolet-visible Spectrophotometry tract with 20 mL of diethyl ether. Wash the diethyl ether ex- <2.24>: it exhibits a maximum between 520 nm and 535 nm tract with two 5-mL portions of sodium hydrogen carbonate (salicylic acid). TS, and extract with 10 mL of dilute sodium hydroxide TS. Shake 1 mL of the extract with 1 mL of sodium TS and Alcohol number <1.01> Not less than 8.8 (Method 2). 1 mL of dilute hydrochloric acid, allow to stand for 10 Assay Measure exactly 10 mL of Salicylic Acid Spirit, add minutes, and add 3 mL of sodium hydroxide TS: a yellow 10 mL of ethanol (95) and water to make exactly 100 mL. color is produced (phenol). Pipet 3 mL of this solution, and dilute with hydrochloric (3) To 0.5 mL of Compound Salicylic Acid Spirit add 5 acid-potassium chloride buŠer solution, pH 2.0, to make ex- mL of dilute hydrochloric acid, extract with 5 mL of chlo- actly 100 mL. Use this solution as the sample solution. Dis- roform, and use the extract as the sample solution (1). To 2 solve about 0.3 g of salicylic acid for assay, previously dried mL of Compound Salicylic Acid Spirit add 5 mL of dilute in a desiccator (silica gel) for 3 hours and accurately weighed, hydrochloric acid, extract with 5 mL of chrloroform, wash in 10 mL of alcohol and water to make exactly 100 mL. Pipet the extract with two 5-mL portions of sodium hydrogen car- 3 mL of this solution, dilute with hydrochloric acid-potassi- bonate TS, and use the chloroform extract as the sample so- um chloride buŠer solution, pH 2.0, to make exactly 100 mL, lution (2). Separately, dissolve 0.01 g each of salicylic acid and use this solution as the standard solution. Pipet 10 mL and phenol in 5 mL each of chloroform, and use both solu- each of the sample solution and standard solution, to each tions as the standard solutions (1) and (2). Perform the test add 5 mL of a solution of iron (III) nitrate enneahydrate (1 in with these solutions as directed under Thin-layer Chro- JP XV O‹cial Monographs / Santonin 1081 matography <2.03>.Spot5mL each of the sample solutions and standard solutions on a plate of silica gel with ‰uorescent indicator for thin-layer chromatography. Develop the plate Santonin with a mixture of chloroform, acetone and acetic acid (100) (45:5:1) to a distance of about 10 cm, and air-dry the plate. サントニン Examine under ultraviolet light (main wavelength: 254 nm): the spots from the sample solution (1) and standard solution (1) show the same Rf value, and the spots from the sample so- lution (2) and standard solution (2) show the same Rfvalue. Spray evenly iron (III) chloride TS upon the plate: the spot from the standard solution (1) and the corresponding spot from the sample solution (1) reveal a purple color. Alcohol number <1.01> Not less than 7.5 (Method 2). C15H18O3: 246.30 (3S,3aS,5aS,9bS)-3,5a,9-Trimethyl-3a,5,5a,9b- Assay Measure accurately 2 mL of Compound Salicylic tetrahydronaphtho[1,2-b]furan-2,8(3H,4H )-dione Acid Spirit, add exactly 5 mL of the internal standard solu- [481-06-1] tionanddilutedmethanol(1in2)tomake100mL,anduse this solution as the sample solution. Weigh accurately about Santonin, when dried, contains not less than 98.5z 0.2 g of salicylic acid for assay, previously dried in a desicca- and not more than 101.0z of C15H18O3. tor (silica gel) for 3 hours, and about 50 mg of phenol for as- say, dissolve in diluted methanol (1 in 2) to make exactly 100 Description Santonin occurs as colorless crystals, or a mL. Pipet 20 mL of this solution, add exactly 5 mL of the in- white, crystalline powder. ternal standard solution and diluted methanol (1 in 2) to It is freely soluble in chloroform, sparingly soluble in make 100 mL, and use this solution as the standard solution. ethanol (95), and practically insoluble in water. Perform the test with 15 mL each of the sample solution and It becomes yellow by light. standard solution as directed under Liquid Chromatography Identiˆcation (1) Determine the absorption spectrum of a <2.01> according to the following conditions, and calculate solution of Cortisone Acetate in ethanol (95) (3 in 250,000) as the ratios, Q Ta and Q Tb, of the peak area of salicylic acid and directed under Ultraviolet-visible Spectrophotometry <2.24>, phenol to that of the internal standard in the sample solution, and compare the spectrum with the Reference Spectrum: and the ratios, Q Sa and Q Sb, of the peak area of salicylic acid both spectra exhibit similar intensities of absorption at the and phenol to that of the internal standard in the standard so- same wavelengths. lution. (2) Determine the infrared absorption spectrum of Corti- sone Acetate as directed in the potassium bromide disk Amount (mg) of salicylic acid (C7H6O3) method under Infrared Spectrophotometry <2.25>,andcom- ＝WSa×(QTa/QSa)×(1/5) pare the spectrum with the Reference Spectrum: both spectra Amount (mg) of phenol (C6H6O) exhibit similar intensities of absorption at the same wave ＝WSb×(QTa/QTb)×(1/5) numbers.

20 WSa: Amount (mg) of salicylic acid for assay Optical rotation <2.49> [a]D : －170 – －1759(0.2 g, chlo- WSb: Amount (mg) of phenol for assay roform, 10 mL, 100 mm). Internal standard solution—A solution of theophylline in Melting point <2.60> 172 – 1759C methanol (1 in 1250). Operating conditions— Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Detector: An ultraviolet absorption photometer Santonin according to Method 4, and perform the test. Pre- (wavelength: 270 nm). pare the control solution with 2.0 mL of Standard Lead Solu- Column: A stainless steel column about 4 mm in inside di- tion (not more than 20 ppm) ameter and 25 to 30 cm in length, packed with octadecyl- (2) Alkaloids—Boil 0.5 g of Santonin with 20 mL of silanized silica gel for liquid chromatography, 5 mminparti- diluted sulfuric acid (1 in 100), cool, and ˆlter. Dilute 10 mL cle diameter. of the ˆltrate with water to 30 mL, add 3 drops of iodine TS, Column temperature: Room temperature. and allow to stand for 3 hours: no turbidity is produced. Mobile phase: A mixture of 0.1 molWL phosphate buŠer (3) Artemisin—Dissolve 1.0 g of powdered Santonin in 2 solution, pH 7.0, and methanol (3:1). mL of chloroform by slight warming: the solution is clear Flow rate: Adjust the ‰ow rate so that the retention time of and colorless, or any yellow color produced is not darker salicylic acid is about 6 minutes. than Matching Fluid A. Selection of column: Dissolve 0.2 g of benzoic acid, 0.2 g (4) —Boil 0.20 g of Santonin with 10 mL of of salicylic acid and 0.05 g of theophylline in 100 mL of dilut- water, cool, and ˆlter. To the ˆltrate add bromine TS until ed methanol (1 in 2). To 10 mL of this solution add 90 mL of the color of the solution becomes yellow: no turbidity is pro- diluted methanol (1 in 2). Proceed with 10 mL of this solution duced. under the above operating conditions. Use a column giving e- (5) Acid-coloring substances—Moisten 10 mg of Santo- lution of benzoic acid, salicylic acid and theophylline in this nin with nitric acid: no color develops immediately. Moisten order, and clearly dividing each peak. Santonin with sulfuric acid, previously cooled to 09C: no color is produced immediately. Containers and storage Containers—Tight containers. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 1082 Scopolamine Butylbromide / O‹cial Monographs JP XV hours). drying, 1 g, water, 10 mL, 100 mm). Residue on ignition <2.44> Not more than 0.2z (1 g). pH <2.54> Dissolve 1.0 g of Scopolamine Butylbromide in 10 mL of water: the pH of this solution is between 5.5 and Assay Weigh accurately about 0.25 g of Santonin, previ- 6.5. ously dried, dissolve in 10 mL of ethanol (95) by warming, add exactly 20 mL of 0.1 mol/L sodium hydroxide VS, and Purity (1) Clarity and color of solution—Dissolve 1.0 g of heat on a water bath under a re‰ux condenser for 5 minutes. Scopolamine Butylbromide in 10 mL of water: the solution is Cool quickly, and titrate <2.50> the excess sodium hydroxide clear, and has no more color than the following control solu- with 0.05 mol/L hydrochloric acid VS (indicator: 3 drops of tion. phenolphthalein TS). Perform a blank determination. Control solution: To 0.5 mL of Matching Fluid F add diluted hydrochloric acid (1 in 40) to make 20 mL. Each mL of 0.1 mol/L sodium hydroxide VS (2) Heavy metals <1.07>—Proceed with 2.0 g of Scopola- ＝24.63 mg of C H O 15 18 3 mine Butylbromide according to Method 1, and perform the Containers and storage Containers—Tight containers. test. Prepare the control solution with 2.0 mL of Standard Storage—Light-resistant. Lead Solution (not more than 10 ppm). (3) Related substances—Dissolve 0.10 g of Scopolamine Butylbromide in the mobile phase to make exactly 10 mL, Scopolamine Butylbromide and use this solution as the sample solution. Separately, dis- solve 10 mg of scopolamine hydrobromide in the mobile ブチルスコポラミン臭化物 phase to make exactly 100 mL. Pipet 10 mL of this solution, add the mobile phase to make exactly 50 mL, and use this so- lution as the standard solution (1). Pipet 5 mL of the stan- dard solution (1), add the mobile phase to make exactly 10 mL, and use this solution as the standard solution (2). Per- form the test with exactly 20 mL each of the sample solution and standard solutions (1) and (2) as directed under Liquid Chromatography <2.01> according to the following condi- C21H30BrNO4:440.37 tions. Determine each peak area of these solutions by the au- (1S,2S,4R,5R,7s)-9-Butyl-7-[(2S)-3-hydroxy-2- tomatic integration method: the peak area of scopolamine phenylpropanoyloxy]-9-methyl-3-oxa-9- from the sample solution is not larger than that from the 2,4 azoniatricyclo[3.3.1.0 ]nonane bromide [149-64-4] standard solution (2), and each area of the peaks other than the peak appearing in the ˆrst elution and the peak of Scopolamine Butylbromide, when dried, contains scopolamine and butylscopolamine from the sample solution not less than 98.5z of C21H30BrNO4. are not larger than the peak area from the standard solution Description Scopolamine Butylbromide occurs as white (1). crystals or crystalline powder. Operating conditions— It is very soluble in water, freely soluble in acetic acid Detector: An ultraviolet absorption photometer (100), soluble in ethanol (95), sparingly soluble in methanol, (wavelength: 210 nm). slightly soluble in acetic anhydride, and practically insoluble Column: A stainless steel column 4.6 mm in inside di- in diethyl ether. ameter and 15 cm in length, packed with octylsilanized silica Melting point: about 1409C (with decomposition). gel for liquid chromatography (10 mminparticlediameter). Column temperature: A constant temperature of about Identiˆcation (1) To 1 mg of Scopolamine Butylbromide 309C. add 3 to 4 drops of fuming nitric acid, and evaporate on a Mobile phase: Dissolve 2 g of sodium lauryl sulfate in 370 water bath to dryness. After cooling, dissolve the residue in 1 mL of water and 680 mL of methanol, and adjust the pH to mL of N,N-dimethylformamide, and add 6 drops of 3.6withdilutedphosphoricacid(1in10). tetraethylammonium hydroxide TS: a red-purple color de- Flow rate: Adjust the ‰ow rate so that the retention time of velops. butylscopolamine is about 7 minutes. (2) Determine the absorption spectrum of a solution of Time span of measurement: About twice as long as the Scopolamine Butylbromide (1 in 1000) as directed under retention time of butylscopolamine. Ultraviolet-visible Spectrophotometry <2.24>,andcompare System suitability— the spectrum with the Reference Spectrum: both spectra ex- System performance: Dissolve 5 mg each of Scopolamine hibit similar intensities of absorption at the same Butylbromide and scopolamine hydrobromide in 50 mL of wavelengths. the mobile phase. When the procedure is run with 20 mLof (3) Determine the infrared absorption spectrum of this solution under the above operating conditions, scopola- Scopolamine Butylbromide, previously dried, as directed in mine and butylscopolamine are eluted in this order with the the potassium bromide disk method under Infrared Spec- resolution between these peaks being not less than 5. trophotometry <2.25>, and compare the spectrum with the System repeatability: When the test is repeated 6 times with Reference Spectrum: both spectra exhibit similar intensities 20 mL of the standard solution (2) under the above operating of absorption at the same wave numbers. conditions, the relative standard deviation of the peak area of (4) A solution of Scopolamine Butylbromide (1 in 20) scopolamine is not more than 2.0z. responds to the Qualitative Tests <1.09> for bromide. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 20 Optical rotation <2.49> [a]D : －18.0 – －20.09 (after JP XV O‹cial Monographs / Serrapeptase 1083

4 hours). (3) Apoatropine—Dissolve 0.20 g of Scopolamine Hydrobromide Hydrate in 20 mL of water, add 0.60 mL of Residue on ignition <2.44> Not more than 0.1z (1 g). 0.002 molWL VS, and allow to Assay Weigh accurately about 0.8 g of Scopolamine Butyl- stand for 5 minutes: the red color in the solution does not dis- bromide, previously dried, dissolve in 40 mL of acetic acid appear. (100) and 30 mL of acetic anhydride, and titrate <2.50> with (4) Related substances—Dissolve 0.15 g of Scopolamine 0.1 molWL perchloric acid VS (potentiometric titration). Per- Hydrobromide Hydrate in 3 mL of water, and use this solu- form a blank determination, and make any necessary correc- tion as the sample solution. tion. (i) To 1 mL of the sample solution add 2 to 3 drops of ammonia TS: no turbidity is produced. Each mL of 0.1 molWL perchloric acid VS (ii) To 1 mL of the sample solution add 2 to 3 drops of ＝ 44.04 mg of C H BrNO 21 30 4 potassium hydroxide TS: a transient white turbidity might be Containers and storage Containers—Tight containers. produced, and disappears clearly in a little while. Loss on drying <2.41> Not more than 13.0z [1.5 g; ˆrst dry in a desiccator (silica gel) for 24 hours, then dry at 1059Cfor Scopolamine Hydrobromide 3 hours]. Hydrate Residue on ignition <2.44> Not more than 0.1z (1 g). スコポラミン臭化水素酸塩水和物 Assay Weigh accurately about 0.5 g of Scopolamine Hydrobromide Hydrate, previously dried in 10 mL of acetic acid (100) by warming. After cooling, add 40 mL of acetic anhydride, and titrate <2.50> with 0.1 molWL perchloric acid VS (potentiometric titration). Perform a blank determina- tion, and make any necessary correction. Each mL of 0.1 molWL perchloric acid VS

＝ 38.43 mg of C17H21NO4.HBr C17H21NO4.HBr.3H2O: 438.31 (1S,2S,4R,5R,7s)-9-Methyl-3-oxa- Containers and storage Containers—Tight containers. 9-azatricyclo-[3.3.1.02,4]non-7-yl (2S)-3-hydroxy- Storage—Light-resistant. 2-phenylpropanoate monohydrobromide trihydrate [6533-68-2] Serrapeptase Scopolamine Hydrobromide Hydrate, when dried, contains not less than 98.5z of scopolamine セラペプターゼ hydrobromide (C17H21NO4.HBr: 384.26). Description Scopolamine Hydrobromide Hydrate occurs as [95077-02-4] colorless or white crystals, or white granules or powder. It is odorless. Serrapeptase is the enzyme preparation having pro- It is freely soluble in water, sparingly soluble in ethanol teolytic activity, produced by the growth of Serratia (95) and in acetic acid (100), and practically insoluble in species. diethyl ether. Usually, it is diluted with Lactose Hydrate. Identiˆcation (1) To 1 mg of Scopolamine Hydrobromide It contains not less than 2000 serrapeptase Units and Hydrate add 3 to 4 drops of fuming nitric acid, evaporate on not more than 2600 serrapeptase Units per mg. a water bath to dryness, and cool. Dissolve the residue in 1 It is hygroscopic. mL of N,N-dimethylformamide, and add 6 drops of Description Serrapeptase occurs as a grayish white to light tetraethylammonium hydroxide TS: a red-purple color is brown powder, having a slight characteristic odor. produced. (2) A solution of Scopolamine Hydrobromide Hydrate (1 Identiˆcation Dissolve 0.4 g of Serrapeptase in 100 mL of in 20) responds to the Qualitative Tests <1.09> for bromide. acetic acid-sodium acetate buŠer solution, pH 5.0, transfer exactly 1 mL each of this solution into three tubes, and refer 20 Optical rotation <2.49> [a]D : －24.0 – －26.09 (after to them as A, B and C. To tube A add exactly 1 mL of water, drying, 0.5 g, water, 10 mL, 100 mm). to tubes B and C add exactly 1 mL of 0.04 mol/L disodium Melting point <2.60> 195–1999C (after drying; previously dihydrogen ethylenediamine tetraacetate TS, mix gently, and heatthebathto1809C). allow them to stand in a water bath at 4±19C for about 1 hour. Then, to the tube B add exactly 2 mL of 0.04 mol/L Purity (1) Clarity and color of solution—Dissolve 0.5 g of zinc chloride TS, to the tubes A and C add exactly 2 mL of Scopolamine Hydrobromide Hydrate in 10 mL of water: the water, mix gently, and allow them to stand in a water bath at solution is clear and colorless. 4±19C for about 1 hour. Pipet 1 mL each of these solutions, (2) Acidity—Dissolve 0.50 g of Scopolamine add borate-hydrochloric acid buŠer solution, pH 9.0 to the Hydrobromide Hydrate in 15 mL of water, and add 0.50 mL solutions A and B to make exactly 200 mL, to the solution C of 0.02 molWL sodium hydroxide and 1 drop of methyl red- to make exactly 50 mL, and use these solutions as the sample TS: a green color develops. solutions. Proceed with these sample solutions as directed in 1084 Sesame Oil / O‹cial Monographs JP XV the Assay: the activities of the solutions A and B are almost make exactly 200 mL. Use after warming to 37±0.59C. Pre- the same, and the activity of the solution C is not more than pare before use. 5z of that of the solution A. (iv) Precipitation reagent: Trichloroacetic acid TS for serrapeptase. Use after warming to 37±0.59C. Activity of solutions A, B or C＝(A /A )×(1/20)×D×176 T S (v) Procedure: Pipet 1 mL of the sample solution, put in

AS: Absorbance of the standard solution a glass-stoppered tube (15×130 mm), allow to stand at 37± AT: Absorbance of the sample solution 0.59C for 5 minutes, add exactly 5 mL of the substrate 20: Reaction time (minute) solution, and mix well immediately. Allow to stand at 37± D: Dilution rate (200 for solution A and B, 50 for solution 0.59C for exactly 20 minutes, add exactly 5 mL of trichloroa- C) cetic acid TS for serrapeptase, mix, allow to stand at 37± 176: Conversion factor (Total volume of enzyme reaction 0.59C for 30 minutes, and ˆlter through a dried ˆlter paper. solution/volume of ˆltrate taken×amount of in 2 Pipet 2 mL of the ˆltrate, add exactly 5 mL of a solution of mL of tyrosine standard solution) anhydrous sodium carbonate (3 in 50), mix, add exactly 1 mL of diluted Folin's TS (1 in 3), mix well, and allow to stand at Purity (1) Heavy metals <1.07>—Put 1.0 g of Serrapep- 37±0.59C for 30 minutes. Determine the absorbance of this tase in a porcelain crucible, add 2 drops each of sulfuric acid solution at 660 nm, A , as directed under Ultraviolet-visible and nitric acid, and incinerate by ignition. After cooling, to 1 Spectrophotometry <2.24> using water as the blank. Separate- the residue add 2 mL of hydrochloric acid, evaporate to dry- ly, pipet 1 mL of the sample solution, add exactly 5 mL of ness on a water bath, add 10 mL of a solution of hydroxyla- trichloroacetic acid TS for serrapeptase, mix, add exactly 5 mine hydrochloride (3 in 100) and 2 mL of dilute acetic acid, mL of the substrate solution, allow to stand at 37±0.59Cfor and heat on a water bath for 5 minutes. After cooling, ˆlter if 30 minutes, and proceed in the same manner as directed necessary, wash the ˆlter paper with 10 mL of water, put the above to determine the absorbance A . Separately, pipet 2 ˆltrate and washing in a Nessler tube, add water to make 50 2 mL of the tyrosine standard solution, add exactly 5 mL of a mL, and use this solution as the test solution. Prepare the solution of anhydrous sodium carbonate (3 in 50), mix, add control solution as follows: Evaporate to dryness 2 drops exactly 1 mL of diluted Folin's TS (1 in 3), mix well, and pro- each of sulfuric acid and nitric acid on a sand bath, add 2 mL ceed in the same manner as directed above to determine the of hydrochloric acid to the residue, evaporate to dryness on a absorbance A . Separately, pipet 2 mL of 0.2 mol/L water bath, add 2.0 mL of Standard Lead Solution, 10 mL of 3 hydrochloric acid TS, and proceed in the same manner as a solution of hydroxylamine hydrochloride (3 in 100) and 2 directed above to determine the absorbance A . mL of dilute acetic acid, and heat on a water bath for 4 5 minutes. Proceed in the same manner as directed for the Serrapeptase Unit per mg of Serrapeptase preparation of the test solution, and add water to make 50 ＝｛(A1－A2)/(A3－A4)｝×(1/20)×200×176 mL (not more than 20 ppm). 20: Reaction time (minute) (2) Arsenic <1.11>—Prepare the test solution with 0.40 g 200: Dilution rate of Serrapeptase according to Method 3, excepting addition of 176: Conversion factor (Total volume of enzyme reaction 5 mL of a solution of nitrate hexahydrate in solution/volume of ˆltrate taken×amount of tyro- ethanol (95) (3 in 10) instead of a solution of magnesium ni- sine in 2 mL of tyrosine standard solution) trate hexahydrate in ethanol (95) (1 in 50), evaporating to One serrapeptase Unit corresponds to the amount of dryness on a water bath, then incinerating with a small ‰ame, serrapeptase which produces 5 mgoftyrosineperminute and perform the test (not more than 5 ppm). from 5 mL of the substrate solution under the above condi- Loss on drying <2.41> Not more than 7.0z (1 g, 1059C, 4 tions. hours). Containers and storage Containers—Tight containers. Residue on ignition <2.44> Not more than 1.5z (1 g). Assay (i) Sample solution: Dissolve exactly 0.100 g of Ser- rapeptase in a solution of ammonium sulfate (1 in 20) to Sesame Oil make exactly 100 mL. Pipet 1 mL of this solution, add borate-hydrochloric acid buŠer solution, pH 9.0 to make Oleum Sesami exactly 200 mL, and use this solution as the sample solution. ゴマ油 (ii) Tyrosine standard solution: Dissolve exactly 0.160 g of Tyrosine Reference Standard, previously dried at 1059C for 3 hours, in 0.2 mol/L hydrochloric acid TS to make Sesame Oil is the ˆxed oil obtained from the seeds of exactly 1000 mL. Pipet 10 mL of this solution, and add Sesamum indicum Linnáe (Pedaliaceae). 0.2 mol/L hydrochloric acid TS to make exactly 100 mL. Description Sesame Oil is a clear, pale yellow oil. It is odor- Prepare before use. less or has a faint, characteristic odor, and has a bland taste. (iii) Substrate solution: Previously determine the loss on It is miscible with diethyl ether and with petroleum ether. drying <2.41> (609C, reduced pressure not exceeding 0.67 It is slightly soluble in ethanol (95). kPa, 3 hours) of milk casein, previously dried. To exactly It congeals between 09Cand－59C. 1.20 g of the milk casein, calculated based on the loss on Congealing point of the fatty acids: 20 – 259C drying, add 160 mL of a solution of sodium borate (19 in 1000), and heat in a water bath to dissolve. After cooling, ad- Identiˆcation To 1 mL of Sesame Oil add 0.1 g of sucrose just the pH to exactly 9.0 with 1 mol/L hydrochloric acid TS, and 10 mL of hydrochloric acid, and shake for 30 seconds: and add borate-hydrochloric acid buŠer solution, pH 9.0 to the acid layer becomes light red and changes to red on stand- JP XV O‹cial Monographs / White Shellac 1085 ing. shaking on a water bath, and continue the heating for 2 hours. After cooling, collect the ‰oating wax by ˆltration, Speciˆc gravity <1.13> d20: 0.914 – 0.921 20 wash the wax and the ˆlter paper with water, transfer to a Acid value <1.13> Not more than 0.2. beaker, and dry at 659C until the water is almost evaporated. Transfer the wax together with the ˆlter paper to an extrac- Saponiˆcation value <1.13> 187 – 194 tion thimble in a Soxhlet extractor. Dissolve the wax remain- Unsaponiˆable matters <1.13> Not more than 2.0z. ing in the beaker with a suitable quantity of chloroform by warming. Pour the solution into the thimble, and extract with Iodine value <1.13> 103 – 118 chloroform for 2 hours. Evaporate the chloroform solution Containers and storage Containers—Tight containers. to dryness, ad dry the residue at 1059C for 3 hours: the mass of the residue is not more than 20 mg. Loss on drying Not more than 2.0z. Weigh accurately Puriˆed Shellac about 1 g of medium powder of Puriˆed Shellac, and dry at 409C for 4 hours, then for 15 hours in a desiccator (calcium 精製セラック chloride for drying). Total ash <5.01> Not more than 1.0z (1 g). Puriˆed Shellac is a resin-like substance obtained Containers and storage Containers—Well-closed contain- from a puriˆed secretion of Laccifer lacca Kerr (Cocci- ers. dae). Description Puriˆed Shellac occurs as light yellow-brown to brown, lustrous, hard, brittle scutella. It has no odor or has a White Shellac faint, characteristic odor. It is freely soluble in ethanol (95) and in ethanol (99.5), and 白色セラック practically insoluble in water and in diethyl ether. It dissolves in sodium hydroxide TS. White Shellac is a resin-like substance obtained from Acid value 60–80 <1.13> Weigh accurately about 1 g of a bleached secretion of Laccifer lacca Kerr (Coccidae). Puriˆed Shellac, add 40 mL of neutralized ethanol, and dis- solve by warming. After cooling, titrate <2.50> with 0.1 molW Description White Shellac occurs as yellowish white to light L potassium hydroxide VS (potentiometric titration). yellow, hard, brittle granules. It is odorless or has a faint, characteristic odor. Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of It is sparingly soluble in ethanol (95), very slightly soluble Puriˆed Shellac according to Method 2, and perform the test. in petroleum ether, and practically insoluble in water. Prepare the control solution with 2.0 mL of Standard Lead It dissolves in sodium hydroxide TS. Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g Acid value <1.13> 65 – 90 Weigh accurately about 0.5 g of of Puriˆed Shellac according to Method 3, and perform the White Shellac, add 50 mL of neutralized ethanol as a solvent, test. Add 10 mL of a solution of magnesium nitrate hexahy- and dissolve by warming. After cooling, perform the test as drate in ethanol (95) (1 in 50), then add 1.5 mL of hydrogen directed in the Acid value under Puriˆed Shellac. peroxide (30), and ˆre to burn (not more than 5 ppm). Purity (1) Chloride <1.03>—Shake and dissolve 0.40 g of (3) Ethanol-insoluble substances—Dissolve about 5 g of White Shellac in 5 mL of ethanol (95) while warming, add 40 Puriˆed Shellac, accurately weighed, in 50 mL of ethanol (95) mL of water, and cool. Add 12 mL of dilute nitric acid and on a water bath while shaking. Pour the ethanol solution into water to make 100 mL, and ˆlter. Perform the test using 50 a tared extraction thimble, previously dried at 1059Cfor2 mL of the ˆltrate as the test solution. Prepare the control so- hours, in a Soxhlet extractor, and extract with ethanol (95) lution as follows: to 0.80 mL of 0.01 molWL hydrochloric for 3 hours: the mass of the residue is not more than 2.0z. acid VS add 2.5 mL of ethanol (95), 6 mL of dilute nitric acid Use a cylindrical weighing bottle for taring the extraction and water to make 50 mL (not more than 0.140z). thimble. (2) Sulfate <1.14>—Shake and dissolve 0.40 g of White (4) Rosin—Dissolve 2.0 g of Puriˆed Shellac in 10 mL of Shellac in 5 mL of ethanol (95) by warming, add 40 mL of ethanol (99.5) with thorough shaking, add gradually 50 mL water, and cool. Add 2 mL of dilute hydrochloric acid and of petroleum ether while shaking, and ˆlter, if necessary. water to make 100 mL, and ˆlter. Perform the test using 50 Wash the solution with two 50-mL portions of water, ˆlter mL of the ˆltrate as the test solution. Prepare the control so- the upper layer, and evaporate the ˆltrate on a water bath to lution as follows: to 0.45 mL of 0.005 molWL sulfuric acid VS dryness. Dissolve the residue in 2 mL of a mixture of carbon add 2.5 mL of ethanol (95), 1 mL of dilute hydrochloric acid tetrachloride and phenol (2:1), transfer the solution to a and water to make 50 mL (not more than 0.110z). depression of a spot plate, and ˆll the neighboring depression (3) Heavy metals <1.07>—Proceed with 2.0 g of White with a mixture of carbon tetrachloride and bromine (4:1). Shellac according to Method 2, and perform the test. Prepare Immediately cover both depressions with a watch glass, and the control solution with 2.0 mL of Standard Lead Solution allow to stand: the solution of the residue exhibits no purple (not more than 10 ppm.) or blue color within 1 minute. (4) Arsenic <1.11>—Prepare the test solution with 0.40 g (5) Wax—Dissolve 10.0 g of Puriˆed Shellac in 150 mL of White Shellac according to Method 3, and perform the of a solution of sodium carbonate decahydrate (9 in 200) with test. Add 10 mL of a solution of magnesium nitrate hexahy- 1086 Siccanin / O‹cial Monographs JP XV drate in ethanol (95) (1 in 50), then add 1.5 mL of hydrogen It contains not less than 980 mg (potency) and not peroxide (30), and ˆre to burn (not more than 5 ppm). more than 1010 mg (potency) per mg, calculated on the (5) Ethanol-insoluble substances—Dissolve about 5 g of dried basis. The potency of Siccanin is expressed as White Shellac, accurately weighed, in 50 mL of ethanol (95) mass (potency) of siccanin (C22H30O3). on a water bath while shaking. Pour the ethanol solution into Description Siccanin occurs as white to light yellow, crys- a tared extraction thimble, previously dried at 1059Cfor2 tals or crystalline powder. hours, in a Soxhlet extractor, and extract with ethanol (95) It is freely soluble in acetone, soluble in methanol and in for 3 hours: the mass of the residue is not more than 2.0z. ethanol (99.5), and practically insoluble in water. Use a cylindrical weighing bottle for taring the extraction thimble. Identiˆcation (1) Determine the absorption spectrum of a (6) Rosin—Dissolve 2.0 g of White Shellac in 10 mL of solution of Siccanin in ethanol (99.5) (1 in 10,000) as directed ethanol (99.5) with thorough shaking, add gradually 50 mL under Ultraviolet-visible Spectrophotometry <2.24>,and of petroleum ether while shaking, and ˆlter, if necessary. compare the spectrum with the Reference Spectrum or the Wash the solution with two 50-mL portions of water, ˆlter spectrum of a solution of Siccanin Reference Standard ob- the upper layer, and evaporate the ˆltrate on a water bath to tained in the same manner as the sample solution: both spec- dryness. Dissolve the residue in 2 mL of a mixture of carbon tra exhibit similar intensities of absorption at the same tetrachloride and phenol (2:1), transfer the solution to a wavelengths. depression of a spot plate, and ˆll the neighboring depression (2) Determine the infrared absorption spectrum of Sicca- with a mixture of carbon tetrachloride and bromine (4:1). nin as directed in the potassium bromide disk method under Immediately cover both depressions with a watch glass, and the Infrared Spectrophotometry <2.25>, and compare the allow to stand: the solution of the residue exhibits no purple spectrum with the Reference Spectrum or the spectrum of or blue color within 1 minute. Siccanin Reference Standard: both spectra exhibit similar in- (7) Wax—Dissolve 10.0 g of White Shellac in 150 mL of tensities of absorption at the same wave numbers. a solution of sodium carbonate decahydrate (9 in 200) with Optical rotation <2.49> [a]20: －165 – －1759(0.1 g, ethanol shaking on a water bath, and continue the heating for 2 D (99.5), 10 mL, 100 mm). hours. After cooling, collect the ‰oating wax by ˆltration, wash the wax and the ˆlter paper with water, transfer to a Melting point <2.60> 138 – 1429C beaker, and dry at 659C until the water is almost evaporated. Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Transfer the wax together with the ˆlter paper to an extrac- Siccanin according to Method 4, and perform the test. Pre- tion thimble in a Soxhlet extractor. Dissolve the wax remain- pare the control solution with 2.0 mL of Standard Lead Solu- ing in the beaker with a suitable quantity of chloroform by tion (not more than 20 ppm). warming. Pour the solution into the thimble, and extract with (2) Related substances—Dissolve 0.20 g of Siccanin in 10 chloroform for 2 hours. Evaporate the chloroform solution mL of acetone, and use this solution as the sample solution. to dryness, and dry the residue at 1059C for 3 hours: the mass Pipet 1 mL of the sample solution, add acetone to make ex- of the residue is not more than 20 mg. actly 200 mL, and use this solution as the standard solution. Loss on drying Not more than 6.0z. Weigh accurately Perform the test with these solutions as directed under Thin- about 1 g of medium powder of White Shellac, and dry at 409 layer Chromatography <2.03>.Spot5mLeachofthesample C for 4 hours, then for 15 hours in a desiccator (calcium chlo- solution and standard solution on a plate of silica gel for ride for drying). thin-layer chromatography. Develop the plate with a mixture of cyclohexane and acetone (5:1) to a distance of about 10 Total ash <5.01> Not more than 1.0z (1 g). cm, and air-dry the plate. Spray evenly 4-chlorobenzenedia- Containers and storage Containers—Well-closed contain- zonium TS on the plate: the number of the spots other than ers. the principal spot obtained from the sample solution is not Storage—In a cold place. more than three, and they are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, reduced Siccanin pressure not exceeding 0.67 kPa, 809C, 3 hours). シッカニン Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately an amount of Siccanin and Sicca- nin Reference Standard, equivalent to about 50 mg (poten- cy), dissolve each in the internal standard solution to make exactly 50 mL, and use these solutions as the sample solution and the standard solution. Perform the test with 10 mLeach of the sample solution and standard solution as directed un- C22H30O3: 342.47 der Liquid Chromatography <2.01> according to the follow- (4aS,6aS,11bR,13aS,13bS )-4,4,6a,9-Tetramethyl- ing conditions, and determine the ratios, Q T and QS,ofthe 1,2,3,4,4a,5,6,6a,11b,13b-decahydro-13H- peak area of siccanin to that of the internal standard. benzo[a]furo[2,3,4-mn]xanthen-11-ol [22733-60-4 ] Amount [ mg (potency)] of C22H30O3 ＝ W × (Q /Q ) × 1000 Siccanin is a substance having activity S T S produced by the growth of Helminthosporium siccans. WS: Amount [mg (potency)] of Siccanin Reference Stand- JP XV O‹cial Monographs / Light Anhydrous Silicic Acid 1087

ard drous Silicic Acid in 20 mL of sodium hydroxide TS by boil- ing, cool, ˆlter if necessary, and wash with 10 mL of water. Internal standard solution—A solution of 1,4-diphenylben- Combine the ˆltrate and washings, add 18 mL of dilute nitric zene in methanol (1 in 30,000). acid, shake, and add water to make 50 mL. Perform the test Operating conditions— using this solution as the test solution. To 0.15 mL of 0.01 Detector: An ultraviolet absorption photometer molWL hydrochloric acid VS add 20 mL of sodium hydroxide (wavelength: 280 nm). TS, 18 mL of dilute nitric acid and water to make 50 mL, and Column: A stainless steel column 4.6 mm in inside di- use this solution as the control solution (not more than ameter and 15 cm in length, packed with octadecylsilanized 0.011z). silica gel for liquid chromatography (5 mminparticledi- (2) Heavy metals <1.07>—Dissolve 0.5 g of Light Anhy- ameter). drous Silicic Acid in 20 mL of sodium hydroxide TS by boil- Column temperature: A constant temperature of about ing, cool, add 15 mL of acetic acid (31), shake, ˆlter if neces- 409C. sary, wash with 10 mL of water, combine the ˆltrate and Mobile phase: A mixture of methanol and phosphate washings, and add water to make 50 mL. Perform the test us- buŠer solution, pH 5.9 (19:6). ing this solution as the test solution. Add acetic acid (31) to Flow rate: Adjust the ‰ow rate so that the retention time of 20 mL of sodium hydroxide TS and 1 drop of siccanin is about 17 minutes. phenolphthalein TS until the color of this solution disap- System suitability— pears, add 2.0 mL of Standard Lead Solution, 2 mL of dilute System performance: When the procedure is run with 10 acetic acid and water to make 50 mL, and use this solution as mL of the standard solution under the above operating condi- the control solution (not more than 40 ppm). tions, siccanin and the internal standard are eluted in this (3) Aluminum—Dissolve 0.5 g of Light Anhydrous Silic- order with the resolution between these peaks being not less ic Acid in 40 mL of sodium hydroxide TS by boiling, cool, than 3. add sodium hydroxide TS to make 50 mL, and ˆlter. Meas- System repeatability: When the test is repeated 6 times with ure 10 mL of the ˆltrate, add 17 mL of acetic acid (31), 10 mL of the standard solution under the above operating shake, add 2 mL of aluminon TS and water to make 50 mL, conditions, the relative standard deviation of the ratios of the and allow to stand for 30 minutes: the color of this solution is peak area of siccanin to that of the internal standard is not not deeper than that of the following control solution. more than 1.0z. Control solution: Dissolve 0.176 g of aluminum potassium Containers and storage Containers—Tight containers. sulfate 12-water in water, and add water to make 1000 mL. To 15.5 mL of this solution add 10 mL of sodium hydroxide TS, 17 mL of acetic acid (31), 2 mL of aluminon TS and Light Anhydrous Silicic Acid water to make 50 mL. (4) Iron <1.10>—To 40 mg of Light Anhydrous Silicic 軽質無水ケイ酸 Acid add 10 mL of dilute hydrochloric acid, and heat for 10 minutes in a water bath while shaking. After cooling, add 0.5 gofL-tartaric acid to dissolve by shaking. Prepare the test so- Light Anhydrous Silicic Acid, calculated on the in- lution with this solution according to Method 2, and perform cinerated basis, contains not less than 98.0z of silicon the test according to Method B. Prepare the control solution dioxide (SiO2: 60.08). with 2.0 mL of Standard Iron Solution (not more than 500 ppm). Description Light Anhydrous Silicic Acid occurs as a white (5) Calcium—Dissolve 1.0 g of Light Anhydrous Silicic to bluish white, light, ˆne power. It is odorless and tasteless, Acid in 30 mL of sodium hydroxide TS by boiling, cool, add andsmoothtothetouch. 20 mL of water, 1 drop of phenolphthalein TS and dilute It is practically insoluble in water, in ethanol (95), and in nitric acid until the color of this solution disappears, immedi- diethyl ether. ately add 5 mL of dilute acetic acid, shake, add water to It dissolves in hydro‰uoric acid, in hot potassium make 100 mL, and obtain a clear liquid by centrifugation or hydroxide TS and in hot sodium hydroxide TS, and does not ˆltration. To 25 mL of this liquid add 1 mL of oxalic acid TS dissolve in dilute hydrochloric acid. and ethanol (95) to make 50 mL, immediately shake, and al- Identiˆcation (1) Dissolve 0.1 g of Light Anhydrous Silic- low to stand for 10 minutes: the turbidity of this solution is ic Acid in 20 mL of sodium hydroxide TS by boiling, and add not deeper than that of the following control solution. 12 mL of ammonium chloride TS: a white, gelatinous Control solution: Dissolve 0.250 g of calcium carbonate, precipitate is produced. The precipitate does not dissolve in previously dried at 1809C for4hours,in3mLofdilute dilute hydrochloric acid. hydrochloric acid, and add water to make 100 mL. To 4 mL (2) To the precipitate obtained in (1) add 10 mL of a solu- of this solution add 5 mL of dilute acetic acid and water to tion of methylene blue trihydrate (1 in 10,000), and wash with make 100 mL. To 25 mL of this solution add 1 mL of oxalic water: the precipitate has a blue color. acid TS and ethanol (95) to make 50 mL, and shake. (3) Prepare a bead by fusing ammonium sodium (6) Arsenic <1.11>—Dissolve 0.40 g of Light Anhydrous hydrogenphosphate tetrahydrate on a platinum loop. Bring Silicic Acid in 10 mL of sodium hydroxide TS by boiling in a the hot, transparent bead into contact with Light Anhydrous porcelain crucible, cool, add 5 mL of water and 5 mL of di- Silicic Acid, and fuse again: an insoluble matter is perceptible lute hydrochloric acid, shake, and perform the test with this in the bead. The resulting bead, upon cooling, becomes solution as the test solution (not more than 5 ppm). opaque and acquires a reticulated appearance. Loss on drying <2.41> Not more than 7.0z (1 g, 1059C, Purity (1) Chloride <1.03>—Dissolve 0.5 g of Light Anhy- 1088 Silver Nitrate / O‹cial Monographs JP XV

4 hours). ＝ 16.99 mg of AgNO3 Loss on ignition <2.43> Not more than 12.0z (1 g, 850 – Containers and storage Containers—Tight containers. 9009C, constant mass). Storage—Light-resistant. Volume test Weigh 5.0 g of Light Anhydrous Silicic Acid, transfer gradually to a 200-mL measuring cylinder, and allow to stand: the volume is not less than 70 mL. Silver Nitrate Ophthalmic Solution Assay Weigh accurately about 1 g of Light Anhydrous Si- 硝酸銀点眼液 licic Acid, add 20 mL of hydrochloric acid, and evaporate to dryness on a sand bath. Moisten the residue with hydrochlor- ic acid, evaporate to dryness, and heat between 1109Cand Silver Nitrate Ophthalmic Solution is an aqueous eye 1209C for 2 hours. Cool, add 5 mL of dilute hydrochloric lotion. acid, and heat. Allow to cool to room temperature, add 20 to It contains not less than 0.95 wWvz and not more 25 mL of hot water, ˆlter rapidly, and wash the residue with than 1.05 wWvz of silver nitrate (AgNO3: 169.87). warm water until the last washing becomes negative to the Method of preparation Qualitative Tests <1.09> (2) for chloride. Transfer the residue together with the ˆlter paper to a platinum crucible, ignite to Silver Nitrate 10 g ash, and continue the ignition for 30 minutes. Cool, weigh Puriˆed Water a su‹cient quantity the crucible, and designate the mass as a (g). Moisten the To make 1000 mL residue in the crucible with water, add 6 mL of hydro‰uoric acid and 3 drops of sulfuric acid, and evaporate to dryness. Prepare as directed under Ophthalmic Solution, with the Heat strongly for 5 minutes, cool, weigh the crucible, and above ingredients. designate the mass as b (g). Description Silver Nitrate Ophthalmic Solution is a clear,

Content (g) of silicon dioxide (SiO2)＝a－b colorless liquid. Containers and storage Containers—Tight containers. Identiˆcation Silver Nitrate Ophthalmic Solution responds to the Qualitative Tests <1.09> for silver salt and for nitrate. Assay Measure accurately 20 mL of Silver Nitrate Silver Nitrate Ophthalmic Solution, add 30 mL of water and 2 mL of nitric acid, and titrate <2.50> with 0.1 molWL ammonium thioc- 硝酸銀 yanate VS (indicator: 2 mL of ammonium iron (III) sulfate TS).

AgNO3: 169.87 Each mL of 0.1 molWL ammonium thiocyanate VS ＝16.99 mg of AgNO3 Silver Nitrate, when dried, contains not less than Containers and storage Containers—Tight containers. 99.8z of AgNO3. Storage—Light-resistant. Description Silver Nitrate occurs as lustrous, colorless or white crystals. It is very soluble in water, soluble in ethanol (95), and prac- Silver Protein tically insoluble in diethyl ether. It gradually turns grayish black by light. プロテイン銀 Identiˆcation A solution of Silver Nitrate (1 in 50) responds to the Qualitative Tests <1.09> forsilversaltandfornitrate. Silver Protein is a compound of silver and proteins. Purity (1) Clarity and color of solution, and acidity or It contains not less than 7.5z and not more than alkalinity—Dissolve 1.0 g of Silver Nitrate in 10 mL of fresh- 8.5z of silver (Ag: 107.87). ly boiled and cooled water: the solution is clear and colorless. Description Silver Protein occurs as a light yellow-brown to It is neutral. brown powder. It is odorless. It (1 g) dissolves slowly in 2 mL (2) Bismuth, copper and lead—To 5 mL of a solution of of water. It is practically insoluble in ethanol (95), in diethyl Silver Nitrate (1 in 10) add 3 mL of ammonia TS: the solution ether and in chloroform. is clear and colorless. The pH of a solution of Silver Protein (1 in 10) is between Loss on drying <2.41> Not more than 0.20z (2 g, silica gel, 7.0 and 8.5. light resistant, 4 hours). It is slightly hygroscopic. It is aŠected by light. Assay Weigh accurately about 0.7 g of Silver Nitrate, previ- ously powdered and dried, dissolve in 50 mL of water, add 2 Identiˆcation (1) To 10 mL of a solution of Silver Protein mL of nitric acid, and titrate <2.50> with 0.1 molWL ammoni- (1 in 100) add 2 mL of dilute hydrochloric acid, shake fre- um thiocyanate VS (indicator: 2 mL of ammonium iron (III) quently for 5 minutes, and ˆlter. To the ˆltrate add 5 mL of a sulfate TS). solution of sodium hydroxide (1 in 10), and add 2 mL of diluted copper (II) sulfate TS (2 in 25): a purple color de- Each mL of 0.1 molWL ammonium thiocyanate VS velops. JP XV O‹cial Monographs / Simple Syrup 1089

(2) To 5 mL of a solution of Silver Protein (1 in 100) add heat cautiously, and evaporate almost to dryness. Then in- dropwise iron (III) chloride TS: the color of the solution cinerate gradually by strong heating, dissolve the residue in 1 fades and a precipitate is gradually formed. mL of nitric acid by warming, and add 10 mL of water: the (3) Incinerate 0.2 g of Silver Protein by strong heating, solution responds to the Qualitative Tests <1.09> (1) for silver dissolve the residue in 1 mL of nitric acid by warming, and salt. add 10 mL of water: this solution responds to the Qualitative Assay Pipet 25 mL of Silver Protein Solution into a 250-mL Tests <1.09> (1) for silver salt. Kjeldahl ‰ask, and heat cautiously until a white gas of glyce- Purity Silver salt—Dissolve 0.10 g of Silver Protein in 10 rin is evolved. After cooling, add 25 mL of sulfuric acid, mL of water, and ˆlter. To the ˆltrate add 1 mL of potassium cover the ‰ask with a funnel, and heat gently for 5 minutes. chromate TS: no turbidity is produced. After cooling, drop gradually 5 mL of nitric acid, heat with occasional shaking in a water bath for 45 minutes, and cool. Assay Transfer about 1 g of Silver Protein, accurately Add 2 mL of nitric acid, boil gently, and repeat this opera- weighed, to a 100-mL decomposition ‰ask, add 10 mL of sul- tion until the solution becomes colorless upon cooling. furic acid, cover the ‰ask with a funnel, and boil for 5 Transfer cautiously the cooled content in the ‰ask into a minutes. Cool, add dropwise 3 mL of nitric acid with cau- 500-mL conical ‰ask with 250 mL of water. Boil gently for 5 tion, and heat for 30 minutes without boiling. Cool, add 1 minutes, cool, and titrate <2.50> with 0.1 molWLammonium mL of nitric acid, boil, and, if necessary, repeat this opera- thiocyanate VS (indicator: 3 mL of ammonium iron (III) sul- tion until the solution becomes colorless. After cooling, fate TS). transfer the solution to a 250-mL conical ‰ask with 100 mL of water, and titrate <2.50> with 0.1 molWL ammonium thioc- Each mL of 0.1 molWL ammonium thiocyanate VS yanate VS (indicator: 3 mL of ammonium iron (III) sulfate ＝10.79 mg of Ag TS). Containers and storage Containers—Tight containers. Each mL of 0.1 molWL ammonium thiocyanate VS Storage—Light-resistant. ＝10.79 mg of Ag Containers and storage Containers—Tight containers. Storage—Light-resistant. Simple Ointment 単軟膏 Silver Protein Solution Method of preparation プロテイン銀液 Yellow Beeswax 330 g Fixed oil a su‹cient quantity Silver Protein Solution contains not less than 0.22 To make 1000 g wWvz and not more than 0.26 wWvz of silver (Ag: Prepare as directed under Ointments, with the above in- 107.87). gredients. Method of preparation Description Simple Ointment is yellow in color. It has a Silver Protein 30 g slight, characteristic odor. Glycerin 100 mL Containers and storage Containers—Tight containers. Mentha Water a su‹cient quantity To make 1000 mL Dissolve and mix the above ingredients. Simple Syrup Description Silver Protein Solution is a clear, brown liquid, 単シロップ having the odor of mentha oil. Identiˆcation (1) To1mLofSilverProteinSolutionadd 10 mL of ethanol (95), mix, and add 2 mL of sodium Simple Syrup is an aqueous solution of Sucrose. hydroxide TS. Add immediately 1 mL of a solution of copper Method of preparation (II) chloride dihydrate in ethanol (95) (1 in 10), shake, and Sucrose 850 g ˆlter: the ˆltrate is blue in color (glycerin). Puriˆed Water a su‹cient quantity (2) To 3 mL of Silver Protein Solution add water to make 10 mL, add 2 mL of dilute hydrochloric acid, shake frequent- To make 1000 mL ly for 5 minutes, and ˆlter. Add 5 mL of a solution of sodium Prepare as directed under Syrups, with the above materi- hydroxide (1 in 10) to the ˆltrate, and add 2 mL of diluted als. copper (II) sulfate TS (2 in 25): a purple color develops (silver protein). Description Simple Syrup is a clear, colorless to pale yel- (3) To 5 mL of the sample solution obtained in (2) add low, viscous liquid. iron (III) chloride TS dropwise: a brown precipitate is formed It is odorless and has a sweet taste. (silver protein). Identiˆcation (1) Evaporate Simple Syrup on a water bath (4) Place 3 mL of Silver Protein Solution in a crucible, 1090 Sisomicin Sulfate / O‹cial Monographs JP XV to dryness. 1 g of the residue so obtained, when ignited, melts (2) Dissolve 15 mg each of Sisomicin Sulfate and Sisomi- to swell, and decomposes, emitting an odor of caramel, to cin Sulfate Reference Standard in 5 mL of water, and use bulky charcoal. these solutions as the sample solution and the standard solu- (2) To 0.1 g of the residue obtained in (1) add 2 mL of di- tion. Perform the test with these solutions as directed under lute sulfuric acid, boil, add 4 mL of sodium hydroxide TS Thin-layer Chromatography <2.03>.Spot5mLeachofthe and 3 mL of Fehling's TS, and heat to boiling: a red to dark sample solution and standard solution on a plate of silica gel red precipitate is produced. for thin-layer chromatography. Develop the plate with a mix- ture of methanol, chloroform, ammonia water (28) and ace- Speciˆc gravity <2.56> d20: 1.310 – 1.325 20 tone (2:2:1:1) to a distance of about 15 cm, and air-dry the Purity (1) Artiˆcial sweetening agents—To 100 mL of plate. Spray evenly 0.2z ninhydrin-water satutated 1- Simple Syrup add 100 mL of water, shake, acidify a 50-mL butabol TS on the plate, and heat at 1009C for 5 minutes: the portion of the solution with dilute sulfuric acid, and make principal spots from the sample solution and standard solu- another 50-mL portion alkaline with sodium hydroxide TS. tion exhibit a red-purple to red-brown color and show the To each portion add 100 mL of diethyl ether, shake, separate same Rfvalue. the diethyl ether layer, and evaporate the combined diethyl (3) A solution of Sisomicin Sulfate (1 in 100) responds to ether extract on a water bath to dryness: the residue has no the Qualitative Tests <1.09> (1) for sulfate. sweet taste. Optical rotation <2.49> [a]20: ＋100 – ＋1109(0.25 g calcu- (2) Salicylic acid—To the residue obtained in (1) add 2 to D latedonthedriedbasis,water,25mL,100mm). 3 drops of dilute iron (III) chloride TS: no purple color de- velops. pH <2.54> Dissolve 0.5 g of Sisomicin Sulfate in 5 mL of water: the pH of the solution is between 3.5 and 5.5. Containers and storage Containers—Tight containers. Purity (1) Clarity and color of solution—Dissolve 0.5 g of Sisomicin Sulfate in 5 mL of water: the solution is clear and Sisomicin Sulfate colorless to light yellow. (2) Heavy metals <1.07>—Proceed with 1.0 g of Sisomi- シソマイシン硫酸塩 cin Sulfate according to Method 2, and perform the test. Pre- pare the control solution with 2.0 mL of Standard Lead Solu- tion (not more than 20 ppm). (3) Related substances—Dissolve 50 mg of Sisomicin Sul- fate, calculated on the dried basis, in water to make 10 mL, and use this solution as the sample solution. Pipet 0.5 mL, 1 mL and 1.5 mL of the sample solution, add water to each to make exactly 50 mL, and use these solutions as the standard solution (1), the standard solution (2) and the standard solu- tion (3), respectively. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>.Spot10 mL each of the sample solution and standard solutions (1), (2) and (3) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol, chloroform, 1 ammonia water (28) and acetone (2:2:1:1) to a distance of C19H37N5O7.2 /2H2SO4:692.72 3-Deoxy-4-C-methyl-3-methylamino-b-L- about 10 cm, and air-dry the plate. Spray evenly 0.2z nin- arabinopyranosyl-(1ª6)-[2,6-diamino-2,3,4,6-tetradeoxy- hydrin-water saturated 1-butabol TS on the plate, and heat at a-D-glycero-hex-4-enopyranosyl-(1ª4)]-2- 1009C for 5 minutes. The spots corresponding to Rf about deoxy-D-streptamine hemipentasulfate [53179-09-2] 0.35 and Rf about 0.30 are not more intense than that of the spot from the standard solution (3), and the spot of gallamine Sisomicin Sulfate is the sulfate of an aminoglycoside corresponding to Rf about 0.25 is not more intense than the substance having antibacterial activity produced by the spot from the standard solution (1). The total amount of the growth of Micromonospora inyoensis. related substances is not more than 6z. It contains not less than 590 mg (potency) and not Loss on drying <2.41> Not more than 15.0z (0.15 g, in more than 700 mg (potency) per mg, calculated on the vacuum not exceeding 0.67 kPa, 1109C, 3 hours). Sampling dried basis. The potency of Sisomicin Sulfate is ex- should be carried out in a manner to avoid moisture absorp- pressed as mass (potency) of sisomicin (C19H37N5O7: tion. 447.53). Residue on ignition <2.44> Not more than 1.0z (1 g). Description Sisomicin Sulfate occurs as a white to light yel- lowish white powder. Assay Perform the test according to the Cylinder-plate It is very soluble in water, and practically insoluble in method as directed under Microbial Assay for Antibiotics ethanol (95). <4.02> according to the following conditions. It is hygroscopic. (i) Test organism—Staphylococcus epidermidis ATCC 12228 Identiˆcation (1) Dissolve 50 mg of Sisomicin Sulfate in 5 (ii) Culture medium—Use the medium ii in 3) Medium mL of water, and add 0.3 mL of bromine TS: the solution is for other organisms under (1) Agar media for seed and base immediately decolorized. layer. Adjust the pH of the medium so that it will be 7.8 to JP XV O‹cial Monographs / Sodium Acetate Hydrate 1091

8.0 after sterilization. (iii) Standard solutions—Weigh accurately an amount of Sisomicin Sulfate Reference Standard equivalent to about 25 Sodium Acetate Hydrate mg (potency), add 0.1 molWL phosphate buŠer solution, pH 8.0 to make exactly 25 mL, and use this solution as the stan- 酢酸ナトリウム水和物 dard stock solution. Keep the standard stock solution at 59C or below and use within 7 days. Take exactly a suitable amount of the standard stock solution before use, add 0.1 molWL phosphate buŠer solution, pH 8.0 to make solutions C2H3NaO2.3H2O: 136.08 so that each mL contains 1 mg (potency) and 0.25 mg (poten- Monosodium acetate trihydrate [6131-90-4 ] cy), and use these solutions as the high concentration stan- dard solution and low concentration standard solution, Sodium Acetate Hydrate, when dried, contains not respectively. less than 99.5z of sodium acetate (C2H3NaO2:82.03). (iv) Sample solutions—Weigh accurately an amount of Description Sodium Acetate Hydrate occurs as colorless Sisomicin sulfate equivalent to about 25 mg (potency), add crystals or a white, crystalline powder. It is odorless or has a 0.1 molWL phosphate buŠer solution, pH 8.0 to make exactly slight, acetous odor. It has a cool, saline and slightly bitter 25 mL. Take exactly a suitable amount of the solution, add taste. 0.1 molWL phosphate buŠer solution, pH 8.0 to make solu- It is very soluble in water, freely soluble in acetic acid tionssothateachmLcontains1mg (potency) and 0.25 mg (100), soluble in ethanol (95), and practically insoluble in (potency), and use these solutions as the high concentration diethyl ether. sample solution and low concentration sample solution, It is eŒorescent in warm, dry air. respectively. Identiˆcation A solution of Sodium Acetate Hydrate (1 in Containers and storage Containers—Tight containers. 10) responds to the Qualitative Tests <1.09> for acetate and Storage—Light-resistant, not exceeding －209C, under for sodium salt. nitrogen or argon atmosphere. Purity (1) Clarity and color of solution—Dissolve 2.0 g of Sodium Acetate Hydrate in 20 mL of water: the solution is Freeze-dried Smallpox Vaccine clear and colorless. (2) Acidity or alkalinity—Dissolve 1.0 g of Sodium 乾燥痘そうワクチン Acetate Hydrate in 20 mL of freshly boiled and cooled water, and add 3 drops of phenolphthalein TS: a red color develops. When cooled to 109C,or1.0mLof0.01molWLhydrochloric Freeze-dried Smallpox Vaccine is a preparation for acid VS is added after cooling to 109C, the red color disap- injection which is dissolved before use. It contains live pears. vaccinia virus. (3) Chloride <1.03>—Perform the test with 1.0 g of Sodi- It conforms to the requirements of Freeze-dried um Acetate Hydrate. Prepare the control solution with 0.30 Smallpox Vaccine in the Minimum Requirements for mL of 0.01 molWL hydrochloric acid VS (not more than 0.011 Biological Products. z). (4) Sulfate <1.14>—Perform the test with 1.0 g of Sodium Description Freeze-dried Smallpox Vaccine becomes a Acetate Hydrate. Prepare the control solution with 0.35 mL white to gray, turbid liquid on addition of solvent. of 0.005 molWL sulfuric acid VS (not more than 0.017z). (5) Heavy metals <1.07>—Proceed with 2.0 g of Sodium Acetate Hydrate according to Method 1, and perform the Freeze-dried Smallpox Vaccine test. Prepare the control solution with 2.0 mL of Standard Prepared in Cell Culture Lead Solution (not more than 10 ppm). (6) Calcium and magnesium—Dissolve 4.0 g of Sodium 乾燥細胞培養痘そうワクチン Acetate Hydrate in 25 mL of water, add 6 g of ammonium chloride, 20 mL of ammonia solution (28) and 0.25 mL of a solution of sodium hydrogensulˆte (1 in 10), and titrate <2.50 Freeze-dried Smallpox Vaccine Prepared in Cell Cul- > with 0.01 molWL disodium dihydrogen ethylenediamine ture is a preparation for injection which is dissolved be- tetraacetate VS until the blue color changes to grayish blue fore use. It contains live vaccinia virus. (indicator: 0.1 g of methylthymol blue-potassium nitrate in- It conforms to the requirements of Freeze-dried dicator): the amount of 0.01 molWL disodium dihydrogen Smallpox Vaccine Prepared in Cell Culture in the Mini- ethylenediamine tetraacetate VS consumed is not more than mum Requirements for Biological Products. 0.5 mL. (7) Arsenic <1.11>—Prepare the test solution with 1.0 g Description Freeze-dried Smallpox Vaccine Prepared in of Sodium Acetate Hydrate, according to Method 1, and per- Cell Culture becomes a reddish clear liquid on addition of form the test (not more than 2 ppm). solvent. (8) Potassium permanganate-reducing substance—Dis- solve 1.0 g of Sodium Acetate Hydrate in 100 mL of water, add 5 mL of dilute sulfuric acid, boil, add 0.50 mL of 0.002 molWL potassium permanganate VS, and further boil for 5 1092 Sodium Aurothiomalate / O‹cial Monographs JP XV minutes: the red color of the solution does not disappear. Sodium Aurothiomalate in 10 mL of water: the solution is clear and light yellow. Loss on drying <2.41> 39.0 – 40.5z (1 g, ˆrst at 809Cfor2 (2) Heavy metals <1.07>—Proceed with 1.0 g of Sodium hours, and then at 1309C for 2 hours). Aurothiomalate according to Method 2, and perform the Assay Weigh accurately about 0.2 g of Sodium Acetate Hy- test. Prepare the control solution with 3.0 mL of Standard drate, previously dried, dissolve in 50 mL of acetic acid (100), Lead Solution (not more than 30 ppm). and titrate <2.50> with 0.1 molWL perchloric acid VS until the (3) Arsenic <1.11>—Prepare the test solution with 1.0 g color of the solution changes from yellow to green (indicator: of Sodium Aurothiomalate according to Method 3, and per- 1mLofp-naphtholbenzein TS). Perform a blank determina- form the test (not more than 2 ppm). tion, and make any necessary correction. (4) Ethanol—Weigh accurately about 0.2 g of Sodium Aurothiomalate, add exactly 3 mL of the internal standard Each mL of 0.1 molWL perchloric acid VS solution and 2 mL of water to dissolve, and use this solution ＝8.203 mg of C H NaO 2 3 2 as the sample solution. Separately, pipet 3 mL of ethanol Containers and storage Containers—Tight containers. (99.5), and add water to make exactly 1000 mL. Pipet 2 mL of this solution, add exactly 3 mL of the internal standard so- lution, and use this solution as the standard solution. Sodium Aurothiomalate Perform the test with 2 mL each of the sample solution and standard solution as directed under Gas Chromatography 金チオリンゴ酸ナトリウム <2.02> according to the following conditions, and determine the ratios of the peak area of ethanol to that of the internal

standard, QT and QS: the amount of ethanol is not more than 3.0z.

Amount (mg) of ethanol＝(QT/QS)×6×0.793 Mixture of C4H3AuNa2O4S: 390.08 and C4H4AuNaO4S: 368.09 0.793: Density (g/mL) of ethanol (99.5) at 209C Monogold monosodium monohydrogen (2RS)-2- Internal standard solution—A solution of 2-propanol (1 in sulˆdobutane-1,4-dioate 500). Monogold disodium (2RS)-2-sulˆdobutane-1,4-dioate Operating conditions— [ , Sodium Aurothiomalate] 12244-57-4 Detector: Hydrogen ‰ame-ionization detector. Column: A column 3 mm in inside diameter and 3 m in Sodium Aurothiomalate contains not less than length, packed with porous styrene-divinylbenzene 49.0z and not more than 52.5z of gold (Au: 196.97), copolymer for gas chromatography (particle diameter: 150 – calculated on the anhydrous basis and corrected by the 180 mm) (average pore size: 0.0085 mm; 300 – 400 m2/g). amount of ethanol. Column temperature: A constant temperature of about Description Sodium Aurothiomalate occurs as white to 1809C. light yellow, powder or granules. Carrier gas: Nitrogen It is very soluble in water, and practically insoluble in Flow rate: Adjust the ‰ow rate so that the retention time of ethanol (99.5). the internal standard is about 7 minutes. It is hygroscopic. System suitability— It changes in color by light to greenish pale yellow. System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi- Identiˆcation (1) To 2 mL of a solution of Sodium tions, ethanol and the internal standard are eluted in this ord- Aurothiomalate (1 in 10) add 1 mL of a solution of calcium er with the resolution between these peaks being not less than nitrate tetrahydrate (1 in 10): a white precipitate is produced, 4. and it dissolves in dilute nitric acid and reappears on the addi- System repeatability: When the test is repeated 6 times with tion of ammonium acetate TS. 2 mL of the standard solution under the above operating con- (2) To 2 mL of a solution of Sodium Aurothiomalate (1 ditions, the relative standard deviation of the ratio of the in 10) add 3 mL of silver nitrate TS: a yellow precipitate is peak area of ethanol to that of the internal standard is not produced, and it dissolves in an excess of ammonia TS. more than 2.0z. (3) Place 2 mL of a solution of Sodium Aurothiomalate (1 in 10) in a porcelain crucible, add 1 mL of ammonia TS Water <2.48> Not more than 5.0z (0.1 g, coulometric titra- and 1 mL of hydrogen peroxide (30), evaporate to dryness, tion). Use a water vaporizer (heating temperature: 1059C; and ignite. Add 20 mL of water to the residue, and ˆlter: the heating time: 30 minutes). residue on the ˆlter paper occurs as a yellow or dark yellow, Assay Weigh accurately about 25 mg of Sodium powder or granules. Aurothiomalate, and dissolve in 2 mL of aqua regia by heat- (4) The ˆltrate obtained in (3) responds to the Qualitative ing. After cooling, add water to make exactly 100 mL. Pipet Tests <1.09> for sodium salt. 2 mL of the solution, add water to make exactly 25 mL, and (5) The ˆltrate obtained in (3) responds to the Qualitative use this solution as the sample solution. Separately, pipet 5 Tests <1.09> for sulfate. mL, 10 mL and 15 mL of Standard Gold Solution for atomic pH <2.54> Dissolve 1.0 g of Sodium Aurothiomalate in 10 absorption spectrophotometry, add water to make exactly 25 mL of water: the pH of this solution is between 5.8 and 6.5. mL, and use these solutions as the standard solutions (1), (2) and (3), respectively. Perform the test with the sample solu- Purity (1) Clarity and color of solution—Dissolve 1.0 g of JP XV O‹cial Monographs / Sodium Bicarbonate 1093 tion and standard solutions (1), (2) and (3) as directed under (5) Arsenic <1.11>—Mix well 1.0 g of Atomic Absorption Spectrophotometry <2.23> under the fol- with 0.40 g of calcium hydroxide, ignite, dissolve the residue lowing conditions. Determine the amount of gold in the sam- in 10 mL of dilute hydrochloric acid, and perform the test us- ple solution using the calibration curve obtained from the ab- ing this solution as the test solution (not more than 2 ppm). sorbances of the standard solutions. (6) Chlorinated compounds—Dissolve 1.0 g of Sodium Gas: Combustible gas—Acetylene Benzoate in 10 mL of water, add 10 mL of dilute sulfuric Supporting gas—Air acid, and extract with two 20-mL portions of diethyl ether. Lamp: Gold hollow-cathode lamp Combine the diethyl ether extracts, and evaporate the diethyl Wavelength: 242.8 nm ether on a water bath. Place 0.5 g of the residue and 0.7 g of calcium carbonate in a crucible, mix with a small amount of Containers and storage Containers—Tight containers. water, and dry. Ignite it at about 6009C, dissolve in 20 mL of Storage—Light-resistant. dilute nitric acid, and ˆlter. Wash the residue with 15 mL of water, combine the ˆltrate and the washing, add water to make 50 mL, and add 0.5 mL of silver nitrate TS: this solu- Sodium Benzoate tion has no more turbidity than the following control solu- tion 安息香酸ナトリウム Control solution: Dissolve 0.7 g of calcium carbonate in 20 mL of dilute nitric acid, and ˆlter. Wash the residue with 15 mL of water, combine the ˆltrate and the washings, add 1.2 mL of 0.01 molWL Hydrochloric acid VS and water to make 50 mL, and add 0.5 mL of silver nitrate TS. C7H5NaO2:144.10 (7) Phthalic acid—To 0.10 g of Sodium Benzoate add 1 Monosodium benzoate [532-32-1] mL of water and 1 mL of resorcinol-sulfuric acid TS, and heat the mixture in an oil bath heated at a temperature be- Sodium Benzoate, when dried, contains not less than tween 1209C and 1259C to evaporate the water, then heat the 99.0z of C7H5NaO2. residue for further 90 minutes, cool, and dissolve in 5 mL of water. To 1 mL of the solution add 10 mL of a solution of so- Description Sodium Benzoate occurs as white granules, dium hydroxide (43 in 500), shake, then examine under light crystals or crystalline powder. It is odorless, and has a sweet at a wavelength between 470 nm and 490 nm: the green and saline taste. ‰uorescence of the solution is not more intense than that of It is freely soluble in water, slightly soluble in ethanol (95), the following control solution. and practically insoluble in diethyl ether. Control solution: Dissolve 61 mg of potassium hydrogen Identiˆcation A solution of Sodium Benzoate (1 in 100) in water to make exactly 1000 mL. Pipet exactly 1 responds to the Qualitative Tests <1.09> for benzoate and the mL of the solution, add 1 mL of resorcinol-sulfuric acid TS, Qualitative Tests <1.09> (1) and (2) for sodium salt. and proceed as directed above. Purity (1) Clarity and color of solution—Dissolve 1.0 g of Loss on drying <2.41> Not more than 1.5z (2 g, 1109C, Sodium Benzoate in 5 mL of water: the solution is clear and 4 hours). colorless. Assay Weigh accurately about 1.5 g of Sodium Benzoate, (2) Acidity or alkalinity—Dissolve 2.0 g of Sodium Ben- previously dried, and transfer to a 300-mL glass-stoppered zoate in 20 mL of freshly boiled and cooled water, and add 2 ‰ask. Dissolve in 25 mL of water, add 75 mL of diethyl ether drops of phenolphthalein TS and 0.20 mL of 0.05 molWLsul- and 10 drops of bromophenol blue TS, and titrate <2.50> with furic acid VS: the solution remains colorless. To this solution 0.5 molWL hydrochloric acid VS, while mixing the aqueous add 0.40 mL of 0.1 molWL sodium hydroxide VS: a red color and diethyl ether layers by vigorous shaking, until a persis- develops. tent, light green color is produced in the aqueous layer. (3) Sulfate <1.14>—Dissolve 0.40 g of Sodium Benzoate in 40 mL of water, add slowly 3.5 mL of dilute hydrochloric Each mL of 0.5 molWL hydrochloric acid VS acid with thorough stirring, allow to stand for 5 minutes, and ＝ 72.05 mg of C7H5NaO2 ˆlter. Discard the ˆrst 5 mL of the ˆltrate, take the subse- Containers and storage Containers—Well-closed contain- quent 20 mL of the ˆltrate, add water to make 50 mL, and ers. perform the test using this solution as the test solution. Pre- pare the control solution with 0.40 mL of 0.005 molWLsul- furicacidVS(notmorethan0.120z). (4) Heavy metals <1.07>—Dissolve 2.0 g of Sodium Ben- Sodium Bicarbonate zoate in 44 mL of water, add gradually 6 mL of dilute 炭酸水素ナトリウム hydrochloric acid with thorough stirring, and ˆlter. Discard the ˆrst 5 mL of the ˆltrate, take the subsequent 25 mL of the ˆltrate, neutralize with ammonia TS, add 2 mL of dilute acet- ic acid and water to make 50 mL, and perform the test using NaHCO3: 84.01 this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 2 mL of Sodium Bicarbonate contains not less than 99.0z of dilute acetic acid and water to make 50 mL (not more than 20 NaHCO3. ppm). Description Sodium Bicarbonate occurs as white crystals or 1094 Sodium Bicarbonate Injection / O‹cial Monographs JP XV crystalline powder. It is odorless, and has a characteristic, sa- It contains not less than 95z and not more than line taste. 105z of the labeled amount of sodium hydrogen car- It is soluble in water, and practically insoluble in ethanol bonate (NaHCO3:84.01). (95) and in diethyl ether. Method of preparation Prepare as directed under Injec- It slowly decomposes in moist air. tions, with Sodium Bicarbonate. Identiˆcation A solution of Sodium Bicarbonate (1 in 30) Description Sodium Bicarbonate Injection is a clear, color- responds to the Qualitative Tests <1.09> for sodium salt and less liquid. for bicarbonate. Identiˆcation To a volume of Sodium Bicarbonate Injec- pH <2.54> Dissolve 1.0 g of Sodium Bicarbonate in 20 mL tion, equivalent to 1 g of Sodium Bicarbonate according to of water: the pH of this solution is between 7.9 and 8.4. the labeled amount, add water to make 30 mL: the solution Purity (1) Clarity and color of solution—Dissolve 1.0 g of responds to the Qualitative Tests <1.09> for sodium salt and Sodium Bicarbonate in 20 mL of water: the solution is clear for bicarbonate. and colorless. pH <2.54> 7.0–8.5 (2) Chloride <1.03>—To 0.40 g of Sodium Bicarbonate add 4 mL of dilute nitric acid, heat to boil, cool, and add 6 Bacterial endotoxins <4.01> Less than 5.0 EUWmEq. mL of dilute nitric acid and water to make 50 mL. Perform Extractable volume <6.05> It meets the requirement. the test using this solution as the test solution. Prepare the control solution with 0.45 mL of 0.01 molWL hydrochloric Assay Measure exactly a volume of Sodium Bicarbonate In- acid VS (not more than 0.040z). jection, equivalent to about 2 g of sodium hydrogen car-

(3) Carbonate—Dissolve 1.0 g of Sodium Bicarbonate in bonate (NaHCO3), titrate with 0.5 molWL sulfuric acid VS, 20 mL of freshly boiled and cooled water with very gentle and proceed as directed in the Assay under Sodium Bicar- swirling at a temperature not exceeding 159C. Add 2.0 mL of bonate. 0.1 mol WL hydrochloric acid VS and 2 drops of Each mL of 0.5 molWL sulfuric acid VS phenolphthalein TS: no red color develops immediately. ＝ 84.01 mg of NaHCO (4) Ammonium—Heat 1.0 g of Sodium Bicarbonate: the 3 gas evolved does not change moistened red litmus paper to Containers and storage Containers—Hermetic containers. blue. Plastic containers for aqueous injections may be used. (5) Heavy metals <1.07>—Dissolve 4.0 g of Sodium Bi- carbonate in 5 mL of water and 4.5 mL of hydrochloric acid, and evaporate on a water bath to dryness. Dissolve the Sodium Bisulfite residue in 2 mL of dilute acetic acid, 35 mL of water and 1 drop of ammonium TS, dilute with water to 50 mL, and per- Sodium Hydrogen Sulfite form the test using this solution as the test solution. Prepare the control solution as follows: evaporate 4.5 mL of 亜硫酸水素ナトリウム hydrochloric acid to dryness, and add 2 mL of dilute acetic acid, 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 5 ppm). NaHSO3:104.06 (6) Arsenic <1.11>—Dissolve 1.0 g of Sodium Bicar- bonate in 3 mL of water and 2 mL of hydrochloric acid, and Sodium Bisulfite is a mixture of sodium hydrogen- perform the test using this solution as the test solution (not sulfite and sodium pyrosulfite. more than 2 ppm). It contains not less than 64.0z and not more than 67.4z of sulfur dioxide (SO2:64.06). Assay Weigh accurately about 2 g of Sodium Bicarbonate, dissolve in 25 mL of water, and titrate with 0.5 molWL sulfur- Description Sodium Bisulfite occurs as white granules or ic acid VS. When the color of the solution changes from blue powder, having the odor of sulfur dioxide. to yellow-green, boil with caution, cool, and continue the Sodium Bisulfite is freely soluble in water, and practically titration <2.50> until a greenish yellow color develops (indica- insoluble in ethanol (95) and in diethyl ether. tor: 2 drops of bromocresol green TS). A solution of Sodium Bisulfite (1 in 20) is acid. Sodium Bisulfite is slowly affected by air or by light. Each mL of 0.5 molWL sulfuric acid VS Identification A solution of Sodium Bisulfite (1 in 20) ＝ 84.01 mg of NaHCO3 responds to the Qualitative Tests <1.09> for sodium salt and Containers and storage Containers—Hermetic containers. for bisulfite. Plastic containers for aqueous injections may be used. Purity (1) Clarity and color of solution—Dissolve 1.0 g of Sodium Bisulfite in 10 mL of water: the solution is clear and Sodium Bicarbonate Injection colorless. (2) Thiosulfate—Dissolve 1.0 g of Sodium Bisulfite in 15 炭酸水素ナトリウム注射液 mL of water, add slowly 5 mL of dilute hydrochloric acid, shake, and allow to stand for 5 minutes: no turbidity is produced. Sodium Bicarbonate Injection is an aqueous solution (3) Heavy metals <1.07>—Dissolve 1.0 g of Sodium Bisul- for injection. fite in 10 mL of water, add 5 mL of hydrochloric acid, and JP XV O‹cial Monographs / Sodium Bromide 1095 evaporateonawaterbathtodryness.Totheresidueadd2 does not eŠervesce. mL of dilute acetic acid and water to make 50 mL, and per- (3) Heavy metals <1.07>—Dissolve 1.5 g of Sodium form the test using this solution as the test solution. Prepare Borate in 25 mL of water and 7 mL of 1 molWL hydrochloric the control solution as follows: evaporate 5 mL of acid TS, add 1 drop of phenolphthalein TS, and add ammo- hydrochloric acid on a water bath to dryness, and add 2 mL nia TS until a pale red color develops. Then add dilute acetic of dilute acetic acid and 2.0 mL of Standard Lead Solution, acid until the solution becomes colorless again, add 2 mL of and dilute with water to make 50 mL (not more than 20 dilute acetic acid, and add water to make 50 mL. Perform the ppm). test using this solution as the test solution. Prepare the con- (4) Iron <1.10>—Prepare the test solution with 1.0 g of trol solution as follows: to 3.0 mL of Standard Lead Solution Sodium Bisulfite according to Method 1, and perform the add 2 mL of dilute acetic acid and water to make 50 mL (not test according to Method A. Prepare the control solution more than 20 ppm). with 2.0 mL of Standard Iron Solution (not more than 20 (4) Arsenic <1.11>—Prepare the test solution with 0.40 g ppm). of Sodium Borate according to Method 1, and perform the (5) Arsenic <1.11>—Dissolve 0.5 g of Sodium Bisulfite in test (not more than 5 ppm). 10 mL of water. Add 1 mL of sulfuric acid, heat on a sand Assay Weigh accurately about 2 g of Sodium Borate, dis- bath until white fumes are evolved, add water to make 5 mL, solve in 50 mL of water, and titrate <2.50> with 0.5 molWL and perform the test with this solution as the test solution hydrochloric acid VS (indicator: 3 drops of methyl red TS). (not more than 4 ppm). Each mL of 0.5 molWL hydrochloric acid VS Assay Weigh accurately about 0.15 g of Sodium Bisulfite, ＝ 95.34 mg of Na B O .10H O and transfer immediately into an iodine flask containing ex- 2 4 7 2 actly50mLof0.05molWL iodine VS, stopper, shake, and al- Containers and storage Containers—Tight containers. low to stand for 5 minutes in a dark place. Add 1 mL of hydrochloric acid, and titrate <2.50> theexcessiodinewith 0.1 molWL sodium thiosulfate VS (indicator: 1 mL of starch Sodium Bromide TS). Perform a blank determination. 臭化ナトリウム Each mL of 0.05 molWLiodineVS＝ 3.203 mg of SO2 Containers and storage Containers—Tight containers. Storage—Light-resistant, preferably well-filled, and not NaBr: 102.89 exceeding 309C. Sodium Bromide, when dried, contains not less than 99.0z of NaBr. Sodium Borate Description Sodium Bromide occurs as colorless or white crystals or crystalline powder. It is odorless. ホウ砂 It is freely soluble in water, and soluble in ethanol (95). It is hygroscopic, but not deliquescent.

Na2B4O7.10H2O: 381.37 Identiˆcation A solution of Sodium Bromide (1 in 10) responds to the Qualitative Tests <1.09> for sodium salt and Sodium Borate contains not less than 99.0z and not for bromide. more than 103.0z of Na B O .10H O. 2 4 7 2 Purity (1) Clarity and color of solution—Dissolve 1.0 g of Description Sodium Borate occurs as colorless or white Sodium Bromide in 3 mL of water: the solution is clear and crystals or a white, crystalline powder. It is odorless, and has colorless. a slightly characteristic, saline taste. (2) Alkalinity—Dissolve 1.0 g of Sodium Bromide in 10 It is freely soluble in glycerin, soluble in water, and practi- mL of water, add 0.10 mL of 0.005 molWL sulfuric acid VS cally insoluble in ethanol (95), in ethanol (99.5) and in diethyl and 1 drop of phenolphthalein TS, heat to boil, and cool: the ether. solution is colorless. When placed in dry air, Sodium Borate eŒoresces and is (3) Chloride—Make a calculation from the result ob- coated with a white powder. tained in the Assay. Not more than 97.9 mL of 0.1 molWLsil- vernitrateVSisconsumedfor1gofSodiumBromide. Identiˆcation A solution of Sodium Borate (1 in 20) (4) Sulfate <1.14>—Perform the test with 2.0 g of Sodium responds to the Qualitative Tests <1.09> for sodium salt and Bromide. Prepare the control solution with 1.0 mL of 0.005 for borate. molWL sulfuric acid VS (not more than 0.024z). pH <2.54> Dissolve 1.0g of Sodium Borate in 20mL of (5) Iodide—Dissolve 0.5 g of Sodium Bromide in 10 mL water: the pH of this solution is between 9.1 and 9.6. of water, add 2 to 3 drops of iron (III) chloride TS and 1 mL of chloroform, and shake: no red-purple color develops in Purity (1) Clarity and color of solution—Dissolve 1.0 g of the chloroform layer. Sodium Borate in 20 mL of water by warming slightly: the so- (6) Bromate—Dissolve 1.0 g of Sodium Bromide in 10 lution is clear and colorless. mLoffreshlyboiledandcooledwater,andadd2dropsof (2) Carbonate or bicarbonate—Dissolve 1.0 g of pow- potassium iodide TS, 1 mL of starch TS and 3 drops of dilute dered Sodium Borate in 20 mL of freshly boiled and cooled sulfuric acid. Shake the mixture gently, and allow to stand water, and add 3 mL of dilute hydrochloric acid: the solution for 5 minutes: no blue color develops. 1096 Sodium Carbonate Hydrate / O‹cial Monographs JP XV

(7) Heavy metals <1.07>—Proceed with 2.0 g of Sodium cetic acid, dilute with water to make 50 mL, and perform the Bromide according to Method 1, and perform the test. Pre- test using this solution as the test solution. Prepare the con- pare the control solution with 2.0 mL of Standard Lead Solu- trol solution as follows: evaporate 8 mL of dilute hydrochlor- tion (not more than 10 ppm). ic acid on a water bath to dryness, add 2 mL of dilute acetic (8) Barium—Dissolve 0.5 g of Sodium Bromide in 10 mL acid and 2.0 mL of Standard Lead Solution, and dilute with of water, add 0.5 mL of dilute hydrochloric acid and 1 mL of water to make 50 mL (not more than 10 ppm). potassium sulfate TS, and allow to stand for 10 minutes: no (4) Arsenic <1.11>—Prepare the test solution with 0.65 of turbidity is produced. Sodium Carbonate Hydrate according to Method 1, and per- (9) Arsenic <1.11>—Prepare the test solution with 1.0 g form the test (not more than 3.1 ppm). of Sodium Bromide according to Method 1, and perform the Loss on drying <2.41> 61.0 ¿ 63.0z (1 g, 1059C, 4 hours). test (not more than 2 ppm). Assay Dissolve about 3 g of Sodium Carbonate Hydrate, Loss on drying <2.41> Not more than 5.0z (1 g, 1109C, weighed accurately, in 25 mL of water, and titrate with 0.5 4 hours). molWL sulfuric acid VS until the color of the solution changes Assay Weigh accurately about 0.4 g of Sodium Bromide, from blue to yellow-green. Boil cautiously, cool, and further previously dried, and dissolve in 50 mL of water. Add 10 mL titrate <2.50> until a greenish yellow color appears (indicator: of dilute nitric acid and 50 mL of 0.1 molWL silver nitrate VS, 2 drops of bromocresol green TS). exactly measured, and titrate <2.50> theexcesssilvernitrate Each mL of 0.5 molWL sulfuric acid VS with 0.1 molWL ammonium thiocyanate VS (indicator: 2 mL ＝143.1 mg of Na CO .10H O of ammonium iron (III) sulfate TS). Perform a blank deter- 2 3 2 mination. Containers and storage Containers—Tight containers. Each mL of 0.1 molWL silver nitrate VS ＝ 10.29 mg of NaBr Dried Sodium Carbonate Containers and storage Containers—Tight containers. 乾燥炭酸ナトリウム Sodium Carbonate Hydrate Na2CO3:105.99 炭酸ナトリウム水和物 Dried Sodium Carbonate, when dried, contains not less than 99.0z of Na2CO3. Na CO .10H O: 286.14 2 3 2 Description Dried Sodium Carbonate occurs as white crys- tals or crystalline powder. Sodium Carbonate Hydrate contains not less than It is freely soluble in water, and practically insoluble in 99.0z and not more than 103.0z of Na CO .10H O. 2 3 2 ethanol (95) and in diethyl ether. Description Sodium Carbonate Hydrate occurs as colorless A solution of Dried Sodium Carbonate (1 in 10) is alkaline. or white crystals. It is hygroscopic. It is freely soluble in water, and practically insoluble in Identiˆcation A solution of Dried Sodium Carbonate (1 in ethanol (95) and in diethyl ether. 20) responds to the Qualitative Tests <1.09> for sodium salt A solution of Sodium Carbonate Hydrate (1 in 10) is alka- and for carbonate. line. It is eŒorescent in air. Purity (1) Clarity and color of solution—Dissolve 1.0 g of Itliqueˆesinitswaterofcrystallizationat349C, and Dried Sodium Carbonate in 10 mL of water: the solution is becomes anhydrous at above 1009C. clear and colorless. (2) Chloride <1.03>—Dissolve 0.5 g of Dried Sodium Identiˆcation A solution of Sodium Carbonate Hydrate (1 Carbonate in 10 mL of water, add 12 mL of dilute nitric acid, in 20) responds to the Qualitative Tests <1.09> for sodium salt dilute with water to make 50 mL, and perform the test using and for carbonate. this solution as the test solution. Prepare the control solution Purity (1) Clarity and color of solution—Dissolve 1.0 g of with 1.0 mL of 0.01 molWL hydrochloric acid VS (not more Sodium Carbonate Hydrate in 5 mL of water: the solution is than 0.071z). clear and colorless. (3) Heavy metals <1.07>—Dissolve 1.0 g of Dried Sodium (2) Chloride <1.03>—Dissolve 0.5 g of Sodium Carbonate Carbonate in 10 mL of water, add 7.5 mL of dilute Hydrate in 10 mL of water, add 7 mL of dilute nitric acid, di- hydrochloric acid, and evaporate on a water bath to dryness. lute with water to make 50 mL, and perform the test using Dissolve the residue in 35 mL of water and 2 mL of dilute this solution as the test solution. Prepare the control solution acetic acid, dilute with water to make 50 mL, and perform with 1.0 mL of 0.01 molWL hydrochloric acid VS (not more the test using this solution as the test solution. Prepare the than 0.071z). control solution as follows: evaporate 7.5 mL of dilute (3) Heavy metals <1.07>—Dissolve 2.0 g of Sodium Car- hydrochloric acid on a water bath to dryness, add 2 mL of di- bonate Hydrate in 10 mL of water, add 8 mL of dilute lute acetic acid and 2.0 mL of Standard Lead Solution, and hydrochloric acid, and evaporate to dryness on a water bath. dilute with water to make 50 mL (not more than 20 ppm). Dissolve the residue in 35 mL of water and 2 mL of dilute a- (4) Arsenic <1.11>—Prepare the test solution with 0.65 g JP XV O‹cial Monographs / Sodium Chloride 1097 of Dried Sodium Carbonate according to Method 1, and per- ing control solution. form the test (not more than 3.1 ppm). Control solution: Dissolve 0.181 g of potassium sulfate in water to make exactly 500 mL. Pipet 5 mL of this solution, Loss on drying <2.41> Not more than 2.0z (2 g, 1069C, add water to make exactly 100 mL, and proceed in the same 4 hours). manner as directed above using this solution instead of the Assay Dissolve about 1.2 g of Dried Sodium Carbonate, sample solution. weighed accurately, in 25 mL of water, and titrate with 0.5 (4) —To 2.0 mL of the sample solution ob- molWL sulfuric acid VS until the color of the solution changes tained in (2) add 5 mL of 2 mol/L sulfuric acid TS and water from blue to yellow-green. Then boil cautiously, cool, and to make 100.0 mL, then add 4 mL of ammonium molybdate- further titrate <2.50> until a greenish yellow color develops sulfuric acid TS and 0.1 mL of tin (II) chloride-hydrochloric (indicator: 2 drops of bromocresol green TS). acid TS, and allow to stand for 10 minutes: the color of the solution is not darker than the following control solution. Each mL of 0.5 molWL sulfuric acid VS Control solution: To 1.0 mL of Standard Phosphoric Acid ＝52.99 mg of Na CO 2 3 Solution add 12.5 mL of 2 mol/L sulfuric acid TS and water Containers and storage Containers—Tight containers. to make exactly 250 mL. To 100 mL of this solution add 4 mL of ammonium molybdate-sulfuric acid TS and 0.1 mL of tin (II) chloride-hydrochloric acid TS, and allow to stand for Sodium Chloride 10 minutes. (5) —To 0.50 mL of the sample solution ob- 塩化ナトリウム tained in (2) add 4.0 mL of water, 2.0 mL of dilute phenol red TS and 1.0 mL of a solution of sodium toluenesulfon- chloramide trihydrate (1 in 10,000), and mix immediately. NaCl: 58.44 After allowing to stand for 2 minutes, add 0.15 mL of 0.1 mol/L sodium thiosulfate VS, mix, add water to make ex- This monograph is harmonized with the European actly 10 mL, and use this solution as the sample solution. Pharmacopoeia and the U.S. Pharmacopeia. The parts Separately, to 5.0 mL of a solution of potassium bromide (3 of the text that are not harmonized are marked with in 1,000,000) add 2.0 mL of dilute phenol red TS and 1.0 mL  symbols ( ). of a solution of sodium toluenesulfonchloramide trihydrate Sodium Chloride contains not less than 99.0z and (1 in 10,000), and mix immediately. Proceed in the same not more than 100.5z of NaCl, calculated on the dried manner as for the preparation of the sample solution, and use basis. the solution so obtained as the standard solution. Perform thetestwiththesamplesolutionandstandardsolutionas Description Sodium Chloride occurs as colorless or white, directed under Ultraviolet-visible Spectrophotometry <2.24> crystals or crystalline powder. using water as the control: the absorbance at 590 nm of the It is freely soluble in water, and practically insoluble in sample solution is not more than that of the standard solu- ethanol (99.5).  tion. Identiˆcation (1) A solution of Sodium Chloride (1 in 20) (6) Iodides—Wet 5 g of Sodium Chloride with drop-wise- responds to the Qualitative Tests <1.09> for sodium salt. ly added 0.15 mL of a freshly prepared mixture of starch TS, (2) A solution of Sodium Chloride (1 in 20) responds to 0.5 mol/L sulfuric acid TS and TS the Qualitative Tests <1.09> for chloride. (1000:40:3), allow to stand for 5 minutes, and examine under daylight: a blue color does not appear. Purity (1) Clarity and color of solution—Dissolve 1.0 g (7) Ferrocyanides—Dissolve 2.0 g of Sodium Chloride in of Sodium Chloride in 5 mL of water: the solution is clear 6 mL of water, and add 0.5 mL of a mixture of a solution of and colorless.  iron (II) sulfate heptahydrate (1 in 100) and a solution of am- (2) Acidity or alkalinity—Dissolve 20.0 g of Sodium monium iron (III) sulfate dodecahydrate in diluted sulfuric Chloride in 100.0 mL of freshly boiled and cooled water, and acid (1 in 400) (1 in 100) (19:1): a blue color does not develop use this solution as the sample solution. To 20 mL of the sam- within 10 minutes. ple solution add 0.1 mL of bromothymol blue TS and 0.5 mL (8) Heavy metals <1.07>—Proceed with 5.0 g of Sodium of 0.01 mol/L hydrochloric acid VS: the color of the solution Chloride according to Method 1, and perform the test. Pre- is yellow. Separately, to 20 mL of the sample solution add 0.1 pare the control solution with 1.5 mL of Standard Lead Solu- mL of bromothymol blue TS and 0.5 mL of 0.01 mol/L sodi- tion (not more than 3 ppm). um hydroxide VS: the color of the solution is blue.  (9) Iron—To 10 mL of the sample solution obtained in (3) —To 7.5 mL of the sample solution obtained (2) add 2 mL of a solution of citric acid monohydrate (1 in 5) in (2) add water to make 30 mL, and use this solution as the and 0.1 mL of mercapto acetic acid, alkalize with ammonia sample solution. Separately, dissolve 0.181 g of potassium TS,addwatertomake20mL,andallowtostandfor5 sulfate in diluted ethanol (99.5) (3 in 10) to make exactly 500 minutes: the solution has not more color than the following mL. Pipet 5 mL of this solution, and add diluted ethanol control solution. (99.5) (3 in 10) to make exactly 100 mL. To 4.5 mL of this so- Control solution: Pipet 1 mL of Standard Iron Solution, lution add 3 mL of a solution of barium chloride dihydrate (1 and add water to make exactly 25 mL. To 10 mL of this solu- in 4), shake, and allow to stand for 1 minutes. To 2.5 mL of tion add 2 mL of a solution of citric acid monohydrate (1 in this solution add 15 mL of the sample solution and 0.5 mL of 5) and 0.1 mL of mercapto acetic acid, and proceed in the acetic acid (31), and allow to stand for 5 minutes: any turbidi- same manner as directed for the sample solution. ty produced does not more than that produced in the follow- (10) Barium—To 5.0 mL of the sample solution obtained 1098 10 Sodium Chloride Injection / O‹cial Monographs JP XV in (2) add 5.0 mL of water and 2.0 mL of dilute sulfuric acid, shaking, with 0.1 molWL silver nitrate VS (indicator: 3 drops and allow to stand for 2 hours: the solution has not more tur- of ‰uorescein sodium TS). bidity than the following control solution. Each mL 0.1 molWL silver nitrate VS Control solution: To 5.0 mL of the sample solution ob- ＝ 5.844 mg of NaCl tained in (2) add 7.0 mL of water, and allow to stand for 2 hours. Containers and storage Containers—Hermetic containers. (11) Magnesium and alkaline-earth materials—To 200 Plastic containers for aqueous injections may be used. mL of water add 0.1 g of hydroxylammonium chloride, 10 mL of ammonium chloride buŠer solution, pH 10, 1 mL of 0.1 mol/L zinc sulfate VS and 0.2 g of eriochrome black T- Isotonic Sodium Chloride Solution sodium chloride indicator, and warm to 409C. Add 0.01 mol/L disodium dihydrogen ethylenediamine tetraacetate VS 0.9 Sodium Chloride Injection dropwise until the red-purple color of the solution changes to Isotonic Salt Solution blue-purple. To this solution add a solution prepared by dis- solving 10.0 g of Sodium Chloride in 100 mL of water, and Isotonic Sodium Chloride Injection add 2.5 mL of 0.01 mol/L disodium dihydrogen ethylenedia- mine tetraacetate VS: the color of the solution is a blue-pur- 生理食塩液 ple. (12) Arsenic <1.11>—Prepare the test solution with 1.0 Isotonic Sodium Chloride Solution is an aqueous so- g of Sodium Chloride according to Method 1, and perform lution for injection. the test (not more than 2 ppm).  It contains not less than 0.85 wWvz and not more Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, than 0.95 wWvz of sodium chloride (NaCl: 58.44). 2 hours). Method of preparation Assay Weigh accurately about 50 mg of Sodium Chloride, Sodium Chloride 9 g dissolve in 50 mL of water, and titrate <2.50> with 0.1 mol/L Water for Injection a su‹cient quantity silver nitrate VS (potentiometric titration). To make 1000 mL Each mL of 0.1 mol/L silver nitrate VS ＝ 5.844 mg of NaCl Prepare as directed under Injections, with the above in- gredients. Containers and storage Containers—Tight containers.  No preservative is added. Description Isotonic Sodium Chloride Solution is a clear, 10 Sodium Chloride Injection colorless liquid. It has a slightly saline taste. Identiˆcation Isotonic Sodium Chloride Solution responds 10 塩化ナトリウム注射液 to the Qualitative Tests <1.09> for sodium salt and for chlo- ride. 10z Sodium Chloride Injection is an aqueous solu- pH <2.54> 4.5–8.0 tion for injection. Purity (1) Heavy metals <1.07>—Concentrate 100 mL of It contains not less than 9.5 wWvz and not more Isotonic Sodium Chloride Solution to about 40 mL on a than 10.5 wWvz of sodium chloride (NaCl: 58.44). water bath, and add 2 mL of dilute acetic acid and water to Method of preparation make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 3.0 mL of Stan- Sodium Chloride 100 g dard Lead Solution and 2 mL of dilute acetic acid, and add Distilled Water for Injection a su‹cient quantity water to make 50 mL (not more than 0.3 ppm). To make 1000 mL (2) Arsenic <1.11>—Prepare the test solution with 20 mL of Isotonic Sodium Chloride Solution, and perform the test Prepare as directed under Injections, with the above in- (not more than 0.1 ppm). gredients. Bacterial endotoxins <4.01> Less than 0.50 EUWmL. Description 10z Sodium Chloride Injection is a clear, colorless liquid. It has a saline taste. Extractable volume <6.05> It meets the requirement. It is neutral. Assay Measure exactly 20 mL of Isotonic Sodium Chloride Identiˆcation 10z Sodium Chloride Injection responds to Solution, add 30 mL of water, and titrate <2.50> with 0.1 the Qualitative Tests <1.09> for sodium salt and for chloride. molWL silver nitrate VS with vigorous shaking (indicator: 3 drops of ‰uorescein sodium TS). Bactetial endotoxins <4.01> Less than 3.6 EUWmL. Each mL of 0.1 molWL silver nitrate VS Extractable volume <6.05> It meets the requirement. ＝ 5.844 mg of NaCl Assay Pipet 10 mL of 10z Sodium Chloride Injection, and Containers and storage Containers—Hermetic containers. add water to make exactly 100 mL. Pipet 20 mL of this solu- Plastic containers for aqueous injections may be used. tion, add 30 mL of water, and titrate <2.50>, with vigorous JP XV O‹cial Monographs / Sodium Citrate Injection for Transfusion 1099

Solution (not more than 10 ppm). (5) Arsenic <1.11>—Prepare the test solution with 1.0 g Sodium Chromate (51Cr) Injection of Sodium Citrate Hydrate according to Method 1, and per- form the test (not more than 2 ppm). クロム酸ナトリウム (51Cr) 注射液 (6) Tartrate—To a solution of 1.0 g of Sodium Citrate Hydrate in 2 mL of water add 1 mL of potassium acetate TS and 1 mL of acetic acid (31): no crystalline precipitate is Sodium Chromate (51Cr) Injection is an aqueous so- formed after the sides of the tube have been rubbed with a g- lution for injection containing chromium-51 (51Cr) in lass rod. the form of sodium chromate. (7) Oxalate—Dissolve 1.0 g of Sodium Citrate Hydrate in It conforms to the requirements of Sodium Chro- a mixture of 1 mL of water and 3 mL of dilute hydrochloric mate (51Cr) Injection in the Minimum Requirements acid, add 4 mL of ethanol (95) and 0.2 mL of calcium chlo- for Radiopharmaceuticals. ride TS, and allow to stand for 1 hour: the solution is clear. Test for Extractable Volume of Parentaral Prepara- (8) Readily carbonizable substances <1.15>—Take 0.5 g tions and Insoluble Particulate Matter Test for Injec- of Sodium Citrate Hydrate, and perform the test by heating tions are not applied to this injection. at 909C for 1 hour: the solution has no more color than Matching Fluid K. Description Sodium Chromate (51Cr) Injection is a clear, light yellow liquid. It is odorless or has an odor of the preser- Loss on drying <2.41> 10.0 – 13.0z (1 g, 1809C, 2 hours). vatives. Assay Weigh accurately about 0.2 g of Sodium Citrate Hy- drate, previously dried, add 30 mL of acetic acid for nona- queous titration, warm to dissolve, and titrate <2.50> with 0.1 Sodium Citrate Hydrate mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any クエン酸ナトリウム水和物 necessary correction. Each mL of 0.1 mol/L perchloric acid VS

＝8.602 mg of C6H5Na3O7 Containers and storage Containers—Tight containers. C6H5Na3O7.2H2O: 294.10 Trisodium 2-hydroxypropane-1,2,3-tricarboxylate dihydrate [6132-04-3] Sodium Citrate Injection for Sodium Citrate Hydrate, when dried, contains not Transfusion less than 99.0z and not more than 101.0z of sodium citrate (C6H5Na3O7: 258.07). 輸血用クエン酸ナトリウム注射液 Description Sodium Citrate Hydrate occurs as colorless crystals, or a white, crystalline powder. It is odorless, and has a cooling, saline taste. Sodium Citrate Injection for Transfusion is an aque- It is freely soluble in water, and practically insoluble in ous solution for injection. ethanol (95) and in diethyl ether. It contains not less than 9.5 wWvz and not more than 10.5 w Wv z of sodium citrate hydrate Identiˆcation A solution of Sodium Citrate Hydrate (1 in (C6H5Na3O7.2H2O: 294.10). 20) responds to the Qualitative Tests <1.09> for citrate and for sodium salt. Method of preparation pH <2.54> Dissolve 1.0 g of Sodium Citrate Hydrate in 20 Sodium Citrate Hydrate 100 g mL of water: the pH of this solution is between 7.5 and 8.5. Water for Injection a su‹cient quantity To make 1000 mL Purity (1) Clarity and color of solution—A solution of 1.0 g of Sodium Citrate Hydrate in 10 mL of water is clear Prepare as directed under Injections, with the above in- and colorless. gredients. (2) Chloride <1.03>—Take 0.6 g of Sodium Citrate Hy- No preservatives may be added. drate, and perform the test. Prepare the control solution with 0.25 mL of 0.01 molWL hydrochloric acid VS (not more than Description Sodium Citrate Injection for Transfusion is a 0.015z). clear, colorless liquid. (3) Sulfate <1.14>—To 0.5 g of Sodium Citrate Hydrate Identiˆcation Sodium Citrate Injection for Transfusion add water to make 40 mL, then add 3.0 mL of dilute responds to the Qualitative Tests <1.09> for sodium salt and hydrochloric acid and water to make 50 mL, and perform the for citrate. test. Prepare the control solution with 0.50 mL of 0.005 molW L sulfuric acid VS (not more than 0.048z). pH <2.54> 7.0–8.5 (4) Heavy metals <1.07>—Proceed with 2.5 g of Sodium Bacterial endotoxins <4.01> Less than 5.6 EUWmL. Citrate Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.5 mL of Standard Lead Extractable volume <6.05> It meets the requirement. Assay Pipet 5 mL of Sodium Citrate Injection for Transfu- 1100 Sodium Cromoglicate / O‹cial Monographs JP XV sion, and add water to make exactly 25 mL. Evaporate 10 mL 1-benzopyran-2-carboxylate) [15826-37-6] of this solution, exactly measured, on a water bath to dry- ness, dry the residue at 1809C for 2 hours, and dissolve in 30 Sodium Cromoglicate contains not less than 98.0z mL of acetic acid (100) by warming. Cool, titrate <2.50> with of C23H14Na2O11, calculated on the dried basis. 0.1 molWL perchloric acid VS (indicator: 3 drops of crystal Description Sodium Cromoglicate occurs as a white, crys- violet TS). Perform a blank determination, and make any talline powder. It is odorless and tasteless at ˆrst, and later necessary correction. develops a slightly bitter taste. Each mL of 0.1 molWL perchloric acid VS It is freely soluble in water, sparingly soluble in propylene

＝ 9.803 mg of C6H5Na3O7.2H2O glycol, very slightly soluble in ethanol (95), and practically in- soluble in 2-propanol and in diethyl ether. Containers and storage Containers—Hermetic containers. It is hygroscopic. It gradually acquires a yellow color by light. Diagnostic Sodium Citrate Solution Identiˆcation (1) Dissolve 0.1 g of Sodium Cromoglicate in2mLofwater,add2mLofsodiumhydroxideTS,and 診断用クエン酸ナトリウム液 boil for 1 minute: a yellow color is produced. After cooling, add 0.5 mL of concentrated diazobenzene sulfonic acid TS: a dark red color is produced. Diagnostic Sodium Citrate Solution contains not less (2) Determine the absorption spectrum of a solution of than 3.3 wWvz and not more than 4.3 w Wvz of sodi- Sodium Cromoglicate in phosphate buŠer solution, pH 7.4 (1 um citrate hydrate (C6H5Na3O7.2H2O: 294.10). in 100,000) as directed under Ultraviolet-visible Spec- The requirements as described for aqueous injec- trophotometry <2.24>, and compare the spectrum with the tions under Injections are applicable. Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Method of preparation (3) Sodium Cromoglicate responds to the Qualitative Sodium Citrate Hydrate 38 g Tests <1.09> for sodium salt. Water for Injection a su‹cient quantity Purity (1) Clarity and color of solution—Dissolve 0.50 g To make 1000 mL of Sodium Cromoglicate in 10 mL of water: the solution is clear and colorless to pale yellow. Prepare as directed under Injections, with the above in- (2) Acidity or alkalinity—Dissolve 2.0 g of Sodium gredients. Cromoglicatein40mLoffreshlyboiledandcooledwater, No preservative may be added. add 6 drops of bromothymol blue TS, and use this solution Description Diagnostic Sodium Citrate Solution is a clear, as the sample solution. To 20 mL of the sample solution add colorless liquid. 0.25 mL of 0.1 molWL sodium hydroxide VS: a blue color is produced. To another 20 mL of the sample solution add 0.25 Identiˆcation Diagnostic Sodium Citrate Solution responds mL of 0.1 molWL hydrochloric acid VS: a yellow color is to the Qualitative Tests <1.09> for sodium salt and for citrate. produced. pH <2.54> 7.0 – 8.5 (3) Heavy metals <1.07>—Proceed with 1.0 g of Sodium Cromoglicate according to Method 2, and perform the test. Assay Pipet 5 mL of Diagnostic Sodium Citrate Solution, Prepare the control solution with 2.0 mL of Standard Lead evaporate on a water bath to dryness, dry the residue at Solution (not more than 20 ppm). 1809Cfor2hours,anddissolvein30mLofaceticacid(100) (4) Oxalate—Dissolve 0.25 g of Sodium Cromoglicate in by warming. Cool, and titrate 2.50 with 0.1 mol Lper- < > W water to make exactly 50 mL, and use this solution as the chloric acid VS (indicator: 3 drops of crystal violet TS). Per- sample solution. Separately, dissolve 0.049 g of oxalic acid form a blank determination, and make any necessary correc- dihydrate, exactly weighed, in water to make exactly 100 mL. tion. Pipet 5 mL of this solution, add water to make exactly 100 Each mL of 0.1 molWL perchloric acid VS mL, and use this solution as the standard solution. Pipet 20

＝ 9.803 mg of C6H5Na3O7.2H2O mL each of the sample solution and standard solution, add exactly 5 mL of iron salicylate TS to each solution, and add Containers and storage Containers—Hermetic containers. water to make 50 mL. Determine the absorbances of these so- lutions as directed under Ultraviolet-visible Spectrophoto- metry <2.24> using water as the blank: the absorbance of the Sodium Cromoglicate sample solution at 480 nm is not smaller than that of the stan- dard solution. クロモグリク酸ナトリウム (5) Related substances—Dissolve 0.20 g of Sodium Cromoglicate 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 10 mL, pipet 1 mL of this solution, add water 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>.Spot10 C H Na O :512.33 23 14 2 11 mL each of the sample solution and standard solution on a Disodium 5,5?-(2-hydroxytrimethylenedioxy)bis(4-oxo-4H- JP XV O‹cial Monographs / Sodium Hydroxide 1101 plate of silica gel with ‰uorescent indicator for thin-layer <1.09> (1) for sodium salt. chromatography. Develop the plate with a mixture of Purity Heavy metals <1.07>—Proceedwith2.0gofSodium methanol, chloroform and acetic acid (100) (9:9:2) to a dis- Fusidate according to Method 4, and perform the test. Pre- tance of about 10 cm, and air-dry the plate. Examine under pare the control solution with 2.0 mL of Standard Lead Solu- ultraviolet light (main wavelength: 254 nm): spots other than tion (not more than 10 ppm). the principal spot from the sample solution is not more in- tense than the spot from the standard solution. Water <2.48> Not more than 2.0z (1 g, volumetric titra- tion, direct titration). Loss on drying <2.41> Not more than 10.0z (1 g, in vacu- um, 1059C, 4 hours). Assay Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics Assay Weigh accurately about 0.18 g of Sodium Cromogli- <4.02> according to the following conditions. cate, and dissolve in a mixture of 25 mL of propylene glycol (i) Test organism— ATCC 6538 P and 5 mL of 2-propanol by warming. After cooling, add 30 (ii) Culture medium—Use the medium ii in 3) Medium mL of 1,4-dioxane, and titrate <2.50> with 0.1 molWLper- for other organisms under (1) Agar media for seed and base chloric acid-dioxane VS (potentiometric titration). Perform a layer. blank determination, and make any necessary correction. (iii) Standard solutions—Weigh accurately an amount of Each mL of 0.1 molWL perchloric acid-dioxane VS Diethanolamine Fusidate Reference Standard, equivalent to

＝ 25.62 mg of C23H14Na2O11 about 20 mg (potency), dissolve in 2 mL of ethanol (95), add water to make exactly 20 mL, and use this solution as the Containers and storage Containers—Tight containers. standard stock solution. Keep the standard stock solution at Storage—Light-resistant. a temperature not exceeding 59C and use within 7 days. Take exactly a suitable amount of the standard stock solution be- fore use, add phosphate buŠer solution, pH 6.0 to make solu- Sodium Fusidate tionssothateachmLcontains4mg (potency) and 1 mg (potency), and use these solutions as the high concentration フシジン酸ナトリウム standard solution and low concentration standard solution, respectively. (iv) Sample solutions—Weigh accurately an amount of Sodium Fusidate, equivalent to about 20 mg (potency), and dissolve in water to make exactly 20 mL. Take exactly a suita- ble amount of this solution, add phosphate buŠer solution, pH 6.0 to make solutions so that each mL contains 4 mg (potency) and 1 mg (potency), and use these solutions as the high concentration sample solution and low concentration sample solution, respectively. Containers and storage Containers—Tight containers. Storage—Light-resistant, and at a temperature 2 to 89C. C31H47NaO6:538.69 Monosodium (17Z)-ent-16a-acetoxy-3b,11b-dihydroxy- 4b,8b,14a-trimethyl-18-nor-5b,10a-cholesta-17(20),24- dien-21-oate [751-94-0 ] Sodium Hydroxide

Sodium Fusidate is the sodium salt of a substance 水酸化ナトリウム having antibacterial activity produced by the growth of Fusidium coccineum. It contains not less than 935 mg (potency) and not NaOH: 40.00 more than 969 mg (potency) per mg, calculated on the anhydrous basis. The potency of Sodium Fusidate is Sodium Hydroxide contains not less than 95.0z of NaOH. expressed as mass (potency) of fusidic acid (C31H48O6: 516.71). Description Sodium Hydroxide occurs as white fused mass- Description Sodium Fusidate occurs as white, crystals of es, in small pellets, in ‰akes, in sticks, and in other forms. It crystalline powder. is hard and brittle, and shows a crystalline fracture. It is freely soluble in water, in methanol and in ethanol It is freely soluble in water and in ethanol (95), and practi- (99.5). cally insoluble in diethyl ether. It rapidly absorbs carbon dioxide in air. Identiˆcation (1) Determine the infrared absorption spec- It deliquesces in moist air. tra of Sodium Fusidate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>,and Identiˆcation (1) A solution of Sodium Hydroxide (1 in compare the spectrum with the Reference Spectrum: both 500) is alkaline. spectra exhibit similar intensities of absorption at the same (2) A solution of Sodium Hydroxide (1 in 25) responds to wave numbers. the Qualitative Tests <1.09> for sodium salt. (2) Sodium Fusidate responds to the Qualitative Tests Purity (1) Clarity and color of solution—Dissolve 1.0 g of 1102 Sodium Iodide / O‹cial Monographs JP XV

Sodium Hydroxide in 20 mL of water: the solution is clear TS, and titrate with 0.5 molWL sulfuric acid VS until the red and colorless. color of the solution disappears. Record the amount, A (2) Chloride < 1.03 > —Dissolve 2.0 g of Sodium (mL), of 0.5 molWL sulfuric acid VS consumed. Then add 2 Hydroxide in water, and add water to make 100 mL. To 25 drops of methyl orange TS to the solution, and further titrate mL of the solution add 10 mL of dilute nitric acid and water <2.50> with 0.5 molWL sulfuric acid VS until the solution to make 50 mL, and perform the test using this solution as shows a persistent light red color. Record the amount, B the test solution. Prepare the control solution with 0.7 mL of (mL), of 0.5 molWL sulfuric acid VS consumed. Calculate the 0.01 molWLhydrochloricacidVS(notmorethan0.050z). amount of NaOH from the diŠerence, A (mL)－B (mL). (3) Heavy metals <1.07>—Dissolve 1.0 g of Sodium Each mL of 0.5 molWL sulfuric acid VS Hydroxide in 5 mL of water, add 11 mL of dilute hydrochlor- ＝40.00 mg of NaOH ic acid, and evaporate on a water bath to dryness. Dissolve the residue in 35 mL of water, add 2 mL of dilute acetic acid Containers and storage Containers—Tight containers. and 1 drop of ammonia TS, add water to make 50 mL, and perform the test using this solution as the test solution. Evaporate 11 mL of dilute hydrochloric acid on a water bath Sodium Iodide to dryness, dissolve the residue in 2 mL of dilute acetic acid and 3.0 mL of Standard Lead Solution, add water to make 50 ヨウ化ナトリウム mL, and use this solution as the control solution (not more than 30 ppm). (4) Potassium—Dissolve 0.10 g of Sodium Hydroxide in NaI: 149.89 water and dilute with water to make 40 mL. Add 1.0 mL of dilute acetic acid to 4.0 mL of this solution, and shake. Add Sodium Iodide, when dried, contains not less than 5.0 mL of a solution of sodium tetraphenylboron (1 in 30), 99.0z of NaI. shake immediately, and allow to stand for 10 minutes: the so- Description Sodium Iodide occurs as colorless crystals or a lution has no more turbidity than the following control solu- white, crystalline powder. It is odorless. tion. Sodium Iodide is very soluble in water, and freely soluble Control solution: Dissolve 9.5 mg of potassium chloride in in glycerin and in ethanol (95). water, and dilute with water to make 1000 mL. Add 1.0 mL It deliquesces in moist air. of dilute acetic acid to 4.0 mL of this solution, shake, and proceed as directed above. Identiˆcation A solution of Sodium Iodide (1 in 20) (5) Sodium carbonate—The amount of sodium car- responds to the Qualitative Tests <1.09> for sodium salt and bonate (Na2CO3: 105.99) is not more than 2.0z,whencalcu- for iodide. lated by the following equation using B (mL) which is ob- Purity (1) Clarity and color of solution—Dissolve 1.0 g of tained in the Assay. Sodium Iodide in 2 mL of water: the solution is clear and Amount (mg) of sodium carbonate＝105.99×B colorless. (2) Alkalinity—Dissolve 1.0 g of Sodium Iodide in 10 mL (6) Mercury—Dissolve 2.0 g of Sodium Hydroxide in 1 of freshly boiled and cooled water, and add 1.0 mL of 0.005 mL of a solution of potassium permanganate (3 in 50) and 30 mol L sulfuric acid VS and 1 drop of phenolphthalein TS: no mL of water, neutralize gradually with puriˆed hydrochloric W color is produced. acid, and add 5 mL of diluted sulfuric acid (1 in 2). To this (3) Chloride, bromide and thiosulfate—Dissolve 0.20 g solution add a solution of hydroxylammonium chloride (1 in of Sodium Iodide in 5 mL of ammonia TS, add 15.0 mL of 5) until the precipitate of manganese dioxide disappears, add 0.1 mol L silver nitrate VS, shake for a few minutes, and water to make exactly 100 mL, and use this solution as the W ˆlter. To 10 mL of the ˆltrate add 15 mL of dilute nitric acid: sample solution. Perform the tests according to the Atomic no brown color appears. The solution has no more turbidity Absorption Spectrophotometry <2.23> (Cold vapor type) with than the following control solution. the sample solution. Place the sample solution in the sample Control solution: To 0.30 mL of 0.01 mol L hydrochloric bottle of an atomic absorption spectrophotometer, add 10 W acid VS add 2.5 mL of ammonia TS, 7.5 mL of 0.1 mol L mL of tin (II) chloride-sulfuric acid TS, connect the bottle W silver nitrate VS and 15 mL of dilute nitric acid. immediately to the atomic absorption spectrophotometer, (4) Nitrate, nitrite and ammonium—Place 1.0 g of Sodi- and circulate air. Read the absorbance A of the sample solu- T um Iodide in a 40-mL test tube, and add 5 mL of water, 5 mL tion when the indication of the recorder rises rapidly and of sodium hydroxide TS and 0.2 g of aluminum wire. Insert a becomes constant at the wavelength of 253.7 nm. On the pledget of absorbent cotton in the mouth of the test tube, and other hand, to 2.0 mL of Standard Mercury Solution add 1 place a piece of moistened red litmus paper on the cotton. mL of a solution of potassium permanganate (3 in 50), 30 mL Heat the test tube on a water bath for 15 minutes: the evolved of water and a volume of puriˆed hydrochloric acid equal to gas does not turn moistened red litmus paper to blue. that used in the preparation of the sample solution, and read (5) Cyanide—Dissolve 0.5 g of Sodium Iodide in 10 mL the absorbance A of the solution obtained by the same S of water. To 5 mL of this solution add 1 drop of iron (II) sul- procedure as used for the sample solution: A is smaller than T fate TS and 2 mL of sodium hydroxide TS, warm, and add 4 A . S mL of hydrochloric acid: no green color develops. Assay Weigh accurately about 1.5 g of Sodium Hydroxide, (6) Iodate—Dissolve 0.5 g of Sodium Iodide in 10 mL of and dissolve in 40 mL of freshly boiled and cooled water. freshly boiled and cooled water, and add 2 drops of dilute Cool the solution to 159C, add 2 drops of phenolphthalein sulfuric acid and 1 drop of starch TS: no blue color develops JP XV O‹cial Monographs / Sodium Iotalamate Injection 1103 immediately. (7) Heavy metals <1.07>—Proceed with 2.0 g of Sodium Iodide according to Method 1, and perform the test. Prepare Sodium Iodide (131I) Solution the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). ヨウ化ナトリウム (131I) 液 (8) Barium—Dissolve 0.5 g of Sodium Iodide in 10 mL of water, add 1 mL of dilute sulfuric acid, and allow to stand for 5 minutes: no turbidity is produced. Sodium Iodide (131I) Solution contains iodine-131 (9) Potassium—Dissolve 1.0 g of Sodium Iodide in (131I) in the form of sodium iodide. water, and add water to make 100 mL. To 4.0 mL of this It conforms to the requirements of Sodium Iodide solution add 1.0 mL of dilute acetic acid, shake, add 5.0 mL (131I) Solution in the Minimum Requirements for of a solution of sodium tetraphenylboron (1 in 30), immedi- Radiopharmaceuticals. ately shake, and allow to stand for 10 minutes: the solution Description Sodium Iodide (131I) Solution is a clear, color- has no more turbidity than the following control solution. less liquid. It is odorless, or has an odor due to the preserva- Control solution: Dissolve 9.5 mg of potassium chloride in tives or stabilizers. water, and add water to make 1000 mL. To 4.0 mL of this so- lution add 1.0 mL of dilute acetic acid, shake, and then pro- ceed as directed above. 131 (10) Arsenic <1.11>—Prepare the test solution with 0.40 g Sodium Iodohippurate ( I) of Sodium Iodide according to Method 1, and perform the Injection test (not more than 5 ppm). 131 Loss on drying <2.41> Not more than 5.0z (2 g, 1209C, ヨウ化ヒプル酸ナトリウム ( I) 注射液 2 hours). Assay Weigh accurately about 0.4 g of Sodium Iodide, Sodium Iodohippurate (131I) Injection is an aqueous previously dried, in an iodine ‰ask, dissolve in 10 mL of solution for injection containing iodine-131 (131I) in the water, add 35 mL of hydrochloric acid and 5 mL of chlo- form of sodium o-iodohippurate. roform, and titrate <2.50> with 0.05 molWL potassium iodate It conforms to the requirements of Sodium Iodohip- VS while shaking vigorously until the red-purple color of the purate (131I) Injection in the Minimum Requirements chloroform layer disappears. The end point is attained when for Radiopharmaceuticals. the red-purple color does not reappear in the chloroform lay- Test for Extractable Volume of Parenteral Prepara- er within 5 minutes after the layer has been decolorized. tions and Insoluble Particulate Matter Test for Injec- Each mL of 0.05 molWL potassium iodate VS tions are not applied to this injection. ＝ 14.99 mg of NaI Description Sodium Iodohippurate (131I) Injection is a Containers and storage Containers—Tight containers. clear, colorless liquid. It is odorless or has an odor of the Storage—Light-resistant. preservatives or stabilizers.

Sodium Iodide (123I) Capsules Sodium Iotalamate Injection

ヨウ化ナトリウム (123I) カプセル イオタラム酸ナトリウム注射液

Sodium Iodide (123I) Capsules contain iodine-123 in Sodium Iotalamate Injection is an aqueous solution the form of sodium iodide. for injection. It conforms to the requirements of Sodium Iodide It contains not less than 95z and not more than (123I) Capsules in the Minimum Requirements for 105z of the labeled amount of iotalamic acid Radiopharmaceuticals. (C11H9I3N2O4: 613.91). Method of preparation Sodium Iodide (131I) Capsules (1) Iotalamic Acid 645 g ヨウ化ナトリウム (131I) カプセル Sodium Hydroxide 42 g Water for Injection a su‹cient quantity To make 1000 mL Sodium Iodide (131I) Capsules are prepared by dis- pensing Sodium Iodide (131I) Solution into capsules and (2) drying it. Iotalamic Acid 772.5 g Sodium Iodide (131I) Capsules conform to the re- Sodium Hydroxide 50.5 g quirements of Sodium Iodide (131I) Capsules in the Water for Injection a su‹cient quantity Minimum Requirements for Radiopharmaceuticals. To make 1000 mL Prepare as directed under Injections, with the above in- 1104 Sodium Lauryl Sulfate / O‹cial Monographs JP XV gredients (1) or (2). WS: Amount (mg) of iotalamic acid for assay Description Sodium Iotalamate Injection is a clear, color- Internal standard solution—A solution of L- in less or pale yellow, slightly viscous liquid. the mobile phase (3 in 2500). It is gradually colored by light. Operating conditions— Identiˆcation (1) To a volume of Sodium Iotalamate In- Detector: An ultraviolet absorption photometer jection, equivalent to 1 g of Iotalamic Acid according to the (wavelength: 240 nm). labeled amount, add 25 mL of water, and add 2.5 mL of Column: A stainless steel column 4.6 mm in inside di- dilute hydrochloric acid with thorough stirring: a white ameter and 15 cm in length, packed with octadecylsilanized precipitate is produced. Filter the precipitate by suction silica gel for liquid chromatography (5 mminparticledi- through a glass ˆlter (G4), wash the precipitate with two ameter). 10-mL portions of water, and dry at 1059C for 1 hour. Pro- Column temperature: A constant temperature of about ceed with the precipitate as directed in the Identiˆcation (2) 209C. under Iotalamic Acid. Mobile phase: To 3.9 g of phosphoric acid and 2.8 mL of (2) Sodium Iotalamate Injection responds to the Qualita- triethylamine add water to make 2000 mL. To this solution tive Tests <1.09> (1) for sodium salt. add 100 mL of acetonitrile. Flow rate: Adjust the ‰ow rate so that the retention time of pH <2.54> 6.5 – 7.7 iotalamic acid is about 6 minutes. Purity (1) Primary aromatic amines—To a volume of So- System suitability— dium Iotalamate Injection, equivalent to 0.20 g of Iotalamic System performance: When the procedure is run with 10 Acid according to the labeled amount, add 15 mL of water, mL of the standard solution under the above operating condi- shake, add 4 mL of a solution of sodium nitrite (1 in 100) un- tions, iotalamic acid and the internal standard are eluted in der ice-cooling, and proceed as directed in the Purity (2) un- this order with the resolution between these peaks being not der Iotalamic Acid: the absorbance is not more than 0.17. less than 5. (2) Iodine and iodide—To a volume of Sodium Iotala- System repeatability: When the test is repeated 6 times with mate Injection, equivalent to 1.5 g of Iotalamic Acid accord- 10 mL of the standard solution under the above operating ingtothelabeledamount,add20mLofwaterand5mLof conditions, the relative standard deviation of the ratios of the dilute sulfuric acid, shake well, and ˆlter the precipitate by peak area of iotalamic acid to that of the internal standard is suction through a glass ˆlter (G4). To the ˆltrate add 5 mL of not more than 1.0z. toluene, and shake vigorously: the toluene layer is colorless. Containers and storage Containers—Hermetic containers, Then add 2 mL of a solution of sodium nitrite (1 in 100), and and colored containers may be used. shake vigorously: the toluene layer has no more color than Storage—Light-resistant. the following control solution. Control solution: Dissolve 0.25 g of potassium iodide in water to make 1000 mL. To 2.0 mL of this solution add 20 mL of water, 5 mL of dilute sulfuric acid, 5 mL of toluene Sodium Lauryl Sulfate and 2 mL of a solution of sodium nitrite (1 in 100), and shake ラウリル硫酸ナトリウム vigorously. Bacterial endotoxins <4.01> Less than 3.4 EUWmL. Extractable volume <6.05> It meets the requirement. Sodium Lauryl Sulfate is a mixture of sodium alkyl sulfate consisting chie‰y of sodium lauryl sulfate Assay Pipet a volume of Sodium Iotalamate Injection, (C12H25NaO4S: 288.38). equivalent to about 4 g of iotalamic acid (C H I N O ), add 11 9 3 2 4 Description Sodium Lauryl Sulfate occurs as white to light watertomakeexactly200mL.Pipet2mLofthissolution, yellow crystals or powder. It has a slightly characteristic add water to make exactly 200 mL. To exactly 5 mL of this odor. solution add exactly 5 mL of the internal standard solution, It is sparingly soluble in methanol and in ethanol (95). add the mobile phase to make 100 mL, and use this solution A solution of Sodium Lauryl Sulfate (1 in 10) is a clear or as the sample solution. Separately, weigh accurately about an opalescent solution, which foams on agitation. 0.4 g of iotalamic acid for assay, previously dried at 1059C for 4 hours, dissolve in 100 mL of water and 1 mL of sodium Identiˆcation (1) To 0.2 g of the residue obtained in Total hydroxide TS, and add water to make exactly 200 mL. Pipet alcohol content add 4 mL of bromine-cyclohexane TS with 5 mL of this solution, add water to make exactly 50 mL. To vigorous shaking, add 0.3 g of N-bromosuccinimide, and exactly 5 mL of this solution add exactly 5 mL of the internal heat in a water bath at 809C for 5 minutes: a red color de- standard solution, add the mobile phase to make 100 mL, velops. and use this solution as the standard solution. Perform the (2) A solution of Sodium Lauryl Sulfate (1 in 10) test with 10 mL each of the sample solution and standard so- responds to the Qualitative Tests <1.09> (1) for sodium salt. lution as directed under Liquid Chromatography <2.01> ac- (3) To a solution of Sodium Lauryl Sulfate (1 in 10) add cording to the following conditions, and calculate the ratios, dilute hydrochloric acid to make acid, boil gently, and cool: QT and Q S, of the peak area of iotalamic acid to that of the the solution responds to the Qualitative Tests <1.09> for sul- internal standard. fate.

Amount (mg) of iotalamic acid (C11H9I3N2O4) Purity (1) Alkalinity—Dissolve 1.0 g of Sodium Lauryl ＝ WS×(QT/QS) Sulfate in 100 mL of water, add 2 drops of phenol red TS and JP XV O‹cial Monographs / Sodium Picosulfate Hydrate 1105

0.60 mL of 0.1 molWL hydrochloric acid VS: the solution ous solution for injection containing technetium-99m remains yellow. (99mTc) in the form of sodium pertechnetate. (2) Sodium chloride—Dissolve about 5 g of Sodium It conforms to the requirements of Sodium Per- Lauryl Sulfate, accurately weighed, in 50 mL of water, neu- technetate (99mTc) Injection in the Minimum Require- tralize the solution with dilute nitric acid, if necessary, add ments for Radiopharmaceuticals. exactly 5 mL of 0.1 molWL sodium chloride TS, and titrate Test for Extractable Volume of Parenteral Prepara- <2.50> with 0.1 molWL silver nitrate VS (indicator: 2 drops of tions and Insoluble Particulate Matter Test for Injec- ‰uorescein sodium TS). Perform a blank determination, and tions are not applied to this injection. make any necessary correction. Description Sodium Pertechnetate (99mTc) Injection is a Each mL of 0.1 molWL silver nitrate VS clear, colorless liquid. ＝5.844 mg of NaCl The combined content of sodium chloride (NaCl: 58.44) Sodium Picosulfate Hydrate and sodium sulfate (Na2SO4: 142.04) obtained in the next paragraph (3) is not more than 8.0z. ピコスルファートナトリウム水和物 (3) Sodium sulfate—Dissolve about 1 g of Sodium Lauryl Sulfate, accurately weighed, in 10 mL of water, add 100 mL of ethanol (95), and heat at a temperature just below the boiling point for 2 hours. Filter through a glass ˆlter (G4) while hot, and wash with 100 mL of boiling ethanol (95). Dis- solve the precipitate by washing with 150 mL of water, col- lecting the washings in a beaker. Add 10 mL of hydrochloric acid, heat to boiling, add 25 mL of barium chloride TS, and allow to stand overnight. Collect the precipitate, and wash C18H13NNa2O8S2.H2O: 499.42 with water until the last washing shows no opalescence with Disodium 4,4?-(pyridin-2-ylmethylene)bis(phenyl sulfate) silver nitrate TS. Dry the precipitate, ignite to a constant monohydrate [10040-45-6,anhydride] mass between 5009C and 6009C by raising the temperature gradually, and weigh as barium sulfate (BaSO4: 233.39). Sodium Picosulfate Hydrate contains not less than 98.5z of sodium picosulfate (C18H13NNa2O8S2: Amount (mg) of sodium sulfate (Na2SO4) 481.41), calculated on the anhydrous basis. ＝amount (mg) of barium sulfate (BaSO4)×0.6086 Description Sodium Picosulfate Hydrate occurs as a white, (4) Unsulfated —Dissolve about 10 g of Sodium crystalline powder. It is odorless and tasteless. Lauryl Sulfate, accurately weighed, in 100 mL of water, add It is very soluble in water, soluble in methanol, slightly 100 mL of ethanol (95), and transfer to a separator. Extract soluble in ethanol (99.5), and practically insoluble in diethyl the solution with three 50-mL portions of petroleum benzin. ether. If an emulsion forms, sodium chloride may be added to pro- It is gradually colored by light. mote separation of the two layers. Combine the petroleum The pH of a solution of Sodium Picosulfate Hydrate (1 in benzinextractsandwashwiththree50-mLportionsofwater. 20) is between 7.4 and 9.4. Evaporate the petroleum benzin on a water bath, and dry the residue at 1059C for 30 minutes. The mass of the dried Identiˆcation (1) Mix 5 mg of Sodium Picosulfate Hy- residue is not more than 4.0z of the mass of the Sodium drate with 0.01 g of 1-chloro-2,4-dinitrobenzene, and melt by Lauryl Sulfate taken. gentle heating for 5 to 6 seconds. After cooling, add 4 mL of potassium hydroxide-ethanol TS: an orange-red color de- Water <2.48> Not more than 5.0z (0.5 g, direct titration). velops. Total alcohol content Dissolve about 5 g of Sodium Lauryl (2) To 0.2 g of Sodium Picosulfate Hydrate add 5 mL of Sulfate, accurately weighed, in 150 mL of water and 50 mL dilute hydrochloric acid, boil for 5 minutes, cool, and add 1 of hydrochloric acid, and boil under a re‰ux condenser for 4 mL of barium chloride TS: a white precipitate is formed. hours. Cool, extract with two 75-mL portions of diethyl (3) Determine the absorption spectrum of a solution of ether, and evaporate the combined diethyl ether extracts on a Sodium Picosulfate Hydrate (1 in 25,000) as directed under water bath. Dry the residue at 1059C for 30 minutes. The Ultraviolet-visible Spectrophotometry <2.24>,andcompare mass of the residue is not less than 59.0z. the spectrum with the Reference Spectrum: both spectra ex- hibit similar intensities of absorption at the same Containers and storage Containers—Well-closed contain- wavelengths. ers. (4) Determine the infrared absorption spectrum of Sodi- um Picosulfate Hydrate, previously dried at 105°Cinvacu- um for 4 hours, as directed in the potassium bromide disk 99m Sodium Pertechnetate ( Tc) method under the Infrared Spectrophotometry <2.25>,and Injection compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same 過テクネチウム酸ナトリウム (99mTc) 注射液 wave numbers. (5) A solution of Sodium Picosulfate Hydrate (1 in 10) responds to the Qualitative Tests <1.09> for sodium salt. Sodium Pertechnetate (99mTc) Injection is an aque- 1z Absorbance <2.24> E 1cm (263 nm): 120 – 130 (calculated on 1106 Sodium Polystyrene Sulfonate / O‹cial Monographs JP XV the anhydrous basis, 4 mg, water, 100 mL). Description Sodium Polystyrene Sulfonate occurs as a yel- Purity (1) Clarity and color of solution—Dissolve 1.0 g of low-brown powder. It is odorless and tasteless. Sodium Picosulfate Hydrate in 10 mL of water: the solution It is practically insoluble in water, in ethanol (95), in ace- is clear and colorless to pale yellow. toneandindiethylether. (2) Chloride <1.03>—Perform the test with 0.5 g of Sodi- Identiˆcation (1) Determine the infrared absorption spec- um Picosulfate Hydrate. Prepare the control solution with trum of Sodium Polystyrene Sulfonate as directed in the 0.40 mL of 0.01 molWL hydrochloric acid VS (not more than potassium bromide disk method under the Infrared Spec- 0.028z). trophotometry <2.25>, and compare the spectrum with the (3) Sulfate <1.14>—Perform the test with 0.40 g of Sodi- Reference Spectrum: both spectra exhibit similar intensities um Picosulfate Hydrate. Prepare the control solution with of absorption at the same wave numbers. 0.35 mL of 0.005 molWL sulfuric acid VS (not more than (2) To 1 g of Sodium Polystyrene Sulfonate add 10 mL of 0.042z). dilute hydrochloric acid, stir, and ˆlter. Add ammonia TS to (4) Heavy metals <1.07>—Proceed with 2.0 g of Sodium the ˆltrate to neutralize: the solution responds to the Qualita- Picosulfate Hydrate according to Method 2, and perform the tive Tests <1.09> for sodium salt. test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Purity (1) Ammonium—Place 1.0 g of Sodium Polysty- (5) Arsenic <1.11>—Prepare the test solution with 2.0 g rene Sulfonate in a ‰ask, add 5 mL of sodium hydroxide TS, of Sodium Picosulfate Hydrate according to Method 3, and cover the ‰ask with a watch glass having a moistened strip of perform the test (not more than 1 ppm). red litmus paper on the underside, and boil for 15 minutes: (6) Related substances—Dissolve 0.25 g of Sodium the gas evolved does not change the red litmus paper to blue. Picosulfate Hydrate in 5 mL of methanol, and use this solu- (2) Heavy metals <1.07>—Proceed with 2.0 g of Sodium tion as the sample solution. Pipet 1 mL of the sample solu- Polystyrene Sulfonate according to Method 2, and perform tion, add methanol to make exactly 500 mL, and use this so- the test. Prepare the control solution with 2.0 mL of Stan- lution as the standard solution. Perform the test with these dard Lead Solution (not more than 10 ppm). solutions as directed under Thin-layer Chromatography (3) Arsenic <1.11>—Prepare the test solution with 2.0 g <2.03>.Spot5mL each of the sample solution and standard of Sodium Polystyrene Sulfonate according to Method 3, and solution on a plate of silica gel with ‰uorescent indicator for perform the test (not more than 1 ppm). thin-layer chromatography. Develop the plate with a mixture (4) Styrene—To 10.0 g of Sodium Polystyrene Sulfonate of 1-butanol, water and acetic acid (100) (74:20:19) to a dis- add 10 mL of acetone, shake for 30 minutes, centrifuge, and tance of about 10 cm, and air-dry the plate. Examine under use the supernatant liquid as the sample solution. Separately, ultraviolet light (main wavelength: 254 nm): the spots other dissolve 10 mg of styrene in acetone to make exactly 100 mL. than the principal spot from the sample solution are not more Pipet 1 mL of this solution, add acetone to make exactly intense than the spot from the standard solution. 100 mL, and use this solution as the standard solution. Per- form the test with exactly 20 mL each of the sample solution Water <2.48> 3.0–4.5z (0.5 g, direct titration). and standard solution as directed under Liquid Chro- Assay Weigh accurately about 0.4 g of Sodium Picosulfate matography <2.01> according to the following conditions,

Hydrate, dissolve in 50 mL of methanol, add 7 mL of acetic and determine peak areas, AT and AS, of styrene in each solu- acid (100), and titrate <2.50> with 0.1 molWL perchloric acid tion: AT is not larger than AS. VS (potentiometric titration). Perform a blank determina- Operating conditions— tion, and make any necessary correction. Detector: An ultraviolet absorption photometer (wavelength: 254 nm). Each mL of 0.1 molWL perchloric acid VS Column: A stainless steel column 4 mm in inside diameter ＝ 48.14 mg of C H NNa O S 18 13 2 8 2 and 15 cm in length, packed with octadecylsilanized silica gel Containers and storage Containers—Tight containers. for liquid chromatography (5 mminparticlediameter). Storage—Light-resistant. Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and acetonitrile (1:1). Sodium Polystyrene Sulfonate Flow rate: Adjust the ‰ow rate so that the retention time of styrene is about 8 minutes. ポリスチレンスルホン酸ナトリウム System suitability— System performance: Dissolve 20 mg each of styrene and butyl parahydroxybenzoate in 100 mL of acetone. To 5 mL Sodium Polystyrene Sulfonate is a cation exchange of this solution add acetone to make 100 mL. When the resin prepared as the sodium form of the sulfonated procedure is run with 20 mL of this solution under the above styrene divinylbenzene copolymer. operating conditions, butyl parahydroxybenzoate and sty- It contains not less than 9.4z and not more than rene are eluted in this order with the resolution between these 11.0z of sodium (Na: 22.99), calculated on the anhy- peaks being not less than 5. drous basis. System repeatability: When the test is repeated 6 times with Each g of Sodium Polystyrene Sulfonate, calculated 20 mL of the standard solution under the above operating on the anhydrous basis, exchanges with not less than conditions, the relative standard deviation of the peak area of 0.110 g and not more than 0.135 g of potassium (K: styrene is not more than 2.0z. 39.10). Water <2.48> Not more than 10.0z (0.2 g, direct titration). JP XV O‹cial Monographs / Sodium Sulfate Hydrate 1107

Assay (1) Sodium—Weigh accurately about 1 g of Sodi- um Polystyrene Sulfonate, calculated on the anhydrous ba- sis, in a glass-stoppered ‰ask, add exactly 50 mL of 3 molWL Sodium Hydrate hydrochloric acid TS, shake for 60 minutes, and ˆlter. Dis- プラステロン硫酸エステルナトリウム水和物 card the ˆrst 20 mL of the ˆltarte, pipet the subsequent 5 mL of the ˆltrate, and add water to make exactly 100 mL. Pipet 20 mL of this solution, add water to make exactly 1000 mL, and use this solution as the sample solution. Separately, pipet a suitable quantity of Standard Sodium Stock Solution, di- lute exactly with water so that each ml of the solution con- tains 1 to 3 mg of sodium (Na: 22.99), and use these solutions as the standard solution. Perform the test with the sample so- lution and the standard solution according to the Atomic Ab- C19H27NaO5S.2H2O: 426.50 sorption Spectrophotometry <2.23> under the following con- Monosodium 17-oxoandrost-5-en-3b-yl sulfate dihydrate ditions, and determine the amount of sodium in the sample [1099-87-2, anhydride] solution using the calibration curve obtained from the stan- dard solutions. Sodium Prasterone Sulfate Hydrate contains not less Gas: Combustible gas—Acetylene than 98.0z of sodium prasterone sulfate (C19H27NaO5 Supporting gas—Air S: 390.47), calculated on the dried basis. Lamp: A sodium hollow-cathode lamp Description Sodium Prasterone Sulfate Hydrate occurs as Wavelength: 589.0 nm white crystals or crystalline powder. It is odorless. (2) Potassium exchange capacity—Weigh accurately It is soluble in methanol, sparingly soluble in water and in about 1.5 g of Sodium Polystyrene Sulfonate, calculated on ethanol (95), and practically insoluble in acetone and in the anhydrous basis, in a glass-stoppered ‰ask, add exactly diethyl ether. 100 mL of Standard Potassium Stock Solution, shake for 15 The pH of a solution of Sodium Prasterone Sulfate Hy- minutes, and ˆlter. Discard the ˆrst 20 mL of the ˆltrate, drate (1 in 200) is between 4.5 and 6.5. pipet the subsequent 10 mL of the ˆltrate, and add water to Melting point: about 1609C (with decomposition, after make exactly 100 mL. Pipet 10 mL of this solution, add drying). water to make exactly 1000 mL, and use this solution as the sample solution. Separately, pipet a suitable quantity of Identiˆcation (1) Dissolve 0.01 g of Sodium Prasterone Standard Potassium Stock Solution, dilute with water so that Sulfate Hydrate in 4 mL of ethanol (95), add 2 mL of 1,3- each mL of the solution contains 1 to 5 mg of potassium (K: dinitrobenzene TS and 2 mL of a solution of sodium 39.10), and use the solution as the standard solution. Per- hydroxide (1 in 8): a red-purple color develops, and gradually form the test with these solutions as directed under Atomic changes to brown. Absorption Spectrophotometry <2.23>, and determine the (2) To 10 mL of a solution of Sodium Prasterone Sulfate amount Y (mg) of potassium in 1000 mL of the sample solu- Hydrate (1 in 200) add 0.5 mL of bromine TS: the color of tion using the calibration curve obtained from the standard bromine TS immediately disappears. solution. The quantity of potassium absorbed on each g of (3) Determine the infrared absorption spectrum of Sodi- Sodium Polystyrene Sulfonate, calculated on the anhydrous um Prasterone Sulfate Hydrate as directed in the potassium basis, is calculated from the following equation: it is between bromide disk method under the Infrared Spectrophotometry 0.110 g and 0.135 g. <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at Quantity (mg) of potassium (K) absorbed on 1 g of Sodium thesamewavenumbers. Polystyrene Sulfonate, calculated on the anhydrous basis (4) A solution of Sodium Prasterone Sulfate Hydrate (1 ＝ (X － 100Y )/W in 200) responds to the Qualitative Tests <1.09> for sodium X: Amount (mg) of potassium in 100 mL of the Standard salt. Potassium Stock Solution before exchange. 20 Optical rotation <2.49> [a]D : ＋10.7 – ＋12.19(0.73 g, cal- W: Mass (g) of Sodium Polystyrene Sulfonate taken, cal- culated on the dried basis, methanol, 20 mL, 100 mm). culated on the anhydrous basis. Gas: Combustible gas—Acetylene Purity (1) Clarity and color of solution—Dissolve 0.25 g Supporting gas—Air of Sodium Prasterone Sulfate Hydrate in 50 mL of water: the Lamp: A potassium hollow-cathode lamp solution is clear and colorless. Wavelength: 766.5 nm (2) Chloride <1.03>—Dissolve 1.0 g of Sodium Praster- one Sulfate Hydrate in 20 mL of acetone and 20 mL of water, Containers and storage Containers—Tight containers. and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Pre- pare the control solution as follows: to 0.30 mL of 0.01 molW L hydrochloric acid VS add 20 mL of acetone, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.011z). (3) Sulfate <1.14>—To 1.2 g of Sodium Prasterone Sul- fate Hydrate add 20 mL of water, shake vigorously for 5 minutes, and ˆlter. To 10 mL of the ˆltrate add 20 mL of ace- 1108 Sodium Pyrosulˆte / O‹cial Monographs JP XV tone, 1 mL of dilute hydrochloric acid and water to make 50 It decomposes slowly on exposure to air. mL. Perform the test using this solution as the test solution. Identiˆcation A solution of Sodium Pyrosulˆte (1 in 20) Prepare the control solution as follows: to 0.40 mL of 0.005 responds to the Qualitative Tests <1.09> for sodium salt and molWL sulfuric acid VS add 20 mL of acetone, 1 mL of dilute for bisulˆte. hydrochloric acid and water to make 50 mL (not more than 0.032z). Purity (1) Clarity and color of solution—Dissolve 1.0 g of (4) Heavy metals <1.07>—Proceed with 2.0 g of Sodium Sodium Pyrosulˆte in 10 mL of water: the solution is clear Prasterone Sulfate Hydrate according to Method 2, and per- and colorless. form the test. Prepare the control solution with 2.0 mL of (2) Thiosulfate—Dissolve 1.0 g of Sodium Pyrosulˆte in Standard Lead Solution (not more than 10 ppm). 15 mL of water, add slowly 5 mL of dilute hydrochloric acid, (5) Related substances—Dissolve 0.10 g of Sodium shake, and allow to stand for 5 minutes: no turbidity is Prasterone Sulfate Hydrate in 10 mL of methanol, and use produced. this solution as the sample solution. Pipet 1 mL of the sample (3) Heavy metals <1.07>—Dissolve 1.0 g of Sodium solution, add methanol to make exactly 200 mL, and use this Pyrosulˆte in 10 mL of water, and evaporate with 5 mL of solution as the standard solution. Perform the test with these hydrochloric acid on a water bath to dryness. Dissolve the solutions as directed under Thin-layer Chromatography residue in 10 mL of water, add 1 drop of phenolphthalein TS, <2.03>.Spot5mL each of the sample solution and standard and add ammonia TS until the solution becomes slightly red. solution on a plate of silica gel with ‰uorescent indicator for Add 2 mL of dilute acetic acid and water to make 50 mL. thin-layer chromatography. Develop the plate with a mixture Perform the test using this solution as the test solution. Pre- of chloroform, methanol and water (75:22:3) to a distance of pare the control solution as follows: evaporate 5 mL of about 10 cm, and air-dry the plate. Spray evenly a mixture of hydrochloric acid on a water bath to dryness, and to the sulfuric acid and ethanol (95) (1:1) on the plate, and heat at residue add 2 mL of dilute acetic acid, 2.0 mL of Standard 809C for 5 minutes: the spots other than the principal spot Lead Solution and water to make 50 mL (not more than 20 from the sample solution are not more intense than the spot ppm). from the standard solution. (4) Iron <1.10>—Prepare the test solution with 1.0 g of Sodium Pyrosulˆte according to Method 1, and perform the Loss on drying <2.41> 8.0–9.0z (0.5 g, in vacuum, phos- test according to Method A. Prepare the control solution phorus (V) oxide, 609C, 3 hours). with 2.0 mL of Standard Iron Solution (not more than 20 Assay Weigh accurately about 0.25 g of Sodium Prasterone ppm). Sulfate Hydrate, dissolve in 30 mL of water. Apply this solu- (5) Arsenic <1.11>—Dissolve 0.5 g of Sodium Pyrosulˆte tion to a chromatographic column 10 mm in inside diameter, in 10 mL of water, heat with 1 mL of sulfuric acid on a sand previously prepared by pouring 5 mL of strongly acidic - bath until white fumes are evolved, and add water to make 5 exchange resin (H type) for column chromatography, and mL. Perform the test with this solution as the test solution elute at the rate of 4 mL per minute. Wash the chromato- (not more than 4 ppm). graphic column with 100 mL of water, combine the washings Assay Weigh accurately about 0.15 g of Sodium Pyrosul- with above eŒuent solution, and titrate <2.50> with 0.05 ˆte, and transfer to an iodine ‰ask containing an exactly molWL sodium hydroxide VS (potentiometric titration). Per- measured 50 mL of 0.05 molWL iodine VS. Stopper tightly, form a blank determination, and make any necessary correc- shake well, and allow to stand for 5 minutes in a dark place. tion. Add 1 mL of hydrochloric acid VS, and titrate <2.50> the ex- Each mL of 0.05 molWLsodiumhydroxideVS cess of iodine with 0.1 molWL sodium thiosulfate VS (indica-

＝ 19.52 mg of C19H27NaO5S tor: 1 mL of starch TS). Perform a blank determination.

Containers and storage Containers—Tight containers. Each mL of 0.05 molWLiodineVS＝4.753 mg of Na2S2O5 Containers and storage Containers—Tight containers. Storage—Light-resistant, preferably well-ˆlled, and not Sodium Pyrosulˆte exceeding 309C. Sodium Metabisulˆte

ピロ亜硫酸ナトリウム Sodium Salicylate

サリチル酸ナトリウム

Na2S2O5:190.11

Sodium Pyrosulˆte contains not less than 95.0z of Na2S2O5.

Description Sodium Pyrosulˆte occurs as white crystals or C7H5NaO3: 160.10 crystalline powder. It has the odor of sulfur dioxide. Monosodium 2-hydroxybenzoate [54-21-7 ] It is freely soluble in water, very slightly soluble in ethanol (95), and practically insoluble in diethyl ether. Sodium Salicylate, when dried, contains not less A solution of Sodium Pyrosulˆte (1 in 20) is acid. than 99.5z of C7H5NaO3. It is hygroscopic. Description Sodium Salicylate occurs as white, crystals or JP XV O‹cial Monographs / Sodium Thiosulfate Hydrate 1109 crystalline powder. It is very soluble in water, freely soluble in acetic acid (100), and soluble in ethanol (95). Dried Sodium Sulˆte It is gradually colored by light. 乾燥亜硫酸ナトリウム Identiˆcation (1) Determine the infrared absorption spec- trum of Sodium Salicylate, previously dried, as directed in the potassium bromide disk method under the Infrared Spec- Na SO :126.04 trophotometry <2.25>, and compare the spectrum with the 2 3 Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dried Sodium Sulˆte contains not less than 97.0z of (2) A solution of Sodium Salicylate (1 in 20) responds to Na2SO3. the Qualitative Tests <1.09> for sodium salt. Description Dried Sodium Sulˆte is white crystals or pow- der. It is odorless. pH <2.54> The pH of a solution of 2.0 g of Sodium Salicy- It is freely soluble in water, and practically insoluble in late in 20 mL of water is between 6.0 and 8.0. ethanol (95) and in diethyl ether. Purity (1) Clarity of solution—Dissolve 1.0 g of Sodium The pH of a solution of Dried Sodium Sulˆte (1 in 10) is Salicylate in 10 mL of water: the solution is clear, and its ab- about 10. sorbance at 420 nm determined as directed under Ultraviolet- It gradually changes in moist air. visible Spectrophotometry <2.24> is not more than 0.02. Identiˆcation An aqueous solution of Dried Sodium Sulˆte (2) Chloride <1.03>—Dissolve 0.5 g of Sodium Salicylate (1 in 20) responds to the Qualitative Tests <1.09> for sodium in 15 mL of water, add 6 mL of dilute nitric acid and ethanol salt and sulˆte. (95) to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.30 Purity (1) Thiosulfate—Dissolve 1.0 g of Dried Sodium mL of 0.01 molWL hydrochloric acid VS, 28 mL of ethanol Sulˆte in 15 mL of water, add gradually 5 mL of hydrochlor- (95), 6 mL of dilute nitric acid and water to make 50 mL (not ic acid, shake, and allow to stand for 5 minutes: no turbidity more than 0.021z). is produced. (3) Sulfate—Dissolve 0.25 g of Sodium Salicylate in 5 mL (2) Heavy metals <1.07>—Dissolve 1.0 g of Dried Sodium of water, and add 0.5 mL of barium chloride TS: the solution Sulˆte in 5 mL of water, add 2 mL of hydrochloric acid grad- shows no change. ually, and evaporate the mixture on a water bath to dryness. (4) Sulˆte and thiosulfate—Dissolve 1.0 g of Sodium Add 3 mL of boiling water and 1 mL of hydrochloric acid to Salicylate in 20 mL of water, add 1 mL of hydrochloric acid, the residue, and again evaporate to dryness on a water bath. and ˆlter. Add 0.15 mL of 0.05 molWLiodineVStothe Dissolve the residue in 2 mL of dilute acetic acid and water to ˆltrate: a yellow color develops. make 50 mL, and perform the test using this solution as the (5) Heavy metals <1.07>—Proceed with 1.0 g of Sodium test solution. Prepare the control solution as follows: Salicylate according to Method 2, and perform the test. Pre- evaporate 3 mL of hydrochloric acid to dryness, and add 2 pare the control solution with 2.0 mL of Standard Lead Solu- mL of dilute acetic acid, 2.0 mL of Standard Lead Solution tion (not more than 20 ppm). and water to make 50 mL (not more than 20 ppm). (6) Arsenic <1.11>—To 1.0 g of Sodium Salicylate in a (3) Arsenic <1.11>—Dissolve 0.5 g of Dried Sodium Sul- decomposition ‰ask add 5 mL of nitric acid and 2 mL of sul- ˆte in 5 mL of water, add 1 mL of sulfuric acid, and furic acid, and heat carefully until white fumes are evolved. evaporate on a sand bath until white fumes are evolved. Add After cooling, add 2 mL of nitric acid, and heat. After cool- watertomake5mL,takethissolutionasthesamplesolu- ing, add several 2-mL portions of hydrogen peroxide (30), tion, and perform the test (not more than 4 ppm). and heat until the solution is colorless to pale yellow. Repeat Assay Weigh accurately about 0.2 g of Dried Sodium Sul- the procedure of adding nitric acid and hydrogen peroxide ˆte, transfer immediately to an iodine ‰ask containing exact- (30) and heating, if necessary. After cooling, add 2 mL of a ly 50 mL of 0.05 mol L iodine VS, stopper, shake, and allow saturated solution of ammonium oxalate monohydrate, and W to stand for 5 minutes in a dark place. Add 1 mL of heat until white fumes are evolved. After cooling, add water hydrochloric acid, and titrate <2.50> theexcessiodinewith tomake5mL,andperformthetestwiththissolution(not 0.1 mol L sodium thiosulfate VS (indicator: 1 mL of starch more than 2 ppm). W TS). Perform a blank determination. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, Each mL of 0.05 mol LiodineVS＝6.302 mg of Na SO 2 hours). W 2 3 Containers and storage Containers—Tight containers. Assay Weigh accurately about 0.3 g of Sodium Salicylate, previously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 molWL perchloric acid VS (potentio- metric titration). Perform a blank determination, and make Sodium Thiosulfate Hydrate any necessary correction. チオ硫酸ナトリウム水和物 Each mL of 0.1 molWL perchloric acid VS

＝ 16.01 mg of C7H5NaO3 Containers and storage Containers—Tight containers. Na2S2O3.5H2O: 248.18 Storage—Light-resistant. Sodium Thiosulfate Hydrate, when dried, contains 1110 Sodium Thiosulfate Injection / O‹cial Monographs JP XV not less than 99.0z and not more than 101.0z of sodi- z of the labeled amount of sodium thiosulfate hydrate um thiosulfate (Na2S2O3:158.11). (Na2S2O3.5H2O: 248.18). Description Sodium Thiosulfate Hydrate occurs as color- Method of preparation Prepare as directed under Injec- less, crystals or crystalline powder. It is odorless. tions, with Sodium Thiosulfate Hydrate. It is very soluble in water, and practically insoluble in Description Sodium Thiosulfate Injection is a clear, color- ethanol (99.5). less liquid. It eŒoresces in dry air, and is deliquescent in moist air. Identiˆcation Sodium Thiosulfate Injection responds to the Identiˆcation (1) A solution of Sodium Thiosulfate Hy- Qualitative Tests <1.09> for sodium salt and for thiosulfate. drate (1 in 10) responds to the Qualitative Tests <1.09> for thiosulfate. Extractable volume <6.05> It meets the requirement. (2) A solution of Sodium Thiosulfate Hydrate (1 in 10) Pyrogen <4.04> Perform the test with Sodium Thiosulfate responds to the Qualitative Tests <1.09> for sodium salt. Injection stored in a container in a volume exceeding 10 mL: pH <2.54> Dissolve 1.0 g of Sodium Thiosulfate Hydrate in it meets the requirements. 10 mL of water: the pH of the solution is between 6.0 and Assay Measure exactly a volume of Sodium Thiosulfate In- 8.0. jection, equivalent to about 0.5 g of sodium thiosulfate hy-

Purity (1) Clarity and color of solution—Dissolve 1.0 g of drate (Na2S2O3.5H2O), add water to make 30 mL, and titrate Sodium Thiosulfate Hydrate in 10 mL of water: the solution <2.50> with 0.05 molWL iodine VS (indicator: 1 mL of starch is clear and colorless. TS). (2) Heavy metals <1.07>—Dissolve 1.0 g of Sodium Each mL of 0.05 molWLiodineVS Thiosulfate Hydrate in 10 mL of water, add slowly 5 mL of ＝ 24.82 mg of Na S O .5H O dilute hydrochloric acid, and evaporate on a water bath to 2 2 3 2 dryness. Add 15 mL of water to the residue, boil gently for 2 Containers and storage Containers—Hermetic containers. minutes, and ˆlter. Heat the ˆltrate to boil, and add bromine TS to the hot ˆltrate to produce a clear solution and provide a slight excess of bromine. Boil the solution to expel the bro- Sodium mine. Cool, add 1 drop of phenolphthalein TS, and add dropwise sodium hydroxide TS until a slight red color is バルプロ酸ナトリウム produced. Add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solu- tion. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and watertomake50mL(notmorethan20ppm). C8H15NaO2:166.19 (3) Calcium—Dissolve 1.0 g of Sodium Thiosulfate in 10 Monosodium 2-propylpentanoate [1069-66-5] mL of water, add 2 mL of ammonium oxalate TS, and allow to stand for 4 minutes: no turbidity is produced. Sodium Valproate, when dried, contains not less (4) Arsenic <1.11>—To 0.40 g of Sodium Thiosulfate add than 98.5z of C8H15NaO2. 3 mL of nitric acid and 5 mL of water, evaporate on a water bath to dryness, and perform the test with the residue. Pre- Description Sodium Valproate occurs as a white, crystalline pare the test solution according to Method 2, and perform powder. It has a characteristic odor and a slightly bitter taste. the test (not more than 5 ppm). It is very soluble in water, freely soluble in formic acid, in ethanol (95), in ethanol (99.5) and in acetic acid (100), and Loss on drying <2.41> 32.0 – 37.0z (1 g, in vacuum, 40 – practically insoluble in chloroform and in diethyl ether. 459C, 16 hours). It is hygroscopic. Assay Weigh accurately about 0.4 g of Sodium Thiosulfate, Identiˆcation (1) To 1 mL of a solution of Sodium Val- previously dried, dissolve in 30 mL of water, and titrate proate in ethanol (99.5) (1 in 200) add 4 mL of hydroxyla- <2.50> with 0.05 mol/L iodine VS (indicator: 1 mL of starch mine perchlorate-dehydrated ethanol TS and 1 mL of N,N?- TS). dicyclohexylcarbodiimide-dehydrated ethanol TS, shake Each mL of 0.05 mol/L iodine VS well, and allow to stand in lukewarm water for 20 minutes. After cooling, add 1 mL of iron (III) perchlorate-dehydrated ＝15.81 mg of Na2S2O3 ethanol TS, and shake: a purple color develops. Containers and storage Containers—Tight containers. (2) To 5 mL of a solution of Sodium Valproate (1 in 20) add 1 mL of a solution of cobalt (II) nitrate hexahydrate (1 in 20), and warm on a water bath: a purple precipitate is Sodium Thiosulfate Injection formed. (3) Dissolve 0.5 g of Sodium Valproate in 5 mL of water, チオ硫酸ナトリウム注射液 add 5 mL of chloroform and 1 mL of 2 mol/L hydrochloric acid TS, and shake vigorously for 1 minute. After allowing to stand, separate the chloroform layer, dehydrate the chlo- Sodium Thiosulfate Injection is an aqueous solution roform with anhydrous sodium sulfate, then ˆlter, and for injection. evaporate the ˆltrate to dryness. Determine the infrared ab- It contains not less than 95z and not more than 105 sorption spectrum of the residue as directed in the liquid ˆlm JP XV O‹cial Monographs / Sorbitan Sesquioleate 1111 method under the Infrared Spectrophotometry <2.25>,and Carrier gas: Nitrogen compare the spectrum with the Reference Spectrum: both Flow rate: Adjust the ‰ow rate so that the retention time of spectra exhibit similar intensities of absorption at the same valproic acid is between 6 and 10 minutes. wave numbers. Selection of column: Mix 1 mL of the sample solution and (4) A solution of Sodium Valproate (1 in 10) responds to 4mLofasolutionofn-valerianic acid in a mixture of formic the Qualitative Tests <1.09> for sodium salt. acid and chloroform (1:1) (1 in 1000). Proceed with 2 mLof this solution under the above operating conditions, and cal- pH <2.54> Dissolve 1.0 g of Sodium Valproate in 20 mL of culate the resolution. Use a column giving elution of n- water: the pH of this solution is between 7.0 and 8.5. valerianic acid and valproic acid in this order with the resolu- Purity (1) Clarity and color of solution—Dissolve 1.0 g of tion between these peaks being not less than 3. Sodium Valproate in 10 mL of water: the solution is clear and Detection sensitivity: Adjust the detection sensitivity so colorless. that the peak height of valproic acid obtained from 2 mLof (2) Chloride <1.03>—Dissolve 0.5 g of Sodium Valproate the standard solution is between 4 mm and 10 mm. in 25 mL of ethanol (95), and add 6 mL of dilute nitric acid Time span of measurement: About twice as long as the and water to make 50 mL. Perform the test using this solu- retention time of valproic acid, beginning after the solvent tion as the test solution. Prepare the control solution as fol- peak. lows: to 0.70 mL of 0.01 molWL hydrochloric acid VS add 25 Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, mL of ethanol (95), 6 mL of dilute nitric acid and water to 3 hours). make 50 mL (not more than 0.050z). (3) Sulfate <1.14>—Dissolve 0.5 g of Sodium Valproate Assay Weigh accurately about 0.2 g of Sodium Valproate, in 25 mL of ethanol (95), and add 1 mL of dilute hydrochlor- previously dried, dissolve in 80 mL of acetic acid (100), and ic acid and water to make 50 mL. Perform the test using this titrate <2.50> with 0.1 molWL perchloric acid VS (potentio- solution as the test solution. Prepare the control solution as metric titration). Perform a blank determination, and make follows: to 0.50 mL of 0.005 molWLsulfuricacidVSadd25 any necessary correction. mL of ethanol (95), 1 mL of dilute hydrochloric acid and Each mL of 0.1 molWL perchloric acid VS watertomake50mL(notmorethan0.048z). ＝ 16.62 mg of C H NaO (4) Heavy metals <1.07>—Dissolve 2.0 g of Sodium Val- 8 15 2 proate in 44 mL of water, shake with 6 mL of dilute Containers and storage Containers—Tight containers. hydrochloric acid, allow to stand for 5 minutes, and ˆlter. Discard the ˆrst 5 mL of the ˆltrate, neutralize the subse- quent 25 mL with ammonia TS, and add 2 mL of dilute acetic Sorbitan Sesquioleate acid and water to make 50 mL. Perform the test using this so- lution as the test solution. Prepare the control solution as fol- ソルビタンセスキオレイン酸エステル lows: to 2.0 mL of Standard Lead Solution add 2 mL of di- lute acetic acid and water to make 50 mL (not more than 20 ppm). Sorbitan Sesquioleate is a mixture of monoester and (5) Arsenic <1.11>—Dissolve 2.0 g of Sodium Valproate diester of anhydride, partially esteriˆed with in 10 mL of water, shake with 10 mL of dilute hydrochloric oleic acid. acid, allow to stand for 5 minutes, and ˆlter. Discard the ˆrst Description Sorbitan Sesquioleate is a pale yellow to light 5 mL of the ˆltrate, and perform the test with the subsequent yellow-brown, viscous oily liquid. It has a faint, characteris- 10 mL (not more than 2 ppm). tic odor and a slightly bitter taste. (6) Related substances—Dissolve 0.10 g of Sodium Val- It is freely soluble in diethyl ether, slightly soluble in proate in 10 mL of a mixture of formic acid and chloroform ethanol (95), and very slightly soluble in methanol. (1:1), and use this solution as the sample solution. Pipet 1 mL It is dispersed as ˆne oily drops in water. of the sample solution, add a mixture of formic acid and chloroform (1:1) to make exactly 200 mL, and use this solu- Identiˆcation (1) To 0.5 g of Sorbitan Sesquioleate add 5 tion as the standard solution. Perform the test with exactly 2 mL of ethanol (95) and 5 mL of dilute sulfuric acid, and heat mL each of the sample solution and the standard solution as on a water bath for 30 minutes. Cool, shake with 5 mL of directed under Gas Chromatography <2.02> according to the petroleum ether, and allow to stand, and separate the upper following conditions. Determine each peak area of both solu- layer and the lower layer. Shake 2 mL of the lower layer with tions by the automatic integration method: the total area of 2 mL of freshly prepared catechol solution (1 in 10), then all peaks other than the area of the valproic acid from the with 5 mL of sulfuric acid: a red to red-brown color develops. sample solution is not larger than the peak area of the val- (2) Heat the upper layer obtained in (1) on a water bath, proic acid from the standard solution. and evaporate petroleum ether. To the residue add 2 mL of Operating conditions— dilutednitricacid(1in2),andthenadd0.5gofpotassiumni- Detector: A hydrogen ‰ame-ionization detector. trite between 309Cand359C with stirring: the solution de- Column: A glass column 3 mm in inside diameter and 2 m velops an opalescence, and, when cooled, crystals are in length, packed with siliceous earth for gas chro- formed. matography (150 to 180 mm in particle diameter) coated with Speciˆc gravity <1.13> d25: 0.960 – 1.020 diethylene glycol adipate ester for gas chromatography and 25 phosphoric acid at the ratios of 5z and 1z, respectively. Saponiˆcation value <1.13> 150 – 168 Column temperature: A constant temperature of about Purity (1) Acidity—To 2.0 g of Sorbitan Sesquioleate add 1459C. 1112 D-Sorbitol / O‹cial Monographs JP XV

50 mL of neutralized ethanol, and heat on a water bath near- Sorbitol. Prepare the control solution with 0.30 mL of 0.01 ly to boiling with stirring once or twice. Cool, add 4.3 mL of molWL hydrochloric acid VS (not more than 0.005z). 0.1 mol WL sodium hydroxide VS and 5 drops of (3) Sulfate <1.14>—Perform the test with 4.0 g of D-Sor- phenolphthalein TS: a red color develops. bitol. Prepare the control solution with 0.50 mL of 0.005 mol (2) Heavy metals <1.07>—Proceed with 1.0 g of Sorbitan WL sulfuric acid VS (not more than 0.006z). Sesquioleate according to Method 2, and perform the test. (4) Heavy metals <1.07>—Proceed with 5.0 g of D-Sor- Prepare the control solution with 2.0 mL of Standard Lead bitol according to Method 1, and perform the test. Prepare Solution (not more than 20 ppm). the control solution with 2.5 mL of Standard Lead Solution (3) Arsenic <1.11>—Prepare the test solution with 1.0 g (not more than 5 ppm). of Sorbitan Sesquioleate according to Method 2, and per- (5) Nickel—Dissolve 0.5 g of D-Sorbitol in 5 mL of form the test (not more than 2 ppm). water, add 3 drops of dimethylglyoxime TS and 3 drops of ammonia TS, and allow to stand for 5 minutes: no red color Water <2.48> Not more than 3.0z (1 g, direct titration, stir develops. for 30 minutes). (6) Arsenic <1.11>—Prepare the test solution with 1.5 g Residue on ignition <2.44> Not more than 1.0z (1 g). of D-Sorbitol according to Method 1, and perform the test (not more than 1.3 ppm). Containers and storage Containers—Tight containers. (7) —Dissolve 20.0 g of D-Sorbitol in 25 mL of water,andboilgentlywith40mLofFehling'sTSfor3 minutes. After cooling, ˆlter the supernatant liquid cautious- D-Sorbitol ly through a glass ˆlter (G4), leaving the precipitate in the ‰ask as much as possible, wash the precipitate with hot water D-ソルビトール until the last washings no longer show an alkali reaction, and ˆlter the washings through the glass ˆlter. Dissolve the precipitate in the ‰ask in 20 mL of iron (III) sulfate TS, ˆlter through the glass ˆlter, and wash with water. Combine the ˆltrate and the washings, heat at 809C, and titrate <2.50> with 0.02 molWL potassium permanganate VS: not more than 6.3 C6H14O6: 182.17 D-Glucitol [50-70-4] mL of volume for titration consumed or consumption is re- quired. (8) Sugars—Dissolve 20.0 g of D-Sorbitol in 25 mL of D-Sorbitol, when dried, contains not less than 97.0z water, and heat with 8 mL of dilute hydrochloric acid under a of C6H14O6. re‰ux condenser in a water bath for 3 hours. After cooling, Description D-Sorbitol occurs as white granules, powder, or add 2 drops of methyl orange TS, followed by sodium crystalline masses. It is odorless, and has a sweet taste with a hydroxide TS until an orange color develops, and add water cold sensation. to make 100 mL. Boil gently 10 mL of this solution with 10 It is very soluble in water, sparingly soluble in ethanol (95), mL of water and 40 mL of Fehling's TS for 3 minutes and and practically insoluble in diethyl ether. proceed as directed in (7). It is hygroscopic. Loss on drying <2.41> Not more than 2.0z (0.5 g, in vacu- Identiˆcation (1) To 1 mL of a solution of D-Sorbitol (7 in um, phosphorus (V) oxide, 809C, 3 hours). 10) add 2 mL of iron (II) sulfate TS and 1 mL of a solution of sodium hydroxide (1 in 5): a blue-green color develops, but Residue on ignition <2.44> Not more than 0.02z (5 g). no turbidity is produced. Assay Weigh accurately about 0.2 g of D-Sorbitol, previ- (2) Shake thoroughly 1 mL of a solution of D-Sorbitol (1 ously dried, dissolve in water and add water to make exactly in 20) with 1 mL of a freshly prepared solution of catechol (1 100 mL. Pipet 10 mL of the solution into an iodine ‰ask, add in 10), add rapidly 2 mL of sulfuric acid, and shake: a reddish exactly 50 mL of potassium periodate TS, and heat for 15 purple to red-purple color immediately develops. minutes in a water bath. Cool, add 2.5 g of potassium iodide, (3) Boil 0.5 g of D-Sorbitol with 10 mL of acetic anhy- immediately stopper tightly, and shake well. Allow to stand dride and 1 mL of pyridine under a re‰ux condenser for 10 for 5 minutes in a dark place, and titrate <2.50> with 0.1 molW minutes, cool, shake with 25 mL of water, and allow to stand L sodium thiosulfate VS (indicator: 3 mL of starch TS). Per- in a cold place. Transfer the solution to a separator, extract form a blank determination. with 30 mL of chloroform, and evaporate the extract on a water bath. Add 80 mL of water to the oily residue, heat for Each mL of 0.1 molWL sodium thiosulfate VS 10 minutes on a water bath, then ˆlter the hot mixture. After ＝ 1.822 mg of C6H14O6 cooling, collect the produced precipitate through a glass ˆlter Containers and storage Containers—Tight containers. (G3), wash with water, recrystallize once from ethanol (95), and dry in a desiccator (in vacuum, silica gel) for 4 hours: the precipitate melts between 979C and 1019C. D-Sorbitol Solution Purity (1) Clarity and color of solution, and acidity or alkalinity—Dissolve 5 g of D-Sorbitol in 20 mL of water by D-ソルビトール液 warming with shaking: the solution is clear, colorless, and neutral. (2) Chloride <1.03>—Perform the test with 2.0 g of D- D-Sorbitol Solution contains not less than 97z JP XV O‹cial Monographs / Soybean Oil 1113 and not more than 103z of the labeled amount of equivalent to 20.0 g of D-Sorbitol according to the labeled D-sorbitol (C6H14O6: 182.17). amount, dilute with water or concentrate to 40 mL of a water bath, if necessary, add 8 mL of dilute hydrochloric acid, and Description D-Sorbitol Solution is a clear, colorless liquid. heat under a re‰ux condenser in a water bath for 3 hours. Af- It is odorless, and has a sweet taste. ter cooling, add 2 drops of methyl orange TS, followed by so- It is miscible with water, with ethanol (95), with glycerin dium hydroxide TS until an orange color develops, and add and with propylene glycol. water to make 100 mL. Boil gently 10 mL of this solution It sometimes separates crystalline masses. with 10 mL of water and 40 mL of Fehling's TS for 3 minutes Identiˆcation (1) To a volume of D-Sorbitol Solution, and proceed as directed in (7). equivalent to 0.7 g of D-Sorbitol according to the labeled Residue on ignition <2.44> Measure exactly a volume of D- amount, add 2 mL of iron (II) sulfate TS and 1 mL of a solu- Sorbitol Solution, equivalent to 5 g of D-Sorbitol according tion of sodium hydroxide (1 in 5): a blue-green color de- to the labeled amount, add 3 to 4 drops of sulfuric acid, and velops, but no turbidity is produced. heat gently to evaporate. Ignite to burn, cool, and perform (2) To a volume of D-Sorbitol Solution, equivalent to 1 g the test with the residue: not more than 1 mg. of D-Sorbitol according to the labeled amount, add water to make 20 mL. To 1 mL of this solution add 1 mL of a freshly Assay Measure exactly a volume of D-Sorbitol Solution, prepared solution of catechol (1 in 10), mix well, then add equivalent to about 5 g of D-sorbitol (C6H14O6) according to rapidly 2 mL of sulfuric acid, and mix: a reddish purple to the labeled amount, and add water to make exactly 250 mL. red-purple color immediately develops. Pipet 10 mL of this solution, add water to make exactly 100 mL. Pipet exactly 10 mL of the solution into an iodine ‰ask, Purity (1) Acidity or alkalinity—D-Sorbitol Solution is add exactly 50 mL of potassium periodate TS, and heat for neutral. 15 minutes in a water bath. Cool, add 2.5 g of potassium (2) Chloride <1.03>—Proceed with a volume of D-Sor- iodide, immediately stopper tightly, and shake well. Allow to bitol Solution, equivalent to 2.0 g of D-Sorbitol according to stand for 5 minutes in a dark place, and titrate <2.50> with 0.1 the labeled amount, and perform the test. Prepare the control molWL sodium thiosulfate VS (indicator: 3 mL of starch TS). solution with 0.30 mL of 0.01 molWL hydrochloric acid VS Perform a blank determination. (not more than 0.005z). (3) Sulfate <1.14>—To a volume of D-Sorbitol Solution, Each mL of 0.1 molWL sodium thiosulfate VS equivalent to 4.0 g of D-Sorbitol according to the labeled ＝ 1.822 mg of C6H14O6 amount, and perform the test. Prepare the control solution Containers and storage Containers—Tight containers. with 0.50 mL of 0.005 molWL sulfuric acid VS (not more than 0.006z). (4) Heavy metals <1.07>—Proceed with a volume of D- Sorbitol Solution, equivalent to 5.0 g of D-Sorbitol according Soybean Oil to the labeled amount, and according to Method 1, perform the test. Prepare the control solution with 2.5 mL of Stan- Oleum Sojae dard Lead Solution (not more than 5 ppm). ダイズ油 (5) Nickel—Take a volume of D-Sorbitol Solution, equivalent to 0.5 g of D-Sorbitol according to the labeled amount, add 3 drops of dimethylglyoxime TS and 3 drops of Soybean Oil is the ˆxed oil obtained from the seeds ammonia TS, and allow to stand for 5 minutes: no red color of max merrill (Leguminosae). develops. Description Soybean Oil is a clear, pale yellow oil. It is (6) Arsenic <1.11>—Take a volume of D-Sorbitol Solu- odorless or has a slight odor, and has a bland taste. tion, equivalent to 1.5 g of D-Sorbitol according to the la- It is miscible with diethyl ether and with petroleum ether. beled amount, dilute with water or concentrate to 5 mL on a It is slightly soluble in ethanol (95), and practically insolu- water bath, if necessary, cool, and perform the test using this ble in water. solution as the test solution (not more than 1.3 ppm). It congeals between －109Cand－179C. (7) Glucose—Take a volume of D-Sorbitol Solution, Congealing point of the fatty acids: 22 – 279C equivalent to 20.0 g of D-Sorbitol according to the labeled 25 amount, dilute with water or concentrate to 40 mL on a water Speciˆc gravity <1.13> d25: 0.916 – 0.922 bath, if necessary, add 40 mL of Fehling's TS, and boil gently Acid value <1.13> Not more than 0.2. for 3 minutes. After cooling, ˆlter the supernatant liquid cau- tiously through a glass ˆlter (G4), leaving the precipitate in Saponiˆcation value <1.13> 188 – 195 the ‰ask as much as possible, wash the precipitate with hot Unsaponiˆable matter <1.13> Not more than 1.0z. water until the last washings no longer show alkalinity, and ˆlter the washings through the glass ˆlter. Dissolve the Iodine value <1.13> 126 – 140 precipitate in the ‰ask in 20 mL of iron (III) sulfate TS, ˆlter Containers and storage Containers—Tight containers. through the glass ˆlter, and wash the ˆlter with water. Com- binetheˆltrateandthewashings,heatat809C, and titrate <2.50> with 0.02 molWL potassium permanganate VS: not more than 6.3 mL of 0.02 molWL potassium permanganate VS is required. (8) Sugars—Take a volume of D-Sorbitol Solution, 1114 Spectinomycin Hydrochloride Hydrate / O‹cial Monographs JP XV

Agar media for seed and base layer. Adjust the pH of the medium so that it will be 7.8 to 8.0 after sterilization. Spectinomycin Hydrochloride (iii) Standard solutions—Weigh accurately an amount of Spectinomycin Hydrochloride Reference Standard, equiva- Hydrate lent to about 20 mg (potency), dissolve in 0.1 mol/L phos- phate buŠer solution, pH 8.0 to make exactly 20 mL, and use スペクチノマイシン塩酸塩水和物 this solution as the standard stock solution. Keep the stan- dard stock solution at a temperature not exceeding 59Cand use within 10 days. Take exactly a suitable amount of the standard stock solution before use, add 0.1 mol/L phosphate buŠer solution, pH 8.0 to make solutions so that each mL contains 200 mg (potency) and 50 mg (potency), and use these solutions as the high concentration standard solution and low concentration standard solution, respectively.

C14H24N2O7.2HCl.5H2O: 495.35 (iv) Sample solutions—Weigh accurately an amount of (2R,4aR,5aR,6S,7S,8R,9S,9aR,10aS )- Spectinomycin Hydrochloride Hydrate, equivalent to about 4a,7,9-Trihydroxy-2-methyl-6,8- 20 mg (potency), and dissolve in 0.1 mol/L phosphate buŠer bis(methylamino)-2,3,4a,5a,6,7,8,9,9a,10a-decahydro- solution, pH 8.0 to make exactly 20 mL. Take exactly a suita- 4H-pyrano[2,3-b][1,4]benzodioxin-4-one dihydrochloride ble amount of this solution, add 0.1 mol/L phosphate buŠer pentahydrate [22189-32-8 ] solution, pH 8.0 to make solutions so that each mL contains 200 mg (potency) and 50 mg (potency), and use these solutions Spectinomycin Hydrochloride Hydrate is the as the high concentration sample solution and low concentra- hydrochloride of a substance having antibacterial ac- tion sample solution, respectively. tivity produced by the growth of Streptomyces spec- Containers and storage Containers—Tight containers. tabilis. It contains not less than 603 mg (potency) and not more than 713 mg (potency) per mg. The potency of Spectinomycin Hydrochloride Hydrate is expressed as Spiramycin Acetate mass (potency) of spectinomycin (C H N O : 14 24 2 7 スピラマイシン酢酸エステル 332.35). Description Spectinomycin Hydrochloride Hydrate occurs as a white to light yellowish white crystalline powder. It is freely soluble in water, and practically insoluble in ethanol (95). Identiˆcation (1) To 5 mL of a solution of Spectinomycin Hydrochloride Hydrate (1 in 100) add gently anthrone TS: a blue to blue-green color is produced at the zone of contact. (2) Determine the infrared absorption spectra of Spec- tinomycin Hydrochloride Hydrate and Spectinomycin Hydrochloride Reference Standard as directed in the paste method under the Infrared Spectrophotometry <2.25>,and compare these spectra: both spectra exhibit similar intensities of absorption at the same wave numbers. (3) To 3 mL of a solution of Spectinomycin Hydrochlo- ride Hydrate (1 in 150) add 1 drop of silver nitrate TS: a white turbidity is produced. Optical rotation <2.49> [a]20: ＋15 – ＋219(2.1 g calculated D (Spiramycin II Acetate (Spiramycin I Acetate)) on the anhydrous basis, water, 25 mL, 200 mm). (3R,4R,5S,6R,8R,9R,10E,12E,15R)-3- pH <2.54> Dissolve 0.10 g of Spectinomycin Hydrochloride Acetoxy-5-[4-O-acetyl-2,6-dideoxy-3-C-methyl-a- Hydrate in 10 mL of water: the pH of the solution is between L-ribo-hexopyranosyl-(1ª4)-3,6-dideoxy-3- 4.0 and 5.6. dimethylamino-b-D-glucopyranosyloxy]-9-(2,3,4,6- tetradeoxy-4-dimethylamino-b-D-erythro- Water <2.48> Not less than 16.0z and not more than hexopyranosyloxy)-6-formylmethyl-9-hydroxy-4-methoxy- 20.0z (0.3 g, volumetric titration, direct titration). 8-methylhexadeca-10,12-dien-15-olide Residue on ignition <2.44> Not more than 1.0z (1 g). (Spiramycin III Acetate) (3R,4R,5S,6R,8R,9R,10E,12E,15R)-5- Assay Perform the test according to the Cylinder-plate [4-O-Acetyl-2,6-dideoxy-3-C-methyl-a-L-ribo- method as directed under Microbial Assay for Antibiotics hexopyranosyl-(1ª4)-3,6-dideoxy-3-dimethylamino-b- <4.02> according to the following conditions. D-glucopyranosyloxy]-9-(2,3,4,6-tetradeoxy-4- (i) Test organism—Klebsiella pneumoniae ATCC 10031 dimethylamino-b-D-erythro-hexopyranosyloxy)-6- (ii) Culture medium—Use the medium i in 3) under (1) formylmethyl-9-hydroxy-4-methoxy-8-methyl-3- JP XV O‹cial Monographs / Spiramycin Acetate 1115 propionyloxyhexadeca-10,12-dien-15-olide System performance: Dissolve 25 mg of Spiramycin [74014-51-0, Spiramycin Acetate] Acetate II Reference Standard in the mobile phase to make 100 mL. When the procedure is run with 5 mLofthissolution Spiramycin Acetate is a derivative of a mixture of under the above operating conditions, the number of theoret- macrolide substances having antibacterial activity pro- ical plates and the symmetry factor of the peak of spiramycin duced by the growth of Streptomyces ambofaciens. acetate II are not less than 14,500 and not more than 2.0, re- It contains not less than 900 mg (potency) and not spectively. more than 1450 mg (potency) per mg, calculated on the System repeatability: When the test is repeated 6 times with dried basis. The potency of Spiramycin Acetate is 5 mL of the sample solution under the above operating condi- expressed as mass (potency) of spiramycin acetate II tions, the relative standard deviation of the peak area of (C47H78N2O16: 927.13). One mg (potency) of Spiramy- spiramycin acetate II is not more than 2.0z. cin Acetate is equivalent to 0.7225 mg of spiramycin Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of acetate II (C H N O ). 47 78 2 16 Spiramycin Acetate according to Method 2, and perform the Description Spiramycin Acetate occurs as a white to light test. Prepare the control solution with 1.0 mL of Standard yellowish white powder. Lead Solution (not more than 10 ppm). It is very soluble in acetonitrile and in methanol, freely (2) Arsenic <1.11>—Prepare the test solution with 2.0 g soluble in ethanol (99.5), and practically insoluble in water. of Spiramycin Acetate according to Method 3, and perform the test (not more than 1 ppm). Identiˆcation (1) Determine the absorption spectrum of a solution of Spiramycin Acetate in methanol (1 in 50,000) as Loss on drying <2.41> Not more than 3.0z (1 g, in vacuum, directed under Ultraviolet-visible Spectrophotometry <2.24>, phosphorus (V) oxide, 609C, 3 hours). and compare the spectrum with the Reference Spectrum: Residue on ignition <2.44> Not more than 0.5z (1 g). both spectra exhibit similar intensities of absorption at the same wavelengths. Assay Perform the test according to the Cylinder-plate (2) Determine the infrared absorption spectrum of method as directed under Microbial Assay for Antibiotics Spiramycin Acetate as directed in the potassium bromide disk <4.02> according to the following conditions. method under Infrared Spectrophotometry <2.25>,andcom- (i) Test organism—Bacillus subtilis ATCC 6633 pare the spectrum with the Reference Spectrum: both spectra (ii) Culture medium—Use the medium i in 1) under (1) exhibit similar intensities of absorption at the same wave Agar media for seed and base layer. numbers. (iii) Standard solutions—Weigh accurately an amount of Spiramycin Acetate II Reference Standard, equivalent to Content ratio of the active principle Dissolve 25 mg of about 50 mg (potency), dissolve in 20 mL of methanol, add Spiramycin Acetate in 25 mL of the mobile phase, and use 0.1 mol/L phosphate buŠer solution for antibiotics, pH 8.0 this solution as the sample solution. Perform the test with 5 to make exactly 50 mL, and use this solution as the standard mL of the sample solution as directed under Liquid Chro- stock solution. Keep the standard stock solution at not matography <2.01> according to the following conditions, exceeding 59C, and use within 3 days. Take exactly a suitable and determine the areas, A , A , A , A , A and A ,of II III IV V VI VII amount of the standard stock solution before use, add the peaks of spiramycin acetate II, spiramycin acetate III, 0.1 mol/L phosphate buŠer solution for antibiotics, pH 8.0 spiramycin acetate IV, spiramycin acetate V, spiramycin to make solutions so that each mL contains 80 mg (potency) acetate VI and spiramycin acetate VII, respectively, by the and 20 mg (potency), and use these solutions as the high automatic integration method, and calculate the ratios of the concentration standard solution and low concentration stan- amounts of A , A and the total of A and A to the total II IV III V dard solution, respectively. amount of all these peaks: the amount of A is30–45z, A II IV (iv) Sample solutions—Weigh accurately an amount of is30–45z, and the total of A and A is not more than III V Spiramycin Acetate, equivalent to about 50 mg (potency), 25z. The relative retention times of spiramycin acetate III, dissolve in 20 mL of methanol, and add 0.1 mol/L phosphate spiramycin acetate IV, spiramycin acetate V, spiramycin buŠer solution for antibiotics, pH 8.0 to make exactly acetate VI and spiramycin acetate VII with respect to 50 mL. Take exactly a suitable amount of this solution, add spiramycin acetate II are 1.3, 1.7, 2.3, 0.85 and 1.4, respec- 0.1 mol/L phosphate buŠer solution for antibiotics, pH 8.0 tively. to make solutions so that each mL contains 80 mg (potency) Operating conditions— and 20 mg (potency), and use these solutions as the high Detector: An ultraviolet absorption photometer (wave- concentration sample solution and low concentration sample length: 231 nm). solution, respectively. Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel Containers and storage Containers—Tight containers. for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of acetonitrile, 0.02 mol/L potas- sium dihydrogen phosphate TS and a solution of dipotassium hydrogen phosphate (87 in 25,000) (26:7:7). Flow rate: Adjust the ‰ow rate so that the retention time of acetylspiramycin II is about 10 minutes. System suitability— 1116 / O‹cial Monographs JP XV

the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with n- Spironolactone butyl acetate to a distance of about 15 cm, and air-dry the plate. Spray evenly a solution of sulfuric acid in methanol (1 スピロノラクトン in 10) on the plate, and heat the plate at 1059Cfor10 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 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 50 mg each of Spironolac- tone and Spironolactone Reference Standard, previously C H O S: 416.57 24 32 4 dried at 1059C for 2 hours, dissolve in methanol to make ex- 7a-Acetylsulfanyl-3-oxo-17a-pregn-4-ene-21,17b- actly 250 mL. Pipet 5 mL each of these solutions, add carbolactone [52-01-7] methanol to make exactly 100 mL, and use these solutions as Spironolactone, when dried, contains not less than the sample solution and standard solution, respectively. Per- 97.0z and not more than 103.0z of C H O S. form the test with these solutions as directed under Ultrav- 24 32 4 iolet-visible Spectrophotometry <2.24>, and determine the ab-

Description Spironolactone occurs as a white to light yel- sorbances, A T and A S, of the sample solution and the stan- low-brown, ˆne powder. dard solution at 238 nm. It is freely soluble in chloroform, soluble in ethanol (95), Amount (mg) of C H O S slightly soluble in methanol, and practically insoluble in 24 32 4 ＝W ×(A /A ) water. S T S

Melting point: 198 – 2079C (Insert the capillary tube into a WS: amount (mg) of Spironolactone Reference Standard bath at about 1259C, and continue the heating so that the Containers and storage Containers—Tight containers. temperature rises at a rate of about 109C per minute in the range between 1409Cand1859C,andwhenthetemperature is near the expected melting range, reduce the heating so that the temperature rises at a rate of about 39C per minute.) Stearic Acid

Identiˆcation (1) Determine the absorption spectrum of a ステアリン酸 solution of Spironolactone in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or Stearic Acid is solid fatty acids obtained from fats, the spectrum of a solution of Spironolactone Reference Stan- and it consists chie‰y of stearic acid (C18H36O2:284.48) dard prepared in the same manner as the sample solution: and palmitic acid (C16H32O2: 256.42). both spectra exhibit similar intensities of absorption at the same wavelengths. Description Stearic Acid occurs as white, unctuous or crys- (2) Determine the infrared absorption spectrum of talline masses or powder. It has a faint, fatty odor. Spironolactone, previously dried, as directed in the potassi- It is freely soluble in diethyl ether, soluble in ethanol (95), um bromide disk method under the Infrared Spectrophoto- and practically insoluble in water. metry <2.25>, and compare the spectrum with the Reference Meltingpoint:56–729C. Spectrum or the spectrum of Spironolactone Reference Stan- Acid value <1.13> 194 – 210 dard: both spectra exhibit similar intensities of absorption at the same wave numbers. If any diŠerence appears between Iodine value <1.13> Not more than 4.0. the spectra, dissolve Spironolactone and Spironolactone Purity (1) Mineral acid—Melt 5 g of Stearic Acid by Reference Standard in methanol, respectively, then evaporate warming, shake with 5 mL of boiling water for 2 minutes, methanol to dryness, and repeat the test on the residues. ˆlter after cooling, and add 1 drop of methyl orange TS to 20 the ˆltrate: no red color develops. Optical rotation <2.49> [a]D : －33 – －379(after drying, 0.25 g, chloroform, 25 mL, 200 mm). (2) Heavy metals <1.07>—Proceed with 1.0 g of Stearic Acid according to Method 2, and perform the test. Prepare Purity (1) Mercapto compounds—Shake 2.0 g of the control solution with 2.0 mL of Standard Lead Solution Spironolactone with 20 mL of water, and ˆlter. To 10 mL of (not more than 20 ppm). the ˆltrate add 1 mL of starch TS and 0.05 mL of 0.01 molWL (3) Fat and para‹n—Boil 1.0 g of Stearic Acid with 0.5 g iodine VS, and mix: a blue color develops. of anhydrous sodium carbonate and 30 mL of water: the so- (2) Related substances—Dissolve 0.20 g of Spironolac- lution, while hot, is clear or not more turbid than the follow- tone in 10 mL of ethanol (95), and use this solution as the ing control solution. sample solution. Pipet 1 mL of this solution, add ethanol (95) Control solution: To 0.70 mL of 0.01 molWL hydrochloric to make exactly 100 mL, and use this solution as the standard acid VS add 6 mL of dilute nitric acid and water to make 30 solution. Perform the test with these solutions as directed un- mL, and add 1 mL of silver nitrate TS. der Thin-layer Chromatography <2.03>.Spot5mLeachof Residue on ignition <2.44> Not more than 0.1z (1 g). JP XV O‹cial Monographs / Streptomycin Sulfate 1117

Containers and storage Containers—Well-closed contain- ers. Streptomycin Sulfate Stearyl Alcohol ストレプトマイシン硫酸塩

ステアリルアルコール

Stearyl Alcohol is a mixture of solid alcohols, and consists chie‰y of stearyl alcohol (C18H38O: 270.49). Description Stearyl Alcohol occurs as a white, unctuous matter. It has a faint, characteristic odor. It is tasteless. It is freely soluble in ethanol (95), in ethanol (99.5), in diethyl ether, and practically insoluble in water. Melting point <1.13> 56–629C Prepare the sample accord- ingtoMethod2,thenattachtightlyacapillarytubetothe bottomofthethermometerbymeansofarubberbandorby any suitable means, and make the bottom of the capillary tube equal in position to the lower end of the thermometer. Insert this thermometer into a test tube 17 mm in inside di- 1 ameter and about 170 mm in height, fasten the thermometer C21O39N7O12.1 /2H2SO4:728.69 with cork stopper so that the lower end of the thermometer is 2-Deoxy-2-methylamino-a-L-glucopyranosyl-(1ª2)- about 25 mm distant from the bottom of the test tube. Sus- 5-deoxy-3-C-formyl-a-L-lyxofuranosyl-(1ª4)-N,N?- pend the test tube in a beaker containing water, and heat the diamidino-D-streptamine sesquisulfate [3810-74-0] beaker with constant stirring until the temperature rises to 59C below the expected melting point. Then regulate the rate Streptomycin Sulfate is the sulfate of an of increase to 19C per minute. The temperature at which the aminoglycoside substance having antibacterial activity sample is transparent and no turbidity is produced is taken as produced by the growth of Streptomyces griseus. the melting point. It contains not less than 740 mg (potency) and not more than 820 mg (potency) per mg, calculated on the Acid value <1.13> Not more than 1.0. dried basis. The potency of Streptomycin Sulfate is Ester value <1.13> Not more than 3.0. expressed as mass (potency) of streptomycin (C H N O :581.57). Hydroxyl value <1.13> 200 – 220 21 39 7 12 Description Streptomycin Sulfate occurs as a white to light Iodine value <1.13> Not more than 2.0. yellowish white powder. Purity (1) Clarity of solution—Dissolve 3.0 g of Stearyl It is freely soluble in water, and very slightly soluble in Alcohol in 25 mL of ethanol (99.5) by warming: the solution ethanol (95). is clear. Identiˆcation (1) Dissolve 50 mg of Streptomycin Sulfate (2) Alkalinity—To the solution obtained in (1) add 2 in 5 mL of water, add 1 mL of ninhydrin TS and 0.5 mL of drops of phenolphthalein TS: no red color develops. pyridine, and heat for 10 minutes: a purple color is devel- Residue on ignition <2.44> Not more than 0.05z (2 g). oped. (2) Dissolve 10 mg each of Streptomycin Sulfate and Containers and storage Containers—Well-closed contain- Streptomycin Sulfate Reference Standard in 10 mL of water, ers. and use these solutions as the sample solution and the stan- dard solution. Perform the test with these solutions as direct- ed under Thin-layer Chromatography <2.03>.Spot10mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a solution of potassium dihydrogen phosphate (7 in 100) to a distance of about 12 cm, and air-dry the plate. Spray evenly a mixture of a solution of 1,3-dihydroxynaphthalene in ethanol (95) (1 in 500) and diluted sulfuric acid (1 in 5) (1:1) on the plate, and heat at about 1509C for about 5 minutes: the principal spots from the sample solution and the standard solution show the same in color tone and Rf value. (3) A solution of Streptomycin Sulfate (1 in 5) responds to the Qualitative Tests <1.09> for sulfate.

20 Optical rotation <2.49> [a]D : －79 – －889(0.5 g calculated on the dried basis, water, 50 mL, 100 mm). 1118 Sucralfate Hydrate / O‹cial Monographs JP XV

the high concentration standard solution and low concentra- pH <2.54> The pH of a solution obtained by dissolving 2.0 g tion standard solution, respectively. of Streptomycin Sulfate in 10 mL of water is between 4.5 and (iv) Sample solutions—Weigh accurately an amount of 7.0. Streptomycin Sulfate, equivalent to about 20 mg (potency), Purity (1) Clarity and color of solution—A solution ob- dissolve in water to make exactly 50 mL. Take exactly a suita- tained by dissolving 1.0 g of Streptomycin Sulfate in 5 mL of ble amount of this solution, add 0.1 mol/L phosphate buŠer waterisclear,andcolorlessorpaleyellow. solution, pH 8.0 to make solutions so that each mL contains (2) Heavy metals <1.07>—Proceed with 2.0 g of Strep- 8 mg (potency) and 2 mg (potency), and use these solutions as tomycin Sulfate according to Method 4, and perform the test. the high concentration sample solution and low concentra- Prepare the control solution with 2.0 mL of Standard Lead tion sample solution, respectively. Solution (not more than 10 ppm). Containers and storage Containers—Tight containers. (3) Arsenic <1.11>—Prepare the test solution with 2.0 g of Streptomycin Sulfate according to Method 3 and perform the test (not more than 1 ppm). (4) Related substances—Dissolve exactly 0.20 g of Strep- Sucralfate Hydrate tomycin Sulfate in a mixture of methanol and sulfuric acid (97:3) to make 5 mL, and heat under a re‰ux condenser for 1 Aluminum Sucrose Sulfate Ester hour. After cooling, wash the inside of the condenser with a スクラルファート水和物 suitable amount of a mixture of methanol and sulfuric acid (97:3), add a mixture of methanol and sulfuric acid (97:3) to make exactly 20 mL, and use this solution as the sample solu- tion. Separately, dissolve exactly 36 mg of D(＋)-mannose in a mixture of methanol and sulfuric acid (97:3) to make 5 mL, and heat under a re‰ux condenser for 1 hour. After cooling, wash the inside of the condenser with a suitable amount of a mixture of methanol and sulfuric acid (97:3), and add a mix- C12H30Al8O51S8.xAl(OH)3.yH2O ture of methanol and sulfuric acid (97:3) to make exactly 50 [54182-58-0] mL. Pipet 5 mL of this solution, add a mixture of methanol andsulfuricacid(97:3)tomakeexactly50mL,andusethis Sucralfate Hydrate contains not less than 17.0z and solution as the standard solution. Perform the test with these not more than 21.0z of aluminum (Al: 26.98) and not solutions as directed under Thin-layer Chromatography less than 34.0z and not more than 43.0z of sucrose <2.03>.Spot10mL each of the sample solution and standard octasulfate ester (C12H22O35S8: 982.80), calculated on solution on a plate of silica gel for thin-layer chro- the dried basis. matography, develop with a mixture of toluene, methanol and acetic acid (100) (2:1:1) to a distance of 13 to 15 cm, and Description Sucralfate Hydrate occurs as a white powder. air-dry the plate. Spray evenly a mixture of a solution of 1,3- It is odorless and tasteless. dihydroxynaphthalene in ethanol (95) (1 in 500) and diluted It is practically insoluble in water, in hot water, in ethanol sulfuric acid (1 in 5) (1:1) on the plate, and heat at 1109Cfor (95) and in diethyl ether. 5 minutes: the spot from the sample solution corresponding It dissolves in dilute hydrochloric acid and in sulfuric acid- to the spot from the standard solution is not more intense sodium hydroxide TS. than the spot from the standard solution. Identiˆcation (1) To 0.05 g of Sucralfate Hydrate in a Loss on drying <2.41> Not more than 5.0z (0.5 g, reduced small test tube add 0.05 g of fresh pieces of sodium, and melt pressure not exceeding 0.67 kPa, 609C, 3 hours). by careful heating. Immerse the test tube immediately in 100 mL of water, break the test tube, shake well, and ˆlter. To 5 Residue on ignition <2.44> Not more than 1.0z (1 g). mL of the ˆltrate add 1 drop of sodium pentacyanonitrosyl- Assay Perform the test according to the Cylinder-plate ferrate (III) TS: a red-purple color develops. method as directed under Microbial Assay for Antibiotics (2) Dissolve 40 mg of Sucralfate Hydrate in 2 mL of di- <4.02> according to the following conditions. lute sulfuric acid, and add gently 2 mL of anthrone TS to (i) Test organism—Bacillus subtilis ATCC 6633 make 2 layers: a blue color develops at the zone of contact, (ii) Culture medium—Use the medium i in 1) Medium for and gradually changes to blue-green. test organism [5] under (1) Agar media for seed and base lay- (3) Dissolve 0.5 g of Sucralfate Hydrate in 10 mL of di- er, having pH 7.8 – 8.0 after sterilization. lute hydrochloric acid: the solution responds to the Qualita- (iii) Standard solutions—Weigh accurately an amount of tive Tests <1.09> for aluminum. Streptomycin Sulfate Reference Standard, previously dried, Purity (1) Clarity and color of solution—Dissolve 1.0 g of equivalent to about 20 mg (potency), dissolve in diluted phos- Sucralfate Hydrate in 10 mL of dilute sulfuric acid: the solu- phate buŠer solution, pH 6.0 (1 in 2) to make exactly 50 mL, tion is clear and colorless. and use this solution as the standard stock solution. Keep the (2) Chloride <1.03>—Dissolve 0.5 g of Sucralfate Hy- standard stock solution between 59Cand159C, and use wi- drate in 30 mL of dilute nitric acid, and heat gently to boil- thin 30 days. Take exactly a suitable amount of the standard ing. After cooling, add water to make 100 mL, and to 10 mL stock solution before use, add 0.1 mol/L phosphate buŠer of this solution add 3 mL of dilute nitric acid and water to solution, pH 8.0 to make solutions so that each mL contains make 50 mL. Perform the test using this solution as the test 8 mg (potency) and 2 mg (potency), and use these solutions as solution. Prepare the control solution with 0.70 mL of 0.01 JP XV O‹cial Monographs / Sucralfate Hydrate 1119 molWL hydrochloric acid VS (not more than 0.50z). Assay (1) Aluminum—Weigh accurately about 1 g of (3) Heavy metals <1.07>—Dissolve 1.0 g of Sucralfate Sucralfate Hydrate, dissolve in 10 mL of dilute hydrochloric Hydrate in 20 mL of a solution of sodium chloride (1 in 5) acid by warming on a water bath, cool, and add water to and 1 mL of dilute hydrochloric acid, and to this solution make exactly 250 mL. Pipet 25 mL of this solution, add ex- add 2 mL of dilute acetic acid and water to make 50 mL. Per- actly 25 mL of 0.05 molWL disodium dihydrogen ethylenedia- form the test using this solution as the test solution. Prepare mine tetraacetate VS and 20 mL of acetic acid-ammonium the control solution as follows: evaporate 1 mL of dilute acetate buŠer solution, pH 4.5, and boil for 5 minutes. After hydrochloric acid on a water bath to dryness, and add 20 mL cooling, add 50 mL of ethanol (95), and titrate <2.50> the ex- of a solution of sodium chloride (1 in 5), 2 mL of dilute acetic cess disodium dihydrogen ethylenediamine tetraacetate with acid, 2.0 mL of Standard Lead Solution and water to make 0.05 molWL zinc acetate VS until the color of the solution 50 mL (not more than 20 ppm). changes from green-purple through purple to red (indicator: (4) Arsenic <1.11>—Dissolve 1.0 g of Sucralfate Hydrate 3 mL of dithizone TS). Perform a blank determination. in 5 mL of dilute hydrochloric acid, use this solution as the test solution, and perform the test (not more than 2 ppm). Each mL of 0.05 molWL disodium dihydrogen ethylenedia- (5) Free aluminum—To 3.0 g of Sucralfate Hydrate add mine tetraacetate VS 50 mL of water, heat in a water bath for 5 minutes, cool, and ＝ 1.349 mg of Al ˆlter. Wash the residue with four 5-mL portions of water, (2) Sucrose octasulfate ester—Weigh accurately about combine the ˆltrate with the washings, add 2 mL of dilute 0.55 g of Sucralfate Hydrate, add exactly 10 mL of sulfuric hydrochloric acid, and heat in a water bath for 30 minutes. acid-sodium hydroxide TS, shake vigorously, and dissolve After cooling, neutralize the solution with sodium hydroxide with ultrasonic wave at below 309C for 5 minutes. To this so- TS, add water to make exactly 100 mL, and use this solution lution add 0.1 molWL sodium hydroxide VS to make exactly as the sample solution. Pipet 50 mL of the sample solution, 25 mL, and use this solution as the sample solution. add exactly 25 mL of 0.05 molWL disodium dihydrogen Separately, weigh accurately about 0.25 g of Potassium Su- ethylenediamine tetraacetate VS and 20 mL of acetic acid- crose Octasulfate Reference Standard, add the mobile phase ammonium acetate buŠer solution, pH 4.5, and boil for 5 to make exactly 25 mL, and use this solution as the standard minutes. After cooling, add 50 mL of ethanol (95), and ti- solution. Prepare rapidly the sample solution and the stan- trate <2.50> the excess disodium dihydrogen ethylenediamine dard solution, and perform the test immediately. Pipet 50 mL tetraacetate with 0.05 molWL zinc acetate VS until the color each of the sample solution and standard solution, and per- of the solution changes from green-purple through purple to form the test as directed under Liquid Chromatography red (indicator: 3 mL of dithizone TS). Perform a blank deter- <2.01> according to the following conditions. Determine the mination (not more than 0.2z). peak areas, AT and AS, of sucrose octasulfate ester from each Each mL of 0.05 molWL disodium dihydrogen solution. ethylenediamine tetraacetate VS Amount (mg) of sucrose octasulfate ester (C H O S ) ＝ 1.349 mg of Al 12 22 35 8 ＝WS×(AT/AS)× 0.7633 (6) Related substances—Proceed with 50 mLofthesam- W : Amount (mg) of Potassium Sucrose Octasulfate ple solution obtained in the Assay (2) Sucrose octasulfate es- S Reference Standard, calculated on the anhydrous basis ter as directed in the Assay (2) Sucrose octasulfate ester, and perform the test as directed under Liquid Chromatography Operating conditions— <2.01>. Determine the peak area of sucrose octasulfate ester Detector: A diŠerential refractometer. from the sample solution and that of a related substance with Column: A stainless steel column about 4 mm in inside di- the relative retention time about 0.7 to the peak of sucrose ameter and about 30 cm in length, packed with aminopropyl- octasulfate ester by the automatic integration method, and silanized silica gel for liquid chromatography (about 8 mmin calculate the ratio of the peak area of the related substance to particle diameter). that of sucrose octasulfate ester: it is not more than 0.1. Column temperature: Room temperature. Detection sensitivity: Adjust the detection sensitivity so Mobile phase: Dissolve a suitable amount (26 to 132 g) of that the peak height of sucrose octasulfate ester from 50 mL ammonium sulfate in 1000 mL of water, and adjust with of the standard solution obtained in the Assay (2) Sucrose oc- phosphoric acid to a pH of 3.5. Allow a solution of Potassi- tasulfate ester composes 60z to 100z of the full scale. um Sucrose Octasulfate Reference Standard in dilute hydrochloric acid (1 in 100) to stand at 609Cfor10minutes, Loss on drying <2.41> Not more than 14.0z (1 g, 1059C, cool, and perform the test immediately. Adjust the amount 3 hours). of ammonium sulfate in the mobile phase so that the peak of Acid-consuming capacity Weigh accurately about 0.25 g of a related substance with the relative retention time being Sucralfate Hydrate, previously dried, place in a 200-mL about 0.7 to that of sucrose octasulfate ester almost returns glass-stoppered conical ‰ask, add exactly 100 mL of 0.1 molW to the base line, and the peak of sucrose octasulfate ester L hydrochloric acid VS, stopper the ‰ask tightly, and shake elutes most rapidly. at 37 ± 29C for exactly 1 hour (150 shakings per minute, am- Flow rate: Adjust the ‰ow rate so that the retention time of plitude: 20 mm). After cooling in water for 5 minutes, pipet sucrose octasulfate ester is between 6 and 11 minutes. 10 mL of the supernatant liquid, and titrate <2.50> the excess Selection of column: Allow a solution of Potassium Su- acid with 0.1 molWL sodium hydroxide VS until the pH crose Octasulfate Reference Standard in dilute hydrochloric becomes 3.5. Perform a blank determination. The amount of acid (1 in 100) to stand at 609C for 10 minutes, cool, and pro- 0.1 molWL hydrochloric acid VS consumed per g of Sucral- ceed immediately with 50 mL of this solution under the above fate is not less than 130 mL. operating conditions. Use a column with a resolution being 1120 Sucrose / O‹cial Monographs JP XV not less than 1.5 between sucrose octasulfate ester and a relat- andblue,andnotchangesonboiling.Thentothissolution ed substance with the relative retention time being about 0.7 add 4 mL of dilute hydrochloric acid, boil, and add 4 mL of 2 to sucrose octasulfate ester. molWL sodium hydroxide TS: orange precipitates are immedi- System repeatability: Repeat the test 6 times with the stan- ately produced. dard solution under the above operating conditions: the rela- Optical rotation <2.49> [a ]20: ＋66.3 – ＋67.09(26 g, water, tive standard deviation of the peak area of sucrose octasul- D 100 mL, 100 mm). fate ester is not more than 2.0z. Purity (1) Clarity and color of solution—The sample so- Containers and storage Containers—Tight containers. lution obtained in the Identiˆcation (2) is clear, and has no more color than the following control solution. Control solution: To exactly 2.4 mL of iron (III) chloride Sucrose colorimetric stock solution and exactly 0.6 mL of cobalt (II) chloride colorimetric stock solution add 7.0 mL of diluted 精製白糖 hydrochloric acid (7 in 250). To 5.0 mL of this solution add 95.0 mL of diluted hydrochloric acid (7 in 250). (2) Acidity or alkalinity—To 10 mL of the sample solu- tion obtained in the Identiˆcation (2) add 0.3 mL of phenolphthalein TS: the solution is colorless, and develops a red color on addition of 0.3 mL of 0.01 molWLsodium hydroxide VS. (3) Sulˆte—Dissolve 5.0 g of Sucrose in 40 mL of water, add 2.0 mL of dilute sodium hydroxide TS and water to make exactly 50 mL, and use this solution as the sample solution. Separately, dissolve 0.076 g of sodium disulˆte in water to C H O :342.30 make exactly 50 mL, then pipet 5 mL of this solution, add 12 22 11 water to make exactly 100 mL. Pipet 3 mL of this solution, b-D-Fructofuranosyl a-D-glucopyranoside [57–50–1 ] add water to make exactly 100 mL, and use this solution as Sucrose contains no additives. the standard solution. Immediately, pipet 10 mL each of the For Sucrose used for preparation of the large volume sample solution and the standard solution, add 1.0 mL of 3 infusions, the label states the purpose. molWL hydrochloric acid, 2.0 mL of decolorized fuchsin TS and 2.0 mL of solution TS, and allow to stand Description Sucrose is a white crystalline powder, or lus- for 30 minutes. Determine the absorbance at 583 nm of these trous colorless or white crystals. solutions as directed under Ultraviolet-visible Spectrophoto- It is very soluble in water, and slightly soluble in ethanol metry <2.24> using the control solution obtained by proceed- (95). ing with 10.0 mL of water in the same manner as above: the absorbance of the sample solution is not larger than that of Identiˆcation (1) To 10 mg each of Sucrose and white soft the standard solution (not more than 15 ppm as SO ). When sugar add diluted methanol (3 in 5) to make 20 mL each, and 2 the standard solution does not show a red-purple to blue-pur- use these solutions as the sample solution and the standard ple color, result of the test is invalid. solution (a), respectively. Separately, to 10 mg each of glu- (4) Lead—Put exactly 50 mg of Sucrose in a poly- cose, lactose hydrate, fructose and white soft sugar add tetrafuruoroethylene decomposition-vessel, add 0.5 mL of methanol (3 in 5) to make 20 mL, and use this solution as the nitric acid to dissolve, seal up the vessel, and heat at 1509C standard solution (b). Perform the test with these solutions as for 5 hours. After cooling, add water to make exactly 5 mL, directed under Thin-layer Chromatography <2.03>.Spot2mL and use this solution as the sample solution. Perform the test each of the sample solution and standard solutions (a) and (b) with more than 3 parts of the sample solution as directed in on a plate of silica gel for thin-layer chromatography, and the standard addition method under Atomic Absorption dry the plate completely. Develop the plate with a mixture of Spectrophotometry <2.23> (electrothermal type) according to 1,2-dichloroethane, acetic acid (100), methanol and water the following conditions. The standard solution is prepared (10:5:3:2) to a distance of about 15 cm, and dry the plate with by adding water to a suitable volume of Standard Lead Solu- a hot air. And immediately repeat the development with tion exactly volumed, and perform a blank determination replaced developing mixture, and dry the plate in the same with a solution prepared by adding water to 10.0 mL of nitric way. Spray evenly a solution of 0.5 g of thymol in 100 mL of acid to make exactly 100 mL, and make any necessary correc- a mixture of ethanol (95) and sulfuric acid (19:1), heat at tion (not more than 0.5 ppm). 1309C for 10 minutes: the principal spot from the sample so- Operating conditions— lution is the same with the principal spot from the standard Lamp: A hollow cathode lamp solution (a) in the Rf, color and size, and four spots from the Wavelengh: 283.3 mm standard solution (b) are apparently distinguishable. Temperature for drying: 1109C (2) Dissolve 50.0 g of Sucrose in recently boiled and Temperature for incineration: 6009C cooled water to make 100 mL, and use this solution as the Temperature for atomization: 21009C sample solution. To 1 mL of the sample solution add water to (5) Invert sugar—Transfer 5 mL of the sample solution make 100 mL, then to 5 mL of this solution add 0.15 mL of obtained in the Identiˆcation (2) to a test-tube about 150 mm freshly prepared copper (II) sulfate TS and 2 mL of freshly long and about 16 mm in diameter, add 5 mL of water, 1.0 prepared 2 molWL sodium hydroxide TS: the solution is clear mL of 1 molWL sodium hydroxide VS and 1.0 mL of methy- JP XV O‹cial Monographs / White Soft Sugar 1121

－1 lene blue TS, mix, and place in a water bath. After exactly 2 x0 ( mS・cm )＝JG0 minutes, take the tube out of the bath, and examine the solu- Determine the corrected conductivity, x ,ofthesample tion immediately: the blue color does not disappear com- C solution by the following expression: not more than pletely (0.04z). Ignore any blue color at the air and solution 35 mS・cm－1. interface. x ( mS・cm－1)＝x －0.35 x Conductivity C T 0 (i) Potassium chloride conductivity calibration standard Loss on drying <2.41> Not more than 0.1z (2 g, 1059C, solution—Dissolve powdered potassium chloride, previously 3 hours). driedat500–6009C for 4 hours, in newly distillated water Dextrins For Sucrose used to prepare large volume aqueous having less conductivity than 2 mS・cm－1 to get three kinds of infusions, to 2 mL of the sample solution obtained in the the standard solution containing 0.7455 g, 0.0746 g and Identiˆcation (2) add 8 mL of water, 0.05 mL of dilute 0.0149 g of potassium chloride in 1000.0 g, respectively. The hydrochloric acid and 0.05 mL of iodine TS: the solution conductivities of these solutions at 209C are shown in the fol- remains yellow. lowing table. Bacterial endotoxins <4.01> Less than 0.25 EU Wmg, for Su- Standard solution Conductivity Resistivity crose exclusively to be used to prepare Injections for in- (gW1000.0 g) ( mS・cm－1)(V・cm) travenous infusion of larger volume. 0.7455 1330 752 Containers and storage Containers—Well-closed contain- 0.0746 133.0 7519 ers. 0.0149 26.6 37594

(ii) Apparatus—Use an appropriate conductivity meter. The conductivity is determined to measure the electrical White Soft Sugar resistance of the column of liquid between the electrodes of 白糖 the immersed measuring device (conductivity cell). The ap- paratus is supplied with alternative current to avoid the eŠects of electrode polarization. It is usually equipped with a temperature compensation device. The conductivity cell con- tains of two parallel platinum electrodes coated with plati- num black, and both electrodes are generally protected by a glass tube which allows good exchange between the solution and the electrodes. Use a cell giving the cell constant of 0.01 to 1 cm－1. (iii) Procedure—Use the suitable potassium chloride con- ductivity calibration standard solution to the measurement. After washing the well with water, rinse 2 to 3 times with the C H O : 342.30 calibration standard solution, ˆll up the cell with the calibra- 12 22 11 b-D-Fructofuranosyl a-D-glucopyranoside [57-50-1] tion standard solution, and determine the conductivity of the calibration standard solution kept at 20 ± 0.1 9C. Description White Soft Sugar is colorless or white crystals Repeat the determination, and measure the conductivity of or crystalline powder. It is odorless and has a sweet taste. It is very soluble in water, very slightly soluble in ethanol the calibration standard solution, Gx0 (mS), after a stable reading of ± 3z is obtained. The cell constant, J,iscalculat- (95), and practically insoluble in diethyl ether. ed by the following: A solution of White Soft Sugar (1 in 10) is neutral. x J＝ KCl Identiˆcation (1) When 1 g of White Soft Sugar is ignited, Gx 0 it melts and swells, and decomposes, emitting an odor of J: Cell constant (cm－1) caramel, to bulky charcoal.

xKCl: Conductivity constant of the potassium chloride con- (2) To 0.1 g of White Soft Sugar add 2 mL of dilute sul- ductivity calibration standard solution ( mS・cm－1) furic acid, boil, add 4 mL of sodium hydroxide TS and 3 mL (209C) ofFehling'sTS,andheattoboiling:aredtodarkred G : Conductivity measured ( mS) precipitate is produced. x0 20 Dissolve 31.3 g of Sucrose in newly distillated water to Optical rotation <2.49> [a]D : ＋65.0 – ＋67.09 (after make exactly 100 mL, and use this solution as the sample so- drying,13g,water,50mL,100mm). lution. After washing well the cell with water, rinse the cell Purity (1) Clarity and color of solution—Dissolve 100 g with the sample solution 2 to 3 times , ˆll up with the sample of White Soft Sugar in 100 mL of water, take 50 mL of this solution, and determine the conductivity of the sample solu- solution in a Nessler tube, and view transversely the Nessler tion, ( S),keptat20± 0.19C, while stirring. Determine GT m tube against a white background: the solution is colorless or the conductivity of the water used for preparation of the sam- only slightly yellow and has no blue color. Fill the solution in ple solution, G ( mS), in the same manner as above, and cal- 0 the Nessler tube, stopper, and allow to stand for 2 days: no culate the conductivity, ( S・cm－1)and ( S・cm－1), by xT m x0 m precipitate is produced. the following expressions: (2) Chloride <1.03>—To 10.0 g of White Soft Sugar add －1 xT ( mS・cm )＝JGT water to make 100 mL, and use this solution as the sample so- 1122 Sulbactam Sodium / O‹cial Monographs JP XV lution. To 20 mL of the sample solution add 6 mL of dilute very slightly soluble in ethanol (99.5), and practically insolu- nitric acid and water to make 50 mL. Perform the test using ble in acetonitrile. this solution as the test solution. Prepare the control solution Identiˆcation (1) Determine the infrared absorption spec- with 0.30 mL of 0.01 molWL hydrochloric acid VS (not more trum of Sulbactam Sodium as directed in the potassium than 0.005z). bromide disk method under the Infrared Spectrophotometry (3) Sulfate <1.14>—To 40 mL of the sample solution ob- <2.25>, and compare the spectrum with the Reference Spec- tained in (2) add 1 mL of dilute hydrochloric acid and water trum: both spectra exhibit similar intensities of absorption at to make 50 mL. Perform the test using this solution as the thesamewavenumbers. test solution. Propare the control solution with 0.50 mL of (2) Sulbactam Sodium responds to the Qualitative Tests 0.005 molWL sulfuric acid VS (not more than 0.006z). <1.09> (1) for sodium salt. (4) Calcium—To 10 mL of the sample solution obtained 20 in (2) add 1 mL of ammonium oxalate TS: this solution Optical rotation <2.49> [a]D : ＋219 – ＋2339(1 g, water, shows immediately no change. 100 mL, 100 mm). (5) Heavy metals <1.07>—Proceed with 5.0 g of White pH <2.54> Dissolve 1.0 g of Sulbactam Sodium in 20 mL of Soft Sugar according to Method 1, and perform the test. Pre- water: the pH of the solution is between 5.2 and 7.2. pare the control solution with 2.5 mL of Standard Lead Solu- tion (not more than 5 ppm). Purity (1) Clarity and color of solution—Dissolve 1.0 g of (6) Arsenic <1.11>—Prepare the test solution with 1.0 g Sulbactam Sodium in 20 mL of water: the solution is clear, of White Soft Sugar according to Method 1, and perform the and colorless to pale yellow. test (not more than 2 ppm). (2) Heavy metals <1.07>—Proceed with 1.0 g of Sulbac- (7) Invert sugar—Dissolve 5.0 g of White Soft Sugar in tam Sodium according to Method 2, and perform the test. water to make 100 mL, ˆlter if necessary, and use this solu- Prepare the control solution with 2.0 mL of Standard Lead tion as the sample solution. Separately place 100 mL of alka- Solution (not more than 20 ppm). line copper (II) sulfate solution in a 300-mL beaker, cover the (3) Arsenic <1.11>—Prepare the test solution with 1.0 g beaker with a watch glass, and boil. Immediately add 50.0 of Sulbactam Sodium as directed in Method 3, and perform mL of the sample solution, boil the mixture exactly for 5 the test (not more than 2 ppm). minutes, add at once 50 mL of freshly boiled and cooled (4) Sulbactam penicillamine—Weigh accurately about water, dip it in a water bath of a temperature below 109Cfor 0.2 g of Sulbactam Sodium, dissolve in the mobile phase to 5 minutes, and collect the precipitate in a tared glass ˆlter make exactly 50 mL, and use this solution as the sample solu- (G4). Wash the residue on the ˆlter with water until the last tion. Separately, weigh accurately about 40 mg of sulbactam washing is neutral, then wash with 10 mL of ethanol (95), add sodium for sulbactam penicillamine, dissolve in 2 mL of 10 mL of diethyl ether, and dry at 1059C for 30 minutes: the water, add 0.5 mL of sodium hydroxide TS, allow to stand mass of the residual precipitate is not more than 0.120 g. for 10 minutes at a room temperature, and add 0.5 mL of 1 molWL hydrochloric acid TS, then add the mobile phase to Loss on drying <2.41> Not more than 1.30z (15 g, 1059C, make exactly 100 mL. Pipet 5 mL of this solution, add the 2 hours). mobile phase to make exactly 50 mL, and use this solution as Residue on ignition <2.44> Not more than 0.1z (2 g). the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed Containers and storage Containers—Well-closed contain- under Liquid Chromatography <2.01> according to the fol- ers. lowing conditions, and determine the peak areas, AT and AS, of sulbactam penicillamine by the automatic integration method: the amount of sulbactam penicillamine is not more Sulbactam Sodium than 1.0z. スルバクタムナトリウム Amount (z) of sulbactam penicillamine ＝(WS/WT)×(A T/A S)× 5

WS: Amount (mg) of sulbactam sodium for sulbactam penicillamine

WT: Amount (mg) of the sample Operating conditions—

C8H10NNaO5S: 255.22 Column, column temperature, mobile phase, and ‰ow Monosodium (2S,5R)-3,3-dimethyl-7-oxo-4-thia- rate: Proceed as directed in the operating conditions in the 1-azabicyclo[3.2.0]heptane-2-carboxylate 4,4-dioxide Assay. [69388-84-7] Detector: An ultraviolet absorption photometer (wavelength: 230 nm). Sulbactam Sodium contains not less than 875 mg System suitability— (potency) per mg, calculated on the anhydrous basis. System performance: Proceed as directed in the system The potency of Sulbactam Sodium is expressed as mass suitability in the Assay. (potency) of sulbactam (C8H11NO5S: 233.24). System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operating Description Sulbactam Sodium occurs as a white to yellow- conditions, the relative standard deviation of the peak areas ish white crystalline powder. of sulbactam penicillamine is not more than 2.0z. It is freely soluble in water, sparingly soluble in methanol, JP XV O‹cial Monographs / Sulbenicillin Sodium 1123

Water <2.48> Not more than 1.0z (0.5 g, volumetric titra- Sulbenicillin Sodium contains not less than 900 mg tion, direct titration). (potency) and not more than 970 mg (potency) per mg, Assay Weigh accurately an amount of Sulbactam Sodium calculated on the anhydrous basis. The potency of Sul- and Sulbactam Reference Standard, equivalent to about 0.1 g benicillin Sodium is expressed as mass (potency) of sul- (potency), dissolve each in a suitable amount of the mobile benicillin (C16H18N2O7S2:414.45). phase, add exactly 10 mL of the internal standard solution, Description Sulbenicillin Sodium occurs as white to light then add the mobile phase to make 100 mL, and use these so- yellowish white powder. lutions as the sample solution and standard solution. Per- It is very soluble in water, freely soluble in methanol, and form the test with 10 mL each of the sample solution and stan- slightly soluble in ethanol (99.5). dard solution as directed under Liquid Chromatography It is hygroscopic. <2.01> according to the following conditions, and calculate the ratios, Q T and Q S, of the peak area of sulbactam to that Identiˆcation (1) Determine the infrared absorption spec- of the internal standard. trum of Sulbenicillin Sodium as directed in the potassium bromide disk method under the Infrared Spectrophotometry Amount [mg (potency)] of sulbactam (C H NO S) 8 11 5 <2.25>, and compare the spectrum with the Reference Spec- ＝W ×(Q /Q )× 1000 S T S trum or the spectrum of Sulbenicillin Sodium Reference

WS: Amount [mg (potency)] of Sulbactam Reference Standard: both spectra exhibit similar intensities of absorp- Standard tion at the same wave numbers. (2) Sulbenicillin Sodium responds to the Qualitative Tests Internal standard solution—A solution of ethyl parahydrox- <1.09> (1) for sodium salt. ybenzoate in the mobile phase (7 in 1000). 20 Operating conditions— Optical rotation <2.49> [a]D : ＋167 – ＋1829(1gcalculated Detector: An ultraviolet absorption photometer on the anhydrous basis, water, 20 mL, 100 mm). (wavelength: 220 nm). pH <2.54> The pH of a solution obtained by dissolving 0.20 Column: A stainless steel column 3.9 mm in inside di- g of Sulbenicillin Sodium in 10 mL of water is between 4.5 ameter and 30 cm in length, packed with octadecylsilanized and 7.0. silica gel for liquid chromatography (10 mminparticledi- ameter). Purity (1) Clarity and color of solution—Dissolve 2.5 g of Column temperature: A constant temperature of about Sulbenicillin Sodium in 5 mL of water: the solution is clear 359C. and colorless to pale yellow. Mobile phase: To 750 mL of 0.005 molWLtetrabutylam- (2) Heavy metals <1.07>—Proceed with 1.0 g of Sul- monium hydroxide TS add 250 mL of acetonitrile for liquid benicillin Sodium according to Method 1, and perform the chromatography. test. Prepare the control solution with 2.0 mL of Standard Flow rate: Adjust the ‰ow rate so that the retention time of Lead Solution (not more than 20 ppm). sulbactam is about 6 minutes. (3) Arsenic <1.11>—Prepare the test solution with 1.0 g System suitability— of Sulbenicillin Sodium according to Method 1, and perform System performance: When the procedure is run with 10 the test (not more than 2 ppm). mL of the standard solution under the above operating condi- (4) Related substances—Dissolve 0.10 g of Sulbenicillin tions, sulbactam and the internal standard are eluted in this Sodium in 15 mL of the mobile phase, and use this solution order with the resolution between these peaks being not less as the sample solution. Perform the test with 10 mLofthe than 1.5. sample solution as directed under Liquid Chromatography System repeatability: When the test is repeated 6 times with <2.01> according to the following conditions, determine each 10 mL of the standard solution under the above operating peak area by the automatic integration method, and calculate conditions, the relative standard deviation of the peak areas the amount of these peaks by the area percentage method: the of sulbactam is not more than 1.0z. amount of the each peak other than the two peaks of sul- benicillin is not more than 2.0z, and the total amount of the Containers and storage Containers—Tight containers. peaks other than the two peaks of sulbenicillin is not more than 5.0z. Operating conditions— Sulbenicillin Sodium Detector: An ultraviolet absorption photometer (wavelength: 254 nm). スルベニシリンナトリウム Column: A stainless steel column 3.9 mm in inside di- ameter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticledi- ameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 10 g of potassium dihydrogen

C16H16N2Na2O7S2: 458.42 phosphate in 750 mL of water, adjust the pH to 6.0 ± 0.1 Disodium (2S,5R,6R)-3,3-dimethyl-7-oxo-6-[(2R)-2-phenyl- with sodium hydroxide TS, and add water to make 1000 mL. 2-sulfonatoacetylamino]-4-thia-1- To 940 mL of this solution add 60 mL of acetonitrile. azabicyclo[3.2.0]heptane-2-carboxylate [28002-18-8] Flow rate: Adjust the ‰ow rate so that the retention time of 1124 Sulfadiazine Silver / O‹cial Monographs JP XV the lately eluted peak of sulbenicillin is about 18 minutes. Time span of measurement: About 1.5 times as long as the retention time of the lately eluted peak of sulbenicillin begin- Sulfadiazine Silver ning after the solvent peak. System suitability— スルファジアジン銀 Test for required detectability: Measure exactly 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitabil- ity test, and add the mobile phase to make exactly 10 mL. Conˆrm that the total area of the two peaks of sulbenicillin obtained from 10 mL of this solution is equivalent to 7 to 13z C10H9AgN4O2S: 357.14 of that from 10 mL of the solution for system suitability test. Monosilver 4-amino-N-(pyrimidin-2-yl)- System performance: When the procedure is run with 10 benzenesulfonamidate [22199-08-2] mL of the sample solution under the above operating condi- tions, the resolution between the two peaks of sulbenicillin is Sulfadiazine Silver, when dried, contains not less not less than 2.0. than 99.0z and not more than 102.0z of System repeatability: When the test is repeated 6 times with C10H9AgN4O2S. 10 mL of the solution for system suitability test under the Description Sulfadiazine Silver occurs as a white to pale above operating conditions, the relative standard deviation yellow, crystalline powder. It is odorless. of the total areas of the two peaks of sulbenicillin is not more It is practically insoluble in water, in ethanol (95) and in than 5.0z. diethyl ether. Water <2.48> Not more than 6.0z (0.5 g, volumetric titra- It dissolves in ammonia TS. tion, direct titration). It is gradually colored by light. Melting point: about 2759C (with decomposition). Assay Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics Identiˆcation Determine the infrared absorption spectrum <4.02> according to the following conditions. of Sulfadiazine Silver, previously dried, as directed in the (i) Test organism—Bacillus subtilis ATCC 6633 paste method under Infrared Spectrophotometry <2.25>,and (ii) Culture medium—Use the medium i in 1) Medium for compare the spectrum with the Reference Spectrum or the test organism [5] under (1) Agar media for seed and base lay- spectrum of previously dried Sulfadiazine Silver Reference er. Adjust the pH of the medium so that it will be 6.4 to 6.6 Standard: both spectra exhibit similar intensities of absorp- after sterilization. tion at the same wave numbers. (iii) Standard solutions—Weigh accurately an amount of Purity (1) Nitrate—To 250 mL of water add 1.0 g of Sul- Sulbenicillin Sodium Reference Standard, equivalent to fadiazine Silver, shake well for 50 minutes, ˆlter, and use this about 50 mg (potency), dissolve in phosphate buŠer solution, ˆltrate as the sample solution. Separately, weigh accurately pH 6.0 to make exactly 50 mL, and use this solution as the 0.25 g of potassium nitrate, and dissolve in water to make ex- standard stock solution. Keep the standard stock solution in actly 2000 mL. Pipet 5 mL of this solution and add water to a freezer, and use within 4 days. Take exactly a suitable make exactly 200 mL, and use this solution as the standard amount of the standard stock solution before use, add phos- solution. Pipet 2.0 mL each of the sample solution and the phate buŠer solution, pH 6.0 to make solutions so that each standard solution, and add 5 mL of a solution of cromotrop- mL contains 40 mg (potency) and 10 mg (potency), and use ic acid in sulfuric acid (1 in 10,000) and sulfuric acid to make these solutions as the high concentration standard solution exactly 10 mL. Determine the absorbances, A and A ,ofthe and low concentration standard solution, respectively. T S sample solution and standard solution at 408 nm as directed (iv) Sample solutions—Weigh accurately an amount of under Ultraviolet-visible Spectrophotometry <2.24>,usinga Sulbenicillin Sodium, equivalent to about 50 mg (potency), solution, prepared with exactly 2.0 mL of water in the same and dissolve in phosphate buŠer solution, pH 6.0 to make ex- manner, as the blank: A is not larger than A (not more than actly 50 mL. Take exactly a suitable amount of this solution, T S 0.05z). add phosphate buŠer solution, pH 6.0 to make solutions so (2) Related substances—Dissolve 50 mg of Sulfadiazine that each mL contains 40 mg (potency) and 10 mg (potency), Silver in 5 mL of a mixture of ethanol (95) and ammonia so- and use these solutions as the high concentration sample solu- lution (28) (3:2), and use this solution as the sample solution. tion and low concentration sample solution, respectively. Pipet 2 mL of the sample solution, and add a mixture of Containers and storage Containers—Hermetic containers. ethanol (95) and ammonia solution (28) (3:2) to make exactly 20 mL. Pipet 2 mL of this solution, add a mixture of ethanol (95) and ammonia solution (28) (3:2) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro- matography <2.03>.Spot5mL each of the sample solution andstandardsolutiononaplateofsilicagelwith‰uorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform, methanol and ammonia solu- tion (28) (10:5:2) to a distance of about 15 cm, and air-dry the JP XV O‹cial Monographs / Sulfamethizole 1125 plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot and spot of the Sulfamethizole, when dried, contains not less than starting point from the sample solution are not more intense 99.0z of C9H10N4O2S2. than the spot from the standard solution. Description Sulfamethizole occurs as white to yellowish Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum, white crystals or crystalline powder. It is odorless. phosphorus (V) oxide, 80z, 4 hours). It is slightly soluble in ethanol (95), and in acetic acid (100) and practical insoluble in water and in diethyl ether. Residue on ignition <2.44> 41.0 – 45.0z (1 g). It dissolves in dilute hydrochloric acid and in sodium Silver content Weigh accurately about 50 mg of Sulfadia- hydroxide TS. zine Silver, previously dried, dissolve in 2 mL of nitric acid, It is gradually colored by light. and add water to make exactly 100 mL. Pipet 1 mL of this so- Identiˆcation Determine the infrared absorption spectrum lution, add water to make exactly 100 mL, and use this solu- of Sulfamethizole, previously dried, as directed in the potas- tion as the sample solution. Measure accurately a suitable sium bromide disk method under Infrared Spectrophotomet- quantity of Standard Silver Solution for Atomic Absorption ry <2.25>, and compare the spectrum with the Reference Spectrophotometry <2.23>, dilute with water to make a solu- Spectrum: both spectra exhibit similar intensities of absorp- tion containing 1.0 to 2.0 mgofsilver(Ag:107.87)perml,and tion at the same wave numbers. use this solution as the standard solution. Perform the test with the sample solution and the standard solution as direct- Melting point <2.60> 208 – 2119C ed under Atomic Absorption Spectrophotometry <2.23> ac- Purity (1) Clarity and color of solution—Dissolve 0.5 g of cording to the following conditions, and calculate the silver Sulfamethizole in 3 mL of sodium hydroxide TS and 20 mL content of the sample solution from the calibration curve ob- of water: the solution is clear and colorless. tained from the absorbance of the standard solution: it con- (2) Acidity—To 1.0 g of Sulfamethizole add 50 mL of tains not less than 28.7z and not more than 30.8z of silver. water, warm at 709C for 5 minutes, allow to stand for 1 hour Gas: Combustible gas—Acetylene in an ice bath, and ˆlter. To 25 mL of the ˆltrate add 2 drops Supporting gas—Air of methyl red TS and 0.60 mL of 0.1 molWLsodium Lamp: A silver hollow cathode lamp hydroxide VS: a yellow color develops. Wavelength: 328.1 nm (3) Heavy metals <1.07>—Proceed with 1.0 g of Sul- Assay Weigh accurately about 0.1 g each of Sulfadiazine famethizole according to Method 2, and perform the test. Silver and Sulfadiazine Silver Reference Standard, each Prepare the control solution with 2.0 mL of Standard Lead previously dried, and add ammonia TS to make exactly 100 Solution (not more than 20 ppm). mL, respectively. Pipet 1 mL each of these solutions, add (4) Arsenic <1.11>—Prepare the test solution with 1.0 g water to make exactly 100 mL, and use these solutions as the of Sulfamethizole according to Method 3, and perform the sample solution and the standard solution, respectively. De- test (not more than 2 ppm). termine the absorbances, AT and AS, of the sample solution (5) Related substances—Dissolve 0.10 g of Sulfamethi- and the standard solution at 255 nm, as directed under zole in acetone to make 10 mL, and use this solution as the Ultraviolet-visible Spectrophotometry <2.24>, using a solu- sample solution. Pipet 1 mL of the sample solution, add ace- tion,preparedwithexactly1mLofammoniaTSanda tone to make exactly 50 mL, then pipet 5 mL of this solution, su‹cient water to make exactly 100 mL, as the blank. add acetone to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions Amount (mg) of C H AgN O S 10 9 4 2 as directed under Thin-layer Chromatography <2.03>.Spot5 ＝W ×(A /A ) S T S mL each of the sample solution and standard solution on a

WS: Amount (mg) of Sulfadiazine Silver plate of silica gel with ‰uorescent indicator for thin-layer Reference Standard chromatography. Develop the plate with a mixture of ethyl acetate and acetic acid (100) (20:1) to a distance of about 10 Containers and storage Containers—Well-closed contain- cm, and air-dry the plate. Examine under ultraviolet light ers. (main wavelength: 254 nm): the spots other than the principal Storage—Light-resistant. spot from the sample solution are not more intense than the spot from the standard solution. Sulfamethizole Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). スルファメチゾール Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.4 g of Sulfamethizole, previously dried, dissolve in 5 mL of hydrochloric acid and 50 mL of water, add 10 mL of a solution of potassium bromide (3 in 10), cool below 159C, and titrate <2.50> with 0.1 molWL sodium nitrite VS according to the potentiometric titration method or the amperometric titration method. C H N O S : 270.33 9 10 4 2 2 Each mL of 0.1 mol LsodiumnitriteVS 4-Amino-N-(5-methyl-1,3,4-thiadiazol-2-yl)- W ＝ 27.03 mg of C H N O S benzenesulfonamide [144-82-1] 9 10 4 2 2 Containers and storage Containers—Well-closed contain- 1126 Sulfamethoxazole / O‹cial Monographs JP XV ers. matography <2.03>.Spot5mL each of the sample solution Storage—Light-resistant. andstandardsolutiononaplateofsilicagelwith‰uorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, acetonitrile and diluted am- Sulfamethoxazole monia solution (28) (7 in 100) (10:8:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light Sulˆsomezole (main wavelength: 254 nm): the spots other than the principal spot from the sample solution are not more intense than the スルファメトキサゾール 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 Sulfamethoxazole, previously dried, dissolve in 30 mL of N,N-dimethylfor-

C10H11N3O3S: 253.28 mamide, add 10 mL of water, and titrate <2.50> with 0.1 mol 4-Amino-N-(5-methylisoxazol-3-yl)benzenesulfonamide WL sodium hydroxide VS until a light blue color is produced [723-46-6] (indicator: 0.5 mL of thymolphthalein TS). Separately, per- form a blank determination in the same manner with a mix- Sulfamethoxazole, when dried, contains not less ture of 30 mL of N,N-dimethylformamide and 26 mL of than 99.0z of C10H11N3O3S. water, and make any necessary correction. Description Sulfamethoxazole occurs as white crystals or Each mL of 0.1 molWL sodium hydroxide VS crystalline powder. It is odorless, and has a slightly bitter ＝ 25.33 mg of C10H11N3O3S taste. Containers and storage Containers—Well-closed contain- It is very soluble in -dimethylformamide, sparingly N,N ers. soluble in ethanol (95), slightly soluble in diethyl ether, and Storage—Light-resistant. very slightly soluble in water. It dissolves in sodium hydroxide TS. It is gradually colored by light. Sulfamonomethoxine Hydrate Identiˆcation Determine the infrared absorption spectrum of Sulfamethoxazole, previously dried, as directed in the スルファモノメトキシン水和物 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. Melting point <2.60> 169 – 1729C Purity (1) Clarity and color of solution—Dissolve 1.0 g of Sulfamethoxazole in 5 mL of sodium hydroxide TS, and add C H N O S.H O: 298.32 20 mL of water: the solution is clear and colorless. 11 12 4 3 2 4-Amino-N-(6-methoxypyrimidin-4-yl)benzenesulfonamide (2) Acidity—To 1.0 g of Sulfamethoxazole add 50 mL of monohydrate [1220-83-3,anhydride] water, heat at 709C for 5 minutes, allow to stand in ice water for 1 hour, and ˆlter. To 25 mL of the ˆltrate add 2 drops of Sulfamonomethoxine Hydrate, when dried, contains methylredTSand0.60mLof0.1molWL sodium hydroxide not less than 99.0z of sulfamonomethoxine (C11H12N4 VS: a yellow color develops. O S: 280.31). (3) Heavy metals <1.07>—Proceed with 1.0 g of Sul- 3 famethoxazole according to Method 2, and perform the test. Description Sulfamonomethoxine Hydrate occurs as white Prepare the control solution with 2.0 mL of Standard Lead to pale yellow crystals, granules or crystalline powder. It is Solution (not more than 20 ppm). odorless. (4) Arsenic <1.11>—Prepare the test solution with 1.0 g It is soluble in acetone, slightly soluble in ethanol (95), very of Sulfamethoxazole according to Method 3, and perform slightly soluble in diethyl ether, and practically insoluble in the test (not more than 2 ppm). water. (5) Related substances—Dissolve 0.20 g of Sulfamethox- It dissolves in dilute hydrochloric acid and in sodium azole in 10 mL of a solution of ammonia solution (28) in hydroxide TS. methanol (1 in 50), and use this solution as the sample solu- It is gradually colored by light. tion. Pipet 1 mL of the sample solution, add a solution of Identiˆcation Determine the infrared absorption spectrum ammonia solution (28) in methanol (1 in 50) to make exactly of Sulfamonomethoxine Hydrate as directed in the potassium 10 mL. Pipet 1 mL of this solution, add a solution of ammo- bromide disk method under Infrared Spectrophotometry nia solution (28) in methanol (1 in 50) to make exactly 20 mL, <2.25>, and compare the spectrum with the Reference Spec- and use this solution as the standard solution. Perform the trum: both spectra exhibit similar intensities of absorption at test with these solutions as directed under Thin-layer Chro- thesamewavenumbers. JP XV O‹cial Monographs / Sulˆnpyrazone 1127

Melting point <2.60> 204 – 2069C Sulˆnpyrazone, when dried, contains not less than Purity (1) Clarity and color of solution—Dissolve 1.0 g of 98.5z of C23H20N2O3S. Sulfamonomethoxine Hydrate in 5 mL of sodium hydroxide Description Sulˆnpyrazone occurs as a white to pale yel- TS and 20 mL of water: the solution is clear and colorless to lowish white powder. It is odorless, and has a bitter taste. pale yellow. Dissolve 0.5 g of Sulfamonomethoxine Hydrate It is freely soluble in acetic acid (100) and in acetone, solu- in 5 mL of sodium hydroxide TS, and heat: no turbidity is ble in ethanol (95), slightly soluble in diethyl ether, and prac- produced. After cooling, add 5 mL of acetone: the solution is tically insoluble in water. clear. It dissolves in sodium hydroxide TS. (2) Heavy metals <1.07>—Proceed with 1.0 g of Sul- Melting point: about 1389C (with decomposition). famonomethoxine Hydrate according to Method 2, and per- form the test. Prepare the control solution with 2.0 mL of Identiˆcation (1) Dissolve 2 mg of Sulˆnpyrazone in 1 mL Standard Lead Solution (not more than 20 ppm). of acetic acid (100), add 1 mL of palladium (II) chloride TS (3) Arsenic <1.11>—Prepare the test solution with 1.0 g and 2 mL of chloroform, and shake: a yellow color develops of Sulfamonomethoxine Hydrate according to Method 3, in the chloroform layer. and perform the test (not more than 2 ppm). (2) Determine the absorption spectrum of a solution of (4) Related substances—Dissolve 0.02 g of Sul- Sulˆnpyrazone in 0.01 mol/L sodium hydroxide TS (1 in famonomethoxine Hydrate in ethanol (95) to make 10 mL, 100,000) as directed under Ultraviolet-visible Spectrophoto- and use this solution as the sample solution. Pipet 1 mL of metry <2.24>, and compare the spectrum with the Reference the sample solution, add ethanol (95) to make exactly 200 Spectrum or the spectrum of a solution of Sulˆnpyrazone mL, and use this solution as the standard solution. Perform Reference Standard prepared in the same manner as the sam- the test with these solutions as directed under Thin-layer ple solution: both spectra exhibit similar intensities of ab- Chromatography <2.03>.Spot5mL each of the sample solu- sorption at the same wavelengths. tionandstandardsolutiononaplateofsilicagelwith‰uores- (3) Determine the infrared absorption spectrum of Sulˆn- cent indicator for thin-layer chromatography. Develop the pyrazone, previously dried, as directed in the potassium plate with a mixture of 1-butanol and ammonia solution (28) bromide disk method under the Infrared Spectrophotometry (4:1) to a distance of about 10 cm, and air-dry the plate. Exa- <2.25>, and compare the spectrum with the Reference Spec- mine under ultraviolet light (main wavelength: 254 nm): the trum: both spectra exhibit similar intensities of absorption at spots other than the principal spot from the sample solution thesamewavenumbers. are not more intense than the spot from the standard solu- Purity (1) Clarity and color of solution—Dissolve 0.5 g of tion. Sulˆnpyrazone in 10 mL of acetone: the solution is clear and Loss on drying <2.41> 4.5–6.5z (1 g, 1059C, 4 hours). colorless. Dissolve 0.5 g of Sulˆnpyrazone in 10 mL of sodi- um hydroxide TS: the solution is clear and colorless. Residue on ignition <2.44> Not more than 0.10z (1 g). (2) Heavy metals <1.07>—Proceed with 2.0 g of Sulˆn- Assay Weigh accurately about 0.5 g of Sul- pyrazone according to Method 4, and perform the test. Pre- famonomethoxine Hydrate, previously dried, dissolve in 5 pare the control solution with 2.0 mL of Standard Lead Solu- mL of hydrochloric acid and 50 mL of water, add 10 mL of a tion (not more than 10 ppm). solution of potassium bromide (3 in 10), cool below 159C, (3) Arsenic <1.11>—Prepare the test solution with 1.0 g and titrate <2.50> with 0.1 molWL sodium nitrite VS (poten- of Sulˆnpyrazone according to Method 3, and perform the tiometric titration or amperometric titration). test (not more than 2 ppm). (4) Related substances—Dissolve 0.10 g of Sulˆnpyra- Each mL of 0.1 molWLsodiumnitriteVS zone in 5 mL of acetone, and use this solution as the sample ＝ 28.03 mg of C H N O S 11 12 4 3 solution. Pipet 1 mL of the sample solution, add acetone to Containers and storage Containers—Well-closed contain- make exactly 100 mL, and use this solution as the standard ers. solution (1). Pipet 1 mL of the sample solution, add acetone Storage—Light-resistant. to make exactly 200 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>.Spotrapidly5mL Sulˆnpyrazone each of the sample solution and standard solutions (1) and (2) on a plate of silica gel with ‰uorescent indicator for thin-layer スルフィンピラゾン chromatography under a stream of nitrogen. Develop the plate with a mixture of chloroform and acetic acid (100) (4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the most intense spot other than the principal spot from the sample so- lution is not more intense than the spot from the standard so- lution (1), and the spots other than the principal and above spots from the sample solution are not more intense than the spot from the standard solution (2).

C23H20N2O3S: 404.48 Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 1,2-Diphenyl-4-[2-(phenylsulˆnyl)ethyl]pyrazolidine-3,5- 2 hours). dione [57-96-5] Residue on ignition <2.44> Not more than 0.1z (1 g). 1128 Sulˆnpyrazone Tablets / O‹cial Monographs JP XV

Assay Weigh accurately about 0.5 g of Sulˆnpyrazone, previously dried, dissolve in 40 mL of acetone, add 40 mL of water, and titrate <2.50> with 0.1 molWL sodium hydroxide Sulˆsoxazole VS (potentiometric titration). Perform a blank determina- tion, and make any necessary correction. Each mL of 0.1 molWL sodium hydroxide VS スルフイソキサゾール

＝ 40.45 mg of C23H20N2O3S Containers and storage Containers—Well-closed contain- ers.

Sulˆnpyrazone Tablets C11H13N3O3S: 267.30 4-Amino-N-(3,4-dimethylisoxazol-5-yl)- スルフィンピラゾン錠 sulfonamide [127-69-5]

Sulˆsoxazole, when dried, contains not less than Sulˆnpyrazone Tablets contain not less than 93z 99.0z of C11H13N3O3S. and not more than 107z of the labeled amount of sul- Description Sulˆsoxazole occurs as white crystals or crys- ˆnpyrazone (C23H20N2O3S: 404.48). talline powder. It is odorless, and has a slightly bitter taste. Method of preparation Prepare as directed under Tablets, It is freely soluble in pyridine and in n-butylamine, soluble with Sulˆnpyrazone. in methanol, sparingly soluble in ethanol (95), slightly soluble Identiˆcation (1) Weigh a portion of powdered Sulˆn- in acetic acid (100), and very slightly soluble in water and in pyrazone Tablets, equivalent to 2 mg of Sulˆnpyrazone ac- diethyl ether. cording to the labeled amount, add 1 mL of acetic acid (100), It dissolves in dilute hydrochloric acid, in sodium and shake. To this solution add 1 mL of palladium (II) chlo- hydroxide TS and in ammonia TS. ride TS and 2 mL of chloroform, and shake: a yellow color It is gradually colored by light. develops in the chloroform layer. Identiˆcation (1) Dissolve 0.01 g of Sulˆsoxazole in 1 mL (2) Determine the absorption spectrum of the sample so- of dilute hydrochloric acid and 4 mL of water: the solution lution obtained in the Assay as directed under Ultraviolet- responds to the Qualitative Tests <1.09> for primary aromatic visible Spectrophotometry <2.24>: it exhibits a maximum be- amines. tween 257 nm and 261 nm. (2) Dissolve 0.02 g of Sulˆsoxazole in 5 mL of water and Dissolution Being speciˆed separately. 1mLofn-butylamine, add 2 to 3 drops of copper (II) sulfate TS, and shake well. Add 5 mL of chloroform, shake, and al- Assay Weigh accurately, and powder not less than 20 Sul- low to stand: a blue-green color develops in the chloroform ˆnpyrazone Tablets. Weigh accurately a portion of the pow- layer. der, equivalent to about 50 mg of sulˆnpyrazone (3) Dissolve 0.01 g of Sulˆsoxazole in 1 mL of pyridine, (C23H20N2O3S), add 25 mL of methanol, and shake for 15 add 2 drops of copper (II) sulfate TS, and shake. Add 3 mL minutes. To this solution add 50 mL of sodium hydroxide of water and 5 mL of chloroform, shake, and allow to stand: TS,shake,cool,andaddwatertomakeexactly200mL.Af- a light yellow-brown color develops in the chloroform layer. ter centrifuging, pipet 4 mL of the supernatant liquid, add (4) To 0.5 g of Sulˆsoxazole add 2 mL of acetic acid water to make exactly 100 mL, and use this solution as the (100), dissolve by heating under a re‰ux condenser, add 1 mL sample solution. Separately, weigh accurately about 0.05 g of of acetic anhydride, and boil for 10 minutes. Add 10 mL of Sulˆnpyrazone Reference Standard, previously dried at water, cool, and alkalize with about 7 mL of a solution of so- 1059C for 2 hours, dissolve in 25 mL of methanol, add 50 mL dium hydroxide (3 in 10). Filter, if necessary, immediately of sodium hydroxide TS, and cool. To this solution add acidify by adding acetic acid (100) dropwise, collect the watertomakeexactly200mL,pipet4mLofthissolution, produced precipitate, recrystallize from methanol, and dry at add water to make exactly 100 mL, and use this solution as 1059C for 1 hour: the crystals melt <2.60> between 2089Cand the standard solution. Determine the absorbances, AT and 2109C. AS, of these solutions at 260 nm as directed under Ultrav- iolet-visible Spectrophotometry <2.24>. Melting point <2.60> 192 – 1969C (with decomposition). Purity (1) Clarity and color of solution—Dissolve 1.0 g of Amount (mg) of sulˆnpyrazone (C23H20N2O3S) Sulˆsoxazole in 5 mL of sodium hydroxide TS and 20 mL of ＝WS×(AT/AS) water: the solution is clear and colorless to pale yellow. WS: Amount (mg) of Sulˆnpyrazone (2) Acidity—To 1.0 g of Sulˆsoxazole add 50 mL of Reference Standard water, warm at 709Cfor5minutes,allowtostandinanice Containers and storage Containers—Well-closed contain- bath for 1 hour, and ˆlter. To 25 mL of the ˆltrate add 2 ers. dropsofmethylredTSand0.20mLof0.1molWLsodium hydroxide VS: a yellow color develops. (3) Heavy metals <1.07>—Proceed with 1.0 g of Sulˆsox- azole according to Method 2, and perform the test. Prepare JP XV O‹cial Monographs / Sulfobromophthalein Sodium 1129 the control solution with 2.0 mL of Standard Lead Solution fobromophthalein Sodium in 20 mL of water is between 4.0 (not more than 20 ppm). and 5.5. Loss on drying <2.41> Not more than 0.5z (2 g, 1059C, 4 Purity (1) Clarity and color of solution—Dissolve 0.5 g of hours). Sulfobromophthalein Sodium in 10 mL of water: the solu- tion is clear and colorless to pale yellow. Residue on ignition <2.44> Not more than 0.1z (1 g). (2) Chloride <1.03>—Perform the test with 2.0 g of Sul- Assay Weigh accurately about 1 g of Sulˆsoxazole, previ- fobromophthalein Sodium. Prepare the control solution with ously dried, dissolve in 50 mL of methanol by warming, cool 0.10 mL of 0.01 molWL hydrochloric acid VS (not more than and titrate <2.50> with 0.2 molWL sodium hydroxide VS (indi- 0.002z). cator: 3 drops of phenolphthalein TS). Perform a blank de- (3) Sulfate—To 10 mL of a solution of Sul- termination using a mixture of 50 mL of methanol and 18 mL fobromophthalein Sodium (1 in 500) add 5 drops of dilute of water, and make any necessary correction. hydrochloric acid, heat to boil, and add 1 mL of hot barium chloride TS: the solution is clear when observed 1 minute af- Each mL of 0.2 molWL sodium hydroxide VS ter the addition of the barium chloride TS. ＝ 53.46 mg of C H N O S 11 13 3 3 (4) Calcium—Weigh accurately about 5 g of Sul- Containers and storage Containers—Well-closed contain- fobromophthalein Sodium, transfer to a porcelain dish, heat ers. gently to char, and heat strongly between 7009Cand7509C until the residue is incinerated. After cooling, add 10 mL of dilute hydrochloric acid, and heat for 5 minutes on a water bath. Transfer the contents to a ‰ask with 50 mL of water, Sulfobromophthalein Sodium and add 5 mL of 8 molWL potassium hydroxide TS and 0.1 g of NN indicator. Titrate <2.50> with 0.01 molWL disodium di- スルホブロモフタレインナトリウム hydrogen ethylenediamine tetraacetate VS until the red-pur- ple color of the solution changes to blue. Each mL of 0.01 molWL disodium dihydrogen ethylenediamine tetraacetate VS ＝ 0.4008 mg of Ca The content of calcium (Ca: 40.08) is not more than 0.05z. (5) Heavy metals <1.07>—Proceed with 1.0 g of Sul- fobromophthalein Sodium according to Method 2, and per- form the test. Prepare the control solution with 2.0 mL of C H Br Na O S :838.00 20 8 4 2 10 2 Standard Lead Solution (not more than 20 ppm). Disodium 5,5?-(4,5,6,7-tetrabromo-3-oxo-1,3-dihydro- (6) Arsenic <1.11>—Transfer 0.65 g of Sulfobromo- isobenzofuran-1,1-diyl)bis(2-hydroxybenzenesulfonate) phthalein Sodium to a crucible, add 10 mL of a solution of [71-67-0] magnesium nitrate hexahydrate in ethanol (95) (1 in 50), ˆre to burn, then heat gently until the residue is incinerated. If Sulfobromophthalein Sodium, when dried, contains any carbon remains, moisten the residue with a small amount not less than 96.0z and not more than 104.0z of of nitric acid, and incinerate again by ignition. After cooling, C H Br Na O S . 20 8 4 2 10 2 add 10 mL of dilute sulfuric acid, and heat until white fumes Description Sulfobromophthalein Sodium occurs as a are evolved. After cooling, add 5 mL of water to the residue, white, crystalline powder. It is odorless. and perform the test with this solution as the test solution It is soluble in water, and practically insoluble in ethanol (not more than 3.1 ppm). (95) and in diethyl ether. Loss on drying <2.41> Not more than 5.0z (0.5 g, 1059C, It is hygroscopic. 3 hours). Identiˆcation (1) Dissolve 0.02 g of Sulfobromo- Residue on ignition <2.44> 14–19z (after drying, 0.5 g, phthaleinSodiumin10mLofwater,andadd1mLofsodi- 700 – 7509C). um carbonate TS: a blue-purple color is produced. Add 1 mL of dilute hydrochloric acid to the solution: the color of the Assay Dissolve about 0.1 g of Sulfobromophthalein Sodi- solution disappears. um, previously dried and accurately weighed, in water to (2) Transfer 0.2 g of Sulfobromophthalein Sodium to a make exactly 500 mL. Pipet 5 mL of this solution, and add a porcelain crucible, mix well with 0.5 g of anhydrous sodium solution of anhydrous sodium carbonate (1 in 100) to make carbonate, and ignite until the mixture is charred. After cool- exactly 200 mL. Perform the test with this solution as direct- ing, add 15 mL of hot water to the residue, heat for 5 minutes ed under Ultraviolet-visible Spectrophotometry <2.24>.De- on a water bath, ˆlter, and render the ˆltrate slightly acid termine the absorbance A of this solution at the wavelength with hydrochloric acid: the solution responds to the Qualita- of maximum absorption at about 580 nm, using water as the tive Tests <1.09> for bromide, and the Qualitative Tests blank. <1.09> (1) and (2) for sulfate. Amount (mg) of C H Br Na O S (3) Sulfobromophthalein Sodium responds to the 20 8 4 2 10 2 ＝(A/881)× 200,000 Qualitative Tests <1.09> (1) for sodium salt. Containers and storage Containers—Tight containers. pH <2.54> The pH of a solution of 1.0 g of Sul- 1130 Sulfobromophthalein Sodium Injection / O‹cial Monographs JP XV

Storage—Light-resistant. der. It is odorless and tasteless. It is freely soluble in carbon disulˆde, and practically in- solubleinwater,inethanol(95)andindiethylether. Sulfobromophthalein Sodium Identiˆcation (1) Ignite Sulfur: it burns with a blue ‰ame Injection and gives a pungent odor of sulfur dioxide. (2) Dissolve 5 mg of Sulfur in 5 mL of sodium hydroxide スルホブロモフタレインナトリウム注射液 TS by heating in a water bath, cool, and add 1 drop of sodi- um pentacyanonitrosylferrate (III) TS: a blue-purple color develops. Sulfobromophthalein Sodium Injection is an aque- (3) Boil 1 mg of sulfur with 2 mL of pyridine and 0.2 mL ous solution for Injection. It contains not less than 94 of sodium hydrogen carbonate TS: a blue color develops. z and not more than 106z of the labeled amount of Purity (1) Clarity of solution—Dissolve 1.0 g of Sulfur in sulfobromophthalein sodium (C20H8Br4Na2O10S2: a mixture of 20 mL of a solution of sodium hydroxide (1 in 6) 838.00). and 2 mL of ethanol (95) by boiling: the solution is clear. Dis- Method of preparation Prepare as directed under Injec- solve 2.0 g of Sulfur in 10 mL of carbon disulˆde: the solu- tions, with Sulfobromophthalein Sodium. tion is almost clear or slightly opalescent. (2) Acidity or alkalinity—Shake 2.0 g of Sulfur with 50 Description Sulfobromophthalein Sodium Injection is a mL of freshly boiled and cooled water, and add 2 drops of clear and colorless or pale yellow liquid. phenolphthalein TS: no red color develops. Further add 1.0 pH:5.0–6.0 mL of 0.1 molWL sodium hydroxide VS: a red color develops. Identiˆcation (1) Measure a volume of Sulfobromo- (3) Arsenic <1.11>—Prepare the test solution with 0.20 g phthalein Sodium Injection, equivalent to 0.02 g of Sul- of Sulfur according to Method 3, and perform the test (not fobromophthalein Sodium according to the labeled amount, more than 10 ppm). and proceed as directed in the Identiˆcation (1) under Sul- Loss on drying <2.41> Not more than 1.0z (1 g, in vacuum, fobromophthalein Sodium. not more than 0.67 kPa, silica gel, 4 hours). (2) Measure a volume of Sulfobromophthalein Sodium Injection, equivalent to 0.1 g of Sulfobromophthalein Sodi- Residue on ignition <2.44> Not more than 0.2z (1 g). um according to the labeled amount, add 0.5 g of anhydrous Assay Weigh accurately about 0.4 g of Sulfur, previously sodium carbonate, and evaporate on a water bath to dryness. dried, dissolve in 20 mL of potassium hydroxide-ethanol TS Ignite the residue until it is charred. Proceed as directed in the and 10 mL of water by boiling, cool, and add water to make Identiˆcation (2) under Sulfobromophthalein Sodium. exactly 100 mL. Transfer exactly 25 mL of the solution to a Extractble volume <6.05> It meets the repequirement. 400-mL beaker, add 50-mL of hydrogen peroxide TS, and heat on a water bath for 1 hour. Acidify the solution with di- Pyrogen <4.04> Add isotonic sodium chloride solution to lute hydrochloric acid, add 200 mL of water, heat to boil, Sulfobromophthalein Sodium Injection to make a 0.5 w Wvz add hot barium chloride TS dropwise until no more solution of Sulfobromophthalein Sodium according to the la- precipitate is formed, and heat on a water bath for 1 hour. beled amount. Inject into each of the rabbits 5 mL of this so- Collect the precipitate, and wash with water until the last lution per kg of body mass: it meets the requirements. washing shows no opalescence with silver nitrate TS. Dry the Assay Measure exactly a volume of Sulfobromophthalein precipitate, heat strongly to constant mass, and weigh as Sodium Injection, equivalent to about 0.1 g of sul- barium sulfate (BaSO4: 233.39). Perform a blank determina- fobromophthalein sodium (C20H8Br4Na2O10S2), add water to tion, and make any necessary correction. make exactly 500 mL, and proceed as directed in the Assay Amount (mg) of S under Sulfobromophthalein Sodium. ＝ amount (mg) of barium sulfate (BaSO4) × 0.13739 Amount (mg) of sulfobromophthalein sodium Containers and storage Containers—Well-closed contain- (C20H8Br4Na2O10S2) ers. ＝(A/881)× 200,000 Containers and storage Containers—Hermetic containers. Storage—Light-resistant. Sulfur and Camphor Lotion

イオウ・カンフルローション Sulfur Method of preparation イオウ Sulfur 60 g d-Camphor or dl-Camphor 5 g S: 32.07 Hydroxypropylcellulose 4 g Calcium Hydroxide 1 g Sulfur, when dried, contains not less than 99.5z Ethanol 4 mL of S. Water or Puriˆed Water a su‹cient quantity Description Sulfur occurs as a light yellow to yellow pow- To make 1000 mL JP XV O‹cial Monographs / 1131

nitrate TS: a purple color is produced (salicylic acid). Dissolve Hydroxypropylcellulose in 200 mL of Water or (2) Shake 1 g of Sulfur, Salicylic Acid and Thianthol Puriˆed Water. Add this solution in small portions to the Ointment with 20 mL of diethyl ether, remove the super- triturate of Sulfur with the Ethanol solution of d-Camphor natant liquid and ‰oating materials. Wash the residue with 10 or dl-Camphor, and triturate again the mixture. Separately, mL of diethyl ether, and remove the diethyl ether by suction. dissolve Calcium Hydroxide in 500 mL of Water or Puriˆed To the residue add 2 mL of pyridine and 0.2 mL of sodium Water, stopper tightly, shake, and allow to stand. Add 300 hydrogen carbonate TS, and boil: a light blue to blue color is mL of this supernatant liquid to the above mixture, then add produced (sulfur). Water or Puriˆed Water to make 1000 mL, and shake (3) To 1 g of Sulfur, Salicylic Acid and Thianthol Oint- thoroughly. ment add 15 mL of ethanol (95), stir well while warming on a Description Sulfur and Camphor Lotion is a light yellow water bath, cool, and ˆlter. Use the ˆltrate as the sample so- suspension. lution. Dissolve 0.01 g each of salicylic acid and thianthol in 5 A part of the components separates out on standing. mL of ethanol (95), and use these solutions as the standard solution (1) and standard solution (2). Perform the test with Identiˆcation (1) To 5 mL of well shaken Sulfur and these solutions as directed under Thin-layer Chromatography Camphor Lotion add 25 mL of water, and centrifuge [use <2.03>.Spot5mL each of the sample solution and standard this supernatant liquid for test (3)]. To 0.02 g of the solutions (1) and (2) on a plate of silica gel with ‰uorescent precipitate add 2 mL of pyridine and 0.2 mL of sodium indicator for thin-layer chromatography. Develop the plate hydrogen carbonate TS, and boil: a blue color develops (sul- with a mixture of chloroform, acetone and acetic acid (100) fur). (45:5:1) to a distance of about 10 cm, and air-dry the plate. (2) To 10 mL of well shaken Sulfur and Comphor Lotion Examine under ultraviolet light (main wavelength: 254 nm): add 5 mL of diethyl ether, and mix. Separate the diethyl ether the spots of each component obtained from the sample solu- layer, and ˆlter through a pledget of cotton. Wash the cotton tion and standard solutions (1) and (2) show the same Rf with a small portion of diethyl ether, combine the washings value. Spray iron (III) chloride TS upon the plate evenly: the with the ˆltrate, and distil cautiously on a water bath to re- spot from the standard solution (1) and that from the cor- move the diethyl ether. Dissolve the residue in 1 mL of responding sample solution reveal a purple color. methanol, add 1 mL of 2,4-dinitrophenylhydrazine TS, and heat for about 2 minutes on a water bath. Cool, dilute with Containers and storage Containers—Tight containers. water to make about 5 mL, and allow to stand. Filter the produced precipitate through a glass ˆlter (G4), and wash the residue on the ˆlter with water until the last washing is color- Sulpiride less. Dissolve the residue in 10 mL of ethanol (95), add 5 mL of sodium hydroxide TS, and allow to stand for 2 minutes: a スルピリド red color develops (d-camphor or dl-camphor). (3) The supernatant liquid obtained in (1) responds to the Qualitative Tests <1.09> (2) and (3) for calcium salt. Containers and storage Containers—Tight containers.

Sulfur, Salicylic Acid and C15H23N3O4S: 341.43 N-(1-Ethylpyrolidin-2-ylmethyl)-2-methoxy-5- Thianthol Ointment sulfamoylbenzamide [15676-16-1] イオウ・サリチル酸・チアントール軟膏 Sulpiride, when dried, contains not less than 98.5z and not more than 101.0z of C15H23N3O4S. Method of preparation Description Sulpiride is a white, crystalline powder. Sulfur 100 g It is freely soluble in acetic acid (100) and in dilute acetic Salicylic Acid, ˆnely powdered 30 g acid, sparingly soluble in methanol, slightly soluble in Thianthol 100 mL ethanol (99.5), and practically insoluble in water. Zinc Oxide, very ˆnely powdered 100 g It is soluble in 0.05 mol/L sulfuric acid TS. Simple Ointment or a suitable A solution of Sulpiride in methanol (1 in 100) shows no op- ointment base a su‹cient quantity tical rotation. To make 1000 g Melting point: about 1789C (with decomposition). Prepare as directed under Ointments, with above in- Identiˆcation (1) Dissolve 0.1 g of Sulpiride in 0.05 gredients. mol/LsulfuricacidTStomake100mL.Dilute5mLofthe solution with water to make 100 mL. Determine the absorp- Description Sulfur, Salicylic Acid and Thianthol Ointment tion spectrum of the solution as directed under Ultraviolet- is light yellow in color. visible Spectrophotometry <2.24>, using water as the blank, Identiˆcation (1) Stir well 0.5 g of Sulfur, Salicylic Acid and compare the spectrum with the Reference Spectrum: and Thianthol Ointment with 10 mL of water while heating, both spectra exhibit similar intensities of absorption at the cool, and ˆlter. To 1 mL of the ˆltrate add 5 mL of iron (III) same wavelengths. (2) Determine the infrared absorption spectrum of Sulpi- 1132 Sulpiride Capsules / O‹cial Monographs JP XV ride as directed in the potassium bromide disk method under sample solution obtained in the Assay as directed under Infrared Spectrophotometry <2.25>, and compare the spec- Ultraviolet-visible Spectrophotometry <2.24>,usingwateras trum with the Reference Spectrum: both spectra exhibit simi- the blank: it exhibits a maximum between 289 nm and 293 lar intensities of absorption at the same wave numbers. nm. Purity (1) Clarity of solution—Dissolve 2.0 g of Sulpiride Uniformity of dosage units <6.02> Perform the test accord- in 7 mL of dilute acetic acid, and add water to make 20 mL: ing to the following method: it meets the requirement of the the solution is clear. Perform the test with the solution as Content uniformity test. directed under Ultraviolet-visible Spectrophotometry <2.24>, To 1 capsule of Sulpiride Capsules add 30 mL of 0.05 using water as the blank: the absorbance at a wavelength of mol/L sulfuric acid TS, shake for 30 minutes, add 0.05 450 nm does not exceed 0.020. mol/LsulfuricacidTStomakeexactlyV mL so that each (2) Heavy metals <1.07>—Proceed with 2.0 g of Sulpiride mL of the solution contains about 1 mg of sulpiride as directed under Method 2, and perform the test. Prepare (C15H23N3O4S), and ˆlter the solution. Discard the ˆrst 20 the control solution with 2.0 mL of Standard Lead Solution mL of the ˆltrate, pipet the subsequent 5 mL of the ˆltrate, (not more than 10 ppm). add water to make exactly 100 mL, and use this solution as (3) Related substances—Dissolve 50 mg of Sulpiride in 10 the sample solution. Proceed as directed in the Assay. mL of methanol, and use this solution as the sample solution. Amount (mg) of sulpiride (C H N O S) Dilute 1 mL of the sample solution, accurately measured, 15 23 3 4 ＝W ×(A /A )×(V/50) with methanol to make exactly 100 mL. Dilute 2 mL of this S T S solution, accurately measured, with methanol to make ex- WS: Amount (mg) of sulpiride for assay actly 10 mL, and use this solution as the standard solution. Dissolution Being speciˆed separately. Perform the test with these solutions as directed under Thin- layer Chromatography <2.03>.Spot20mLeachofthesample Assay Cut the capsule of not less than 20 Sulpiride Cap- solution and standard solution on a plate of silica gel with sules, weigh accurately the mass of the contents, and powder. ‰uorescent indicator for thin-layer chromatography. Develop Weigh accurately a portion of the powder, equivalent to the plate with a mixture of 1-butanol, water and acetic acid about 0.1 g of sulpiride (C15H23N3O4S), add 70 mL of 0.05 (100) (4:2:1) to a distance of about 10 cm, and air-dry the mol/L sulfuric acid TS, shake for 30 minutes, and add 0.05 plate. Examine under ultraviolet light (main wavelength: 254 mol/L sulfuric acid TS to make exactly 100 mL, and ˆlter. nm): the number of the spots other than the principal spot Discard the ˆrst 20 mL of the ˆltrate, pipet the subsequent 5 from the sample solution is not more than 2, and they have mL of the ˆltrate, add water to make exactly 100 mL, and use no more color than the spot from the standard solution. this solution as the sample solution. Separately, weigh ac- When the plate is exposed to iodine vapor for 30 minutes, the curately about 50 mg of sulpiride for assay, previously dried number of the spots other than the principal spot from the at 1059C for 3 hours, and dissolve in 0.05 m/L sulfuric acid sample solution is not more than 2, and they have no more TS to make exactly 50 mL. Pipet 5 mL of this solution, add color than the spot from the standard solution. water to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, A and A ,of Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, T S the sample solution and standard solution at 291 nm as 3 hours). directed under Ultraviolet-visible Spectrophotometry <2.24>, Residue on ignition <2.44> Not more than 0.1z (1 g). using water as the blank.

Assay Dissolve about 0.4 g of Sulpiride, previously dried Amount (mg) of sulpiride (C15H23N3O4S)＝WS×(AT/AS)×2 and accurately weighed, in 80 mL of acetic acid (100), and ti- W : Amount (mg) of sulpiride for assay trate <2.50> with 0.1 molWL perchloric acid VS (indicator: 2 S drops of crystal violet TS) until the color of the solution Containers and storage Containers—Tight containers. changes from violet through blue to bluish green. Perform a blank determination, and make any necessary correction. Each mL of 0.1 molWL perchloric acid VS Sulpiride Tablets ＝ 34.14 mg of C H N O S 15 23 3 4 スルピリド錠 Containers and storage Containers—Well-closed contain- ers. Sulpiride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of sulpi- Sulpiride Capsules ride (C15H23N3O4S: 341.43). Method of preparation Prepare as directed under Tablets, スルピリドカプセル with Sulpiride. Identiˆcation Determine the absorption spectrum of the Sulpiride Capsules contain not less than 95.0z and sample solution obtained in the Assay as directed under not more than 105.0z of the labeled amount of sulpi- Ultraviolet-visible Spectrophotometry <2.24>,usingwateras ride (C15H23N3O4S: 341.43). the blank: it exhibits a maximum between 289 nm and 293 nm. Method of preparation Prepare as directed under Capsules, with Sulpiride. Uniformity of dosage units <6.02> Perform the test accord- ing to the following method: it meets the requirement of the Identiˆcation Determine the absorption spectrum of the JP XV O‹cial Monographs / Sulpyrine Hydrate 1133

Content uniformity test. Amount (mg) of sulpiride (C H N O S)＝W ×(A /A )×2 To 1 tablet of Sulpiride Tablets add 30 mL of 0.05 mol/L 15 23 3 4 S T S sulfuric acid TS, shake for 30 minutes, add 0.05 mol/L sul- WS: Amount (mg) of sulpiride for assay furicacidTStomakeexactlyVmLsothateachmLofthe Containers and storage Containers—Tight containers. solution contains about 1 mg of sulpiride (C15H23N3O4S), and ˆlter the solution. Discard the ˆrst 20 mL of the ˆltrate, pipet the subsequent 5 mL of the ˆltrate, add water to make exactly 100 mL, and use this solution as the sample solution. Proceed Sulpyrine Hydrate as directed in the Assay. スルピリン水和物 Amount (mg) of sulpiride (C15H23N3O4S) ＝WS×(AT/AS)×(V/50)

WS: Amount (mg) of sulpiride for assay Dissolution <6.10> Perform the test according to the follow- ing method: it meets the requirement. Perform the test with 1 tablet of Sulpiride Tablets at 50 revolutions per minute according to the Paddle method, us- C13H16N3NaO4S.H2O: 351.35 ing 900 mL of 2nd ‰uid for dissolution test as the dissolution Monosodium [(1,5-dimethyl-3-oxo-2-phenyl-2,3-dihydro- medium. Withdraw 20 mL or more of the dissolution medi- 1H-pyrazol-4-yl)(methyl)amino]methanesulfonate um 30 minutes after starting the test for a 50-mg tablet and 45 monohydrate [5907-38-0] minutes after for a 100-mg or a 200-mg tablet, and ˆlter through a membrane ˆlter with pore size of not more than 0.5 Sulpyrine Hydrate contains not less than 98.5z of mm. Discard the ˆrst 10 mL of the ˆltrate, pipet the subse- sulpyrine (C13H16N3NaO4S: 333.34), calculated on the quent V mL, add 2nd ‰uid for dissolution test to make ex- dried basis. actly V? mL so that each mL contains about 56 mg of sulpi- Description Sulpyrine Hydrate occurs as white to light yel- ride (C H N O S) according to the labeled amount, and use 15 23 3 4 low crystals or crystalline powder. It is odorless, and has a this solution as the sample solution. Separately, weigh ac- bitter taste. curately about 28 mg of sulpiride for assay, previously dried It is very soluble in water, slightly soluble in ethanol (95), at 1059C for 3 hours, and dissolve in 2nd ‰uid for dissolution and practically insoluble in diethyl ether. test to make exactly 100 mL. Pipet 5 mL of this solution, add It is colored by light. 2nd ‰uid for dissolution test to make exactly 25 mL, and use this solution as the standard solution. Perform the test with Identiˆcation (1) Add 2 drops of dilute sulfuric acid and 1 the sample solution and the standard solution as directed un- mL of chlorinated lime TS to 3 mL of a solution of Sulpyrine der Ultraviolet-visible Spectrophotometry <2.24>,anddeter- Hydrate (1 in 15): a deep blue color develops at ˆrst, but the mine the absorbances, AT and AS,at291nm.Thedissolution color immediately turns red, then gradually changes to yel- rate of a 50-mg tablet in 30 minutes is not less than 80z,that low. ofa100-mgtabletin45minutesisnotlessthan75z,and (2) Boil 5 mL of a solution of Sulpyrine Hydrate (1 in 25) that of a 200-mg tablet in 45 minutes is not less than 70z. with 3 mL of dilute hydrochloric acid: the odor of sulfur di- oxide is perceptible at ˆrst, and on further boiling, the odor Dissolution rate (z) with respect to the labeled amount of of formaldehyde is perceptible. sulpiride (C H N O S) 15 23 3 4 (3) A solution of Sulpyrine Hydrate (1 in 10) responds to ＝WS×(AT/AS)×(V?/V)×(1/C)×180 the Qualitative Tests <1.09> for sodium salt.

WS: Amount (mg) of sulpiride for assay Purity (1) Clarity of solution, and acidity or C: Labeled amount (mg) of sulpiride (C15H23N3O4S) in 1 alkalinity—Dissolve 1.0 g of Sulpyrine Hydrate in 10 mL of tablet water: the solution is clear and neutral. (2) Sulfate <1.14>—Dissolve 0.20 g of Sulpyrine Hydrate Assay Weigh accurately, and powder not less than 20 Sulpi- in 0.05 mol L hydrochloric acid VS to make 50 mL, and per- ride Tablets. Weigh accurately a portion of the powder, W form the test using this solution as the test solution. Prepare equivalent to about 0.1 g of sulpiride (C H N O S), add 70 15 23 3 4 the control solution with 0.50 mL of 0.005 mol L sulfuric mL of 0.05 mol/L sulfuric acid TS, shake for 30 minutes, W acid VS and 0.05 mol LhydrochloricacidVStomake50mL and add 0.05 mol/L sulfuric acid TS to make exactly 100 mL, W (not more than 0.120z). and ˆlter. Discard the ˆrst 20 mL of the ˆltrate, pipet the sub- (3) Heavy metals <1.07>—Proceed with 1.0 g of Sulpyrine sequent 5 mL of the ˆltrate, add water to make exactly 100 Hydrate according to Method 2, and perform the test. Pre- mL, and use this solution as the sample solution. Separately, pare the control solution with 2.0 mL of Standard Lead Solu- weigh accurately about 50 mg of sulpiride for assay, previ- tion (not more than 20 ppm). ously dried at 1059C for 3 hours, and dissolve in 0.05 mL sul- (4) Merbuline—Transfer 0.10 g of Sulpyrine Hydrate furicacidTStomakeexactly50mL.Pipet5mLofthissolu- with 2 mL of water and 1 mL of dilute sulfuric acid into a tion, add water to make exactly 100 mL, and use this solution ‰ask, cover with a funnel, and boil gently for 15 minutes. as the standard solution. Determine the absorbances, A and T Cool, add 2 mL of a solution of sodium acetate trihydrate (1 A , of the sample solution and standard solution at 291 nm as S in 2) and water to make 5 mL, shake this solution with 5 mL directed under Ultraviolet-visible Spectrophotometry<2.24>, of benzaldehyde-saturated solution, and allow to stand for 5 using water as the blank. minutes: the solution is clear. 1134 Sulpyrine Injection / O‹cial Monographs JP XV

(5) Chloroform-soluble substances—Mix, by frequent the sample solution and standard solution into separate shaking, 1.0 g of Sulpyrine Hydrate and 10 mL of chlo- 25-mL volumetric ‰asks, add 5 mL of ethanol (95), 2 mL of a roform for 30 minutes. Collect the precipitate, wash with two solution of 4-dimethylaminocinnamaldehyde in ethanol (95) 5-mL portions of chloroform, combine the washings with the (1 in 250) and 2 mL of acetic acid (100) to each of these solu- ˆltrate, and evaporate on a water bath to dryness. Dry the tions, shake well, allow to stand for 15 minutes, and add residue at 1059C for 4 hours: the mass of the residue is not water to exactly 25 mL. Perform the test with these solutions more than 5.0 mg. as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution prepared with 2 mL of water in the Loss on drying <2.41> Not more than 6.0z (1 g, 1059C, same manner as the blank. Determine the absorbances, A 4 hours). T and AS, of the subsequent solutions of the sample solution Assay Weigh accurately about 0.25 g of Sulpyrine Hydrate, and the standard solution at 510 nm. dissolve in 100 mL of diluted hydrochloric acid (1 in 20), Amount (mg) of sulpyrine hydrate (C H N NaO S.H O) previously cooled below 109C. Titrate <2.50> immediately 13 16 3 4 2 in 1 mL of Sulpyrine Injection with 0.05 molWL iodine VS while keeping the temperature be- ＝W ×(A /A )×(50/V)× 1.0540 tween 59Cand109C, until the color of the solution remains S T S blue upon shaking vigorously for 1 minute after the addition WS: Amount (mg) of sulpyrine for assay, calculated of 0.05 molWL iodine VS (indicator: 1 mL of starch TS). on the dried basis Each mL of 0.05 molWLiodineVS Containers and storage Containers—Hermetic containers,

＝ 16.67 mg of C13H16N3NaO4S and colored containers may be used. Storage—Light-resistant, and under nitrogen atmosphere. Containers and storage Containers—Tight containers. Storage—Light-resistant. Sultamicillin Tosilate Hydrate

Sulpyrine Injection スルタミシリントシル酸塩水和物 スルピリン注射液

Sulpyrine Injection is an aqueous solution for injec- tion. It contains not less than 95z and not more than 105z of the labeled amount of sulpyrine hydrate (C13H16N3NaO4S.H2O: 351.35). Method of preparation Prepare as directed under Injec- tions, with Sulpyrine Hydrate. C25H30N4O9S2.C7H8O3S.2H2O: 802.89 Description Sulpyrine Injection is a clear, colorless or pale (2S,5R)-(3,3-Dimethyl-4,4,7-trioxo-4-thia-1-azabicy- yellow liquid. clo[3.2.0]hept-2-ylcarbonyloxy)methyl (2S,5R,6R)-6-[(2R)- pH:5.0–8.5 2-amino-2-phenylacetylamino]-3,3-dimethyl- 7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylate Identiˆcation (1) To a volume of Sulpyrine Injection, monotosylate dihydrate [83105-70-8, anhydride] equivalent to 0.2 g of Sulpyrine Hydrate according to the la- beled amount, add water to make 3 mL, then add 2 drops of Sultamicillin Tosilate Hydrate contains not less than dilute sulfuric acid and 1 mL of chlorinated lime TS: a deep 698 mg (potency) per mg, calculated on the anhydrous blue color develops at ˆrst, and the color immediately turns basis and corrected by the amount of residual solvent. red and gradually changes to yellow. The potency of Sultamicillin Tosilate is expressed as (2) To a volume of Sulpyrine Injection, equivalent to 0.2 mass (potency) of sultamicillin (C H N O S : g of Sulpyrine according to the labeled amount, add water to 25 30 4 9 2 594.66). make 5 mL, and boil this solution with 3 mL of dilute hydrochloric acid: the odor of sulfur dioxide is perceptible at Description Sultamicillin Tosilate Hydrate occurs as a ˆrst, and on further boiling the odor of formaldehyde is per- white to yellowish white crystalline powder. ceptible. It is freely soluble in acetonitrile, in methanol and in ethanol (99.5), and very slightly soluble in water. Assay Pipet 2 mL of Sulpyrine Injection, dilute with water to exactly 100 mL. Measure exactly a volume (V mL) of this Identiˆcation Determine the infrared absorption spectrum solution, equivalent to about 0.05 g of sulpyrine hydrate of Sultamicillin Tosilate Hydrate as directed in the paste

(C13H16N3NaO4S.H2O), and add water to make exactly 100 method under Infrared Spectrophotometry <2.25>,andcom- mL. Pipet 5 mL of this solution, add water to exactly 100 pare the spectrum with the Reference Spectrum or the spec- mL, and use this solution as the sample solution. Weigh ac- trum of Sultamicillin Tosilate Reference Standard: both spec- curately about 0.05 g of sulpyrine for assay (previously deter- tra exhibit similar intensities of absorption at the same wave mine the loss on drying <2.41> inthesamemannerasSulpy- numbers. rine Hydrate), and dissolve in water to make exactly 100 mL. Optical rotation <2.49> [a]20: ＋173 – ＋1879(0.5 g calcu- Pipet 5 mL of this solution, add water to exactly 100 mL, and D lated on the anhydrous bases, a mixture of water and acetoni- use this solution as the standard solution. Pipet 2 mL each of JP XV O‹cial Monographs / Sultamicillin Tosilate Hydrate 1135 trile (3:2), 25 mL, 100 mm). Proceed as directed in the operating conditions in the Puri- ty (3). Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of System suitability— Sultamicillin Tosilate Hydrate according to Method 2, and Proceed as directed in the system suitability in the Purity perform the test. Prepare the control solution with 2.0 mL (3). Standard Lead Solution (not more than 20 ppm). (5) Penicilloic acids—Weigh accurately about 25 mg of (2) Arsenic <1.11>—Prepare the test solution with 1.0 g Sultamicillin Tosilate Hydrate, dissolve in 1 mL of acetoni- of Sultamicillin Tosilate Hydrate, according to Method 3, trile, and add 25 mL of 0.02 molWL phosphate buŠer solu- and perform the test (not more than 2 ppm). tion, pH 3.0, in a 100-mL ‰ask with stopper. Add exactly 5 (3) Ampicillin—Perform the procedure rapidly. Weigh mL of 0.005 molWL iodine VS, and allow to stand the stop- accurately about 20 mg of Sultamicillin Tosilate Hydrate, pered‰askfor5minutes.Titrate<2.50> with 0.005 molWLso- dissolve in the mobile phase to make exactly 50 mL, and use dium thiosulfate VS (indicator: 1.0 mL of starch TS). Per- this solution as the sample solution. Separately, weigh ac- form a blank determination in the same manner, and make curately about 20 mg (potency) of Ampicillin Reference Stan- any necessary correction. Calculate the amount of penicilloic dard, dissolve in the mobile phase to make exactly 100 mL. acid (C H N O S : 630.70) by using the following equa- Pipet 6 mL of this solution, add the mobile phase to make ex- 25 34 4 11 2 tion: it is not more than 3.0z. actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solu- Each mL of 0.005 molWL sodium thiosulfate VS tion and standard solution as directed under Liquid Chro- ＝ 0.2585 mg of C25H34N4O11S2 matography <2.01> according to the following conditions, (6) Residual solvent <2.46>—Weigh accurately about 0.1 and calculate the area of the peak of ampicillin by the auto- g of Sultamicillin Tosilate Hydrate, dissolve in 2 mL of matic integration method: the peak area from the sample so- methanol, add water to make exactly 20 mL, and use this so- lution is not more than that from the standard solution. lution as the sample solution. Separately, weigh accurately Operating conditions— about 1 g of ethyl acetate, and mix with water to make exact- Detector, column, and column temperature: Proceed as ly 200 mL. Pipet 2 mL of this solution, add 10 mL of directed in the operating conditions in the Assay. methanol and water to make exactly 100 mL, and use this so- Mobile phase: Dissolve 3.12 g of sodium dihydrogen lution as the standard solution. Perform the test with exactly phosphate dihydrate in about 750 mL of water, adjust to pH 5 mL each of the sample solution and standard solution as 3.0withdilutedphosphoricacid(1in10),andaddwaterto directed under Gas Chromatography <2.02> according to the make following conditions, and determine the peak areas, A and 1000 mL. To 80 mL of acetonitrile for liquid chromato- T A , of ethyl acetate of these solutions. Calculate the amount graphy add this solution to make 1000 mL. S of ethyl acetate by the following equation: not more than 2.0 Flow rate: Adjust the ‰ow rate so that the retention time of z. ampicillin is about 14 minutes. System suitability— Amount (z) of ethyl acetate

System performance: Dissolve 12 mg of Ampicillin Refer- ＝(WS/WT)×(A T/A S)× (1/5) ence Standard, 4 mg of Sulbactam Reference Standard and 4 W : Amount (mg) of ethyl acetate mg of p-toluenesulfonic acid monohydrate in 1000 mL of the S W :Amount(mg)ofthesample mobile phase. When the procedure is run with 25 mLofthis T solution under the above operating conditions, sulbactam, p- Operating conditions— toluenesulfonic acid and ampicillin are eluted in this order Detector: A hydrogen ‰ame-ionization detector. with the resolution between these peaks being not less than Column: A column 3 mm in inside diameter and 1 m in 2.0. length, packed with porous stylene-divinylbenzene System repeatability: When the test is repeated 6 times with copolymer for gas chromatography (0.0085 mm in average 25 mL of the standard solution under the above operating pore size and 300 – 400 m2/g in speciˆc surface area) (150 to conditions, the relative standard deviation of the peak areas 180 mminparticlediameter). of ampicillin is not more than 2.0z. Column temperature: A constant temperature of about (4) Sulbactam—Perform the procedure rapidly. Weigh 1559C. accurately about 20 mg of Sultamicillin Tosilate Hydrate, Carrier gas: Nitrogen dissolve in the mobile phase to make exactly 50 mL, and use Flow rate: Adjust the ‰ow rate so that the retention time of this solution as the sample solution. Separately, weigh ac- ethyl acetate is about 6 minutes. curately an amount of Sulbactam Reference Standard, e- System suitability— quivalent to about 20 mg (potency), dissolve in the mobile System performance: When the procedure is run with 5 mL phase to make exactly 100 mL. Pipet 2 mL of this solution, of the standard solution under the above operating condi- add the mobile phase to make exactly 100 mL, and use this tions, the number of theoretical plates and the symmetry fac- solution as the standard solution. Perform the test with ex- tor of the peak of ethyl acetate are not less than 500 steps and actly 25 mL each of the sample solution and standard solution not more than 1.5, respectively. as directed under Liquid Chromatography <2.01> according System repeatability: When the test is repeated 6 times with to the following conditions, and calculate the area of the 5 mL of the standard solution under the above operating con- peak of sulbactam by the automatic integration method: the ditions, the relative standard deviation of the peak areas of peak area from the sample solution is not more than that ethyl acetate is not more than 5z. from the standard solution. Water <2.48> 4.0–6.0z (0.5 g, volumetric titration, direct Operating conditions— titration). 1136 Sultiame / O‹cial Monographs JP XV

2-yl)benzenesulfonamide S,S-dioxide [61-56-3] Residue on ignition <2.44> Not more than 0.2z (1 g). Assay Perform the procedure rapidly. Weigh accurately an Sultiame, when dried, contains not less than 98.5z amount of Sultamicillin Tosilate Hydrate and Sultamicillin of C10H14N2O4S2. Tosilate Reference Standard, equivalent to about 50 mg Description Sultiame occurs as white crystals or crystalline (potency), dissolve each in the mobile phase to make exactly powder. It is odorless, and has a slightly bitter taste. 50 mL. Pipet 5 mL each of these solutions, add exactly 5 mL It is very soluble in N,N-dimethylformamide, freely solu- of the internal standard solution, add the mobile phase to ble in n-butylamine, slightly soluble in methanol and in make 25 mL, and use these solutions as the sample solution ethanol (95), very slightly soluble in water, and practically in- and standard solution, respectively. Perform the test with 10 soluble in diethyl ether. mLeachofthesesolutionsasdirectedunderLiquidChro- It dissolves in sodium hydroxide TS. matography <2.01> according to the following conditions, and calculate the ratios, Q T and QS, of the peak area of sul- Identiˆcation (1) Dissolve 0.02 g of Sultiame in 5 mL of tamicillin to that of the internal standard of each solution. water and 1 mL of n-butylamine, add 2 to 3 drops of copper (II) sulfate TS, and shake well. To this solution add 5 mL of Amount [mg (potency)] of sultamicillin (C H N O S ) 25 30 4 9 2 chloroform, shake, and allow to stand: a green color de- ＝W ×(Q /Q )×1000 S T S velops in the chloroform layer.

WS: Amount [mg (potency)] of Sultamicillin Tosilate (2) Mix 0.1 g of Sultiame with 0.5 g of sodium carbonate Reference Standard decahydrate, and melt carefully: the gas evolved changes moistened red litmus paper to blue. After cooling, crush the Internal standard solution—A solution of isopropyl-4- fused substance with a glass rod, stir with 10 mL of water, aminobenzoate in the mobile phase (1 in 2500). andˆlter.To4mLoftheˆltrateadd2dropsofhydrogen Operating conditions— peroxide (30), 5 mL of diluted hydrochloric acid (1 in 5) and 2 Detector: An ultraviolet absorption photometer to 3 drops of barium chloride TS: a white precipitate is (wavelength: 215 nm). formed. Column: A stainless steel column 3.9 mm in inside di- (3) Determine the absorption spectrum of a solution of ameter and 30 cm in length, packed with octadecylsilanized Sultiame in methanol (1 in 100,000) as directed under Ultrav- silica gel for liquid chromatography (10 mminparticledi- iolet-visible Spectrophotometry <2.24>, and compare the ameter). spectrum with the Reference Spectrum: both spectra exhibit Column temperature: A constant temperature of about similar intensities of absorption at the same wavelengths. 359C. Mobile phase: Dissolve 3.12 g of sodium dihydrogen- Melting point <2.60> 185 – 1889C phosphate in about 750 mL of water, adjust to pH 3.0 with Purity (1) Chloride <1.03>—Dissolve 1.0 g of Sultiame in diluted phosphoric acid (1 in 10), and add water to make 1000 20 mL of sodium hydroxide TS by warming, cool, and add 2 mL. To 400 mL of acetonitrile for liquid chromatography mL of acetic acid (100) and water to make 100 mL. After add this solution to make 1000 mL. shaking, ˆlter, and discard the ˆrst 10 mL of the ˆltrate. To Flow rate: Adjust the ‰ow rate so that the retention time of the subsequent 40 mL add 6 mL of dilute nitric acid and sultamicillin is about 4 minutes. water to make 50 mL, and perform the test using this solution System suitability— as the test solution. Prepare the control solution as follows: System performance: When the procedure is run with 10 to 0.25 mL of 0.01 molWL hydrochloric acid VS add 8 mL of mL of the standard solution under the above operating condi- sodium hydroxide TS, 0.8 mL of acetic acid (100), 6 mL of tions, p-toluenesulfonic acid, sultamicillin and the internal dilute nitric acid and water to make 50 mL (not more than standard are eluted in this order with the resolution between 0.022z). these peaks being not less than 2.0. (2) Sulfate <1.14>—Dissolve 1.0 g of Sultiame in 20 mL System repeatability: When the test is repeated 6 times with of sodium hydroxide TS by warming, cool, and add 8 mL of 10 mL of the standard solution under the above operating dilute hydrochloric acid and water to make 100 mL. After conditions, the relative standard deviation of the peak area of shaking, ˆlter, and discard the ˆrst 10 mL of the ˆltrate. To sultamicillin is not more than 2.0z. the subsequent 40 mL add 1 mL of dilute hydrochloric acid Containers and storage Containers—Tight containers. and water to make 50 mL, and perform the test using this so- lution as the test solution. Prepare the control solution as fol- lows: to 0.40 mL of 0.005 molWL sulfuric acid VS add 8 mL Sultiame of sodium hydroxide TS, 4.2 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.048z). スルチアム (3) Heavy metals <1.07>—Proceed with 2.0 g of Sultiame 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). (4) Arsenic <1.11>—Prepare the test solution with 1.0 g of Sultiame according to Method 3, and perform the test (not more than 2 ppm). (5) Related substances—Dissolve 0.10 g of Sultiame in C10H14N2O4S2:290.36 methanol to make exactly 20 mL, and use this solution as the 4-(3,4,5,6-Tetrahydro-2H-1,2-thiazin- sample solution. Separately, dissolve 10 mg of sulfanilamide JP XV O‹cial Monographs / Suxamethonium Chloride for Injection 1137 in methanol to make exactly 100 mL. Pipet 10 mL of this so- Purity (1) Clarity and color of solution—Dissolve 1.0 g of lution, add methanol to make exactly 100 mL, and use this Suxamethonium Chloride Hydrate in 10 mL of water: the so- solution as the standard solution. Perform the test with these lution is clear and colorless. solutions as directed under Thin-layer Chromatography (2) Related substances—Dissolve 0.25 g of Suxamethoni- <2.03>.Spot20mL each of the sample solution and standard um Chloride Hydrate in 5 mL of water, and use this solution solution on a plate of silica gel with ‰uorescent indicator for as the sample solution. Pipet 1 mL of the sample solution, thin-layer chromatography. Develop the plate with a mixture add water to make exactly 200 mL, and use this solution as of chloroform, methanol and ammonia solution (28) (30:8:1) the standard solution. Perform the test with these solutions to a distance of about 12 cm, and air-dry the plate. Examine as directed under Thin-layer Chromatography <2.03>.Spot1 under ultraviolet light (main wavelength: 254 nm): the spots mL each of the sample solution and standard solution on a other than the principal spot from the sample solution are not plate of cellulose for thin-layer chromatography. Develop the more intense than the spot from the standard solution. plate with a mixture of a solution of ammonium acetate (1 in Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 100), acetone, n-butanol and formic acid (20:20:20:1) to a 3 hours). distance of about 10 cm, and dry the plate at 1059Cfor15 minutes. Spray evenly platinic chloride-potassium iodide TS Residue on ignition <2.44> Not more than 0.1z (1 g). on the plate, and allow to stand for 15 minutes: the spots Assay Weigh accurately about 0.8 g of Sultiame, previously other than the principal spot from the sample solution are not dried, dissolve in 70 mL of N,N-dimethylformamide, and ti- more intense than the spot from the standard solution. trate < 2.50 > with 0.2 mol WL tetramethylammonium Water <2.48> 8.0–10.0z (0.4 g, direct titration). hydroxide VS (potentiometric titration). Perform a blank de- termination, and make any necessary correction. Residue on ignition <2.44> Not more than 0.1z (1 g). Each mL of 0.2 molWL tetramethylammonium Assay Weigh accurately about 0.4 g of Suxamethonium hydroxide VS Chloride Hydrate, dissolve in 80 mL of a mixture of acetic

＝ 58.07 mg of C10H14N2O4S2 anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 molWL perchloric acid VS (potentiometric titration). Containers and storage Containers—Well-closed contain- ers. Each mL of 0.1 molWL perchloric acid VS

＝ 18.07 mg of C14H30Cl2N2O4 Suxamethonium Chloride Hydrate Containers and storage Containers—Tight containers. スキサメトニウム塩化物水和物 Suxamethonium Chloride for Injection

注射用スキサメトニウム塩化物

C14H30Cl2N2O4.2H2O: 397.34 2,2?-Succinyldioxybis(N,N,N-trimethylethylaminium) Suxamethonium Chloride for Injection is a prepara- dichloride dihydrate [6101-15-1] tion for injection which is dissolved before use. It con- tains not less than 93z and not more than 107z of the Suxamethonium Chloride Hydrate contains not less labeled amount of suxamethonium chloride than 98.0z of suxamethonium chloride (C14H30Cl2N2 (C14H30Cl2N2O4:361.31). O4: 361.31), calculated on the anhydrous basis. The concentration of Suxamethonium Chloride for Injection should be stated as the amount of sux- Description Suxamethonium Chloride Hydrate occurs as a amethonium chloride (C H Cl N O ). white, crystalline powder. 14 30 2 2 4 It is freely soluble in water, in methanol and in acetic acid Method of preparation Prepare as directed under Injec- (100), slightly soluble in ethanol (95), very slightly soluble in tions, with Suxamethonium Chloride Hydrate. acetic anhydride, and practically insoluble in diethyl ether. Description Suxamethonium Chloride for Injection occurs Identiˆcation (1) Determine the infrared absorption spec- as a white, crystalline powder or mass. trum of Suxamethonium Chloride Hydrate as directed in the Identiˆcation Take an amount of Suxamethonium Chloride potassium bromide disk method under Infrared Spec- for Injection, equivalent to 0.05 g of Suxamethonium Chlo- trophotometry <2.25>, and compare the spectrum with the ride Hydrate according to the labeled amount, dissolve in Reference Spectrum: both spectra exhibit similar intensities water to make 10 mL, and use this solution as the sample so- of absorption at the same wave numbers. lution. Separately, dissolve 0.05 g of suxamethonium chlo- (2) A solution of Suxamethonium Chloride Hydrate (1 in ride for thin-layer chromatography in 10 mL of water, and 20) responds to the Qualitative Tests <1.09> for chloride. use this solution as the standard solution. Perform the test pH <2.54> The pH of a solution of Suxamethonium Chlo- with these solutions as directed under Thin-layer Chro- ride Hydrate (1 in 100) is between 4.0 and 5.0. matography <2.03>.Spot1mL each of the sample solution and standard solution on a plate of cellulose for thin-layer Melting point <2.60> 159 – 1649C (hydrate form). chromatography. Develop the plate with a mixture of a solu- 1138 Suxamethonium Chloride Injection / O‹cial Monographs JP XV tion of ammonium acetate (1 in 100), acetone, 1-butanol and tralization with 0.1 molWL sodium hydroxide VS in the As- formic acid (20:20:20:1) to a distance of about 10 cm, and say: not more than 0.7 mL of 0.1 molWL sodium hydroxide dry the plate at 1059C for 15 minutes. Spray evenly platinic VS is required for each 200 mg of Suxamethonium Chloride chloride-potassium iodide TS on the plate: the spots obtained (C14H30Cl2N2O4)taken. from the sample solution and standard solution are blue-pur- Extractable volume <6.05> It meets the requiremeut. ple in color and have similar Rf. Assay Transfer to a separator an accurately measured pH <2.54> The pH of a solution of Suxamethonium Chlo- volume of Suxamethonium Chloride Injection, equivalent to ride for Injection (1 in 100) is between 4.0 and 5.0. about 0.2 g of suxamethonium chloride (C14H30Cl2N2O4), Purity Related substances—Take an amount of Sux- add 30 mL of freshly boiled and cooled water, and wash the amethonium Chloride for Injection, equivalent to 0.25 g of solution with ˆve 20-mL portions of diethyl ether. Combine Suxamethonium Chloride Hydrate according to the labeled the diethyl ether washings, and extract the combined diethyl amount, and proceed as directed in the Purity (2) under Sux- ether layer with two 10-mL portions of freshly boiled and amethonium Chloride Hydrate. cooled water. Wash the combined water extracts with two 10-mL portions of diethyl ether. Combine the solution and Assay Weigh accurately the contents of not less than 10 the water extracts, add 2 drops of bromothymol blue TS, and preparations of Suxamethonium Chloride for Injection. neutralize with 0.1 molWL sodium hydroxide VS. Add ac- Weigh accurately about 0.5 g of the contents, and proceed as curately measured 25 mL of 0.1 molWL sodium hydroxide directed in the Assay under Suxamethonium Chloride Hy- VS, and boil for 40 minutes under a re‰ux condenser, and drate. cool. Titrate <2.50> the excess sodium hydroxide with 0.1 mol Each mL of 0.1 molWL perchloric acid VS WL hydrochloric acid VS. Transfer 50 mL of the freshly

＝ 18.07 mg of C14H30Cl2N2O4 boiled and cooled water to a ‰ask, add 2 drops of bromothymol blue TS, neutralize the solution with 0.1 molW Containers and storage Containers—Hermetic containers. L sodium hydroxide VS, and perform a blank determination. Each mL of 0.1 molWL sodium hydroxide VS Suxamethonium Chloride Injection ＝ 18.07 mg of C14H30Cl2N2O4 Containers and storage Containers—Hermetic containers. スキサメトニウム塩化物注射液 Storage—Not exceeding 59C, and avoid freezing. Expiration date 12 months after preparation. Suxamethonium Chloride Injection is an aqueous solution for injection. It contains not less than 93z and not more than 107z of the labeled amount of sux- Talampicillin Hydrochloride amethonium chloride (C14H30Cl2N2O4: 361.31). The concentration of Suxamethonium Chloride In- Ampicillinphthalidyl Hydrochloride jection should be stated as the amount of suxamethoni- タランピシリン塩酸塩 um chloride (C14H30Cl2N2O4). Method of preparation Prepare as directed under Injec- tions, with Suxamethonium Chloride Hydrate. Description Suxamethonium Chloride Injection is a clear, colorless liquid. Identiˆcation Take a volume of Suxamethonium Chloride Injection, equivalent to 0.05 g of Suxamethonium Chloride Hydrate according to the labeled amount, add water to make

10 mL, and use this solution as the sample solution. C24H23N3O6S.HCl: 517.98 Separately, dissolve 0.05 g of suxamethonium chloride for 3-Oxo-1,3-dihydroisobenzofuran-1-yl (2S,5R,6R)-6-[(2R)-2- thin-later chromatography in 10 mL of water, and use this amino-2-phenylacetylamino]-3,3-dimethyl-7-oxo-4-thia-1- solution as the standard solution. Perform the test with these azabicyclo[3.2.0]heptane-2-carboxylate solutions as directed under Thin-layer Chromatography monohydrochloride [47747-56-8] <2.03>.Spot1mL each of the sample solution and standard solution on a plate of cellulose for thin-layer chro- Talampicillin Hydrochloride is the hydrochloride of matography. Develop the plate with a mixture of a solution ampicillin phthalidyl ester. of ammonium acetate (1 in 100), acetone, 1-butanol and for- It contains not less than 600 mg (potency) and not mic acid (20:20:20:1) to a distance of about 10 cm, and dry more than 700 mg (potency) per mg, calculated on the the plate at 1059C for 15 minutes. Spray evenly platinic chlo- anhydrous basis. The potency of Talampicillin ride-potassium iodide TS on the plate: the spots obtained Hydrochloride is expressed as mass (potency) of am- from the sample solution and the standard solution are blue- picillin (C16H19N3O4S: 349.40). purple in color and have similar Rf. Description Talampicillin Hydrochloride occurs as a white pH <2.54> 3.0 – 5.0 to light yellowish white powder. It is very soluble in methanol, and freely soluble in water Purity Hydrolysis products—Perform the preliminary neu- JP XV O‹cial Monographs / Talc 1139 and in ethanol (99.5). acid obtained from the sample solution is not more intense than that obtained from the standard solution. Identiˆcation (1) To 1 mL of a solution of Talampicillin Hydrochloride (1 in 30) add 1 mL of sodium hydroxide TS, Water <2.48> Not more than 3.0z (0.5 g, volumetric titra- mix, allow to stand for 5 minutes, and add 2 mL of dilute sul- tion, direct titration). furic acid and 2 to 3 drops of 2,4-dinitrophenylhydrazine TS: Assay Weigh accurately an amount of Talampicillin an orange-yellow precipitate is formed. Hydrochloride and Talampicillin Hydrochloride Reference (2) Determine the infrared absorption spectrum of Standard, equivalent to about 20 mg (potency), dissolve in Talampicillin Hydrochloride as directed in the potassium water to make exactly 20 mL each, and use these solutions as bromide disk method under Infrared Spectrophotometry the sample solution and the standard solution. The standard <2.25>, and compare the spectrum with the Reference Spec- solution should be prepared before use. Pipet 2 mL each of trum or the spectrum of Talampicillin Hydrochloride Refer- the sample solution and the standard solution in separate ence Standard: both spectra exhibit similar intensities of ab- 100-mL glass-stoppered ‰asks, add 2.0 mL of sodium sorption at the same wave numbers. hydroxide TS, and allow them to stand for exactly 15 (3) To 10 mL of a solution of Talampicillin Hydrochlo- minutes.Add2.0mLofdilutedhydrochloricacid(1in10) ride (1 in 300) add 1 mL of dilute nitric acid, and add silver and exactly 10 mL of 0.005 mol/L iodine VS, allow them to nitrate TS: a white precipitate is formed. stand for exactly 15 minutes, and titrate <2.50> with 0.01 20 Optical rotation <2.49> [a]D : ＋151 – ＋1719(0.2 g calcu- mol/L sodium thiosulfate VS until the color of the solution is lated on the anhydrous basis, ethanol (99.5), 20 mL, 100 disappeared. If necessary, add 0.2 to 0.5 mL of starch TS. mm). Separately, pipet 2 mL each of the sample solution and the standard solution in separate 100-mL glass-stoppered ‰asks, Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of add exactly 10 mL of 0.005 mol/L iodine VS, titrate <2.50> Talampicillin Hydrochloride according to Method 2, and with 0.01 mol/L sodium thiosulfate VS until the color of the perform the test. Prepare the control solution with 2.0 mL of solution is disappeared, and make any necessary correction. Standard Lead Solution (not more than 20 ppm). Forthistitration,add0.2to0.5mLofstarchTS,ifnecessa- (2) Arsenic <1.11>—Prepare the test solution with 1.0 g ry. Calculate the amount (mL) of 0.005 mol/L iodine VS, V of Talampicillin Hydrochloride according to Method 4, and T and V , consumed by the sample solution and the standard perform the test (not more than 2 ppm). S solution, respectively. (3) Related substances—Dissolve 50 mg of Talampicillin

Hydrochloride in ethanol (99.5) to make exactly 10 mL, and Amount [ mg (potency)] of ampicillin (C16H19N3O4S) use this solution as the sample solution. Pipet 1 mL, 2 mL ＝WS×(VT/VS)×1000 and 3 mL of the sample solution, add ethanol (99.5) to each W : Amount [mg (potency)] of Talampicillin Hydrochlo- to make exactly 100 mL, and use these solutions as the stan- S ride Reference Standard dard solution (1), the standard solution (2) and the standard solution (3), respectively. Perform the test with these solu- Containers and storage Containers—Tight containers. tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu- tions (1), (2) and (3) on a plate of silica gel for thin-layer Talc chromatography. Develop the plate with a mixture of tetra- hydrofuran, ethyl acetate, water and ethanol (95) (4:4:2:1) to タルク a distance of about 13 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in ethanol (99.5) (1 in 500) on the plate, and heat at 1109C for 5 minutes: the spots other than Talc is a native, hydrous magnesium silicate, some- the principal spot from the sample solution is not more in- times containing a small portion of aluminum silicate. tense than the spot from the standard solution (3), and the Description Talc occurs as a white to grayish white, ˆne, total of the amount of each spot other than the principal spot crystalline powder. It is odorless and tasteless. from the sample solution, which is calculated by the compari- It is unctuous, and adheres readily to the skin. son with the spots obtained from the standard solutions (1), It is practically insoluble in water, in ethanol (95) and in (2) and (3), is not more than 5z. diethyl ether. (4) 2-Formylbenzoic acid—Dissolve 50 mg of Talampicil- lin Hydrochloride in ethanol (99.5) to make exactly 10 mL, Identiˆcation (1) Mix 0.2 g of Talc with 0.9 g of anhy- and use this solution as the sample solution. Separately, dis- drous sodium carbonate and 1.3 g of potassium carbonate, solve 10 mg of 2-formylbenzoic acid in ethanol (99.5) to and heat the mixture in a platinum or nickel crucible until fu- make exactly 100 mL. Pipet 5 mL of this solution, add sion is complete. Cool, and transfer the fused mixture to a ethanol (99.5) to make exactly 10 mL, and use this solution as beaker with the aid of 50 mL of hot water. Add hydrochloric the standard solution. Perform the test with these solutions acid until it ceases to cause eŠervescence, add 10 mL of as directed under Thin-layer Chromatography <2.03>.Spot hydrochloric acid, and evaporate the mixture on a water bath 10 mLeachofthesamplesolutionandstandardsolutionona to dryness. Cool, add 20 mL of water, boil, and ˆlter. Add 10 plate of silica gel for thin-layer chromatography, develop the mL of a solution of methylene blue trihydrate (1 in 10,000) to plate with a mixture of chloroform and acetic acid (100) (4:1) the residue, and wash with water: the precipitate is blue in to a distance of about 13 cm, and air-dry the plate. Spray color. evenly a solution of 2,4-dinitrophenylhydrazine in diluted (2) Dissolve 2 g of ammonium chloride and 5 mL of am- sulfuric acid (6 in 25) (1 in 500): the spot of 2-formylbenzoic monia TS in the ˆltrate obtained in (1), ˆlter if necessary, and 1140 Tamsulosin Hydrochloride / O‹cial Monographs JP XV add disodium hydrogenphosphate TS: a white, crystalline directed under Ultraviolet-visible Spectrophotometry <2.24>, precipitate is produced. and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the Purity (1) Acid-soluble substances—Weigh accurately same wavelengths. about 1 g of Talc, heat with 20 mL of dilute hydrochloric (2) Determine the infrared absorption spectrum of Tam- acid at 509C for 15 minutes with stirring. Cool, add water to sulosin Hydrochloride as directed in the potassium chloride make exactly 50 mL, and ˆlter. Centrifuge, if necessary, until disk method under Infrared Spectrophotometry <2.25>,and theˆltratebecomesclear.To25mLofthisˆltrateadd1mL compare the spectrum with the Reference Spectrum: both of dilute sulfuric acid, evaporate to dryness, and ignite to spectra exhibit similar intensities of absorption at the same constant mass at 800 ± 259C: the amount of the residue is wave numbers. not more than 2.0z. (3) To 5 mL of an ice cooled solution of Tamsulosin (2) Acid or alkali, and water-soluble substances—To 10.0 Hydrochloride (3 in 400) add 3 mL of dilute nitric acid, shake g of Talc, add 50 mL of water, weigh, and boil for 30 well, allow to stand at room temperature for 30 minutes, and minutes, supplying water lost by evaporation. Cool, add ˆlter: the ˆltrate responds to the Qualitative Tests <1.09> for water to restore the original mass, and ˆlter. Centrifuge, if chloride. necessary, until the ˆltrate becomes clear: the ˆltrate is neu- 20 tral. Evaporate 20 mL of the ˆltrate to dryness, and dry the Optical rotation <2.49> [a]D : －17.5 – －20.59 (after residue at 1059C for 1 hour: the mass of the residue is not drying,0.15g,water,warming,aftercooling,20mL,100 more than 4.0 mg. mm). (3) Water-soluble iron—Make 10 mL of the ˆltrate ob- Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of tained in (2) weakly acidic with hydrochloric acid, and add Tamsulosin Hydrochloride according to Method 4, and per- dropwise potassium hexacyanoferrate (II) TS: the liquid does form the test. Prepare the control solution with 2.0 mL of not acquire a blue color. Standard Lead Solution (not more than 20 ppm). (4) Arsenic <1.11>—To0.5gofTalcadd5mLofdilute (2) Related substances— sulfuric acid, and heat gently to boiling with shaking. Cool (i) Dissolve 50 mg of Tamsulosin Hydrochloride in 10 immediately, ˆlter, and wash the residue with 5 mL of dilute mL of the mobile phase, and use this solution as the sample sulfuric acid, then with 10 mL of water. Combine the ˆltrate solution. Pipet 2 mL of the sample solution, and add the mo- and the washings, evaporate to 5 mL on a water bath, and bile phase to make exactly 50 mL. Pipet 2.5 mL of this solu- perform the test with this solution as the test solution (not tion, add the mobile phase to make exactly 100 mL, and use more than 4 ppm). this solution as the standard solution. Perform the test with Loss on ignition <2.43> Not more than 5.0z (1 g, 450 – 10 mLeachofthesamplesolutionandstandardsolutionas 5509C, 3 hours). directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by Containers and storage Containers—Well-closed contain- the automatic integration method: the area of the peak other ers. than tamsulosin obtained from the sample solution is not larger than the peak area of tamsulosin from the standard so- lution. Tamsulosin Hydrochloride Operating conditions— Detector: An ultraviolet absorption photometer タムスロシン塩酸塩 (wavelength: 225 nm). Column: A stainless steel column 4 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 4.4 mL of perchloric acid and 1.5 g C20H28N2O5S.HCl: 444.97 5-｛(2R)-2-[2-(2-Ethoxyphenoxy)ethylamino]propyl｝- of sodium hydroxide in 950 mL of water, adjust the pH to 2.0 2-methoxybenzenesulfonamide monohydrochloride with sodium hydroxide TS, and add water to make 1000 mL. [106463-17-6] To 700 mL of this solution add 300 mL of acetonitrile for liq- uid chromatography. Tamsulosin Hydrochloride, when dried, contains Flow rate: Adjust the ‰ow rate so that the retention time of not less than 98.5z and not more than 101.0z of tamsulosin is about 6 minutes. Time span of measurement: Until tamsulosin is eluted, be- C20H28N2O5S.HCl. ginning after the solvent peak. Description Tamsulosin Hydrochloride occurs as white System suitability— crystals. Test for required detectability: Measure exactly 1 mL of It is freely soluble in formic acid, sparingly soluble in the standard solution, and add the mobile phase to make ex- water, slightly soluble in acetic acid (100), and very slightly actly 50 mL. Conˆrm that the peak area of tamsulosin ob- soluble in ethanol (99.5). tained from 10 mL of this solution is equivalent to 1.4 to Melting point: about 2309C (with decomposition). 2.6z of that from 10 mL of the standard solution. Identiˆcation (1) Determine the absorption spectrum of a System performance: Dissolve 5 mg of Tamsulosin solution of Tamsulosin Hydrochloride (3 in 160,000) as Hydrochloride and 10 mg of propyl parahydroxybenzoate in 20 mL of the mobile phase. To 2 mL of this solution add the JP XV O‹cial Monographs / Tartaric Acid 1141 mobilephasetomake20mL.Whentheprocedureisrunwith 10 mL of this solution under the above operating conditions, tamsulosin and propyl parahydroxybenzoate are eluted in Tannic Acid this order with the resoluton between these peaks being not less than 12. タンニン酸 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 Tannic Acid is the tannin usually obtained from nut- tamsulosin is not more than 4.0z. galls or rhusgalls. (ii) Perform the test with 10 mL each of the sample solu- Description Tannic Acid occurs as a yellowish white to light tionandstandardsolutionwhichareobtainedinabove(i)as brown, amorphous powder, glistening lea‰ets, or spongy directed under Liquid Chromatography <2.01> according to masses. It is odorless or has a faint, characteristic odor, and the following conditions, and determine each peak area by has a strongly astringent taste. the automatic integration method: the area of the peak other It is very soluble in water and in ethanol (95), and practi- than tamsulosin obtained from the sample solution is not cally insoluble in diethyl ether. larger than the peak area of tamsulosin from the standard so- lution. Identiˆcation (1) To 5 mL of a solution of Tannic Acid (1 Operating conditions— in 400) add 2 drops of iron (III) chloride TS: a blue-black Detector, column, and column temperature: Proceed as color develops. Allow the solution to stand: a blue-black directed in the operating conditions in the Purity (2) (i). precipitate is produced. Mobile phase: Dissolve 4.4 mL of perchloric acid and 1.5 g (2) To 5 mL of a solution of Tannic Acid (1 in 20) add 1 of sodium hydroxide in 950 mL of water, adjust the pH to 2.0 drop each of albumin TS, gelatin TS, or 1 mL of starch TS: a with sodium hydroxide TS, and add water to make 1000 mL. precipitate is produced in each solution. To this solution add 1000 mL of acetonitrile for liquid chro- Purity (1) Gum, dextrin and sucrose—Dissolve 3.0 g of matography. Tannic Acid in 15 mL of boiling water: the solution is clear or Flow rate: Adjust the ‰ow rate so that the retention time of slightly turbid. Cool, and ˆlter the solution. To 5 mL of the tamsulosin is about 2.5 minutes. ˆltrate add 5 mL of ethanol (95): no turbidity is produced. Time span of measurement: About 5 times as long as the Add further 3 mL of diethyl ether to this solution: no turbidi- retention time of tamsulosin, beginning after the peak of ty is produced. tamsulosin. (2) Resinous substances—To 5 mL of the ˆltrate ob- System suitability— tained in (1) add 10 mL of water: no turbidity is produced. Test for required detectability: Measure exactly 1 mL of the standard solution, and add the mobile phase used in the Loss on drying <2.41> Not more than 12.0z (1 g, 1059C, Purity (2) (i) to make exactly 50 mL. Conˆrm that the peak 2 hours). area of tamsulosin obtained from 10 mLofthissolutionis Residue on ignition <2.44> Not more than 1.0z (0.5 g). equivalent to 1.4 to 2.6z of that from 10 mL of the standard solution. Containers and storage Containers—Tight containers. System performance: Proceed as directed in the system Storage—Light-resistant. suitability in the Purity (2) (i). System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operating Tartaric Acid conditions, the relative standard deviation of the peak area of tamsulosin is not more than 4.0z. 酒石酸 Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.7 g of Tamsulosin C H O : 150.09 Hydrochloride, previously dried, dissolve in 5 mL of formic 4 6 6 (2R,3R )-2,3-Dihydroxybutanedioic acid [87-69-4 ] acid, add 75 mL of a mixture of acetic acid (100) and acetic anhydride (3:2), and immediately titrate <2.50> with 0.1 Tartaric Acid, when dried, contains not less than mol/L perchloric acid VS (potentiometric titration). Perform 99.7z of C H O . a blank determination in the same manner, and make any 4 6 6 necessary correction. Description Tartaric Acid occurs as colorless crystals or a white, crystalline powder. It is odorless, and has a strong acid Each mL of 0.1 mol/L perchloric acid VS taste. ＝44.50 mg of C H N O S.HCl 20 28 2 5 It is very soluble in water, freely soluble in ethanol (95), Containers and storage Containers—Well-closed contain- and slightly soluble in diethyl ether. ers. A solution of Tartaric Acid (1 in 10) is dextrorotatory. Identiˆcation (1) Ignite Tartaric Acid gradually: it decomposes and an odor of burning sugar is perceptible. (2) AsolutionofTartaricAcid(1in10)changesbluelit- 1142 / O‹cial Monographs JP XV mus paper to red, and responds to the Qualitative Tests (2) Chloride<1.03>—Perform the test with 1.0 g of Tau- <1.09> for tartrate. rine. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.011z). Purity (1) Sulfate <1.14>—Perform the test with 0.5 g of (3) Sulfate<1.14>—Perform the test with 2.0 g of Tau- Tartaric Acid. Prepare the control solution with 0.50 mL of rine. Prepare the control solution with 0.40 mL of 0.005 0.005 molWL sulfuric acid VS (not more than 0.048z). mol/L sulfuric acid VS (not more than 0.010z). (2) Oxalate—Dissolve 1.0 g of Tartaric Acid in 10 mL of (4) Ammonium<1.02>—Perform the test with 0.25 g of water, and add 2 mL of calcium chloride TS: no turbidity is Taurine. Prepare the control solution with 5.0 mL of Stan- produced. dard Ammonium Solution (not more than 0.02z). (3) Heavy metals <1.07>—Proceed with 2.0 g of Tartaric (5) Heavy metals <1.07>—Proceed with 2.0 g of Taurine Acid according to Method 4, and perform the test. Prepare according to Method 1, and perform the test. Prepare the the control solution with 2.0 mL of Standard Lead Solution control solution with 2.0 mL of Standard Lead Solution (not (not more than 10 ppm). more than 10 ppm). (4) Calcium—Neutralize a solution of 1.0 g of Tartaric (6) Iron<1.10>—Prepare the test solution with 2.0 g of Acid in 10 mL of water with ammonia TS, and add 1 mL of Taurine according to Method 1, and perform the test accord- ammonium oxalate TS: no turbidity is produced. ing to Method A. Prepare the control solution with 2.0 mL of (5) Arsenic <1.11>—Prepare the test solution with 2.0 g Standard Iron Solution (not more than 10 ppm). of Tartaric Acid according to Method 1, and perform the test (7) Related substances—Dissolve 1.0 g of Taurine in 50 (not more than 1 ppm). mL of water, and use this solution as the sample solution. Loss on drying <2.41> Not more than 0.5z (3 g, silica gel, Pipet 1 mL of the sample solution, and add water to make ex- 3 hours). actly 50 mL. Pipet 1 mL of this solution, add water to make exactly 10 mL, and use this solution as the standard solution. Residue on ignition <2.44> Not more than 0.05z (1 g). Perform the test with these solutions as directed under Thin- Assay Weigh accurately about 1.5 g of Tartaric Acid, previ- layer Chromatography <2.03>.Spot5mLeachofthesample ously dried, dissolve in 40 mL of water, and titrate <2.50> solution and standard solution on a plate of silica gel for with 1 molWL sodium hydroxide VS (indicator: 2 drops of thin-layer chromatography. Develop the plate with a mixture phenolphthalein TS). of water, ethanol (99.5), 1-butanol and acetic acid (100) (150:150:100:1) to a distance of about 10 cm, and air-dry the Each mL of 1 molWL sodium hydroxide VS plate. Spray evenly ninhydrin-butanol TS on the plate, and ＝75.04 mg of C H O 4 6 6 heat at 1059C for 5 minutes: the spot other than the principle Containers and storage Containers—Well-closed contain- spot with the sample solution is not more than one spot, and ers. it is not more intense than the spot with the standard solu- tion. Loss on drying <2.41> Not more than 0.20z (1 g, 1059C, 2 Taurine hours). タウリン Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.2 g of Taurine, previously dried, dissolve in 50 mL of water, add 5 mL of formaldehyde solution, and titrate <2.50> with 0.1 mol/L sodium hydroxide C2H7NO3S: 125.15 VS (potentiometric titration). Perform a blank determination 2-Aminoethanesulfonic acid in the same manner, and make any necessary correction. [107-35-7] Each mL of 0.1 mol/L sodium hydroxide VS Taurine, when dried, contains not less than 99.0z ＝12.52 mg of C2H7NO3S and not more than 101.0z of C2H7NO3S. Containers and storage Containers—Well-closed contain- Description Taurine occurs as colorless or white crystals, or ers. a white crystalline powder. It is soluble in water, and practically insoluble in ethanol (99.5). The pH of a solution prepared by dissolving 1.0 g of Tau- rine in 20 mL of freshly boiled and cooled water is between 4.1 and 5.6. Identiˆcation Determine the infrared absorption spectrum of Taurine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>,andcomparethe spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Purity (1) Clarity and color of solution—A solution ob- tained by dissolving 1.0 g of Taurine in 20 mL of water is clear and colorless. JP XV O‹cial Monographs / Teceleukin (Genetical Recombination) 1143

and use one as the sample (1). To the other, add 50 mLofa mixture of formic acid and hydrogen peroxide (30) (9:1) that Teceleukin (Genetical has been left at room temperature for one hour, cool for 4 hours in ice, add 0.5 mL of water, and then evaporate to dry- Recombination) ness under vacuum to give the sample (2). To 1.3 mL of methanesulfonic acid add 3.7 mL of water, mix well, add and テセロイキン（遺伝子組換え） dissolve 10 mg of 3-(2-aminoethyl)indole, to make a 4 mol/L methanesulfonic acid solution. Dissolve 39.2 g of trisodium citrate dihydrate, 33 mL of hydrochloric acid, 40 mL of thio- diglycol, and 4 mL of lauromacrogol solution (1 in 4) in 700 mL of water, adjust the pH to 2.2, add water to 1000 mL, add 100 mL of capric acid, and mix to make a sodium citrate solution for dilution. Add 50 mL of freshly prepared 4 mol/L methanesulfonic acid to the sample (1) and sample (2), cool to －709C, and then deaerate under vacuum. Heat to 1159C ±29C for 24 hours after sealing these test tubes under reduced pressure. After cooling, unseal, add 50 mLof4 C H N O 0 S : 15547.01 698 1127 179 2 4 8 mol/L sodium hydroxide TS followed by 0.4 mL of sodium [136279-32-8] citrate solution for dilution to make the sample solution (1) and sample solution (2). Separately, accurately measure 0.25 The desired product of Teceleukin (Genetical mmol amounts of L-, L-threonine, L-, L- Recombination) is a protein consisting of 134 amino , L-, glycine, L-, L-valine, L- acid residues manufactured by E. coli through expres- , L-isoleucine, L-leucine, L-tyrosine, L-phenylala- sion of human -2 cDNA. It is a solution and nine, L-lysine hydrochloride, ammonium chloride, L-histi- possesses a T-lymphocyte activating eŠect. dine hydrochloride monohydrate, and L-arginine hydrochlo- It contains potency between 7.7×106 and 1.54×107 ride as well as 0.125 mmol of L- and then dissolve in units/mL, and not less than 7.7×106 units per mg of 0.1 mol/L hydrochloric acid TS to make exactly 100 mL. protein. This is the amino acid standard stock solution. Accurately Description Teceleukin (Genetical Recombination) occurs measure 1 mL of this solution, and add sodium citrate solu- as a clear and colorless liquid. tion for dilution to make exactly 25 mL. This is solution A. Accurately weigh approximately 20 mg of L-tryptophan and Identiˆcation (1) Measure accurately an appropriate dissolve in water to make exactly 1000 mL. This is solution B. amount of Teceleukin (Genetical Recombination), add ac- Accurately measure 10 mL of both solution A and solution curately to a concentration of 200 units per mL of culture B, combine together, and add sodium citrate solution for di- medium for assay of teceleukin, and use this solution as the lution to make exactly 50 mL. This is the amino acid standard sample stock solution. Dilute reference anti-interleukin-2 an- solution. Separately, accurately weigh approximately 17 mg tibody for teceleukin with culture medium for assay of of L-cysteic acid and dissolve in sodium citrate solution for teceleukin to a concentration of approximately 200 neutral dilution to make exactly 50 mL. Accurately measure 1 mL of units/mL and use this solution as the interleukin-2 neutral this solution and add sodium citrate solution for dilution to antibody solution. Accurately add an equivalent volume of make exactly 100 mL. This is the cysteic acid standard solu- the interleukin-2 neutral antibody solution to the sample tion. Accurately measure 0.25 mL of the sample solution (1), stock solution, shake, and then leave for 1 hour in a 379Cin- the sample solution (2), amino acid standard solution, and cubator in air containing 5z carbon dioxide. This solution is the cysteic acid standard solution. When the test is conducted the sample solution. Prepare a standard solution by accurate- by Liquid Chromatography <2.01> under the following condi- ly adding an equivalent volume of culture medium for assay tions, peaks for the 18 amino acids are observed in the chro- of teceleukin to the sample stock solution, mixing, and then matogram obtained from the sample solution (1). Also, processing in the same way. Process the sample and standard measure the peak area of each amino acid in the sample solu- solutions according to the assay method, determine their tion (1) and the amino acid standard solution, and taking the respective dilution coe‹cients, D and D , and then deter- N T molar number of alanine in the sample solution (1) as 5.0, de- mine the neutralization rate, which should be at least 90z, termine the concentrations of aspartic acid, glutamic acid, using the following formula. proline, glycine, methionine, leucine, tyrosine, phenylala-

Neutralization rate (z)＝｛(DT－DN)/DT｝×100 nine, lysine, histidine, tryptophan, and arginine and then cal- culate the molar ratio for each amino acid. Also, measure the However, please note if the mean values of the absorbance of cysteic acid peak areas of the sample solution (2) and the cys- the maximum uptake control solution and absorbance of the teic acid standard solution, determine the concentration of minimum uptake control solution do not ˆt the standard cysteine, and, taking the molar number of alanine in the sam- curve, the neutralization coe‹cient is to be determined within ple solution (2) as 5.0, calculate the molar ratio of cysteine. the following range. When determining the molar ratios of the respective amino

Neutralization coe‹cient (z)À｛(DT－2)/DT｝×100 acids, aspartic acid is 11.4 to 12.6, glutamic acid 17.1 to 18.9, proline 4.5 to 5.5, glycine 1.8 to 2.2, cysteine 2.7 to 3.3, (2) Place a volume of Teceleukin (Genetical Recombina- methionine 4.5 to 5.5, leucine 20.9 to 23.1, tyrosine 2.7 to tion) corresponding to approximately 50 mg of protein into 2 3.3, 5.4 to 6.6, lysine 10.5 to 11.6, histidine 2.7 test tubes for hydrolysis, evaporate to dryness under vacuum, to 3.3, tryptophan 0.7 to 1.2, and arginine 3.6 to 4.4. 1144 Teceleukin (Genetical Recombination) / O‹cial Monographs JP XV

Operating conditions— (3) Dissolve 0.242 g of 2-amino-2-hydroxymethyl-1,3- Detector: Visible absorption photometer [wavelengths: 440 propanediol, 5.0 g of sodium lauryl sulfate, and 74 mg of dis- nm (proline) and 570 nm (amino acids other than proline)] odium dihydrogen ethylenediamine tetraacetate dihydrate in Column: A stainless steel column with an inside diameter 60 mL of water. After adjusting the pH to 8.0 using 1 mol/L of 4 mm and length of 25 cm packed with a strongly acidic hydrochloric acid TS, add water to make 100 mL. This is the ion exchange resin for liquid chromatography consisting of molecular weight determination buŠer solution. Accurately polystyrene to which sulphonate group binds. measure 20 mL of Teceleukin (Genetical Recombination), add Column temperature: A constant temperature of about exactly 20 mL of the molecular weight determination buŠer 509C when the sample is injected. After a certain time, in- solution and 2 mL of 2-mercaptoethanol, and then heat for 5 crease the temperature to a constant temperature of about minutes on a 90 to 1009C water bath without allowing any 629C water evaporation from the mixture. After cooling, add ex- Reaction temperature: A constant temperature of about actly 1 mL of bromophenol blue solution (1 in 2000) and then 989C. shake. This is the sample solution. Separately, accurately Time for color formation: Approximately 2 minutes measure 5 mL of molecular weight marker for teceleukin, and Mobile phase: After preparing mobile phases A, B, and C add exactly 50 mLofwater,55mLofthemolecularweightde- according to the following table, add 0.1 mL of capric acid to termination buŠer solution, and 5 mL of 2-mercaptoethanol, each. and then heat for 5 minutes on a 90 to 1009Cwaterbath without allowing any water evaporation from the mixture. Mobile phase Mobile phase Mobile phase After cooling, add exactly 1 mL of bromophenol blue solu- A B C tion (1 in 2000), and shake well. This is the molecular weight Citric acid standard solution. When conducting a test using SDS- 18.70 g 10.50 g 7.10 g monohydrate polyacrylamide gel electrophoresis with 1 mLeachofthesam- Trisodium citrate 7.74 g 14.71 g 26.67 g ple solution and the molecular weight standard solution, the dihydrate Sodium chloride 7.07 g 2.92 g 54.35 g molecular weight of the main band is between the range of Ethanol (99.5) 60 mL — — 14,000 and 16,000. Benzyl alcohol — — 10 mL Operating conditions— Thiodiglycol 5 mL 5 mL — Equipment: Horizontal electrophoresis vessel equipped Lauromacrogol 4mL 4mL 4mL with a cooling unit, a device that accumulates load voltage solution (1 in 4) Water Appropriate Appropriate Appropriate over time, and a direct current power source device that con- amount amount amount trols the amperage, voltage, wattage. pH 3.2 4.3 4.7 Spotting of solutions: Solutions are spotted on concentrat- Total volume 1000 mL 1000 mL 1000 mL ing gel of polyacrylamide gel sheets. Electrophoresis conditions Changing mobile phases and column temperature: When Polyacrylamide gel sheet: Polyester sheet to which a operating under the above conditions using 0.25 mL of ami- polyacrylamide gel (width, about 43 mm, length, about 50 no acid standard solution, the amino acids will elute in the mm, and thickness, about 0.5 mm) is closely adhered. The following order; aspartic acid, threonine, serine, glutamic polyacrylamide gel consists of a concentrating gel with a acid, proline, glycine, alanine, cystine, valine, methionine, gel support concentration of 7.5z and a 3z degree of isoleucine, leucine, tyrosine, phenylalanine, lysine, ammo- crosslinking and a separating gel with corresponding values nia, histidine, tryptophan, and arginine. Switchover to mo- of 20z and 2z. The gel contains pH 6.5 Tris-acetate bile phase A, mobile phase B, and mobile phase C, in se- buŠer. quence so that the resolution between the peaks of cystine BuŠer solution for electrode: Prepared by dissolving and valine is 2.0 or more and that between ammonia and 35.83 g of tricine, 24.23 g of 2-amino-2-hydroxymethyl- histidine is 1.5 or more. Also, increase the temperature after 1,3-propanediol, and 5.5 g of sodium lauryl sulfate in a constant length of time so that the resolution between the water to make 1000 mL. peaks of glutamic acid and proline is at least 2.0. Cooling temperature of gel support plate: 159C Reaction reagents: Dissolve 408 g of lithium acetate dihy- Running conditions drate in water, and add 100 mL of acetic acid (100) and water Pre-electrophoresis and electrophoresis: The voltage, to make 1000 mL. To this solution add 1200 mL of dimethyl- amperage, and wattage should not exceed 250 V, 10 mA, sulfoxide and 800 mL of 2-methoxyethanol. This is solution and 3 W, respectively. The amperage and wattage should (I). Separately, mix together 600 mL of dimethylsulfoxide be proportional to the number of polyacrylamide sheets. and 400 mL of 2-methoxyethanol and then add 80 g of nin- Immediately after adding sample: The voltage, am- hydrin and 0.15 g of sodium borohydride. This is solution perage, and wattage should not exceed 250 V, 1 mA, and 3 (II). After gassing 3000 mL of the solution (I) for 20 minutes W, respectively. The amperage and wattage should be with nitrogen, rapidly add 1000 mL of the solution (II) and proportional to the number of polyacrylamide sheets. then mix by gassing for 10 minutes with nitrogen. Electrophoresis time Mobile phase ‰ow rate: About 0.275 mL every minute Before adding sample: Until value of load voltage in- Reaction reagent ‰ow rate: About 0.3 mL every minute tegrated with respect to time reaches 60 V･h. System suitability— Immediately after adding sample: Until value of load System performance: When 0.25 mL of the amino acid voltage integrated with respect to time reaches 1 V･h. standard solution is run under the above conditions, the reso- Main electrophoresis: Until value of load voltage in- lution between the peaks of threonine and serine is at least tegrated with respect to time reaches 140 V･h. 1.5. Fixation and JP XV O‹cial Monographs / Teceleukin (Genetical Recombination) 1145

Dissolve 25 g of anhydrous sodium carbonate anhydride Dissolve 28.75 g of trichloroacetic acid and 8.65 g of 5- and 0.8 mL of formaldehyde solution in water to make sulfosalicylic acid dihydrate in 75 mL of methanol and 175 1000 mL. This is the developing solution. After immersing mL of water. Immerse the gel in this solution for 60 the polyacrylamide gel sheet in a mixture of ethanol (99.5), minutes to ˆx the protein to the gel. After ˆxation, im- water and acetic acid (100) (5:4:1) for 2 minutes, immerse merse for 10 minutes in a mixture of water, ethanol (99.5) for 2 minutes in a mixture of water, ethanol (99.5) and a- and acetic acid (100) (67:25:8). cetic acid (100) (17:2:1). Change the mixture, immerse for Staining and decolorization another 4 minutes, immerse in water for 2 minutes to rinse Dissolve 0.11 g of Coomassie brilliant blue G-250 in 25 the polyacrylamide gel sheet, and change the water to im- mL of ethanol (99.5), and add 8 mL of acetic acid (100) merse for 2 minutes. This procedure is carried out with and water to make 100 mL. This is the staining solution. heating to 509C. Next, while heating at 409C, immerse for Immerse the gel for 10 minutes while heating at 609Cin 10 to 15 minutes in diluted silver nitrate TS (1 in 7), warm freshly ˆltered staining solution. After staining, decolorize to 309C, and gently rinse the polyacrylamide gel sheet with by immersing in a mixture of water, ethanol (99.5) and water. While warming at 309C, immerse the poly- acetic acid (100) (67:25:8). acrylamide gel sheet in freshly prepared developing solu- Determination of isoelectric point tion. After obtaining adequate color formation, immerse Plot the protein isoelectric points and the distance from the polyacrylamide gel sheet in diluted acetic acid (100) (1 the cathode of each band obtained from the isoelectric in 20) to terminate the color formation. markers for teceleukin. Determine the isoelectric point Estimation of molecular weight from the corresponding position of the major bands ob- Plot graphs for each band obtained from the molecular tained from the sample solution. weight standard solution, distance from the border of the pH <2.54> 2.7 to 3.5 concentrating gel and separating gel, and the logarithm of the molecular weight of proteins in each band. Determine Purity (1) Host cell-derived protein—Take an appropriate the molecular mass by reading the corresponding position amount of Teceleukin (Genetical Recombination) and add an of the major band obtained from the sample solution on exact amount of diluted acetic acid (100) (1 in 350) to make a the graph. solution containing between 0.68 and 0.72 mg of protein in (4) The isoelectric point determined from the electropho- one mL. This is the sample stock solution. Dissolve 1.52 g of resis position is 7.4 to 7.9 when 3 mL of Teceleukin (Genetical 2-amino-2-hydroxymethyl-1,3-propanediol hydrochloride Recombination) and 8 mL of isoelectric marker for teceleukin and 10.94 g of 2-amino-2-hydroxymethyl-1,3-propanediol in are tested by the polyacrylamide gel isoelectric method. water to make 200 mL. Dissolve 0.5 g of bovine serum albu- Operating conditions— min in 25 mL of this solution. This is 2 w/vz bovine serum Equipment: Horizontal electrophoretic vessel with cooling albumin-Tris-hydrochloride buŠer solution. Accurately unit and direct current power source that can perform con- measure 0.5 mL of the sample stock solution, add exactly 30 stant wattage control. mL of sodium carbonate TS, stir, and immediately add ex- Preparation of polyacrylamide gel: Dissolve 1.62 g of poly- actly 0.47 mL of the 2 w/vz bovine serum albumin-Tris- acrylamide and 50 mg of N,N?-methylenebisacrylamide in hydrochloride buŠer solution to make the sample solution. water to make 25 mL. Accurately measure 7.5 mL of this so- Accurately measure 10 mL of dilute acetic acid (100) (1 in lution, 2 mL of a 10 mL solution prepared by adding water to 350), add 0.6 mL of sodium carbonate TS, and then add 2 5 g of glycerin, and 0.64 mL of a pH 3 to pH 10 amphoteric w/vz bovine serum albumin-Tris-hydrochloride buŠer solu- electrolyte solution, and degas under reduced pressure while tion to make exactly 20 mL. This is the dilution solution. stirring thoroughly. Next, accurately measure 74 mL of fresh- Add the E. coli protein stock solution to this dilution solution ly prepared ammonium peroxodisulfate solution (1 in 50), 3 to make a solution containing 0.015 mg of E. coli protein in mLofN,N,N?,N?-tetramethylethylenediamine, and 50 mLof one mL. This is standard solution (1). Accurately dilute this freshly prepared ribo‰avin sodium phosphate solution (1 in solution serially two-fold with the dilution solution to make 1000), stir well, immediately pour on a gel preparation plate standard solutions (2) to (8) having diŠerent concentrations (10 cm wide, 11 cm long, and 0.8 mm thick), and then expose of E. coli protein. Dissolve 0.5 g of bovine serum albumin in to a ‰uorescent light source for 60 minutes to gelate. 100 mL of 0.01 mol/L phosphate buŠer-sodium chloride TS, Spotting pH 7.4. This is the wash solution. Accurately measure 0.1 mL Add Teceleukin (Genetical Recombination) or isoelectric of the sample solution, standard solutions (1) to (8), and dilu- marker for teceleukin 30 minutes after starting electropho- tion solution as a blank standard solution and place each in 3 resis to wells in gel plates to which plastic tape (3.5 mm wells in solid phase plates (place dilution solution in 6 wells), wide, 3.5 mm long, 0.4 mm thick) has been applied in ad- cover with plastic wrap, mix by shaking in a horizontal direc- vance and that have undergone gelation. tion, and then leave standing at a constant temperature of Electrophoresis conditions about 259C for 5 to 16 hours. Next, remove the solution from Cathode solution: Sodium hydroxide TS each well by aspiration, add 0.25 mL of the the wash solu- Anode solution: DL-aspartic acid solution (133 in 25,000) tion, mix again by shaking in a horizontal direction, and then Cooling temperature of gel support plate: 2±19C remove by aspiration. Repeat this procedure 2 more times by Running conditions: After starting the electrophoresis, a adding 0.25 mL of the wash solution to each well. Freshly di- constant wattage of 10 W for 20 minutes and 20 W there- lute peroxidase marker antibody stock solution with 1 w/vz after. However, the voltage should be 3000 V or less. bovine serum albumin-phosphate buŠer-sodium chloride TS, Running time: 120 to 140 minutes while blowing Nitrogen add exactly 0.1 mL to each well, cover with plastic wrap, mix into the electrophoresis vessel. by shaking in a horizontal direction, and then leave standing Fixation and washing at a constant temperature of about 259C for 16 to 24 hours. 1146 Teceleukin (Genetical Recombination) / O‹cial Monographs JP XV

Next, remove the solution in the wells by aspiration, add 0.25 propriate amount of Hydrochloride Reference mL of the wash solution, mix by shaking in a horizontal Standard and dilute accurately with water to make a clear so- direction, and then remove the solution by aspiration. Using lution with a concentration of 1 mg (potency)/mL. Accurate- 0.25 mL of the wash solution, repeat this procedure 2 more ly measure an appropriate amount of this solution and dilute times for each well. To each well, accurately add 0.1 mL of precisely with water to make standard solutions with concen- teceleukin chromophore solution, stir gently, and then shield trations of 4, 2, 1 and 0.5 mg (potency)/mL. Separately, di- from light and leave standing for 30 minutes at a constant lute Teceleukin (Genetical Recombination) with diluted acet- temperature of about 259C. Add exactly 0.1 mL of diluted ic acid (100) (3 in 1000) if needed, or alternatively concentrate sulfuric acid (3 in 50) to each well and then mix by gently under reduced pressure, to make a sample solution with a shaking horizontally. Perform the test with these solutions as protein concentration of 0.8 to 1.2 mg/mL. Accurately meas- directed under Ultraviolet-visible spectrophotometry <2.24>, ure 25 mL of the sample solution and each standard solution, and measure the absorbances AT2 and AS2 at 450 nm and AT1 and add each to the wells in the same test plate. Repeat the and AS1 at 510 nm. Prepare a standard curve by plotting the same procedure for at least 3 more test plates. Leave the test valuesobtainedfromeachstandardsolution(AS2 – AS1)ona plates at room temperature for 30 to 60 minutes and then in- graph having the concentration of E. coli protein (ng/mL) in cubate for 16 to 18 hours at 35 to 379C. Measure the inhibi- logarithmic scale on the horizontal axis and the absorbance tions zones to a diameter of 0.25 mm. Determine the mean values on the vertical axis. Match the values obtained from among the test plates for each of the solutions. the sample solution (AT2 – AT1) to the standard curve, deter- Prepare a standard curve by plotting a graph with the con- mine the concentration A of E. coli protein in the sample so- centrationofeachstandardsolutioninlogarithmicscaleon lution, and take the mean. The amount of E. coli protein is the horizontal axis and the diameter of the inhibition zone on not more than 5 ng when the amount of E. coli protein per the vertical axis. Match the diameter of the inhibition zone of mg of protein is determined using the following formula. teceleukin from the standard curve and determine A,thecon- centration of tetracycline hydrochloride. When the amount Amount (ng) of E. coli-derived protein per mg protein of tetracycline hydrochloride per mg of protein is determined ＝A/C by the following formula, the amount is not more than 0.7 C: Protein concentration (mg/mL) in sample solution mg. However, if an inhibition zone is not seen, or is seen but the diameter is smaller than 0.5 mg/mL on the standard The test is valid if the E. coli protein concentration is 0.3 curve, A is taken as being 0.5 mg/mL or less. ng/mL or less when the concentration is obtained by ˆtting the absorbance value at detection limit, calculated from the Amount [mg (potency)] of tetracycline hydrochloride following formula using absorvance value of the dilution so- (C22H24N2O8.HCl) per mg of protein＝A/P lution, to the standard curve. P: The protein concentration (mg/mL) of the sample solu- Absorbance at detection limit tion. 6 (3) Desmethionyl form—Add water to an appropriate ＝X˜ ＋3.3× (Xi－X˜ )2 /(6－1) «S $ amount of teceleukin to make a sample solution with a pro- i＝1 tein concentration of about 0.17 mg/mL. Perform the test Xi: Individual absorbance values obtained from the dilu- with 1.2 mL of this solution as directd under Liquid Chro- tion solution matography <2.01> under the following conditions. Deter- X˜ : The mean of absorbance values obtained from the dilu- mine using automatic integration the peak area, A ,of tion solution 2 teceleukin and the peak area of the desmethionyl form with a 6: The number of wells in the microplate containing dilu- relative retention time of about 0.8 relative to teceleukin, A . tion solution 1 The content of the desmethionyl form is not more than 1.0z (2) Tetracycline hydrochloride—Serially subculture when determined using the following formula. through 2 passages at 35 to 379C the test Micrococ- Amount (z) of desmethionyl form＝｛A1/(A1＋A2)｝×100 cus luteus ATCC9341 in a slant culture of test bacteria inocu- lation media for teceleukin and then dilute this 100-fold by Operating conditions— adding sterilized puriˆed water. This is the test bacteria solu- Detector: Ultraviolet absorption photometer (wavelength: tion. Store the test bacteria solution at 59C or less and use the 280 nm) solution within 5 days. Dilute the test bacteria solution serial- Columns: Two stainless steel columns with inside di- ly by adding sterilized puriˆed water, add an appropriate ameters of 7.5 mm and lengths of 7.5 cm connected in se- amount to 100 mL of normal agar medium for teceleukin, quenceandpackedwith10mm synthetic polymer bound to conduct a preliminary test, and determine the amount of diethylaminoethyl base for liquid chromatography. tetracycline hydrochloride that shows an inhibition zone cor- Column temperature: A constant temperature of about responding to standard solution containing 0.5 mg (potency) 259C. in 1 mL. Add this amount to 100 mL of normal agar medium Mobile phase A: Mix 0.658 g of diethanolamine in 400 mL for teceleukin dissolved and then cooled to 45 to 509Cand of water, adjust the pH to 9.0 by adding 1 mol/L hydrochlor- mix. Pipet 25 mL of this solution into square Petri dishes ic acid TS, and then add water to make 500 mL. (135×95 mm) and spread horizontally to solidify. Prepare Mobile phase B: Add 300 mL of water to 2.6 mL of a pH 6 plates for testing by making an appropriate number of wells to 9 amphoteric electrolyte solution and 0.5 mL of a pH 8 to in this agar medium. The volume of the test bacteria solution 10.5 amphoteric electrolyte solution, adjust to pH 7 with to which 100 mL of normal agar medium for teceleukin has dilutedhydrochloricacid(9in100),andthenaddwaterto been added is 0.25 to 1.0 mL. Accurately measure an ap- make 400 mL. JP XV O‹cial Monographs / Teceleukin (Genetical Recombination) 1147

Switching mobile phases and sample injection: Inject the (5) Other related proteins—Perform the test on 5 mLof sample solution while running the mobile phase A. Repeated- Teceleukin (Genetical Recombination) as directed under Liq- ly inject 10 times a sample solution volume of 0.11 mL fol- uid Chromatography <2.01> under the following conditions, lowed by a single injection of 100 mL. After injecting the en- and measure the area of each peak using automatic integra- tire volume and running mobile phase A for 60 minutes, tion. When the amounts are determined by the area percent switch to mobile phase B. After measuring the sample solu- method, the total amount of peaks other than the teceleukin tion and after running 1 mol/L sodium chloride TS for 10 and solvent peaks is not more than 1.0z. minutes for postreatment and cleaning of the columns, inject Operating conditions— 100 mL of sodium hydroxide TS while running the mobile Detector: Ultraviolet absorption photometer (wavelength: phase A and then 55 minutes later start injection of the next 220 nm) sample solution. Column: A stainless steel column with an inside diameter Flow: Adjust the ‰ow of the mobile phase B so that the of 4.6 mm and 15 cm in length, packed with octadecyl- retention time for teceleukin is 45 to 65 minutes. Measure the silanized silica gel for liquid chromatography (5 mminparti- retention time from the point at which the mobile phase is cle diameter) switched to the mobile phase B. Column temperature: A constant temperature of about System suitability— 309C. System performance: Dissolve in water a mixture of two Mobile phase A: A solution of tri‰uoroacetic acid in a mix- kinds of equine heart-derived myoglobin whose isoelectric ture of water and acetonitrile (19:1) (1 in 1000) points are 6.76 and 7.16 to make a concentration of approxi- Mobile phase B: A solution of tri‰uoroacetic acid in mately 0.5 mg/mL. Mix together 50 mL of this solution, 50 acetonitrile (7 in 10,000) mL of Teceleukin (Genetical Recombination), and 1.47 mL of Mobile phase ‰ow: Control the concentration gradient by water. When 1.2 mL of this solution is run under the above changing the mobile phase A and mobile phase B as shown in conditions, myoglobin and teceleukin are eluted in this order, the table below. and their respective peaks are completely separated. (4) —Prepare a sample solution by adding 20 mL Time after injection Mobile phase Mobile phase of 0.2z sodium laurylsulfate TS to 20 mL of Teceleukin (Ge- of sample (min) A(volz) B(volz) netical Recombination). Perform the test as directed under Liquid Chromatography <2.01> using 20 mLofthissolution 0¿12 60ª50 40ª50 under the following conditions. Determine using automated 12¿25 50 50 integration the teceleukin peak area, A ,andthepeakarea, 2 25¿45 50ª050ª100 A , of the dimer with a relative retention time of 0.8 to 0.9 in 1 45¿50 0 100 relation to teceleukin. The amount of the dimer is not more than 1.0z by the following formula. Flow rate: 1.0 mL/min Amount (z)ofdimer＝[A /(A ＋A )]×100 1 1 2 Time span of measurement: a range that is approximately Operating conditions— 1.2-fold the retention time of teceleukin. Detector: Ultraviolet absorption photometer (wavelength: System suitability— 220 nm) System performance: Add 3.8 mL of water and 16.6 mLof Column: A stainless steel column with an inside diameter polysorbet 80 solution (1 in 100) to 83.6 mL of Teceleukin of 7.5 mm and 60 cm in length, packed with gycol etheriˆzed (Genetical Recombination) and let stand for at least one silica gel for liquid chromatography (particle diameter: 10 hour. When 5 mL of this solution is tested by running under mm) the above conditions, there is complete separation between Column temperature: A constant temperature of about the teceleukin peak and the peak with a relative retention 259C. time of about 0.98 in relation to the teceleukin peak. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in 0.1 (6) Acetic acid—Measure exactly 0.25 mL of Teceleukin mol/L sodium phosphate buŠer, pH 7.0, to make 1000 mL. (Genetical Recombination) and add exactly 0.25 mL of the Flow rate: Adjust the ‰ow rate so that the retention time of internal standard solution to make the sample solution. teceleukin is between 30 and 40 minutes. Separately, measure exactly 3 mL of acetic acid (100) and add System suitability— water to make exactly 100 mL. Take exactly 10 mL of this so- System performance: Add 20 mLof0.2z sodium lauryl lution and add water to make exactly 100 mL. Measure ex- sulfateTSto20mL of a solution consisting of 5 mg of car- actly 2 mL of this solution and add exactly 2 mL of the inter- bonic anhydrase and 5 mg of a-lactoalbumin dissolved in nal standard solution to make the standard solution. Perform 100 mL of water. When 20 mL of this solution is tested under the test with 1 mL each of the sample solution and the stan- the above conditions, carbonic anhydrase and a-lactoalbu- dard solution by Gas Chromatography <2.02> under the fol- min are eluted in this order with the resolution between these lowing conditions. Determine the ratios of the peak area of peaks being not less than 1.5. acetic acid to that of the internal standard, QT and QS,and System repeatability: Measure exactly 1.0 mL of the sam- the amount of acetic acid (C2H4O2) in 1 mL of Teceleukin ple solution, add the mobile phase to exactly 20 mL. To ex- (Genetical Recombination) determined by the following for- actly 1 mL of this solution add the mobile phase to make ex- mula is between 2.85 and 3.15 mg. actly 10 mL. When the test is repeated 3 times with 20 mLof Amount (mg) of acetic acid (C H O ) in 1 mL of Teceleukin this solution under the above conditions, the relative stan- 2 4 2 (Genetical Recombination) dard deviation of the teceleukin peak area is not more than ＝(Q /Q )×1.5×1.049×2 7z. T S 1148 Teceleukin for Injection (Genetical Recombination) / O‹cial Monographs JP XV

of culture medium for assay of teceleukin to all but 8 wells in 1.5: Concentration (mL/mL) of acetic acid (100) in the a microtest plate. Add 50 mL of the sample solution and the standard solution standard solution to 2 wells each containing culture medium 1.049: Density (mg/mL) of acetic acid (100) at 259C for assay of teceleukin. From these 4 wells, remove exactly 50 2: Dilution coe‹cient mL and add to 4 other wells containing culture medium for Internal standard solution—Diluted propionic acid (1 in 500) assay of teceleukin. From these 4 wells, remove exactly 50 mL Operating conditions— and add to 4 other wells containing culture medium for assay Detector: Hydrogen ‰ame ionization detector of teceleukin and repeat this procedure to prepare 2 wells that Column: A glass column with an inside diameter of 1.2 contain each of 1/2, 1/4, 1/8, 1/16, 1/32, 1/64, 1/128, and mm and 40 m in length, whose inside is covered with 1/256 dilutions of the sample and standard solutions. Add 50 chemically-bound polyethylene glycol for gas chro- mL of the standard solution to each of the 8 empty wells to matography 1.0 mminthickness. make maximum uptake controls. Eight wells containing only Column temperature: A constant temperature of about 110 culture medium for assay of teceleukin serve as the minimum 9C. uptake controls. After adding exactly 50 mL of cell suspen- Carrier gas: Helium sion solution for teceleukin to each well in a microtest plate, Flow rate: Adjust the ‰ow rate so that the retention time of leave for 15 to 17 hours in an incubator at 379C ˆlled with air acetic acid is about 8 minutes. containing 5z carbon dioxide. After adding 25 mLofMTT System suitability— TS to each of the wells in the plate, leave for 4 hours in an in- System performance: When 1 mL of the standard solution cubator at 379Cˆlledwithaircontaining5z carbon dioxide. is run under the above conditions, acetic acid and the internal Transfer the culture medium in all of the wells to empty wells standard are eluted in this order with the resolution between in another microtest plate. To each of the empty wells from these peaks being not less than 3.0. which the culture medium was removed, add 100 mLof System repeatability: When the test is repeatedly run 6 hydrochloric acid-2-propanol TS, and then shake the plates times under the above conditions using 1 mLofstandardso- horizontally for 5 minutes to elute the pigment. After return- lution, the relative standard deviation of the ratio of the acet- ing the transferred culture medium to each original well, per- ic acid peak area to the internal standard peak area is not form the test with the solution in each well, determine the more than 5z. diŠerence in absorption at wavelengths of 560 nm and 690 nm, and calculate the mean values of the identical respective Bacterial endotoxins <4.01> Less than 5EU per mg of pro- solutions in the two wells (dilution solutions of the sample so- tein lution and standard solutions) as well as the 8 wells contain- Speciˆc activity Accurately measure an appropriate amount ing the maximum or minimum uptake controls. Prepare stan- of Teceleukin (Genetical Recombination), and add water ac- dard curves by plotting the values obtained from each dilu- curately so that 1 mL contains about 0.1 mg. This is the sam- tion solution of the sample solution, with the dilution ple solution. Separately, measure precisely about 25 mg of coe‹cient of the sample solution on the microtest plates in human serum albumin for assay, dissolve in water, and add logarithmic scale on the horizontal axis and the absorbance water to make 50 mL. Measure exactly an appropriate on the vertical axis. Determine the mean absorbance values amount of this solution, and accurately dilute with water to of the maximum and minimum uptake controls, ˆnd the make standard solutions with concentrations of 0.05, 0.10, values on the standard curve, and then determine the dilution and 0.15 mg/mL. Accurately measure 1 mL each of the sam- coe‹cient, DT. Perform the same plot for the dilution solu- ple solution, the standard solutions, and water, add 2.5 mL tion of the standard solution, determine the dilution of alkaline copper solution, mix, leave for at least 10 minutes coe‹cient, DS, and then determine the potency in 1 mL by to dissolve, add exactly 2.5 mL of water and 0.5 mL of dilut- the following formula. ed Folin reagent (1 in 2), immediately shake vigorously, and Teceleukin potency (units) in 1 mL of Teceleukin (Genetical then leave for 30 minutes at 379C. Perform the test on these Recombination) solutions, with water as a control, as directd under Ultrav- ＝S×(D /D )×d iolet-visible Spectrophotometry <2.24>, and measure the ab- T S sorbance at 750 nm. With the concentration of the standard S: Concentration of standard solution (units/mL) solution as the x-axis and the absorbance as the y-axis, per- d: Dilution coe‹cient when sample solution prepared form linear regression using their respective reciprocals, and Containers and storage Containers—Tight containers determine the protein content. Storage—Store at －709Corbelow. Determine the ratio of the potency determined by Assay and the protein content. Assay Accurately measure an appropriate amount of Teceleukin for Injection (Genetical Teceleukin (Genetical Recombination) and, depending on the cell sensitivity, dilute precisely by adding culture medium for Recombination) assay of teceleukin to a constant concentration of 10 to 50 units/mL (estimated value). This is the sample solution. 注射用テセロイキン（遺伝子組換え） Separately, dissolve Interleukin-2 Reference Substance in 1 mL of sterilized puriˆed water, and, depending on the cell Teceleukin for Injection (Genetical Recombination) sensitivity, dilute precisely by adding culture medium for as- is a preparation for injection which is dissolved before say of teceleukin to a constant concentration of 10 to 50 use. units/mL. This is the standard solution. Add exactly 50 mL It contains not less than 70.0z and not more than JP XV O‹cial Monographs / Tegafur 1149

150.0z of the labeled amount of teceleukin (genetical Recombination), and calculate the amount (unit) of teceleu- recombination) (C698H1127N179O204S8: 15547.01). kin in 1 vial by the following formula.

Method of preparation Prepare as directed under Injection, Amount (unit) of teceleukin in 1 vial＝S×(DT/DS)×d×1 with Teceleukin (Genetical Recombination). S: Concentration (unit/mL) of the standard solution Description Teceleukin for Injection (Genetical Recombi- d: Rate of dilution to prepare the sample solution nation)occursasawhite,lightmassorpowder. l: Volume (mL) of the sample solution Identiˆcation (1) Dissolve the content of 1 vial of Teceleu- Containers and storage Containers—Hermetic containers. kin for Injection (Genetical Recombination) in 1 mL of steri- Storage—Light-resistant, not exceeding 109C, avoiding lized puriˆed water, dilute exactly with culture medium for freezing. assay of teceleukin to make the sample stock solution con- taining about 200 units/mL (estimate). Proceed as directed in the Identiˆcation (1) and (3) under Teceleukin (Genetical Tegafur Recombination). (2) Dissolve 0.242 g of 2-amino-2-hydroxymethyl-1,3- テガフール propanediol, 5.0 g of sodium lauryl sulfate and 74 mg of dis- odium dihydrogen ethylenediamine tetraacetate dihydrate in 60 mL of water. Adjust to pH 8.0 with 1 mol/L hydrochloric acid TS, add water to make 100 mL, and use this solution as the buŠer solution for molecular mass determination. Separately, dissolve the content of 1 vial of Teceleukin for In- jection (Genetical Recombination) in exactly 1 mL of water. To exactly 100 mL of this solution add exactly 100 mLofthe buŠer solution for molecular mass determination and 10 mL C8H9FN2O3: 200.17 of 2-mercaptoethanol, and heat on a water bath for 5 minutes 5-Fluoro-1-[(2RS)-tetrahydrofuran-2-yl]uracil without allowing any water evaporation from the mixture. [17902-23-7] After cooling, add exactly 1 mL of bromophenol blue solu- tion (1 in 2000), mix, and use this solution as the sample solu- Tegafur, when dried, contains not less than 98.0z tion. Proceed as directed in Identiˆcation (3) under Teceleu- of C8H9FN2O3. kin (Genetical Recombination): a band appears in the range Description Tegafur occurs as a white, crystalline powder. of molecular mass between 14,000 and 16,000. It is soluble in methanol and in acetone, and sparingly pH <2.54> Dissolve the content of one vial of Teceleukin for solubleinwaterandinethanol(95). Injection (Genetical Recombination) in 1 mL of water: the It dissolves in dilute sodium hydroxide TS. pH of the solution is between 7.0 and 7.7. A solution of Tegafur in methanol (1 in 50) shows no opti- cal rotation. Purity Clarity and color of solution—Dissolve the content of one vial of Teceleukin for Injection (Genetical Recombi- Identiˆcation (1) Prepare the test solution with 0.01 g of nation) in 1 mL of water: the solution is clear and colorless. Tegafur as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 molWLsodium Loss on drying <2.41> Transfer the content of the vial of hydroxide TS and 20 mL of water as an absorbing liquid: the Teceleukin for Injection (Genetical Recombination) to a test solution responds to the Qualitative Tests <1.09> (2) for weighing bottle under the atmosphere not exceeding 10z rel- ‰uoride. ative humidity, and perform the test as directed in the Water (2) Determine the absorption spectrum of a solution of content determination described in the Minimum Require- Tegafur in 0.01 mol/L sodium hydroxide TS (1 in 100,000) as ments for Biological Products: not more than 3z. directed under Ultraviolet-visible Spectrophotometry <2.24>, Bacterial endotoxins <4.01> Less than 5 EU/350,000 units. and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the Uniformity of dosage units <6.02> It meets the requirement same wavelengths. of the Mass variation test. Calculate as － ＝0. `M A` (3) Determine the infrared absorption spectrum of Foreign insoluble matter <6.06> Perform the test according Tegafur, previously dried, as directed in the potassium to Method 2: it meets the requirement. bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- Insoluble particulate matter <6.07> Perform the test accord- trum: both spectra exhibit similar intensities of absorption at ing to Method 1: it meets the requirement. thesamewavenumbers. Sterility <4.06> Perform the test according to the Membrane pH <2.54> Dissolve 0.5 g of Tegafur in 50 mL of water: the ˆltration method: it meets the requirement. pH of this solution is between 4.2 and 5.2. Assay Dissolve the content of 1 vial of Teceleukin for Injec- Melting point <2.60> 166 – 1719C tion (Genetical Recombination) in exactly 1 mL of sterilized puriˆed water, dilute exactly with culture medium for assay Purity (1) Clarity and color of solution—Dissolve 0.2 g of of teceleukin to make the sample solution containing a Tegafur in 10 mL of dilute sodium hydroxide TS: the solu- deˆnite concentration of 10 to 50 units/mL (estimate). Pro- tion is clear and colorless. ceed as directed in the Assay under Teceleukin (Genetical (2) Chloride <1.03>—Dissolve 0.8 g of Tegafur in 40 mL 1150 Teicoplanin / O‹cial Monographs JP XV of water by warming, cool, ˆlter if necessary, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the con- Teicoplanin trol solution with 0.25 mL of 0.01 molWLhydrochloricacid VS (not more than 0.011z). テイコプラニン (3) Heavy metals <1.07>—Dissolve 1.0 g of Tegafur in 40 mL of water by warming, cool, ˆlter if necessary, and add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (4) Arsenic <1.11>—Prepare the test solution in a plati- num crucible with 1.0 g of Tegafur according to Method 4, incinerating by ignition between 7509Cand8509C, and per- form the test (not more than 2 ppm). (5) Related substances—Dissolve 0.10 g of Tegafur in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make ex- actly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin- layer Chromatography <2.03>.Spot5mLeachofthesample solution and standard solution on a plate of silica gel with ‰uorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform and ethanol (95) (5:1) to a distance of about 10 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. 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, plati- num crucible). Assay Weigh accurately about 0.15 g of Tegafur, previous- ly dried, place in an iodine bottle, dissolve in 75 mL of water, and add exactly 25 mL of 1W60 molWL potassium bromate VS. Add rapidly 1.0 g of potassium bromide and 12 mL of hydrochloric acid, stopper the bottle tightly at once, and al- low to stand for 30 minutes with occasional shaking. To this Teicoplanin A2-1 solution add 1.6 g of potassium iodide, shake gently, allow to C88H95Cl2N9O33: 1877.64 stand for exactly 5 minutes, and titrate <2.50> the liberated io- (3S,15R,18R,34R,35S,38S,48R,50aR)-34-(2-Acetylamino-2- dine with 0.1 molWL sodium thiosulfate VS (indicator: 2 mL deoxy-b-D-glucopyranosyloxy)-15-amino-22,31-dichloro- of starch TS). Perform a blank determination. 56-[2-(4Z)-dec-4-enoylamino-2-deoxy-b-D-glucopyranosylox- y]-6,11,40,44-tetrahydroxy-42-(a-D-mannopyranosyloxy)- Each mL of 1/60 molWL potassium bromate VS 2,16,36,50,51,59-hexaoxo- ＝ 10.01 mg of C8H9FN2O3 2,3,16,17,18,19,35,36,37,38,48,49,50,50a-tetradecahydro- Containers and storage Containers—Tight containers. 1H,15H,34H-20,23:30,33-dietheno-3,18:35,48- bis(iminomethano)-4,8:10,14:25,28:43,47-tetrametheno- 28H-[1,14,6,22]dioxadiazacyclooctacosino[4,5- m][10,2,16]benzoxadiazacyclotetracosine-38- carboxylic acid [91032-34-7]

Teicoplanin A2-2 C88H97Cl2N9O33: 1879.66 (3S,15R,18R,34R,35S,38S,48R,50aR)-34- (2-Acetylamino-2-deoxy-b-D-glucopyranosyloxy)- 15-amino-22,31-dichloro-56-[2-deoxy- 2-(8-methylnonanoylamino)-b-D-glucopyranosyloxy]- 6,11,40,44-tetrahydroxy-42-(a-D-mannopyranosyloxy)- 2,16,36,50,51,59-hexaoxo- 2,3,16,17,18,19,35,36,37,38,48,49,50,50a-tetradecahydro- 1H,15H,34H-20,23:30,33-dietheno-3,18:35,48- JP XV O‹cial Monographs / Teicoplanin 1151 bis(iminomethano)-4,8:10,14:25,28:43,47-tetrametheno- 28H-[1,14,6,22]dioxadiazacyclooctacosino[4,5- [61036-62-2, Teicoplanin] m][10,2,16]benzoxadiazacyclotetracosine-38- carboxylic acid [91032-26-7] Teicoplanin is a mixture of glycopeptide substances having antibacterial activity produced by the growth of Actinoplanes teichomyceticus. Teicoplanin A 2-3 It contains not less than 900 mg (potency) per mg, C H Cl N O : 1879.66 88 97 2 9 33 calculated on the anhydrous, de-sodium chloride and (3S,15R,18R,34R,35S,38S,48R,50aR)-34-(2-Acetylamino- de-residual solvents basis. The potency of Teicoplanin 2-deoxy-b-D-glucopyranosyloxy)-15-amino-22,31-dichloro- is expressed as mass (potency) of teicoplanin 56-(2-decanoylamino-2-deoxy-b-D-glucopyranosyloxy)- (C H Cl N O ). 6,11,40,44-tetrahydroxy-42-(a-D-mannopyranosyloxy)- 72-89 68-99 2 8-9 28-33 2,16,36,50,51,59-hexaoxo- Description Teicoplanin occurs as a white to light yellowish 2,3,16,17,18,19,35,36,37,38,48,49,50,50a-tetradecahydro- white powder. 1H,15H,34H-20,23:30,33-dietheno-3,18:35,48- It is freely soluble in water, sparingly soluble in N,N- bis(iminomethano)-4,8:10,14:25,28:43,47-tetrametheno- dimethylformamide, and practically insoluble in acetonitrile, 28H-[1,14,6,22]dioxadiazacyclooctacosino[4,5- in methanol, in ethanol (95), in acetone, in acetic acid (100) m][10,2,16]benzoxadiazacyclotetracosine-38- andindiethylether. carboxylic acid [91032-36-9] Identiˆcation (1) To 1 mL of a solution of Teicoplanin (1 in 100) add 2 mL of ninhydrin TS, and warm for 5 minutes: a

Teicoplanin A2-4 blue-purple color develops. C89H99Cl2N9O33: 1893.68 (2) To 1 mL of a solution of Teicoplanin (3 in 100) add (3S,15R,18R,34R,35S,38S,48R,50aR)-34- slowly 2 mL of anthrone TS, and shake gently: a dark brown (2-Acetylamino-2-deoxy-b-D-glucopyranosyloxy)- color develops. 15-amino-22,31-dichloro-56-[2-deoxy- (3) Determine the infrared absorption spectra of Tei- 2-(8-methyldecanoylamino)-b-D-glucopyranosyloxy]- coplanin and Teicoplanin Reference Standard as directed in 6,11,40,44-tetrahydroxy-42-(a-D-mannopyranosyloxy)- the potassium bromide disk method under Infrared Spec- 2,16,36,50,51,59-hexaoxo- trophotometry <2.25>, and compare the spectrum with the 2,3,16,17,18,19,35,36,37,38,48,49,50,50a-tetradecahydro- spectrum of Teicoplanin Reference Standard: both spectra 1H,15H,34H-20,23:30,33-dietheno-3,18:35,48- exhibit similar intensities of absorption at the same wave bis(iminomethano)-4,8:10,14:25,28:43,47-tetrametheno- numbers. 28H-[1,14,6,22]dioxadiazacyclooctacosino[4,5- pH <2.54> Dissolve 0.5 g of Teicoplanin in 10 mL of water: m][10,2,16]benzoxadiazacyclotetracosine-38- the pH of the solution is between 6.3 and 7.7. carboxylic acid [91032-37-0] Content ratio of the active principle Dissolve about 20 mg of Teicoplanin in water to make 10 mL, and use this solution Teicoplanin A 2-5 as the sample solution. Perform the test with 20 mLofthe C H Cl N O : 1893.68 89 99 2 9 33 sample solution as directed under Liquid Chromatography (3S,15R,18R,34R,35S,38S,48R,50aR)-34- <2.01> according to the following conditions, and calculate (2-Acetylamino-2-deoxy-b-D-glucopyranosyloxy)- the sum of peak areas of teicoplanin A group, S ,thesumof 15-amino-22,31-dichloro-56-[2-deoxy- 2 a peak areas of teicoplanin A3 group, Sb, and the sum of peak 2-(9-methyldecanoylamino)-b-D-glucopyranosyloxy]- areas of other contents, Sc from the sample solution by the 6,11,40,44-tetrahydroxy-42-(a-D-mannopyranosyloxy)- automatic integration method. Calculate the content ratio of 2,16,36,50,51,59-hexaoxo- them by the formula given below: teicoplanin A group, tei- 2,3,16,17,18,19,35,36,37,38,48,49,50,50a-tetradecahydro- 2 coplanin A group, and the other are not less than 80.0z,not 1H,15H,34H-20,23:30,33-dietheno-3,18:35,48- 3 more than 15.0z and not more than 5.0z, respectively. bis(iminomethano)-4,8:10,14:25,28:43,47-tetrametheno- The elution order of each content and the relative retention 28H-[1,14,6,22]dioxadiazacyclooctacosino[4,5- time of each content to the retention time of teicoplanin A m][10,2,16]benzoxadiazacyclotetracosine-38- 2-2 are shown in the following table. carboxylic acid [91032-38-1] Elution Relative Name of content order retention time Teicoplanin A3-1 C72H68Cl2N8O28: 1564.25 teicoplanin A3 group ≦0.42 (3S,15R,18R,34R,35S,38S,48R,50aR)-34-(2-Acetylamino-2- teicoplanin A3-1 10.29 deoxy-b-D-glucopyranosyloxy)-15-amino-22,31-dichloro- teicoplanin A2 group 0.42＜,≦1.25 6,11,40,44,56-pentahydroxy-42-(a-D-mannopyranosyloxy)- teicoplanin A2-1 20.91 2,16,36,50,51,59-hexaoxo- teicoplanin A2-2 31.00 2,3,16,17,18,19,35,36,37,38,48,49,50,50a-tetradecahydro- teicoplanin A2-3 41.04 1H,15H,34H-20,23:30,33-dietheno-3,18:35,48- teicoplanin A2-4 51.17 bis(iminomethano)-4,8:10,14:25,28:43,47-tetrametheno- teicoplanin A2-5 61.20 28H-[1,14,6,22]dioxadiazacyclooctacosino[4,5- others 1.25＜ m][10,2,16]benzoxadiazacyclotetracosine-38- carboxylic acid [93616-27-4] 1152 Teicoplanin / O‹cial Monographs JP XV

Content ratio (z) of teicoplanin A2 group (4) Arsenic <1.11>—Being speciˆed separately. ＝{Sa/(Sa＋0.83Sb＋Sc)}×100 (5) Residual solvents <2.46>—Weigh accurately about 0.1 g of Teicoplanin, dissolve in N,N-dimethylformamide to Content ratio (z) of teicoplanin A group 3 make exactly 10 mL, and use this solution as the sample solu- ＝{0.83S /(S ＋0.83S ＋S )}×100 b a b c tion. Separately, weigh accurately about 1 g each of methanol Content ratio (z)ofothers and acetone, and add N,N-dimethylformamide to make ex-

＝{Sc/(Sa＋0.83Sb＋Sc)}×100 actly 100 mL. Pipet 1 mL of this solution, add N,N-dimethyl- formamide to make exactly 100 mL, and use this solution as Operating conditions— the standard solution. Perform the test with exactly 4 mLeach Detector: An ultraviolet absorption photometer (wave- of the sample solution and standard solution as directed un- length: 254 nm). der Gas Chromatography <2.02> according to the following Column: A stainless steel column 4.6 mm in inside di- condition. Calculate the peak area of methanol, A ,andthe ameter and 25 cm in length, packed with octadecylsilanized 1 peak area of acetone, A , obtained from the sample solution, silica gel for liquid chromatography (5 mminparticledi- 2 and the peak area of methanol, A , and the peak area of ace- ameter). S1 tone, A , obtained from the standard solution by the auto- Column temperature: A constant temperature of about S2 matic integration method, and calculate the amounts of 259C. methanol and acetone by the following formula: not more Mobile phase A: Dissolve 7.80 g of sodium dihydrogen than 0.5z and not more than 1.0z, respectively. phosphate dihydrate in 1650 mL of water, add 300 mL of acetonitrile, adjust pH to 6.0 with sodium hydroxide TS, and Amount (z) of methanol add water to make 2000 mL. ＝WS1×(A1/AS1)×0.001×(1/WT1)× 100 Mobile phase B: Dissolve 7.80 g of sodium dihydrogen W : Amount (g) of methanol phosphate dihydrate in 550 mL of water, add 1400 mL of S1 W : Amount (g) of Teicoplanin acetonitrile, adjust the pH to 6.0 with sodium hydroxide TS, T1 and add water to make 2000 mL. Amount (z) of acetone

Flowing of the mobile phase: Flow mobile phase A for 10 ＝WS2×(A2/AS2)×0.001×(1/WT2)×100 minutes before injection. After injection, control the W : Amount (g) of acetone gradient by mixing the mobile A and B as directed in the fol- S2 W : Amount (g) of Teicoplanin lowing table. T2 Operating conditions— Time after injection Mobile phase Mobile phase Detector: Hydrogen ‰ame-ionization detector. of sample (min) A (volz)B(volz) Column: A glass column 2 mm in inside diameter and 3 m 0–32 100→ 70 0 → 30 in length, packed with graphite carbon for gas chro- 32–40 70→ 50 30 → 50 matography, 150 to 180 mm in particle diameter, coated with 40–42 50→ 100 50 → 0 0.1z of polyethylene glycol esteriˆed. Column temperature: Inject the sample at a constant tem- Flow rate: 1.8 mL per minute. perature of about 709C, maintain the temperature for 4 Time span of measurement: About 1.7 times as long as the minutes, then program to increase the temperature at the rate retention time of teicoplanin A2-2, beginning after the solvent of 89C per minute to 2109C. peak. Detector temperature: A constant temperature of about System suitability— 2409C. Test for required detection: Conˆrm that peak height of Carrier gas: Nitrogen teicoplanin A2-2 obtained from the sample solution is equiva- Flow rate: Adjust the ‰ow rate so that the retention times lent to 90z of the full scale. of methanol and acetone are about 2 minutes and 5 minutes, System performance: When the procedure is run with 20 respectively. mL of the sample solution under the above operating condi- System suitability— tions, the symmetry factor of the peak of teicoplanin A3-1 is Test for required detection: Conˆrm that the peak height not more than 2.2. of acetone obtained from 4 mL of the standard solution is e- System repeatability: When the test is repeated 3 times with quivalent to about the full scale. 20 mL of the sample solution under the above operating con- System performance: When the procedure is run with 4 mL ditions, the relative standard deviation of the peak area of of the standard solution under the above operating condi- teicoplanin A2-2 is not more than 2.0z. tions, methanol and acetone are eluted in this order with the resolution between these peaks being not less than 2.0. Purity (1) Clarity and color of solution—Being speciˆed System repeatability: When the test is repeated 3 times with separately. 4 L of the standard solution under the above operating con- (2) Sodium chloride—Weigh accurately about 0.5 g of m ditions, the relative standard deviation of the peak areas of Teicoplanin, dissolve in 50 mL of water, titrate <2.50> with acetone is not more than 3z. 0.1 molWL silver nitrate VS (indicator: 1 mL of potassium chromate TS), and calculate an amount of sodium chloride: Water <2.48> Not more than 15.0z (0.2 g, volumetric titra- not more than 5.0z. tion, direct titration). Each mL of 0.1 molWL silver nitrate VS Bacterial endotoxins <4.01> Less than 0.75 EUWmg (poten- ＝ 5.844 mg of NaCl cy). (3) Heavy metals <1.07>—Being speciˆed separately. Blood pressure depressant Being speciˆed separately. JP XV O‹cial Monographs / Terbutaline Sulfate 1153

10,000) as directed under Ultraviolet-visible Spectrophoto- Assay Perform the test according to the Cylinder-plate metry <2.24>, and compare the spectrum with the Reference method as directed under Microbial Assay for Antibiotics Spectrum: both spectra exhibit similar intensities of absorp- <4.02> according to the following conditions. tion at the same wavelengths. This maximum can be biphasic. (i) Test organism—Bacillus subtilis ATCC 6633 (3) A solution of Terbutaline Sulfate (1 in 50) responds to (ii) Culture medium—Use the medium i in 1) under (1) the Qualitative Tests <1.09> for sulfate. Agar media for seed and base layer. (iii) Standard solutions—Weigh accurately an amount of pH <2.54> Dissolve Terbutaline Sulfate in 10 mL of water: Teicoplanin Reference Standard equivalent to about 50 mg the pH of this solution is between 4.0 and 4.8. (potency), dissolve in phosphate buŠer solution, pH 6.0 to Purity (1) Clarity and color of solution—Dissolve 0.10 g make exactly 50 mL, and use this solution as the standard of Terbutaline Sulfate in 10 mL of water: the solution is clear stock solution. Keep the standard stock solution at not ex- and colorless or slightly yellow. ceeding 59C and use within 14 days. Take exactly a suitable (2) Chloride <1.03>—Perform the test with 2.0 g of Ter- amount of this solution before use, add phosphate buŠer so- butaline Sulfate. Prepare the control solution with 0.25 mL lution, pH 6.0 to make solutions so that each mL contains of 0.01 molWLhydrochloricacidVS(notmorethan0.004z). 160 mg (potency) and 40 mg (potency), and use these solutions (3) Acetic acid—Dissolve 0.50 g of Terbutaline Sulfate in asthehighconcentrationstandardsolutionandlowconcen- a solution of phosphoric acid (59 in 1000) to make exactly 10 tration standard solution, respectively. mL, and use this solution as the sample solution. Separately, (iv) Sample solutions—Weigh accurately an amount of dissolve 1.50 g of acetic acid (100) in a solution of phosphoric Teicoplanin equivalent to about 50 mg (potency), dissolve in acid (59 in 1000) to make exactly 100 mL. Dilute 2 mL of this phosphate buŠer solution, pH 6.0 to make exactly 50 mL. solution, accurately measured, with a solution of phosphoric Take exactly a suitable amount of this solution, add phos- acid (59 in 1000) to make exactly 200 mL, and use this solu- phate buŠer solution, pH 6.0 to make solutions so that each tion as the standard solution. Perform the test with exactly 2 mL contains 160 mg (potency) and 40 mg (potency), and use mL each of the sample solution and standard solution as these solutions as the high concentration sample solution and directed under Gas Chromatography <2.02> according to the low concentration sample solution, respectively. following operating conditions. Measure the peak areas, AT Containers and storage Containers—Tight containers. and AS, of acetic acid for the two solutions: AT is not larger Storage—Light-resistant, and not exceeding 59C. than AS. Operating conditions— Detector: A hydrogen ‰ame-ionization detector. Terbutaline Sulfate Column: A glass column 3 mm in inside diameter and 1 m in length, packed with 10z of macrogol 6000 on 180- to テルブタリン硫酸塩 250-mm terephthalic acid for gas chromatography. Column temperature: A constant temperature at about 1209C. Carrier gas: Nitrogen. Flow rate: Adjust the ‰ow rate so that the retention time of acetic acid is about 5 minutes. System suitability— System performance: Mix 0.05 g each of acetic acid (100)

(C12H19NO3)2.H2SO4: 548.65 and propionic acid in 100 mL of diluted phosphoric acid (59 5-[(1RS)-2-(1,1-Dimethylethylamino)- in 1000). When the procedure is run with 2 mL of this solution 1-hydroxyethyl]benzene-1,3-diol hemisulfate [23031-32-5] under the above conditions, acetic acid and propionic acid are eluted in this order with the resolution between these Terbutaline Sulfate contains not less than 98.5z of peaks being not less than 2.0. (C12H19NO3)2.H2SO4, calculated on the anhydrous ba- System repeatability: When the test is repeated 6 times with sis. 2 mL of the standard solution under the above operating con- ditions, the relative standard deviation of the peak areas of Description Terbutaline Sulfate is white to slightly acetic acid is not more than 3.0z. brownish white crystals or crystalline powder It is odorless or (4) 3,5-Dihydroxy-v-tert-butylaminoacetophenone sul- hasafaintodorofaceticacid. fate—Dissolve 0.50 g of Terbutaline Sulfate in 0.01 molWL It is freely soluble in water, and practically insoluble in hydrochloric acid TS to make exactly 25 mL, and perform acetonitrile, in ethanol (95), in acetic acid (100), in chlo- the test as directed under Ultraviolet-visible Spectrophoto- roform, and in diethyl ether. metry <2.24>: the absorbance at a wavelength of 330 nm does It is gradually colored by light and by air. not exceed 0.47. Melting point: about 2559C (with decomposition). (5) Heavy metals <1.07>—Proceed with 2.0 g of Terbuta- Identiˆcation (1) Dissolve 1 mg of Terbutaline Sulfate in line Sulfate as directed under Method 2, and perform the test. 1 mL of water, and add 5 mL of Tris buŠer solution, pH 9.5, Prepare the control solution with 2.0 mL of Standard Lead 0.5 mL of 4-aminoantipyrine solution (1 in 50) and 2 drops of Solution (not more than 10 ppm). potassium hexacyanoferrate (III) solution (2 in 25): a reddish (6) Arsenic <1.11>—Prepare the test solution with 1.0 g purple color is produced. of Terbutaline Sulfate according to method 3, and perform (2) Determine the absorption spectrum of a solution of the test (not more than 2 ppm). Terbutaline Sulfate in 0.01 mol/L hydrochloric acid TS (1 in Water <2.48> Not more than 0.5z (1 g, direct titration). 1154 Enanthate / O‹cial Monographs JP XV

exactly 100 mL. Measure exactly 10 mL of this solution, and Residue on ignition <2.44> Not more than 0.2z (1 g). dilute with ethanol (95) to make exactly 100 mL. Measure ex- Assay Weigh accurately about 0.5 g of Terbutaline Sulfate, actly 10 mL of this solution, and dilute with ethanol (95) to dissolve in 50 mL of a mixture of acetonitrile and acetic acid make exactly 100 mL. Perform the test as directed under (100) (1:1) by stirring and warming. Allow to cool, and titrate Ultraviolet-visible Spectrophotometry <2.24> with this solu- <2.50> with 0.1 molWL perchloric acid VS (potentiometric tion. Read the absorbance A of this solution at the titration, substituting a saturated solution of potassium chlo- wavelength of maximum absorption at about 241 nm. ride in methanol for the internal ‰uid). Amount (mg) of C26H40O3 Each mL of 0.1 molWL perchloric acid VS ＝(A/426)×100,000 ＝ 54.87 mg of (C H NO ) .H SO 12 19 3 2 2 4 Containers and storage Containers—Tight containers. Containers and storage Containers—Tight containers. Storage—Light-resistant, and not exceeding 309C. Storage—Light-resistant. Injection Testosterone Enanthate テストステロンエナント酸エステル注射液 テストステロンエナント酸エステル

Testosterone Enanthate Injection is an oily solution for injection. It contains not less than 90z and not more than 110 z of the labeled amount of testosterone enanthate (C26 H40O3:400.59). Method of preparation Prepare as directed under Injec- tions, with Testosterone Enanthate. C26H40O3:400.59 3-Oxoandrost-4-en-17b-yl heptanoate [315-37-7] Description Testosterone Enanthate Injection is a clear, colorless or pale yellow oily liquid. Testosterone Enanthate, when dried, contains not Identiˆcation Measure a volume of Testosterone Enanthate less than 95.0z and not more than 105.0z of Injection, equivalent to 0.05 g of Testosterone Enanthate ac- C26H40O3. cordingtothelabeledamount,add8mLofpetroleumether, Description Testosterone Enanthate occurs as white to pale and extract with three 10-mL portions of diluted acetic acid yellow crystals, crystalline powder or a pale yellow-brown, (31) (7 in 10). Combine the extracts, wash with 10 mL of viscous liquid. It is odorless or has a slight, characteristic petroleum ether, add 0.5 mL of diluted sulfuric acid (7 in 10) odor. to 0.1 mL of the extract, and heat on a water bath for 5 It is very soluble in ethanol (95), in 1,4-dioxane and in minutes. Cool, and add 0.5 mL of iron (III) chloride-acetic diethyl ether, and practically insoluble in water. acid TS: the color of the solution is blue. Melting point: about 369C Extractable volume <6.05> It meets the requirement. Identiˆcation Heat 25 mg of Testosterone Enanthate with 2 Assay Measure accurately a volume of Testosterone Enan- mL of a solution of potassium hydroxide in methanol (1 in thate Injection, equivalent to about 25 mg of testosterone 100) under a re‰ux condenser on a water bath for 1 hour, enanthate (C26H40O3), and dissolve in chloroform to make ex- cool, and add 10 mL of water. Collect the produced actly 25 mL. Pipet 3 mL of this solution, add chloroform to precipitate by suction, wash with water until the last washing make exactly 50 mL, and use this solution as the sample solu- is neutral, and dry the precipitate in a desiccator (in vacuum, tion. Separately, weigh accurately about 25 mg of Testoster- phosphorus (V) oxide) for 4 hours: the precipitate melts be- one Propionate Reference Standard, proceed in the same tween 1519Cand1579C. manner as for the sample solution, and use this solution as 20 the standard solution. Pipet 5 mL each of the sample solution Optical rotation <2.49> [a]D : ＋77 – ＋889(after drying, 0.1 g, 1,4-dioxane, 10 mL, 100 mm). and standard solution, add exactly 10 mL of TS, add methanol to make exactly 20 mL, and allow to stand for Purity Acidity—Dissolve 0.5 g of Testosterone Enanthate 45 minutes. Determine the absorbances, A T and A S,ofthese in 10 mL of ethanol (95) which has previously been rendered solutions at 380 nm, respectively, as directed under Ultrav- neutral to bromothymol blue TS, and add 2 drops of iolet-visible Spectrophotometry <2.24>, using a solution ob- bromothymol blue TS and 0.50 mL of 0.01 molWLsodium tained by proceeding with 5 mL of chloroform as the blank. hydroxide VS: the color of the solution is light blue. Amount (mg) of testosterone enanthate (C26H40O3) Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu- ＝WS×(A T/A S)×1.1629 um, phosphorus (V) oxide, 4 hours). WS: Amount (mg) of Residue on ignition <2.44> Not more than 0.1z (0.5 g). Reference Standard Assay Weigh accurately about 0.1 g of Testosterone Enan- Containers and storage Containers—Hermetic containers. thate, previously dried, and dissolve in ethanol (95) to make Storage—Light-resistant. JP XV O‹cial Monographs / Testosterone Propionate Injection 1155

Propionate and Testosterone Propionate Reference Standard, previously dried, and dissolve in methanol to make Testosterone Propionate exactly 100 mL. To exactly 5 mL of these solutions add ex- actly 5 mL of the internal standard solution and methanol to テストステロンプロピオン酸エステル make 20 mL, and use these solutions as the sample solution and standard solution. Perform the test with 5 mLeachofthe sample solution and standard solution as directed under Liq- uid Chromatography <2.01> according to the following con-

ditions, and determine the ratios, QT and QS, of the peak area of testosterone propionate to that of the internal standard.

Amount (mg) of C22H32O3 ＝WS×(QT/QS)

WS: Amount (mg) of Testosterone Propionate Reference C22H32O3:344.49 3-Oxoandrost-4-en-17b-yl propanoate [57-85-2] Standard Internal standard solution—A solution of in Testosterone Propionate, when dried, contains not methanol (9 in 100,000). less than 97.0z and not more than 103.0z of Operating conditions— C22H32O3. Detector: An ultraviolet absorption photometer Description Testosterone Propionate occurs as white to (wavelength: 241 nm). pale yellow crystals or crystalline powder. Column: A stainless steel column 4.6 mm in inside di- It is freely soluble in methanol and in ethanol (95), and ameter and 15 cm in length, packed with octadecylsilanized practically insoluble in water. silica gel for liquid chromatography (5 mminparticledi- ameter). Identiˆcation (1) Determine the absorption spectrum of a Column temperature: A constant temperature of about solution of Testosterone Propionate in ethanol (95) (1 in 359C. 100,000) as directed under Ultraviolet-visible Spectrophoto- Mobile phase: A mixture of acetonitrile and water (7:3). metry <2.24>, and compare the spectrum with the Reference Flow rate: Adjust the ‰ow rate so that the retention time of Spectrum or the spectrum of a solution of Testosterone testosterone propionate is about 10 minutes. Propionate Reference Standard prepared in the same manner System suitability— as the sample solution: both spectra exhibit similar intensities System performance: When the procedure is run with 5 mL of absorption at the same wavelengths. of the standard solution under the above operating condi- (2) Determine the infrared absorption spectrum of tions, the internal standard and testosterone propionate are Testosterone Propionate, previously dried, as directed in the eluted in this order with the resolution between these peaks potassium bromide disk method under Infrared being not less than 9. Spectrophotometry <2.25>, and compare the spectrum with System repeatability: When the test is repeated 6 times with the Reference Spectrum or the spectrum of Testosterone 5 mL of the standard solution under the above operating con- Propionate Reference Standard: both spectra exhibit similar ditions, the relative standard deviation of the ratio of the intensities of absorption at the same wave numbers. peak area of testosterone propionate to that of the internal 20 standard is not more than 1.0z. Optical rotation <2.49> [a]D : ＋83 – ＋909(after drying, 0.1 g, ethanol (95), 10 mL, 100 mm). Containers and storage Containers—Tight containers. Melting point <2.60> 118 – 1239C Storage—Light-resistant. Purity Related substances—Dissolve 40 mg of Testosterone Propionate in 2 mL of ethanol (95), and use this solution as Testosterone Propionate Injection the sample solution. Pipet 1 mL of this solution, add ethanol (95) to make exactly 100 mL, and use this solution as the テストステロンプロピオン酸エステル注射液 standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>.Spot10 mLeachofthesamplesolutionandstandardsolutionona Testosterone Propionate Injection is an oily solution plate of silica gel with ‰uorescent indicator for thin-layer for injection. chromatography. Develop the plate with a mixture of chlo- It contains not less than 92.5z and not more than roform and diethylamine (19:1) to a distance of about 15 cm, 107.5z of the labeled amount of testosterone and air-dry the plate. Examine under ultraviolet light (main propionate (C22H32O3:344.49). wavelength: 254 nm): the spot other than the principal spot from the sample solution is not more intense than the spot Method of preparation Prepare as directed under Injec- from the standard solution. tions, with Testosterone Propionate. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu- Description Testosterone Propionate Injection is a clear, um, phosphorus (V) oxide, 4 hours). colorless or pale yellow oily liquid. Residue on ignition <2.44> Not more than 0.1z (0.5 g). Identiˆcation Dissolve the residue obtained as directed in the procedure in the Assay in exactly 20 mL of methanol, and Assay Weigh accurately each about 10 mg of Testosterone 1156 Freeze-dried Tetanus Antitoxin, Equine / O‹cial Monographs JP XV use this solution as the sample solution. Separately, dissolve 1 methanol (9 in 100,000). mg of Testosterone Propionate Reference Standard in 10 mL Containers and storage Containers—Hermetic containers. of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin- layer Chromatography <2.03>.Spot10mLeachofthesample solution and standard solution on a plate of silica gel with Freeze-dried Tetanus Antitoxin, ‰uorescent indicator for thin-layer chromatography. Develop Equine the plate with a mixture of chloroform and diethylamine (19:1) to a distance of about 15 cm, and air-dry the plate. Ex- 乾燥破傷風ウマ抗毒素 amine under ultraviolet light (main wavelength: 254 nm): the Rf values of the principal spot with the sample solution and of the spot with the standard solution are not diŠerent each Freeze-dried Tetanus Antitoxin, Equine, is a prepa- other. ration for injection which is dissolved before use. Extractable volume <6.05> It meets the requirement. It contains tetanus antitoxin in immunoglobulin of horse origin. Foreign insoluble matter <6.06> Perform the test according It conforms to the requirements of Freeze-dried to Method 1: it meets the requirement. Tetanus Antitoxin, Equine, in the Minimum Require- Sterility <4.06> Perform the test according to the Membrane ments for Biological Products. ˆltration method: it meets the requirement. Description Freeze-dried Tetanus Antitoxin, Equine, Assay (i) Chromatographic tube A glass tube about 1 becomes a clear, colorless to light yellow-brown liquid or cm in inside diameter and about 18 cm in length, with a glass slightly white-turbid liquid on addition of solvent. ˆlter(G3)atthelowerend. (ii) Chromatographic column To about 2 g of silica gel for liquid chromatography add 5 mL of dichloromethane, Adsorbed Tetanus Toxoid and mix gently. Transfer and wash into the chromatographic tube with the aid of dichloromethane, allow to elute the 沈降破傷風トキソイド dichloromethane through the column, and put a ˆlter paper on the upper end of the silica gel. (iii) Standard solution Weigh accurately about 10 mg Adsorbed Tetanus Toxoid is a liquid for injection of Testosterone Propionate Reference Standard, previously containing tetanus toxoid prepared by treating tetanus dried at 1059C for 4 hours, and dissolve in methanol to make toxin with formaldehyde by a method involving no exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 appreciable loss of the immunogenicity and rendered mL of the internal standard solution and methanol to make insoluble by the addition of aluminum salt. exactly 20 mL. It conforms to the requirements of Adsorbed (iv) Sample stock solution To exactly a volume of Tetanus Toxoid in the Minimum Requirements for Testosterone Propionate Injection, equivalent to about Biological Products. 20 mg of testosterone propionate (C22H32O3), add Description Adsorbed Tetanus Toxoid becomes a uniform dichloromethanetomakeexactly20mL. white-turbid liquid on shaking. (v) Procedure Transfer exactly 2 mL of the sample stock solution into the chromatographic column, and elute to the upper surface of the silica gel. Wash the inner surface of Tetracaine Hydrochloride the chromatographic tube with 15 mL of dichloromethane, elute to the upper surface of the silica gel, and discard the テトラカイン塩酸塩 eŒuent. Elute 15 mL of a mixture of dichloromethane and methanol (39:1), discard the ˆrst 5 mL of the eŒuent, and collect the subsequent eŒuent. Wash the lower part of the column with a few amount of dichloromethane, combine the washings and the eŒuent, and evaporate the solvent under reduced pressure. Dissolve the residue so obtained with methanol to make exactly 20 mL. Pipet 5 mL of this solution, C15H24N2O2.HCl: 300.82 add exactly 5 mL of the internal standard solution and 2-(Dimethylamino)ethyl 4-(butylamino)benzoate methanol to make exactly 20 mL, and use this solution as the monohydrochloride [136-47-0] sample solution. Perform the test with 5 mLeachofthesam- ple solution and standard solution as directed in the Assay Tetracaine Hydrochloried, when dried, contains not under Testosterone Propionate. less than 98.5z of C15H25N2O2.HCl. Amount (mg) of testosterone propionate (C22H32O3) Description Tetracaine Hydrochloride occurs as white crys- ＝WS×(QT/QS)×2 tals or crystalline powder. It is odorless, and has a slightly bitter taste followed by a sense of numbness on the tongue. WS: Amount (mg) of Testosterone Propionate Reference Standard It is very soluble in formic acid, freely soluble in water, soluble in ethanol (95), sparingly soluble in ethanol (99.5), Internal standard solution—A solution of Progesterone in slightly soluble in acetic anhydride, and practically insoluble JP XV O‹cial Monographs / Tetracycline Hydrochloride 1157 in diethyl ether. duced by the growth of Streptomyces aureofaciens. A solution of Tetracaine Hydrochloride (1 in 10) is neutral. It contains not less than 950 mg (potency) and not Melting point: about 1489C more than 1010 mg (potency) per mg, calculated on the dried basis. The potency of Tetracycline Hydrochlo- Identiˆcation (1) Dissolve 0.5 g of Tetracaine Hydrochlo- ride is expressed as mass (potency) of tetracycline ride in 50 mL of water, add 5 mL of ammonia TS, shake, hydrochloride (C H N O .HCl). and allow to stand in a cold place. Collect the precipitate, 22 24 2 8 wash with water until the washings is neutral, and dry in a Description Tetracycline Hydrochloride occurs as a yellow desiccator (silica gel) for 24 hours: it melts <2.60> between 42 to pale brownish yellow crystalline powder. 9Cand449C. It is freely soluble in water, and sparingly soluble in (2) Dissolve 0.1 g of Tetracaine Hydrochloride in 8 mL ethanol (95). of water, and add 3 mL of ammonium thiocyanate TS: a Identiˆcation (1) Determine the absorption spectrum of a crystalline precipitate is produced. Collect the precipitate, solution of Tetracycline Hydrochloride (1 in 62,500) as recrystallize from water, and dry at 809C for 2 hours: it directed under Ultraviolet-visible Spectrophotometry <2.24>, melts <2.60> between 1309Cand1329C. and compare the spectrum with the Reference Spectrum or (3) Determine the absorption spectrum of a solution of the spectrum of a solution of Tetracycline Hydrochloride Tetracaine Hydrochloride in ethanol (99.5) (1 in 200,000) as Reference Standard prepared in the same manner as the sam- directed under Ultraviolet-visible Spectrophotometry <2.24>, ple solution: both spectra exhibit similar intensities of ab- and compare the spectrum with the Reference Spectrum: sorption at the same wavelengths. both spectra exhibit similar intensities of absorption at the (2) Determine the infrared absorption spectrum of same wavelengths. Tetracycline Hydrochloride as directed in the potassium chlo- (4) A solution of Tetracaine Hydrochloride (1 in 10) ride disk method under Infrared Spectrophotometry <2.25>, responds to the Qualitative Tests <1.09> for chloride. and compare the spectrum with the Reference Spectrum or Purity Heavy metals <1.07>—Proceed with 1.0 g of Tetra- the spectrum of Tetracycline Hydrochloride Reference Stan- caine Hydrochloride according to Method 1, and perform the dard: both spectra exhibit similar intensities of absorption at test. Prepare the control solution with 2.0 mL of Standard thesamewavenumbers. Lead Solution (not more than 20 ppm). (3) A solution of Tetracycline Hydrochloride (1 in 100) responds to the Qualitative Tests <1.09> (2) for chloride. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 4 hours). pH <2.54> Dissolve 1.0 g of Tetracycline Hydrochloride in 100 mL of water: the pH of the solution is between 1.8 and Residue on ignition <2.44> Not more than 0.1z (1 g). 2.8. Assay Weigh accurately about 0.5 g of Tetracaine Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Hydrochloride, previously dried, dissolve in 2 mL of formic Tetracycline Hydrochloride according to Method 2, and per- acid, add 80 mL of acetic anhydride, allow to stand at 309C form the test. Prepare the control solution with 1 mL of Stan- on a water bath for 15 minutes, cool, and titrate <2.50> with dard Lead Solution (not more than 10 ppm). 0.1 molWL perchloric acid VS (potentiometric titration). Per- (2) Arsenic <1.11>—Prepare the test solution with 1.0 g form a blank determination, and make any necessary correc- of Tetracycline Hydrochloride according to Method 4, and tion. perform the test (not more than 2 ppm). Each mL of 0.1 molWL perchloric acid VS (3) Related substances—Dissolve 25 mg of Tetracycline

＝ 30.08 mg of C15H24N2O2.HCl Hydrochloride in 50 mL of 0.01 molWL hydrochloric acid TS, and use this solution as the sample solution. Pipet 3 mL of Containers and storage Containers—Tight containers. the sample solution, add 0.01 molWL hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mLeachofthesam- Tetracycline Hydrochloride ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi- テトラサイクリン塩酸塩 tions, and calculate the areas of each peak by the automatic integration method: the peak areas other than tetracycline from the sample solution is not more than the peak area of tetracycline from the standard solution, and the total of the peak areas other than tetracycline from the sample solution is not more than 3 times of the peak area of tetracycline from the standard solution. Operating conditions—

C22H24N2O8.HCl: 480.90 Detector, column, column temperature, mobile phase, and (4S,4aS,5aS,6S,12aS)-4-Dimethylamino- ‰ow rate: Proceed as directed in the operating conditions in 3,6,10,12,12a-pentahydroxy-6-methyl-1,11-dioxo- the Assay. 1,4,4a,5,5a,6,11,12a-octahydrotetracene-2- Time span of measurement: About 7 times as long as the carboxamide monohydrochloride [64-75-5] retention time of tetracycline beginning after the solvent peak. Tetracycline Hydrochloride is the hydrochloride of a System suitability— tetracycline substance having antibacterial activity pro- Test for required detection: Pipet 3 mL of the standard so- 1158 Thallium (201Tl) Chloride Injection / O‹cial Monographs JP XV lution, add 0.1 molWL hydrochloric acid TS to make exactly Storage—Light-resistant. 100 mL, and conˆrm that the peak area of tetracycline ob- tained from 20 mL of this solution is equivalent to 1 to 5z of that of tetracycline obtained from 20 mL of the standard solu- Thallium (201Tl) Chloride Injection tion. System performance: Proceed as directed in the system 塩化タリウム (201Tl) 注射液 suitability in the Assay. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operating Thallium (201Tl) Chloride Injection is an aqueous conditions, the relative standard deviation of the peak areas solution for injection of tetracycline is not more than 1.0z. It contains thallium-201 (201Tl) in the form of thal- Loss on drying <2.41> Not more than 2.0z (1 g, in vacuum, lous chloride. 201 609C, 3 hours). It conforms to the requirements of Thallium ( Tl) Chloride Injection in the Minimum Requirements for Residue on ignition <2.44> Not more than 0.3z (1.0 g). Radiopharmaceuticals. Assay Weigh accurately an amount of Tetracycline Test for Extractable Volume of Parenteral Prepara- Hydrochloride and Tetracycline Hydrochloride Reference tions and Insoluble Particulate Matter Test for Injec- Standard, equivalent to about 25 mg (potency), and dissolve tions are not applied to this injection. each in 0.1 molWLhydrochloricacidTStomakeexactly50 Description Thallium (201Tl) Chloride Injection is a clear, mL, and use these solutions as the sample solution and stan- colorless liquid. dard solution, respectively. Perform the test with exactly 20 mLeachofthesesolutionsasdirectedunderLiquidChro- matography <2.01> according to the following conditions, Theophylline and calculate the peak area, AT and A S, of tetracycline of each solution. テオフィリン

Amount [mg (potency)] of C22H24N2O8.HCl ＝WS×(A T/A S)×1000

WS: Amount [mg (potency)] of Tetracycline Hydrochloride Reference Standard Operating conditions— Detector: An ultraviolet absorption photometer C7H8N4O2: 180.16 (wavelength: 254 nm). 1,3-Dimethyl-1H-purine-2,6(3H,7H)-dione [58-55-9] Column: A stainless steel column 4.6 mm in inside di- ameter and 25 cm in length, packed with stylene-divinylben- Theophylline, when dried, contains not less than zene copolymer for liquid chromatography (0.01 mminpore 99.0z of C7H8N4O2. diameter). Column temperature: A constant temperature of about Description Theophylline occurs as white crystals or crys- 609C. talline powder. Mobile phase: Dissolve 3.5 g of dipotassium hydrogen- It is soluble in N,N-dimethylformamide, and slightly solu- phosphate, 2.0 g of tetrabutylammonium hydrogensulfate ble in water and in ethanol (99.5). and 0.4 g of disodium dihydrogen ethylenediamine tetraa- It dissolves in 0.1 mol/L hydrochloric acid TS. cetate dihydrate in 300 mL of water, adjust to pH 9.0 with so- Identiˆcation (1) Determine the absorption spectrum of a dium hydroxide TS, add 90.0 g of t-butanol, and add water solution of Theophylline in 0.1 mol/L hydrochloric acid TS to make 1000 mL. (1 in 200,000) as directed under Ultraviolet-visible Spec- Flow rate: Adjust the ‰ow rate so that the retention time of trophotometry <2.24>, and compare the spectrum with the tetracycline is about 5 minutes. Reference Spectrum: both spectra exhibit similar intensities System suitability— of absorption at the same wavelengths. System performance: Dissolve 0.05 g of Tetracycline (2) Determine the infrared absorption spectrum of The- Hydrochloride Reference Standard in water to make 25 mL. ophylline, previously dried, as directed in the potassium Heat5mLofthissolutiononawaterbathfor60minutes, bromide disk method under Infrared Spectrophotometry then add water to make 25 mL. When the procedure is run <2.25>, and compare the spectrum with the Reference Spec- with 20 mL of this solution under the above operating condi- trum: both spectra exhibit similar intensities of absorption at tions, the retention time of 4-epitetracycline is about 3 thesamewavenumbers. minutes, and 4-epitetracycline and tetracycline are eluted in this order with the resolution between these peaks being not Melting point <2.60> 271 – 2759C less than 2.5. Purity (1) Acidity—To 0.5 g of Theophylline add 75 mL System repeatability: When, the test is repeated 6 times of water, 2.0 mL of 0.01 molWL sodium hydroxide VS and 1 with 20 mL of the standard solution under the above operat- drop of methyl red TS: a yellow color develops. ing conditions, the relative standard deviation of the peak (2) Heavy metals <1.07>—Proceed with 1.0 g of The- areas of tetracycline is not more than 1.0z. ophylline according to Method 4, and perform the test. Pre- Containers and storage Containers—Tight containers. JP XV O‹cial Monographs / Thiamazole 1159 pare the control solution with 2.0 mL of Standard Lead Solu- changes to blue. tion (not more than 20 ppm). (2) To 2 mL of a solution of Thiamazole (1 in 200) add 1 (3) Arsenic <1.11>—Prepare the test solution with 1.0 g mL of sodium carbonate TS and 1 mL of diluted Folin's TS of Theophylline according to Method 3, and perform the test (1 in 5): a deep blue color develops. (not more than 2 ppm). Melting point <2.60> 144 – 1479C (4) Related substances—Dissolve 0.10 g of Theophylline in 3 mL of N,N-dimethylformamide, add 10 mL of Purity (1) Selenium—Proceed with 0.10 g of Thiamazole methanol, and use this solution as the sample solution. Pipet as directed under Oxygen Flask Combustion Method <1.06>, 1 mL of the sample solution, add methanol to make exactly using 25 mL of diluted nitric acid (1 in 30) as the absorbing 200 mL, and use this solution as the standard solution. Per- liquid, and prepare the test solution. Apply a small amount form the test with these solutions as directed under Thin-lay- of water to the upper part of apparatus A, pull out C careful- er Chromatography <2.03>.Spot10mLeachofthesampleso- ly, and transfer the test solution to a beaker. Wash C, B and lution and standard solution on a plate of silica gel with the inner side of A with 25 mL of water, and combine the fluorescent indicator for thin-layer chromatography. De- washings with the test solution. Boil gently for 10 minutes, velop the plate with a mixture of acetone, chloroform, cool to room temperature, add water to make exactly 50 mL, methanol, 1-butanol and ammonia solution (28) (3:3:2:2:1) and use this solution as the sample solution. Separately, to a distance of about 10 cm, and air-dry the plate. Examine weighexactly40mgofselenium,dissolvein100mLofdilut- under ultraviolet light (main wavelength: 254 nm): the spots ed nitric acid (1 in 2), heat to dissolve on a water bath if other than the principal spot from the sample solution are not necessary, and add water to make exactly 1000 mL. Pipet 5 more intense than the spot from the standard solution. mL of this solution, and add water to make exactly 200 mL. To 2 mL of this solution, exactly measured, add diluted nitric Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, acid (1 in 60) to make exactly 50 mL, and use this solution as 4 hours). the standard solution. Pipet 40 mL each of the sample solu- Residue on ignition <2.44> Not more than 0.1z (1 g). tion and standard solution into separate beakers, and adjust each solution with ammonia solution (28) to a pH of 1.8 to Assay Weigh accurately about 0.25 g of Theophylline, 2.2. To each solution add 0.2 g of hydroxylammonium chlo- previously dried, and dissolve in 100 mL of water, add ex- ride, shake gently to dissolve, then add 5 mL of 2,3-di- actly 20 mL of 0.1 mol/L silver nitrate VS, shake the mix- aminonaphthalene TS, shake, and allow to stand for 100 ture, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS minutes. Transfer these solutions to corresponding separa- (potentiometric titration). Perform a blank determination, tors, rinse the beakers with 10 mL of water, combine the rins- and make any necessary correction. ings in the respective separators, shake well with 5.0 mL of Each mL of 0.1 mol/L sodium hydroxide VS cyclohexane for 2 minutes, and extract. Centrifuge the cyclo-

＝ 18.02 mg of C7H8N4O2 hexane extracts to remove any water remaining in these solu- tions. Perform the test with these solutions as directed under Containers and storage Containers—Well-closed contain- Ultraviolet-visible Spectrophotometry <2.24>,usingasolu- ers. tion prepared with 40 mL of diluted nitric acid (1 in 60) in the same manner as the blank. The absorbance of of the sample solution at the wavelength of maximum absorbance at about Thiamazole 378 nm does not exceed the absorbance of the standard solu- tion. チアマゾール (2) Heavy metals <1.07>—Proceed with 1.0 g of Thiama- zole 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). (3) Arsenic <1.11>—Prepare the test solution with 1.0 g of Thiamazole according to Method 1, and perform the test

C4H6N2S: 114.17 (not more than 2 ppm). 1-Methyl-1H--2-thiol [60-56-0] Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Thiamazole, when dried, contains not less than 98.0z of C4H6N2S. Residue on ignition <2.44> Not more than 0.1z (1 g). Description Thiamazole occurs as white to pale yellowish Assay Weigh accurately about 0.25 g of Thiamazole, previ- white crystals or crystalline powder. It has a faint, charac- ously dried, dissolve in 75 mL of water, add 15 mL of 0.1 teristic odor, and has a bitter taste. molWL sodium hydroxide VS from a burette, and add 30 mL It is freely soluble in water and in ethanol (95), and slightly of 0.1 molWL silver nitrate VS with stirring. Add 1 mL of soluble in diethyl ether. bromothymol blue TS, and titrate <2.50> with 0.1 molWLso- The pH of the solution (1 in 50) is between 5.0 and 7.0. dium hydroxide VS, until a persistent blue-green color is produced. Determine the total volume of 0.1 molWLsodium Identification (1) Dissolve 5 mg of Thiamazole in 1 mL of hydroxide VS consumed. water, shake with 1 mL of sodium hydroxide TS, and add 3 drops of sodium pentacyanonitrosylferrate (III) TS: a yellow Each mL of 0.1 molWL sodium hydroxide VS color develops, and it gradually changes to yellow-green to ＝ 11.42 mg of C4H6N2S green. To this solution add 1 mL of acetic acid (31): it Containers and storage Containers—Well-closed contain- 1160 Thiamazole Tablets / O‹cial Monographs JP XV ers. Storage—Light-resistant. Chloride Hydrochloride

Thiamazole Tablets Vitamin B1 Hydrochloride

チアマゾール錠 チアミン塩化物塩酸塩

Thiamazole Tablets contain not less than 94z and not more than 106z of the labeled amount of thiamazole (C4H6N2S: 114.17). C H ClN OS.HCl: 337.27 Method of preparation Prepare as directed under Tablets, 12 17 4 3-(4-Amino-2-methylpyrimidin-5-ylmethyl)-5-(2- with Thiamazole. hydroxyethyl)-4-methylthiazolium chloride Identification (1) To a quantity of powdered Thiamazole monohydrochloride [67-03-8] Tablets, equivalent to 0.05 g of Thiamazole according to the labeled amount, add 20 mL of hot ethanol (95), shake for 15 Thiamine Chloride Hydrochloride contains not less minutes, filter, and evaporate the filtrate on a water bath to than 98.5z of C12H17ClN4OS.HCl,calculatedonthe dryness. Dissolve the residue in 10 mL of water, filter if anhydrous basis. necessary, and use this solution as the sample solution. To 1 Description Thiamine Chloride Hydrochloride occurs as mL of the sample solution add 1 mL of sodium hydroxide white crystals or crystalline powder. It is odorless or has a TS, shake, and add 3 drops of sodium pentacyanonitrosylfer- slight, characteristic odor. rate (III) TS: a yellow color develops, and it gradually It is freely soluble in water, sparingly soluble in methanol, changestoyellow-greentogreen.Tothissolutionadd1mL slightly soluble in ethanol (95), and practically insoluble in of acetic acid (31): it changes to blue. diethyl ether. (2) With 2 mL of the sample solution obtained in (1), Melting point: about 2459C (with decomposition). proceed as directed in the Identification (2) under Thiama- zole. Identification (1) To 5 mL of a solution of Thiamine Chloride Hydrochloride (1 in 500) add 2.5 mL of sodium Assay Weigh accurately and powder not less than 20 hydroxide TS and 0.5 mL of potassium hexacyanoferrate Thiamazole Tablets. Weigh accurately a quantity of the pow- (III) TS. Then add 5 mL of 2-methyl-1-propanol, shake the der, equivalent to about 0.15 g of thiamazole (C H N S), add 4 6 2 mixture vigorously for 2 minutes, allow to stand, and exa- 80 mL of water, shake for 15 minutes, add water to make ex- mine under ultraviolet light (main wavelength: 365 nm): the actly 100 mL, and centrifuge. Filter, discard the first 20 mL 2-methyl-1-propanol layer shows a blue-purple fluorescence. of the filtrate, pipet 50 mL of the subsequent filtrate, add 1 This fluorescence disappears when the mixture is acidified, mL of bromothymol blue TS, and if a blue color develops, but reappears when it is again made alkaline. neutralize with 0.1 molWL hydrochloric acid VS until the (2) Determine the absorption spectrum of a solution of colorofthesolutionchangestogreen.Tothissolutionadd Thiamine Chloride Hydrochloride (1 in 100,000) as directed 4.5 mL of 0.1 molWL sodium hydroxide VS from a burette, under Ultraviolet-visible Spectrophotometry <2.24>,and add 15 mL of 0.1 molWL silver nitrate VS while stirring, and compare the spectrum with the Reference Spectrum or the titrate <2.50> with 0.1 molWL sodium hydroxide VS. Continue spectrum of a solution of Thiamine Chloride Hydrochloride the titration until a persistent blue-green color is produced, Reference Standard prepared in the same manner as the sam- and determine the total volume of 0.1 molWLsodium ple solution: both spectra exhibit similar intensities of ab- hydroxide VS consumed. sorption at the same wavelengths. Each mL of 0.1 molWL sodium hydroxide VS (3) Determine the infrared absorption spectrum of Thia- ＝ 11.42 mg of C4H6N2S mine Chloride Hydrochloride, previously dried at 1059Cfor 2 hours, as directed in the potassium bromide disk method Containers and storage Containers—Well-closed contain- under the Infrared Spectrophotometry <2.25>,andcompare ers. the spectrum with the Reference Spectrum, or the spectrum Storage—Light-resistant. of Thiamine Chloride Hydrochloride Reference Standard previously driedat 1059C for 2 hours: both spectra exhibit similar intensities of absorption at the same wave numbers. In case when some differences are found between the spectra, repeat the test with residues obtained by dissolving these sub- stances in water, evaporating to dryness, and drying at 1059C for 2 hours. (4) A solution of Thiamine Chloride Hydrochloride (1 in 500) responds to the Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve1.0gofThiamineChlorideHydrochlo- ride in 100 mL of water: the pH of this solution is between 2.7 and 3.4. JP XV O‹cial Monographs / Thiamine Chloride Hydrochloride Injection 1161

as directed under Liquid Chromatography <2.01> according Purity (1) Clarity and color of solution—Dissolve 1.0 g of to the following conditions, and calculate the ratios, Q and Thiamine Chloride Hydrochloride in 10 mL of water: the so- T Q , of the peak area of thiamine to that of the internal stan- lution is clear, and has no more color than the following con- S dard. trol solution.

Control solution: To 1.5 mL of 1/60 molWL potassium Amount (mg) of C12H17ClN4OS.HCl dichromate VS add water to make 1000 mL. ＝WS×(QT/QS) (2) Sulfate <1.14>—Weigh 1.5 g of Thiamine Chloride W : Amount (mg) of Thiamine Chloride Hydrochloride Hydrochloride, and perform the test. Prepare the control so- S Reference Standard, calculated on the anhydrous lution with 0.35 mL of 0.005 molWL sulfuric acid VS (not basis more than 0.011z). (3) Nitrate—Dissolve 0.5 g of Thiamine Chloride Internal standard solution—A solution methyl benzoate in Hydrochloride in 25 mL of water. Add 2 mL of sulfuric acid methanol (1 in 50). to 2 mL of this solution, shake, cool, and superimpose iron Operating conditions— (II) sulfate TS: no dark brown ring is produced at the junc- Detector: An ultraviolet absorption photometer tion of the two layers. (wavelength: 254 nm). (4) Heavy metals <1.07>—Proceed with 1.0 g of Thiamine Column: A stainless steel column 4.6 mm in inside Chloride Hydrochloride according to Method 1, and perform diameter and 15 cm in length, packed with octadecylsilanized the test. Prepare the control solution with 2.0 mL of Stan- silica gel for liquid chromatography (5 mminparticledi- dard Lead Solution (not more than 20 ppm). ameter). (5) Related substances—Dissolve 0.10 g of Thiamine Column temperature: A constant temperature of about Chloride Hydrochloride in 100 mL of the mobile phase, and 259C. use this solution as the sample solution. Pipet 1 mL of the Mobile phase: Dissolve 1.1 g of sodium 1-octanesulfonate sample solution, add the mobile phase to make exactly 100 in 1000 mL of diluted acetic acid (100) (1 in 100). To 600 mL mL, and use this solution, as the standard solution. Perform of this solution add 400 mL of a mixture of methanol and the test with exactly 10 mL each of the sample solution and acetonitrile (3:2). standard solution as directed under Liquid Chromatography Flow rate: Adjust the ‰ow rate so that the retention time of <2.01> according to the following conditions, and determine thiamine is about 12 minutes. the area of each peak by the automatic integration method: System suitability— the total area of the peaks other than thiamine is not larger System performance: When the procedure is run with than the peak area of thiamine from the standard solution. 10 mL of the standard solution under the above operating Operating conditions— conditions, thiamine and the internal standard are eluted in Detector, column, column temperature, mobile phase, and this order with the resolution between these peaks being not ‰ow rate: Proceed as directed in the operating conditions in less than 6. the Assay. System repeatability: When the test is repeated 6 times with Time span of measurement: About 3 times as long as the 10 mL of the standard solution under the above operating retention time of thiamine. conditions, the relative standard deviation of the ratio of the System suitability— peak area of thiamine to that of the internal standard is not Test for required detectability: To exactly 5 mL of the stan- more than 1.0z. dard solution add water to make exactly 50 mL. Conˆrm that Containers and storage Containers—Tight containers. the peak area of thiamine obtained from 10 mLofthissolu- Storage—Light-resistant. tion is equivalent to 7 to 13z of that of thiamine obtained from 10 mL of the standard solution. System performance: Proceed as directed in the system suitability in the Assay. Thiamine Chloride Hydrochloride System repeatability: When the test is repeated 6 times with Injection 10 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of Vitamin B1 Hydrochloride Injection thiamine is not more than 1.0z. チアミン塩化物塩酸塩注射液 Water <2.48> Not more than 5.0z (30 mg, coulometric titration). Thiamine Chloride Hydrochloride Injection is an Residue on ignition <2.44> Not more than 0.2z (1 g). aqueous solution for injection. Assay Weigh accurately about 0.1 g each of Thiamine It contains not less than 95z and not more than Chloride Hydrochloride and Thiamine Chloride Hydrochlo- 115z of the labeled amount of thiamine Chloride ride Reference Standard (separately determine the water hydrochloride (C12H17ClN4OS.HCl: 337.27). <2.48> in the same manner as Thiamine Chloride Hydrochlo- Method of preparation Prepare as directed under Injec- ride), and dissolve them in the mobile phase to make exactly tions, with Thiamine Chloride Hydrochloride. 50 mL. To 10 mL each of the solutions, accurately measured, add exactly 5 mL each of the internal standard solution, add Description Thiamine Chloride Hydrochloride Injection is the mobile phase to make 50 mL, and use these solutions as a clear, colorless liquid. the sample solution and standard solution. Perform the test pH:2.5–4.5 with 10 mL each of the sample solution and standard solution Identification To a volume of Thiamine Chloride 1162 Thiamine Chloride Hydrochloride Powder / O‹cial Monographs JP XV

Hydrochloride Injection, equivalent to 0.05 g of Thiamine Purity Rancidity—Thiamine Chloride Hydrochloride Pow- Chloride Hydrochloride according to the labeled amount, der has no unpleasant or rancid odor. It is tasteless. addwatertomake25mL.Proceedwith5mLofthissolution as directed in the Identification (1) under Thiamine Chloride Assay Weigh accurately a quantity of Thiamine Chloride Hydrochloride. Hydrochloride Powder, equivalent to about 20 mg of thia- mine chloride hydrochloride (C H ClN OS.HCl), add 60 Extractable volume <6.05> It meets the requirement. 12 17 4 mL of 0.01 molWLhydrochloricacidTS,andheatonawater Assay Dilute with 0.001 molWL hydrochloric acid TS if bath for 30 minutes. Shake vigorously for 10 minutes, cool, necessary, then measure exactly a volume of Thiamine Chlo- add methanol to make exactly 100 mL, and centrifuge. Pipet ride Hydrochloride Injection, equivalent to about 20 mg of 25 mL of the supernatant, add exactly 5 mL of the internal thiamine chloride hydrochloride (C12H17ClN4OS.HCl), and standard solution, add water to make 50 mL, and use this so- add 20 mL of methanol and 0.001 molWLhydrochloricacid lution as the sample solution. Separately, weigh accurately TS to make 100 mL. To 25 mL of this solution, exactly meas- about 0.1 g of Thiamine Chloride Hydrochloride Reference ured, add exactly 5 mL of the internal standard solution, add Standard (separately determine the water <2.48> in the same 0.001 molWL hydrochloric acid TS to make 50 mL, and use manner as Thiamine Chloride Hydrochloride), and dissolve this solution as the sample solution. Separately, weigh ac- in 0.01 molWL hydrochloric acid TS to make exactly 50 mL. curately about 0.1 g of Thiamine Chloride Hydrochloride To 10 mL of this solution, exactly measured, add 50 mL of Reference Standard (separately determine the water <2.48> in 0.01 molWL hydrochloric acid TS, and add methanol to make the same manner as Thiamine Chloride Hydrochloride), and exactly 100 mL. To 25 mL of this solution, exactly measured, dissolve in 0.001 molWL hydrochloric acid TS to make exactly add exactly 5 mL of the internal standard solution, add water 50 mL. To 10 mL of this solution, exactly measured, add 20 to make 50 mL, and use this solution as the standard solu- mL of methanol and 0.001 molWL hydrochloric acid TS to tion. Proceed as directed in the Assay under Thiamine Chlo- make exactly 100 mL. To 25 mL of this solution, exactly ride Hydrochloride. measured, add exactly 5 mL of the internal standard solu- Amount (mg) of thiamine chloride hydrochloride tion, add 0.001 molWLhydrochloricacidTStomake50mL, (C H ClN OS.HCl) and use this solution as the standard solution. Proceed as 12 17 4 ＝ W ×(Q /Q )×(1/5) directed in the Assay under Thiamine Chloride Hydrochlo- S T S ride. WS: Amount (mg) of Thiamine Chloride Hydrochloride Reference Standard, calculated on the anhydrous ba- Amount (mg) of thiamine chloride hydrochloride sis (C12H17ClN4OS.HCl) ＝ WS×(QT/QS)×(1/5) Internal standard solution—A solution of methyl benzoate in methanol (1 in 200). WS: Amount (mg) of Thiamine Chloride Hydrochloride Reference Standard, calculated on the anhydrous Containers and storage Containers—Tight containers. basis Storage—Light-resistant. Internal standard solution—A solution of methyl benzoate in methanol (1 in 200). Thiamine Nitrate Containers and storage Containers—Hermetic containers. Storage—Light-resistant. Vitamin B1 Nitrate

チアミン硝化物 Thiamine Chloride Hydrochloride Powder

Vitamin B1 Hydrochloride Powder

チアミン塩化物塩酸塩散 C12H17N5O4S: 327.36 3-(4-Amino-2-methylpyrimidin-5-ylmethyl)-5-(2- hydroxyethyl)-4-methylthiazolium nitrate [532-43-4] Thiamine Chloride Hydrochloride Powder contains not less than 95z and not more than 115z of the la- Thiamine Nitrate, when dried, contains not less than beled amount of thiamine chloride hydrochloride 98.0 z and not more than 102.0 z of (C12H17ClN4OS.HCl: 337.27). C12H17N5O4S. Method of preparation Prepare as directed under Powders, Description Thiamine Nitrate occurs as white crystals or with Thiamine Chloride Hydrochloride. crystalline powder. It is odorless or a slight, characteristic Identification To a portion of Thiamine Chloride odor. Hydrochloride Powder, equivalent to 0.02 g of Thiamine It is sparingly soluble in water, and very slightly soluble in Chloride Hydrochloride according to the labeled amount, ethanol (95), and practically insoluble in diethyl ether. add 50 mL of water and 10 mL of dilute acetic acid, shake, Melting point: about 1939C (with decomposition). and filter. Proceed with 5 mL of the filtrate as directed in the Identification (1) Take 2-mL portions of a solution of Identification (1) under Thiamine Chloride Hydrochloride. Thiamine Nitrate (1 in 500), and add 2 to 3 drops of iodine JP XV O‹cial Monographs / Sodium 1163

TS: a red-brown precipitate or turbidity is produced. Upon Internal standard solution—A solution of methyl benzoate in further addition of 1 mL of 2,4,6-trinitrophenol TS, a yellow methanol (1 in 50). precipitate or turbidity is produced. Operating conditions— (2) To 1 mL of a solution of Thiamine Nitrate (1 in 500) Detector: An ultraviolet spectrophotometer (wavelength: add 1 mL of lead (II) acetate TS and 1 mL of a solution of so- 254 nm). dium hydroxide (1 in 10), and warm: the color of the solution Column: A stainless steel column 4.6 mm in inside di- changes through yellow to brown, and on standing, a black- ameter and 15 cm in length, packed with octadecylsilanized brown precipitate is produced. silica gel for liquid chromatography (5 mminparticledi- (3) To 5 mL of a solution of Thiamine Nitrate (1 in 500) ameter). add 2.5 mL of sodium ;hydroxide TS and 0.5 mL of potassi- Column temperature: A constant temperature of about um hexacyanoferrate (III) TS. Then add 5 mL of 2-methyl-1- 309C. propanol, shake the mixture vigorously for 2 minutes, allow Mobile phase: Dissolve 1.1 g of sodium l-octanesulfonate to stand, and examine under ultraviolet light (main in 1000 mL of diluted acetic acid (100) (1 in 100). To 600 mL wavelength: 365 nm): the 2-methyl-1-propanol layer shows a of this solution add 400 mL of a mixture of methanol and blue-purple fluorescence. This fluorescence disappears when acetonitrile (3:2). the mixture is acidified, but reappears when it is again made Flow rate: Adjust the ‰ow rate so that the retention time of alkaline. thiamine is about 12 minutes. (4) A solution of Thiamine Nitrate (1 in 50) responds to System suitability— the Qualitative Tests <1.09> (1) and (2) for nitrate. System performance: When the procedure is run with 10 pH <2.54> Dissolve 1.0 g of Thiamine Nitrate in 100 mL of mL of the standard solution under the above operating condi- water: the pH of this solution is between 6.5 and 8.0. tions, thiamine and the internal standard are eluted in this order with the resolution between these peaks being not less Purity (1) Chloride <1.03>—Perform the test with 0.20 g than 6. of Thiamine Nitrate. Prepare the control solution with 0.30 System repeatability: When the test is repeated 6 times with mL of 0.01 molWL hydrochloric acid VS (not more than 10 mL of the standard solution under the above operating 0.053z). conditions, the relative standard deviation of the ratios of the (2) Sulfate <1.14>—Dissolve 1.5 g of Thiamine Nitrate in peak area of thiamine to that of the internal standard is not 30 mL of water and 2 mL of dilute hydrochloric acid, and more than 1.0z. add water to make 50 mL. Perform the test using this solu- tion as the test solution. Prepare the control solution with Containers and storage Containers—Tight containers. 0.35 mL of 0.005 molWL sulfuric acid VS and 2 mL of dilute Storage—Light-resistant. hydrochloric acid, and add water to make 50 mL (not more than 0.011z). (3) Heavy metals <1.07>—Dissolve 1.0 g of Thiamine Ni- Thiamylal Sodium trate in 30 mL of water by warming, cool, and add 12 mL of 6molWL acetic acid TS and water to make 50 mL. Perform チアミラールナトリウム the test with this solution as the test solution. 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 1.0z (0.5 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.2z (1 g). Assay Weigh accurately about 0.1 g each of Thiamine Ni- C12H17N2NaO2S: 276.33 trate, previously dried, and Thiamine Chloride Hydrochlo- Monosodium 5-allyl-5-[(1RS)-1-methylbutyl]-4,6- ride Reference Standard (separately determine the water dioxo-1,4,5,6-tetrahydropyrimidine-2-thiolate [337-47-3] <2.48> in the same manner as Thiamine Chloride Hydrochlo- ride), and dissolve them in the mobile phase to make exactly Thiamylal Sodium contains not less than 97.5z and 50 mL. To 10 mL each of the solutions, accurately measured, not more than 101.0z of C12H17N2NaO2S, calculated add exactly 5 mL each of the internal standard solution, add on the dried basis. the mobile phase to make 50 mL, and use these solutions as Description Thiamylal Sodium occurs as light yellow crys- the sample solution and standard solution. Perform the test tals or powder. with 10 mL each of the sample solution and standard solution It is very soluble in water, and freely soluble in ethanol as directed under Liquid Chromatography <2.01> according (95). to the following conditions and calculate the ratios, Q T and The pH of a solution of Thiamylal Sodium (1 in 10) is QS, of the peak area of thiamine to that of the internal stan- between 10.0 and 11.0. dard. It is hygroscopic. It is gradually decomposed by light. Amount (mg) of C12H17N5O4S ＝ WS × (Q T/QS) × 0.9706 Its solution in ethanol (95) (1 in 10) shows no optical WS: Amount (mg) of Thiamine Chloride Hydrochloride rotation. Reference Standard, calculated on the anhydrous ba- sis Identiˆcation (1) Determine the absorption spectrum of a solution of Thiamylal Sodium in ethanol (95) (7 in 1,000,000) 1164 Thiamylal Sodium for Injection / O‹cial Monographs JP XV

as directed under Ultraviolet-visible Spectrophotometry ＝WS×(QT/QS)×10×1.0864 <2.24>, and compare spectrum with the Reference Spectrum: W : Amount (mg) of Thiamylal Reference Standard both spectra exhibit similar intensities of absorption at the S same wavelengths. Internal standard solution—A solution of phenyl benzoate in (2) Determine the infrared absorption spectrum of methanol (3 in 500). Thiamylal Sodium, previously dried, as directed in the potas- Operating conditions— sium bromide disk method under Infrared Detector: An ultraviolet absorption photometer Spectrophotometry <2.25>, and compare the spectrum with (wavelength: 289 nm). the Reference Spectrum: both spectra exhibit similar intensi- Column: A stainless steel column about 4 mm in inside ties of absorption at the same wave numbers. diameter and about 15 cm in length, packed with octadecyl- (3) A solution of Thiamylal Sodium (1 in 10) responds to silanized silica gel for liquid chromatography (5 mmin Qualitative Tests <1.09> for sodium salt. particle diameter). Column temperature: A constant temperature of about Purity (1) Clarity and color of solution—To 1.0 g of 259C. Thiamylal Sodium in a 11- to 13-mL glass-stoppered test tube Mobile phase: A mixture of methanol and 0.05 mol/L add10mLoffreshlyboiledandcooledwater,stoppertight- acetic acid-sodium acetate buŠer solution, pH 4.6 (13:7). ly, allow to stand, and dissolve by occasional gentle shaking: Flow rate: Adjust the ‰ow rate so that the retention time of the solution is clear and light yellow. thiamylal is about 6 minutes. (2) Heavy metals <1.07>—Proceed with 1.0 g of System suitability— Thiamylal Sodium according to Method 2, and perform the System performance: When the procedure is run with test. Prepare the control solution with 2.0 mL of Standard 20 mL of the standard solution under the above operating Lead Solution (not more than 20 ppm). conditions, thiamylal and the internal standard are eluted in (3) Related substances—Dissolve 0.10 g of Thiamylal So- this order with the resolution between these peaks being not dium in 10 mL of ethanol (95), and use this solution as the less than 12. sample solution. Pipet 1 mL and 3 mL of the sample solu- System repeatability: When the test is repeated 6 times with tion, add ethanol (95) to make exactly 200 mL, and use these 20 mL of the standard solution under the above operating solutions as the standard solution (1) and the standard solu- conditions, the relative standard deviation of the ratio of the tion (2). Perform the test with these solutions as directed peak area of thiamylal to that of the internal standard is not under Thin-layer Chromatography <2.03>.Spot10mLeach more than 1.0z. of the sample solution and standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography, develop Containers and storage Containers—Tight containers. with a mixture of toluene, methanol and ethyl acetate Storage—Light-resistant. (40:7:3) to a distance of about 12 cm, and air-dry the plate. Allow the plate to stand in iodine vapor for a night: the spot appeared around Rf 0.1 obtained with the sample solution is Thiamylal Sodium for Injection not more intense than the spot with the standard solution (2), and the spot other than the principal spot, the spot at origin 注射用チアミラールナトリウム and the spot mentioned above obtained with the sample solu- tion is not more intense than the spot with the standard solu- tion (1). Thiamylal Sodium for Injection is a preparation for injection which is dissolved before use. Loss on drying <2.41> Not more than 2.0z (1 g, 1059C, It contains not less than 93.0z and not more than 1hour). 107.0z of the labeled amount of thiamylal sodium Assay Weigh accurately about 0.25 g of Thiamylal Sodium, (C12H17N2NaO2S: 276.33). dissolve in 50 mL of methanol and 5 mL of dilute Method of preparation Prepare as directed under Injec- hydrochloric acid, and add methanol to make exactly 100 tions, with 100 parts of Thiamylal Sodium and 7 parts of mL. Pipet 10 mL of this solution, and add methanol to make Dried Sodium Carbonate in mass. exactly 100 mL. Pipet 5 mL of this solution, add exactly 10 Description Thiamylal Sodium for Injection occurs as light mL of the internal standard solution and the mobile phase to yellow crystals, powder or masses. make 200 mL, and use this solution as the sample solution. It is hygroscopic. Separately, weigh accurately about 23 mg of Thiamylal It is gradually decomposed by light. Reference Standard, previously dried at 1059Cfor1hour, dissolve in 50 mL of methanol and 0.5 mL of dilute Identiˆcation (1) To 1.0 g of Thiamylal Sodium for hydrochloric acid, and add methanol to make exactly 100 Injection add 20 mL of ethanol (95), shake vigorously, and mL. Pipet 5 mL of this solution, add exactly 10 mL of the in- ˆlter. Dissolve the precipitate so obtained in 1 mL of water, ternal standard solution and the mobile phase to make 200 and add 1 mL of barium chloride TS: a white precipitate is mL, and use this solution as the standard solution. Perform produced. Centrifuge this solution, take oŠ the supernatant the test with 20 mL each of the sample solution and standard liquid, and to the precipitate add dilute hydrochloric acid solution as directed under Liquid Chromatography <2.01> ac- dropwise: the precipitate dissolves with eŠervescence. cording to the following conditions, and determine the ratios, (2) To 50 mg of Thiamylal Sodium for Injection add QT and QS, of the peak area of thiamylal to that of the inter- 100 mL of ethanol (95), shake vigorously, and ˆlter. To 3 mL nal standard. of the ˆltrate add ethanol (95) to make 200 mL. Determine the absorption spectrum of this solution as directed under Amount (mg) of C12H17N2NaO2S JP XV O‹cial Monographs / Compound Thianthol and Salicylic Acid Solution 1165

Ultraviolet-visible Spectrophotometry <2.24>: it exhibits max- Identiˆcation To 0.1 g of Thianthol add cautiously 5 mL of ima between 236 nm and 240 nm, and between 287 nm and sulfuric acid: a blue-purple color develops. Add 5 to 6 drops 291 nm. of nitric acid to the solution: the color of the solution changes pH <2.54> The pH of a solution obtained by dissolving 1.0 g to yellow-red with evolution of gas. of Thiamylal Sodium for Injection in 40 mL of water is be- Purity (1) Acidity or alkalinity—Shake 10 g of Thianthol tween 10.5 and 11.5. with 20 mL of water, allow to stand, and separate the water Purity Related substances—To 0.10 g of Thiamylal Sodium layer. The solution is neutral. for Injection add 10 mL of ethanol (95), shake vigorously, (2) Sulfate—To 10 mL of the water layer obtained in (1) ˆlter, and use the ˆltrate as the sample solution. Proceed as add 2 to 3 drops of barium chloride TS: no opalescence is diected in the Purity (3) under Thiamylal Sodium. produced. Bacterial endotoxins <4.01> Less than 1.0 EU/mg. Residue on ignition <2.44> Not more than 0.1z (1 g). Uniformity of dosage units <6.02> It meets the requirement Assay Weigh accurately about 10 mg of Thianthol, and of the Mass variation test. proceed as directed in the sulfur determination of Oxygen Flask Combustion Method <1.06>, using a mixture of 5 mL Foreign insoluble matter <6.06> Perform the test according of diluted sodium hydroxide TS (1 in 10) and 1.0 mL of to Method 2: it meets the requirement. hydrogen peroxide TS as an absorbing liquid. Insoluble particulate matter <6.07> Perform the test accord- Containers and storage Containers—Tight containers. ing to Method 1: it meets the requirement. Sterility <4.06> Perform the test according to the Membrane ˆltration method: it meets the requirement. Compound Thianthol and Assay Open carefully 10 containers of Thiamylal Sodium Salicylic Acid Solution for Injection, dissolve the contents with water, wash out the inside of each container with water, combine them, and add 複方チアントール・サリチル酸液 water to make exactly V mL so that each mL contains about 5 mg of thiamylal sodium (C12H17N2NaO2S). Pipet 5 mL of this solution, and add 0.5 mL of dilute hydrochloric acid and Compound Thianthol and Salicylic Acid Solution methanol to make exactly 100 mL. Pipet 5 mL of this solu- contains not less than 1.8 wWvz and not more than 2.2 tion, add exactly 10 mL of the internal standard solution and wWvz of salicylic acid (C7H6O3: 138.12), and not less the mobile phase to make 200 mL, and use this solution as the than 1.8 w Wvz and not more than 2.2 w Wvz of phenol sample solution. Proceed the test with the sample solution as (C6H6O: 94.11). directed in the Assay under Thiamylal Sodium. Method of preparation Amount (mg) of thiamylal sodium (C12H17N2NaO2S) in 1 container Thianthol 200 mL Salicylic Acid 20 g ＝WS×(QT/QS)×(V/50)×1.0864 Phenol 20 g WS: Amount (mg) of Thiamylal Reference Standard Olive Oil 50 mL Internal standard solution—A solution of phenyl benzoate in Ether 100 mL methanol (3 in 500). Petroleum Benzin a su‹cient quantity To make 1000 mL Containers and storage Containers—Hermetic containers. Storage—Light-resistant. Dissolve Salicylic Acid and Phenol in Ether, add Thian- thol, Olive Oil and Petroleum Benzin to this solution, mix and dissolve to make 1000 mL. Thianthol Description Compound Thianthol and Salicylic Acid Solu- tion is a light yellow liquid, having a characteristic odor. チアントール Identiˆcation (1) Place 1 mL of Compound Thianthol and Salicylic Acid Solution to a porcelain dish, and evaporate Thianthol consists of dimethylthianthrene and ditol- on a water bath to dryness. To the residue add cautiously 5 uene disulˆde. mL of sulfuric acid: the color of the solution changes to yel- It contains not less than 23.5z and not more than low-red with evolution of gas (thianthol). 26.5z of sulfur (S: 32.07). (2) Shake 10 mL of Compound Thianthol and Salicylic Acid Solution with 10 mL of sodium hydrogen carbonate TS, Description Thianthol is a yellowish, viscous liquid. It has a and separate the water layer. To 0.5 mL of the water layer faint, agreeable odor. add hydrochloric acid-potassium chloride buŠer solution, pH It is freely soluble in diethyl ether, slightly soluble in 2.0, to make 50 mL, and to 5 mL of this solution add 5 mL of ethanol (95), and practically insoluble in water. a solution of iron (III) nitrate enneahydrate (1 in 200): a red- It, when cold, may separate crystals, which melt on warm- purple color is produced (salicylic acid). ing. (3) Wash the upper phase obtained in (2) with 10 mL of Speciˆc gravity d20: 1.19 – 1.23 20 sodium hydrogen carbonate TS, and extract with 10 mL of 1166 Thiopental Sodium / O‹cial Monographs JP XV dilute sodium hydroxide TS. Shake 1 mL of the extract with 1 Flow rate: Adjust the ‰ow rate so that the retention time of mL of sodium nitrate TS and 1 mL of dilute hydrochloric salicylic acid is about 6 minutes. acid, and add 3 mL of sodium hydroxide TS: a yellow color is Selection of column: Dissolve 0.2 g of benzoic acid, 0.2 g produced (phenol). of salicylic acid and 0.05 g of theophylline in 100 mL of dilut- (4) To 1 mL of Compound Thianthol and Salicylic Acid ed methanol (1 in 2). To 10 mL of this solution add 90 mL of Solution add 10 mL of ethanol (95), mix, and use this solu- diluted methanol (1 in 2). Proceed with 10 mL of this solution tion as the sample solution. Dissolve 0.01 g each of salicylic under the above operating conditions. Use a column giving acid, phenol and thianthol in 5 mL each of ethanol (95), and elution of benzoic acid, salicylic acid and theophylline in this use each solution as standard solutions (1), (2) and (3). Per- order, and clearly dividing each peak. form the test with these solutions as directed under Thin-lay- Containers and storage Containers—Tight containers. er Chromatography <2.03>.Spot5mLeachofthesampleso- Storage—Light-resistant, and not exceeding 259C. lution and standard solutions on a plate of silica gel with ‰uorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform, acetone and acetic acid (100) (45:5:1) to a distance of about 10 cm, and air-dry Thiopental Sodium the plate. Examine under ultraviolet light (main wavelength: チオペンタールナトリウム 254 nm): three spots obtained from the sample solution and the corresponding spots of standard solutions (1), (2) and (3) show the same Rf value. Spray evenly iron (III) chloride TS on the plate: the spot from standard solution (1) and the cor- responding spot from the sample solution reveal a purple color. Assay Measure exactly 2 mL of Compound Thianthol and C H N NaO S: 264.32 Salicylic Acid Solution, add exactly 10 mL of the internal 11 17 2 2 Monosodium 5-ethyl-5-[(1RS)-1-methylbutyl]-4,6- standard solution, then add 70 mL of diluted methanol (1 in dioxo-1,4,5,6-tetrahydropyrimidine-2-thiolate [71-73-8] 2),mixwell,andadddilutedmethanol(1in2)tomake100 mL. Filter, discard the ˆrst 10 mL of the ˆltrate, and use the Thiopental Sodium, when dried, contains not less subsequent ˆltrate as the sample solution. Weigh accurately than 97.0z of C H N NaO S. about 0.2 g of salicylic acid for assay, previously dried in a 11 17 2 2 desiccator (silica gel) for 3 hours, and about 0.2 g of phenol Description Thiopental Sodium occurs as a light yellow for assay, dissolve in diluted methanol (1 in 2) to make exact- powder. It has a faint, characteristic odor. ly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of It is very soluble in water, freely soluble in ethanol (95), the internal standard solution and diluted methanol (1 in 2) to and practically insoluble in diethyl ether. make 100 mL, and use this solution as the standard solution. A solution of Thiopental Sodium (1 in 10) is alkaline. With 5 mL each of the sample solution and standard solution, It is hygroscopic. perform the test as directed under Liquid Chromatography Its solution gradually decomposes on standing. <2.01> according to the following conditions, and calculate Identification (1) Dissolve 0.2 g of Thiopental Sodium in the ratios, Q and Q , of the peak area of salicylic acid and Ta Tb 5 mL of sodium hydroxide TS, and add 2 mL of lead (II) phenol to that of the internal standard in the sample solution, acetate TS: a white precipitate, which dissolves upon heating, and the ratios, Q and Q , of the peak area of salicylic acid Sa Sb is produced. Boil the solution thus obtained: a black and phenol to that of the internal standard in the standard so- precipitate forms gradually, and the precipitate responds to lution. the Qualitative Tests <1.09> for . Amount (mg) of salicylic acid (C7H6O3) (2) Dissolve 0.5 g of Thiopental Sodium in 15 mL of ＝WSa×(QTa/QSa)×(1/5) water, add 10 mL of dilute hydrochloric acid to produce white precipitate, and extract with four 25-mL portions of Amount (mg) of phenol (C H O) 6 6 chloroform. Combine the chloroform extracts, evaporate on ＝W ×(Q /Q )×(1/5) Sb Tb Sb a water bath, and dry at 1059C for 2 hours: the residue melts WSa: Amount (mg) of salicylic acid for assay <2.60> between 1579Cand1629C. WSb: Amount (mg) of phenol for assay (3) A solution of Thiopental Sodium (1 in 10) responds to the Qualitative Tests <1.09> (1) and (2) for sodium salt. Internal standard solution—A solution of theophylline in methanol (1 in 1000). Purity (1) Clarity and color of solution—Dissolve 1.0 g of Operating conditions— Thiopental Sodium in 10 mL of freshly boiled and cooled Detector: An ultraviolet absorption photometer water: the solution is clear and light yellow. (wavelength: 270 nm). (2) Heavy metals <1.07>—Dissolve 2.0 g of Thiopental Column: A stainless steel column about 4 mm in inside di- Sodium in 76 mL of water, add 4 mL of dilute hydrochloric ameter and 25 to 30 cm in length, packed with octadecyl- acid, shake, and filter through a glass filter (G4). To 40 mL silanized silica gel for liquid chromatography (5 mminparti- of the filtrate add 2 mL of ammonium acetate TS, dilute with cle diameter). water to 50 mL, and perform the test using this solution as Column temperature: Room temperature. the test solution. Prepare a control solution as follows: to 2.0 Mobile phase: A mixture of 0.1 molWL phosphate buŠer mL of Standard Lead Solution add 2 mL of dilute acetic acid, solution, pH 7.0, and methanol (3:1). 2 mL of ammonium acetate TS and water to make 50 mL JP XV O‹cial Monographs / Thiopental Sodium for Injection 1167

(not more than 20 ppm). and extract the washings with two 10-mL portions of chlo- (3) Neutral and basic substances—Weigh accurately roform. Filter the combined chloroform extracts into a coni- about 1 g of Thiopental Sodium, dissolve in 10 mL of water cal flask, and wash the filter paper with three 5-mL portions and 5 mL of sodium hydroxide TS, and shake vigorously of chloroform. Combine the filtrate and the washings, and with 40 mL of chloroform. Separate the chloroform layer, add 10 mL of ethanol (95). Titrate <2.50> with 0.1 molWL wash with two 5-mL portions of water, filter, and evaporate potassium hydroxide-ethanol VS until the color of the solu- the filtrate on a water bath to dryness. Dry the residue at tion changes from yellow through light blue to purple (indica- 1059C for 1 hour: the amount of the residue is not more than tor: 2 mL of alizarin yellow GG-thymolphthalein TS). Per- 0.50z. form a blank determination with a mixture of 160 mL of (4) Related substances—Dissolve 50 mg of Thiopental chloroform and 30 mL of ethanol (95), and make any neces- Sodium in 50 mL of the mobile phase, and use this solution sary correction. as the sample solution. Pipet 1 mL of the sample solution, Each mL of 0.1 molWL potassium hydroxide-ethanol VS add the mobile phase to make exactly 200 mL, and use this ＝ 26.43 mg of C H N NaO S solution as the standard solution. Perform the test with ex- 11 17 2 2 actly 20 mL each of the sample solution and the standard so- Containers and storage Containers—Tight containers. lution as directed under Liquid Chromatography <2.01> ac- Storage—Light-resistant. cording to the following conditions. Measure each peak area of each solution by the automatic integration method: the total area of peaks other than those of thiopental in the sam- Thiopental Sodium for Injection ple solution is not larger than the peak area of thiopental in the standard solution. 注射用チオペンタールナトリウム Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 254 nm). Thiopental Sodium for Injection is a preparation for Column: A stainless steel column 4.6 mm in inside di- injection which is dissolved before use. ameter and 15 cm in length, packed with octadecylsilanized It contains not less than 93z and not more than silica gel (5 mm in particle diameter). 107z of the labeled amount of thiopental sodium Column temperature: A constant temperature of about (C11H17N2NaO2S: 264.32). 409C. Method of preparation Prepare as directed under Injec- Mobile phase: Dissolve 1 g of potassium dihydrogen phos- tions, with 100 parts of Thiopental Sodium and 6 parts of phate in 1000 mL of water, and adjust the pH to 3.0 with Dried Sodium Carbonate in mass. phosphoric acid. To 700 mL of this solution add 300 mL of acetonitrile. Description Thiopental Sodium for Injection is a light yel- Flow rate: Adjust the ‰ow rate so that the retention time of low powder or mass, and has a slight, characteristic odor. thiopental is about 15 minutes. It is very soluble in water, and practically insoluble in de- Time span of measurement: About 1.5 times as long as the hydrated diethyl ether. retention time of thiopental. It is hygroscopic. System suitability— Identification (1) Dissolve 0.1 g of Thiopental Sodium for Test for required detection: To exactly 2 mL of the stan- Injection in 10 mL of water, and add 0.5 mL of barium chlo- dard solution add the mobile phase to make exactly 10 mL. ride TS: a white precipitate is formed. Collect the precipitate, Conˆrm that the peak area of thiopental obtained from 20 and add dilute hydrochloric acid dropwise: the precipitate mL of this solution is equivalent to 15 to 25z of that of dissolves with effervescence. thiopental obtained from 20 mL of the standard solution. (2) Proceed as directed in the Identification under System performance: Dissolve 5 mg each of isopropyl Thiopental Sodium. parahydroxybenzoate and propyl parahydroxybenzoate in 50 mL of acetonitrile, and add water to make 100 mL. When the pH <2.54> Dissolve 1 g of Thiopental Sodium for Injection procedure is run with 20 mL of this solution under the above in 40 mL of water: the pH of this solution is between 10.2 and operating conditions, isopropyl parahydroxybenzoate and 11.2. propyl parahydroxybenzoate are eluted in this order with the Purity Proceed as directed in the Purity under Thiopental resolution between these peaks being not less than 1.9. Sodium. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operating Loss on drying <2.41> Not more than 2.0z (1 g, in vacuum, conditions, the relative standard deviation of the peak area of 809C, 4 hours). thiopental is not more than 2.0z. Sterility <4.06> Perform the test according to the Membrane Loss on drying <2.41> Not more than 2.0z (1 g, in vacuum, filtration method: it meets the requirement. 809C, 4 hours). Assay Take 10 samples of Thiopental Sodium for Injection, Assay Weigh accurately about 0.5 g of Thiopental Sodium, and open each container carefully. Dissolve each content previously dried, transfer to a separator, dissolve in 20 mL of with water, wash each container with water, combine the water, add 5 mL of ethanol (95) and 10 mL of dilute washings with the former solution, and add water to make hydrochloric acid, and extract with 50 mL of chloroform, exactly 1000 mL. Pipet 10 mL of this solution, and add water then with three 25-mL portions of chloroform. Combine the to make exactly 100 mL. Measure exactly a volume (V mL) chloroform extracts, wash with two 5-mL portions of water, of this solution, equivalent to about 15 mg of thiopental sodi- 1168 Hydrochloride / O‹cial Monographs JP XV

um (C11H17N2NaO2S), and add water to make exactly 1000 Reference Spectrum: both spectra exhibit similar intensities mL. Pipet 10 mL of this solution, add 15 mL of diluted dilute of absorption at the same wave numbers. sodium hydroxide TS (1 in 100), add water to make exactly 30 (4) To 5 mL of a solution of Thioridazine Hydrochloride mL, and use this solution as the sample solution. Separately, (1 in 100) add 2 mL of ammonia TS, and heat on a water bath weigh accurately about 46 mg of thiopental for assay, previ- for 5 minutes. After cooling, filter, and acidify the filtrate ously dried at 1059C for 3 hours, dissolve in 50 mL of dilute with dilute nitric acid: the solution responds to the Qualita- sodium hydroxide TS, and add water to make exactly 200 tive Tests <1.09> (2) for chloride. mL. Pipet 2 mL of this solution, add water to make exactly Melting point <2.60> 159 – 1649C 100 mL, and use this solution as the standard solution. Per- form the test with the sample solution and standard solution Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of as directed under Ultraviolet-visible Spectrophotometry Thioridazine Hydrochloride according to Method 2, and per-

<2.24>, and determine the absorbances, AT and AS,at304 form the test. Prepare the control solution with 2.0 mL of nm. Standard Lead Solution (not more than 20 ppm). (2) Arsenic <1.11>—Prepare the test solution with 1.0 g Amount (mg) of thiopental sodium (C H N NaO S) 11 17 2 2 of Thioridazine Hydrochloride, according to Method 3, and in each sample of Thiopental Sodium for Injection perform the test (not more than 2 ppm). ＝ W ×(A /A )×(300/V)×1.0907 S T S (3) Related substances—Conduct this procedure under

WS: Amount (mg) of thiopental sodium for assay the protection from the sunlight. Dissolve 0.10 g of Thiorida- zine Hydrochloride in 10 mL of methanol, and use this solu- Containers and storage Containers—Hermetic containers. tion as the sample solution. Pipet 1 mL of the sample solu- Storage—Light-resistant. tion, and add methanol to make exactly 20 mL. Pipet 2 mL of this solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test Thioridazine Hydrochloride with these solutions as directed under Thin-layer Chro- matography <2.03>.Spot5mL each of the sample solution チオリダジン塩酸塩 and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform, 2-propanol and ammonia so- lution (28) (74:25:1) to a distance of about 10 cm, and air-dry the plate. Examine the plate 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. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). C21H26N2S2.HCl: 407.04 10-｛2-[(2RS)-1-Methylpiperidin-2-yl]ethyl｝-2- Residue on ignition <2.44> Not more than 0.1z (1 g). methylsulfanyl-10H- monohydrochloride [130-61-0] Assay Weigh accurately about 0.35 g of Thioridazine Hydrochloride, previously dried, dissolve in 80 mL of a mix- Thioridazine Hydrochloride, when dried, contains ture of acetic anhydride and acetic acid (100) (1:1), and titrate <2.50> with 0.1 molWL perchloric acid VS (potentiometric not less than 99.0z of C21H26N2S2.HCl. titration). Perform a blank determination, and make any Description Thioridazine Hydrochloride occurs as a white necessary correction. to pale yellow, crystalline powder. It is odorless, and has a bitter taste. Each mL of 0.1 molWL perchloric acid VS It is freely soluble in water, in methanol, in ethanol (95) ＝ 40.70 mg of C21H26N2S2.HCl and in acetic acid (100), sparingly soluble in acetic anhydride, Containers and storage Containers—Tight containers. and practically insoluble in diethyl ether. Storage—Light-resistant. The pH of a solution of Thioridazine Hydrochloride (1 in 100) is between 4.2 and 5.2. It is gradually colored by light. Thiotepa Identification (1) Dissolve 0.01 g of Thioridazine Hydrochloride in 2 mL of sulfuric acid: a deep blue color de- チオテパ velops. (2) Dissolve 0.01 g of Thioridazine Hydrochloride in 2 mL of water, and add 1 drop of cerium (IV) tetraammonium sulfate TS: a blue color develops, and the color disappears on the addition of excess of the reagent.

(3) Determine the infrared absorption spectrum of C6H12N3PS: 189.22 Thioridazine Hydrochloride, previously dried, as directed in Tris(aziridin-1-yl)phosphine sulˆde [52-24-4] the potassium chloride disk method under Infrared Spec- trophotometry <2.25>, and compare the spectrum with the Thiotepa, when dried, contains not less than 98.0z JP XV O‹cial Monographs / L-Threonine 1169 of C6H12N3PS. 98.5z of C4H9NO3.

Description Thiotepa occurs as colorless or white crystals, Description L-Threonine occurs as white crystals or crystal- or white, crystalline powder. It is odorless. line powder. It is odorless or has a slight, characteristic odor, It is freely soluble in water, in ethanol (95) and in diethyl and has a slightly sweet taste. ether. It is freely soluble in formic acid, soluble in water, and A solution of Thiotepa (1 in 10) is neutral. practically insoluble in ethanol (95). Identification (1) To 5 mL of a solution of Thiotepa (1 in Identification Determine the infrared absorption spectrum 100) add 1 mL of hexaammonium heptamolybdate TS, and of L-Threonine, previously dried, as directed in the potassium allow to stand: a dark blue color develops slowly when the so- bromide disk method under the Infrared Spectrophotometry lution is cold, or quickly when warm. <2.25>, and compare the spectrum with the Reference Spec- (2) To 5 mL of a solution of Thiotepa (1 in 100) add 1 mL trum: both spectra exhibit similar intensities of absorption at of nitric acid: this solution responds to the Qualitative Tests thesamewavenumbers. <1.09> (2) for phosphate. Optical rotation <2.49> [a]20: －26.0 – －29.09 (after (3) Dissolve 0.1 g of Thiotepa in a mixture of 1 mL of D drying, 1.5 g, water, 25 mL, 100 mm). lead (II) acetate TS and 10 mL of sodium hydroxide TS, and boil: the gas evolved changes moistened red litmus paper to pH <2.54> Dissolve 0.20 g of L-Threonine in 20 mL of blue, and the solution shows a grayish red color. water: the pH of this solution is between 5.2 and 6.2. Melting point <2.60> 52–579C Purity (1) Clarity and color of solution—Dissolve 1.0 g of L-Threonine in 20 mL of water: the solution is clear and Purity (1) Clarity and color of solution—Dissolve 1.0 g of colorless. Thiotepa in 20 mL of water: the solution is clear and color- (2) Chloride <1.03>—Perform the test with 0.5 g of L- less. Threonine. Prepare the control solution with 0.30 mL of 0.01 (2) Heavy metals <1.07>—Proceed with 1.0 g of Thiotepa molWLhydrochloricacidVS(notmorethan0.021z). in a platinum crucible according to Method 2, and perform (3) Sulfate <1.14>—Perform the test with 0.6 g of L- the test. Prepare the control solution with 2.0 mL of Stan- Threonine. Prepare the control solution with 0.35 mL of dard Lead Solution (not more than 20 ppm). 0.005 molWL sulfuric acid VS (not more than 0.028z). (3) Arsenic <1.11>—Dissolve 0.20 g of Thiotepa in 5 mL (4) Ammonium <1.02>—Perform the test with 0.25 g of of water, and add 1 mL of nitric acid and 1 mL of sulfuric L-Threonine. Prepare the control solution with 5.0 mL of acid. Take this solution, prepare the test solution according Standard Ammonium Solution (not more than 0.02z). to Method 2, and perform the test (not more than 10 ppm). (5) Heavy metals <1.07>—Proceed with 1.0 g of L-Threo- Loss on drying <2.41> Not more than 0.20z (1 g, in vacu- nine according to Method 1, and perform the test. Prepare um, silica gel, 4 hours). the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Residue on ignition <2.44> Not more than 0.1z (1 g, plati- (6) Arsenic <1.11>—Dissolve 1.0 g of L-Threonine in 5 num crucible). mL of dilute hydrochloric acid, and perform the test with this Assay Weigh accurately about 0.1 g of Thiotepa, previous- solution as the test solution (not more than 2 ppm). ly dried, dissolve in 50 mL of a solution of potassium thioc- (7) Related substances—Dissolve 0.30 g of L-Threonine yanate (3 in 20), add 25 mL of 0.05 molWL sulfuric acid VS, in 50 mL of water, and use this solution as the sample solu- exactly measured, and allow to stand for 20 minutes with oc- tion. Pipet 1 mL of this solution, and add water to make ex- casional shaking. Titrate <2.50> the excess sulfuric acid with actly 50 mL. Pipet 5 mL of this solution, add water to make 0.1 molWL sodium hydroxide VS until the color of the solu- exactly 20 mL, and use this solution as the standard solution. tion changes from red to light yellow (indicator: 3 drops of Perform the test with these solutions as directed under Thin- methyl red TS). Perform a blank determination. layer Chromatography <2.03>.Spot5mLeachofthesample solution and standard solution on a plate of silica gel for Each mL of 0.05 molWL sulfuric acid VS thin-layer chromatography. Develop the plate with a mixture ＝ 6.307 mg of C H N PS 6 12 3 of 1-butanol, water and acetic acid (100) (3:1:1) to a distance Containers and storage Containers—Tight containers. of about 10 cm, and dry the plate at 809C for 30 minutes. Storage—Light-resistant, and in a cold place. Spray evenly the plate with a solution of ninhydrin in acetone (1 in 50), and heat the plate at 809C for 5 minutes: the spots other than the principal spot from the sample solution are not L-Threonine more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.20z (1 g, 1059C, L-トレオニン 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).

Assay Weigh accurately about 0.12 g of L-Threonine, previ- ously dried, dissolve in 3 mL of formic acid, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol L perchloric C H NO : 119.12 W 4 9 3 acid VS (potentiometric titration). Perform a blank determi- (2S,3R)-2-Amino-3-hydroxybutanoic acid [72-19-5] nation, and make any necessary correction. L-Threonine, when dried, contains not less than Each mL of 0.1 molWL perchloric acid VS 1170 Thrombin / O‹cial Monographs JP XV

＝ 11.91 mg of C4H9NO3 to contain about 5 Units in each mL, treat 0.10 mL of the so- lution with the same reagents in the same manner ˆve times, Containers and storage Containers—Tight containers. determine the clotting times, and calculate the average value. Plot the average values of the clotting times of the four kinds of the standard solutions on a logarithmic graph, using Units Thrombin as the abscissa and clotting times as the ordinate, and draw a calibration line which best ˆts the four plotted points. Using トロンビン this line, read the Units U from the average value of the clot- ting times of the sample solution. Thrombin is prepared from prothrombin obtained UnitsofasinglecontainerofThrombin＝U×10×V from blood of man or bull, through interaction with V: The number of mL of the volume in which the contents added thromboplastin in the presence of calcium , of a single container of Thrombin has been dissolved. sterilized and lyophilized. It contains not less than 80z and not more than Calculate the units for 1 mg of the contents. 150z of the labeled Units of thrombin. Containers and storage Containers—Hermetic containers. Each mg contains not less than 10 Units of throm- Storage—Not exceeding 109C. bin. Expiration date Use within 36 months after the date of Description Thrombin is a white to light yellow, amor- manufacture. phous substance. Thrombin (500 Units) dissolves in 1.0 mL of isotonic sodi- um chloride solution clearly or with slight turbidity within 1 minute. Thymol Loss on drying <2.41> Not more than 3z (50 mg, in vacu- チモール um, phosphorus (V) oxide, 4 hours). Sterility <4.06> It meets the requirement. Assay (i) Fibrinogen solution—Weigh accurately about 30 mg of ˆbrinogen, and dissolve in 3 mL of isotonic sodium chloride solution. Allow the solution to clot su‹ciently with C H O: 150.22 frequent shaking after the addition of about 3 Units of 10 14 5-Methyl-2-(1-methylethyl)phenol [89-83-8] thrombin. Wash the precipitated clot thoroughly until the washings yield no turbidity on addition of silver nitrate TS, Thymol contains not less than 98.0z of C H O. weigh the clot after drying at 1059C for 3 hours, and calcu- 10 14 late the percentage of the clot in the ˆbrinogen. Dissolve the Description Thymol occurs as colorless crystals or white, ˆbrinogen in isotonic sodium chloride solution so that the crystalline masses. It has an aromatic odor, and has a burn- clot should be 0.20z, adjust the pH of the solution between ing taste. 7.0and7.4byadditionof0.05molWL dibasic sodium phos- It is very soluble in acetic acid (100), freely soluble in hateTS(orifnecesary,use0.5molWL disodium hydrogen- ethanol (95) and in diethyl ether, and slightly soluble in phosphate TS), and dilute with isotonic sodium chloride solu- water. tion to make a 0.10z solution. It sinks in water, but when warmed, it melts and rises to the (ii) Procedure—Dissolve Thrombin Reference Standard surface of water. in isotonic sodium chloride solution, and prepare four kinds Identiˆcation (1) To 1 mL of a solution of Thymol in of standard solutions which contain 4.0, 5.0, 6.2, and 7.5 acetic acid (100) (1 in 300) add 6 drops of sulfuric acid and 1 Units in 1 mL. Transfer accurately 0.10 mL each of the stan- drop of nitric acid: a blue-green color develops by re‰ected dard solutions maintained at a given degree ±19C between light and a red-purple color develops by transmitted light. 209Cand309C to a small test tube, 10 mm in inside diameter, (2) Dissolve 1 g of Thymol in 5 mL of a solution of sodi- 100 mm in length, blow out 0.90 mL of the ˆbrinogen solu- um hydroxide (1 in 10) by heating in a water bath, and con- tion at the same temperature into the test tube from a pipet, tinue heating for several minutes: a light yellow-red color start a stop watch simultaneously, shake the tube constantly, slowly develops. Allow this solution to stand at room temper- and determine the time for the ˆrst appearance of clot. Cal- ature: the color changes to dark yellow-brown. Shake this so- culate the average values of ˆve determinations for the four lution with 2 to 3 drops of chloroform: a purple color grad- kinds of standard solutions, respectively. If the deviation be- ually develops. tween the maximum and the minimum values of ˆve determi- (3) Triturate Thymol with an equal mass of camphor or nations is more than 10z of the average value, reject the menthol: the mixture liqueˆes. whole run, and try the experiment again. The concentration of the standard solution may be changed appropriately wi- Melting point <2.60> 49–519C thin the range between 14 and 60 seconds of the clotting time. Purity (1) Non-volatile residue—Volatilize 2.0 g of The determination proceeds at the same temperature Thymol by heating on a water bath, and dry the residue at described above. Next, weigh accurately the whole contents 1059C for 2 hours: the mass is not more than 1.0 mg. of a single container of Thrombin, dissolve it in isotonic sodi- (2) Other phenols—Shake vigorously 1.0 g of Thymol um chloride solution to provide a solution which is presumed with 20 mL of warm water for 1 minute, and ˆlter. To 5 mL JP XV O‹cial Monographs / Tiaramide Hydrochloride 1171 of the ˆltrate add 1 drop of iron (III) chloride TS: a green diethyl ether extract, and dry the residue at 1059Ctoconstant color may develop, but no blue to purple color develops. mass: the mass of the residue is not more than 0.030 g. Assay Weigh accurately about 0.5 g of Thymol, dissolve in Loss on drying <2.41> Not more than 6.0z (1 g, 1059C, 10 mL of sodium hydroxide TS, and add water to make ex- constant mass). actly 100 mL. Measure exactly 10 mL of the solution into an Total ash <5.01> Not more than 5.0z (0.5 g). iodine ‰ask, add 50 mL of water and 20 mL of dilute sulfuric acid, and cool in ice water for 30 minutes. Add exactly 20 mL Assay Transfer about 1 g of Dried Thyroid, accurately of 0.05 molWL bromine VS, stopper tightly immediately, al- weighed, to a crucible, add 7 g of potassium carbonate, mix low to stand for 30 minutes in ice water with occasional shak- carefully, and gently tap the crucible on the table to compact ing in a dark place, add 14 mL of potassium iodide TS and 5 the mixture. Overlay with 10 g of potassium carbonate, and mL of chloroform, stopper tightly, shake vigorously, and ti- compact again thoroughly by tapping. Place the crucible in a trate <2.50> the liberated iodine with 0.1 molWLsodium muŒe furnace preheated to a temperature between 6009C thiosulfate VS (indicator: 3 mL of starch TS). Stopper tight- and 7009C, and ignite the mixture for 25 minutes. Cool, add ly, shake vigorously near the end point, and continue the 20 mL of water, heat gently to boiling, and ˆlter into a ‰ask. titration until the blue color in the chloroform layer disap- To the residue add 20 mL of water, boil, and ˆlter into the pears. Perform a blank determination. same ‰ask. Rinse the crucible and the char on the funnel with boiling water until the ˆltrate measures 200 mL. Add slowly 7 Each mL of 0.05 molWLbromineVS mL of freshly prepared bromine TS, 40 mL of diluted phos- ＝3.755 mg of C H O 10 14 phoric acid (1 in 2), and boil until starch iodide paper is no Containers and storage Containers—Tight containers. longer colored blue by the evolved gas. Wash down inside of Storage—Light-resistant. the ‰ask with water, and continue boiling for 5 minutes. Dur- ingtheboilingaddwaterfromtimetotimetomaintaina volume at not less than 200 mL. Cool, add 5 mL of a solution Dried Thyroid of phenol (1 in 20), again rinse inside of the ‰ask with water, and allow to stand for 5 minutes. Add 2 mL of diluted phos- 乾燥甲状腺 phoric acid (1 in 2) and 5 mL of potassium iodide TS, and ti- trate <2.50> immediately the liberated iodine with 0.01 molWL sodium thiosulfate VS (indicator: 3 mL of starch TS). Per- Dried Thyroid is the fresh thyroid gland, previously form a blank determination, and make any necessary correc- deprived of connective tissue and fat, minced, dried tion. rapidly at a temperature not above 509C, and pow- Each mL of 0.01 molWL sodium thiosulfate VS dered, or diluted with suitable diluents. It is obtained ＝0.2115 mg of I from domesticated animals that are used for food by man. Containers and storage Containers—Tight containers. It contains not less than 0.30z and not more than 0.35z of iodine (I: 126.90) in the form of organic com- pounds peculiar to the thyroid gland. Tiaramide Hydrochloride Description Dried Thyroid occurs as a light yellow to チアラミド塩酸塩 grayish brown powder. It has a slight, characteristic, meat- like odor. Identiˆcation Mount Dried Thyroid in diluted formalde- hyde solution (1 in 10), stain in hematoxylin TS for 10 to 30 minutes, wash with water, soak in a mixture of 1 mL of hydrochloric acid and 99 mL of diluted ethanol (7 in 10) for 5 to 10 seconds, and again wash with water for about 1 hour. C H ClN O S.HCl: 392.30 StaininasolutionofeosinY(1in100)for1to5minutes, 15 18 3 3 5-Chloro-3-｛2-[4-(2-hydroxyethyl)piperazin-1-yl]- wash with water, dehydrate, and soak successively in diluted 2-oxoethyl｝-1,3-benzothiazol-2(3H)-one monohydrochloride ethanol (7 in 10) for 5 to 10 seconds, in diluted ethanol (4 in [35941-71-0] 5) for 5 to 10 seconds, in diluted ethanol (9 in 10) for 1 to 2 minutes, in ethanol (95) for 1 to 5 minutes then in ethanol Tiaramide Hydrochloride, when dried, contains not (99.5) for 1 to 5 minutes. Interpenetrate in , seal with less than 98.5z of C H ClN O S.HCl. balsam, and examine under a : epithelial nuclei 15 18 3 3 forming follicles peculiar to the thyroid gland are observed. Description Tiaramide Hydrochloride occurs as a white, crystalline powder. It is odorless. Purity (1) Inorganic iodides—Mix 1.0 g of Dried Thyroid It is freely soluble in water, slightly soluble in ethanol (95) with 10 mL of a saturated solution of zinc sulfate heptahy- and in acetic acid (100), and practically insoluble in acetic an- drate, shake for 5 minutes, and ˆlter. To 5 mL of the ˆltrate hydride and in diethyl ether. add 0.5 mL of starch TS, 4 drops of sodium nitrite TS and 4 The pH of a solution of Tiaramide Hydrochloride (1 in 20) drops of dilute sulfuric acid with thorough shaking: no blue is between 3.0 and 4.5. color is produced. Melting point: about 2659C (with decomposition). (2) Fat—Extract 1.0 g of Dried Thyroid with diethyl ether for 2 hours using a Soxhlet extractor. Evaporate the Identification (1) Dissolve 5 mg of Tiaramide Hydrochlo- 1172 Tiaramide Hydrochloride Tablets / O‹cial Monographs JP XV ride in 5 mL of 0.1 molWL hydrochloric acid TS, and add 3 drops of Dragendorff's TS: an orange precipitate is formed. (2) Determine the infrared absorption spectrum of Tiaramide Hydrochloride Tablets Tiaramide Hydrochloride, previously dried, as directed in the potassium chloride disk method under the Infrared Spec- チアラミド塩酸塩錠 trophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Tiaramide Hydrochloride Tablets contain not less (3) A solution of Tiaramide Hydrochloride (1 in 50) than 95.0z and not more than 105.0z of the labeled responds to the Qualitative Tests <1.09> for chloride. amount of tiaramide (C15H18ClN3O3S: 355.84). Method of preparation Prepare as directed under Tablets, Purity (1) Clarity and color of solution—Dissolve 0.5 g of with Tiaramide Hydrochloride. Tiaramide Hydrochloride in 10 mL of water: the solution is clear and colorless. Identiˆcation (1) Determine the absorption spectrum of (2) Heavy metals <1.07>—Proceed with 2.0 g of the sample solution obtained in the Assay as directed under Tiaramide Hydrochloride according to Method 2, and per- Ultraviolet-visible Spectrophotometry <2.24>:itexhibits form the test. Prepare the control solution with 2.0 mL of maxima between 285 nm and 289 nm, and between 292 nm Standard Lead Solution (not more than 10 ppm). and 296 nm. (3) Arsenic <1.11>—Prepare the test solution with 1.0 g (2) To a quantity of powdered Tiaramide Hydrochloride of Tiaramide Hydrochloride according to Method 1, and per- Tablets, equivalent to 0.1 g of tiaramide according to the form the test. In the procedure, add 20 mL of diluted labeled amount, add 10 mL of diluted ethanol (7 in 10), shake hydrochloric acid (1 in 2) (not more than 2 ppm). well, ˆlter, and use the ˆltrate as the sample solution. (4) Related substances—Dissolve 0.20 g of Tiaramide Separately, dissolve 0.11 g of tiaramide hydrochloride for as- Hydrochloride in 10 mL of diluted ethanol (99.5) (7 in 10), say in 10 mL of diluted ethanol (7 in 10), and use this solution and use this solution as the sample solution. Pipet 1 mL of as the standard solution. Perform the test with these solu- the sample solution, and add diluted ethanol (99.5) (7 in 10) tions as directed under Thin-layer Chromatography <2.03>. to make exactly 100 mL. Pipet 2 mL of this solution, add Spot 20 mLeachofthesamplesolutionandstandardsolution diluted ethanol (99.5) (7 in 10) to make exactly 10 mL, and on a plate of silica gel for thin-layer chromatography, de- use this solution as the standard solution. Perform the test velop with a mixture of 1-butamol, water and acetic acid with these solutions as directed under Thin-layer Chro- (100) (4:2:1) to a distance of about 10 cm, and dry the plate at matography <2.03>.Spot5mL each of the sample solution 1009C for 30 minutes. Spray evenly DragendorŠ's TS for and standard solution on a plate of silica gel with fluorescent spraying followed by diluted nitric acid (1 in 50) on the plate: indicator for thin-layer chromatography. After air-drying, the principal spot obtained with the sample solution and the immediately develop the plate with a mixture of 1-butanol, spot with the standard solution are yellow-red in color and water and acetic acid (100) (4:2:1) to a distance of about 10 have the same Rf value. cm, air-dry the plate, and then dry at 1009Cfor30minutes. Uniformity of dosage units <6.02> Perform the test accord- After cooling, examine under ultraviolet light (main ing to the following method: it meets the requirement of the wavelength: 254 nm): the spots other than the principal spot Content uniformity test. and the spot of the starting point from the sample solution To 1 tablet of Tiaramide Hydrochloride Tablets add a are not more intense than the spot from the standard solu- volume of 0.1 mol/L hydrochloric acid TS, equivalent to 3/5 tion. Allow the plate to stand in iodine vapor for 30 minutes: volume of mLwhichmakesasolutionsothateachmL the spots other than the principal spot and the spot of the V contains about 1 mg of tiaramide (C H ClN O S) according starting point from the sample solution are not more intense 15 18 3 3 to the labeled amount, shake for 60 minutes, then add 0.1 than the spot from the standard solution. mol/LhydrochloricacidTStomakeexactlyV mL, and Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, ˆlter. Discard the ˆrst 20 mL of the ˆltrate, pipet 5 mL of the 3 hours). subsequent ˆltrate, add water to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh ac- Residue on ignition <2.44> Not more than 0.1z (1 g). curately about 55 mg of tiaramide hydrochloride for assay, Assay Weigh accurately about 0.5 g of Tiaramide previously dried at 1059C for 3 hours, and dissolve in 0.1 Hydrochloride, previously dried, dissolve in 50 mL of a mix- mol/LhydrochloricacidTStomakeexactly50mL.Pipet5 ture of acetic anhydride and acetic acid (100) (7:3) by warm- mL of this solution, add water to make exactly 100 mL, and ing, cool, and titrate <2.50> with 0.1 molWL perchloric acid use this solution as the standard solution. Determine the ab-

VS until the color of the solution changes from red through sorbances, AT and AS, of the sample solution and standard purple to blue-purple (indicator: 3 drops of TS). solution at 294 nm as directed under Ultraviolet-visible Spec- Perform a blank determination, and make any necessary cor- trophotometry <2.24>. rection. Amount (mg) of tiaramide (C15H18ClN3O3S) Each mL of 0.1 molWL perchloric acid VS ＝WS×(AT/AS)×(V/50)×0.907 ＝ 39.23 mg of C15H18ClN3O3S.HCl WS: Amount (mg) of tiaramide hydrochloride for assay Containers and storage Containers—Well-closed contain- Dissolution <6.10> Perform the test according to the follow- ers. ing method: it meets the requirement. Perform the test with 1 tablet of Tiaramide Hydrochloride JP XV O‹cial Monographs / Ticlopidine Hydrochloride 1173

Tablets at 50 revolutions per minute according to the Paddle tetrahydrothieno[3,2-c]pyridine monohydrochloride method, using 900 mL of water as the dissolution medium. [53885-35-1] Withdraw 20 mL or more of the dissolution medium 15 minutes after starting the test for a 50-mg tablet or 30 Ticlopidine Hydrochloride contains not less than minutes after starting the test for a 100-mg tablet, and ˆlter 99.0z of C14H14ClNS.HCl, calculated on the anhy- through a membrane ˆlter with pore size of not more than 0.5 drous basis. mm. Discard the ˆrst 10 mL of the ˆltrate, pipet V mL of the Description Ticlopidine Hydrochloride occurs as a white to subsequent ˆltrate, add water to make exactly V? mL so that pale yellowish white crystalline powder. each mL contains about 56 mgoftiaramide(C H ClN O S) 15 18 3 3 It is freely soluble in acetic acid (100), soluble in water and according to the labeled amount, and use this solution as the in methanol, sparingly soluble in ethanol (95), and practically sample solution. Separately, weigh accurately about 15 mg of insoluble in diethyl ether. tiaramide hydrochloride for assay, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 50 mL. Identification (1) Determine the infrared absorption spec- Pipet 5 mL of this solution, add water to make exactly 25 trum of Ticlopidine Hydrochloride as directed in the potassi- mL, and use this solution as the standard solution. Determine um bromide disk method under Infrared Spectrophotometry the absorbances, AT and AS, at 294 nm of the sample solution <2.25>, and compare the spectrum with the Reference Spec- and standard solution as directed under Ultraviolet-visible trum: both spectra exhibit similar intensities of absorption at Spectrophotometry <2.24>. The dissolution rates for a 50-mg thesamewavenumbers. tablet in 15 minutes and a 100-mg tablet in 30 minutes are not (2) A solution of Ticlopidine Hydrochloride (1 in 20) less than 80z, respectively. responds to the Qualitative Tests <1.09> (2) for chloride. Dissolution rate (z) with respect to the labeled amount of Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of tiaramide (C15H18ClN3O3S) Ticlopidine Hydrochloride according to Method 3, and per- ＝WS×(AT/AS)×(V?/V)×(1/C)×360×0.907 form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). W : Amount (mg) of tiaramide hydrochloride for assay S (2) Arsenic <1.11>—Prepare the test solution with 1.0 g C: Labeled amount (mg) of tiaramide (C H ClN O S) in 15 18 3 3 of Ticlopidine Hydrochloride according to Method 4, and 1tablet perform the test (not more than 2 ppm). Assay Weigh accurately the mass of more than 20 (3) Related substances—Dissolve 0.5 g of Ticlopidine Tiaramide Hydrochloride Tablets, and powder. Weigh ac- Hydrochloride in 20 mL of a solution of hydrochloric acid in curately an amount of the powder, equivalent to about 0.1 g methanol (1 in 20,000), and use this solution as the sample so- of tiaramide (C15H18ClN3O3S), add 60 mL of 0.1 mol/L lution. To exactly 5 mL of the sample solution add a solution hydrochloric acid TS, shake for 30 minutes, add 0.1 mol/L of hydrochloric acid in methanol (1 in 20,000) to make exact- hydrochloric acid TS to make exactly 100 mL, and ˆlter. Dis- ly 200 mL, and use this solution as the standard solution (1). card the ˆrst 20 mL of the ˆltrate, pipet 5 mL of the subse- Separately, pipet 1 mL of the sample solution, add a solution quent ˆltrate, add water to make exactly 100 mL, and use this of hydrochloric acid in methanol (1 in 20,000) to make exact- solution as the sample solution. Separately, weigh accurately ly 50 mL, and use this solution as the standard solution (2). about 0.11 g of tiaramide hydrochloride for assay, previously Perform the test with these solutions as directed under Thin- dried at 1059C for 3 hours, and dissolve in 0.1 mol/L layer Chromatography <2.03>.Spot10mLeachofthesample hydrochloric acid TS to make exactly 100 mL. Pipet 5 mL of solution and standard solution (1) on a plate of silica gel for this solution, add water to make exactly 100 mL, and use this thin-layer chromatography (Plate 1), and spot 10 mLeachof solution as the standard solution. Determine the absor- the sample solution and standard solution (2) on another bances, AT and AS, of the sample solution and standard solu- plate of silica gel for thin-layer chromatography (Plate 2). tion at 294 nm as directed under Ultraviolet-visible Spec- Develop the plates with an upper layer of a mixture of water, trophotometry <2.24>. 1-butanol and acetic acid (100) (5:4:1) to a distance of about 15 cm, and air-dry the plates. Spray evenly a solution of nin- Amount (mg) of tiaramide (C H ClN O S) 15 18 3 3 hydrin in acetone (1 in 50) on Plate 1, and heat at 1009Cfor ＝W ×(A /A )×0.907 S T S 20 minutes: the spots other than the principal spot from the

WS: Amount (mg) of tiaramide hydrochloride for assay sample solution are not more intense than the spot from the standard solution (1). Allow Plate 2 to stand in an iodine Containers and storage Containers—Tight containers. vapor for 30 minutes: the spots other than the principal spot from the sample solution are not more intense than the spot from the standard solution (2). Ticlopidine Hydrochloride (4) Formaldehyde—Dissolve 0.80 g of Ticlopidine Hydrochloride in 19.0 mL of water, add 1.0 mL of 4 molWL チクロピジン塩酸塩 sodium hydroxide TS, shake well, centrifuge, and filter the supernatant liquid. To 5.0 mL of the filtrate add 5.0 mL of acetylacetone TS, mix, and warm at 409C for 40 minutes: the solution has no more color than the following control solu- tion. Control solution: Weigh accurately 0.54 g of formalde- C14H14ClNS.HCl: 300.25 hyde solution, and add water to make exactly 1000 mL. To 5-(2-Chlorobenzyl)-4,5,6,7- exactly 10 mL of this solution add water to make exactly 1000 1174 Timepidium Bromide Hydrate / O‹cial Monographs JP XV mL. Prepare before use. To 8.0 mL of this solution add water thesamewavenumbers. to make 20.0 mL, and filter. To 5.0 mL of the filtrate add 5.0 (4) A solution of Timepidium Bromide Hydrate (1 in 100) mL of acetylacetone TS, and proceed in the same manner. responds to the Qualitative Tests <1.09> (1) for Bromide. Water <2.48> Not more than 1.0z (0.3 g, direct titration). Purity (1) Clarity and color of solution—Dissolve 0.10 g of Timepidium Bromide Hydrate in 10 mL of water: the solu- Residue on ignition <2.44> Not more than 0.1z (1 g). tion is clear and colorless. Assay Weigh accurately about 0.4 g of Ticlopidine (2) Heavy metals <1.07>—Proceed with 1.0 g of Hydrochloride, dissolve in 20 mL of acetic acid (100), add 40 Timepidium Bromide Hydrate according to Method 2, and mL of acetic anhydride, and titrate <2.50> with 0.1 molWL perform the test. Prepare the control solution with 2.0 mL of perchloric acid VS (potentiometric titration). Perform a Standard Lead Solution (not more than 20 ppm). blank determination, and make any necessary correction. (3) Related substances—Dissolve 0.10 g of Timepidium Bromide Hydrate in 10 mL of methanol, and use this solu- Each mL of 0.1 molWL perchloric acid VS tion as the sample solution. Pipet 1 mL of the sample solu- ＝ 30.03 mg of C H ClNS.HCl 14 14 tion, and add methanol to make exactly 100 mL. Pipet 1 mL Containers and storage Containers—Well-closed contain- of this solution, add methanol to make exactly 10 mL, and ers. use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro- matography <2.03>.Spot10mL each of the sample solution Timepidium Bromide Hydrate and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate チメピジウム臭化物水和物 with a mixture of chloroform, methanol, water, acetic acid (100) and ethyl acetate (5:4:1:1:1) to a distance of about 13 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. Water <2.48> 3.5–5.0z (0.4 g, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g). C17H22BrNOS2.H2O: 418.41 (5RS)-3-(Dithien-2-ylmethylene)-5-methoxy-1,1- Assay Weigh accurately about 0.6 g of Timepidium dimethylpiperidinium bromide monohydrate BromideHydrate,dissolvein60mLofamixtureofacetic [35035-05-3, anhydride] anhydride and acetic acid (100) (2:1), and titrate <2.50> with 0.1 molWL perchloric acid VS (potentiometric titration). Per- Timepidium Bromide Hydrate contains not less than form a blank determination, and make any necessary correc- tion. 98.5z of timepidium bromide (C17H22BrNOS2: 400.40), calculated on the anhydrous basis. Each mL of 0.1 molWL perchloric acid VS Description Timepidium Bromide Hydrate occurs as white ＝ 40.04 mg of C17H22BrNOS2 crystals or crystalline powder. Containers and storage Containers—Tight containers. It is very soluble in methanol and in acetic acid (100), freely Storage—Light-resistant. soluble in ethanol (99.5), sparingly soluble in water and in acetic anhydride, and practically insoluble in diethyl ether. The pH of a solution of Timepidium Bromide Hydrate in Timolol Maleate freshly boiled and cooled water (1 in 100) is between 5.3 and 6.3. チモロールマレイン酸塩 A solution of Timepidium Bromide Hydrate in methanol (1 in 20) shows no optical rotation. Identification (1) To 1 mL of a solution of Timepidium Bromide Hydrate (1 in 100) add 1 mL of ninhydrin-sulfuric acid TS: a red purple color develops. (2) Determine the absorption spectrum of a solution of Timepidium Bromide Hydrate (1 in 50,000) as directed under C H N O S･C H O : 432.49 Ultraviolet-visible Spectrophotometry <2.24>,andcompare 13 24 4 3 4 4 4 (2S)-1-[(1,1-Dimethylethyl)amino]-3-(4-morpholin-4-yl- the spectrum with the Reference Spectrum: both spectra ex- 1,2,5-thiadiazol-3-yloxy)propan-2-ol monomaleate hibit similar intensities of absorption at the same [26921-17-5] wavelengths. (3) Determine the infrared absorption spectrum of Timolol Maleate, when dried, contains not less than Timepidium Bromide Hydrate as directed in the potassium 98.0 z and not more than 101.0 z of bromide disk method under the Infrared Spectrophotometry C H N O S.C H O . <2.25>, and compare the spectrum with the Reference Spec- 13 24 4 3 4 4 4 trum: both spectra exhitit similar intensities of absorption at Description Timolol Maleate occurs as a white to pale yel- JP XV O‹cial Monographs / Tinidazole 1175 lowish white crystalline powder. Flow rate: Adjust the ‰ow rate so that the retention time of It is freely soluble in acetic acid (100), and soluble in water timolol is about 18 minutes. and in ethanol (99.5). Time span of measurement: About 2 times as long as the It dissolves in 0.1 mol/L hydrochloric acid TS. retention time of timolol beginning after the solvent peak. Melting point: about 1979C (with decomposition). System suitability— Test for required detectability: To exactly 1 mL of the stan- Identiˆcation (1) Determine the absorption spectrum of a dard solution add the mobile phase to make 10 mL. Conˆrm solution of Timolol Maleate in 0.1 mol/L hydrochloric acid that the peak area of timolol obtained from 25 mL of this so- TS (3 in 100,000) as directed under Ultraviolet-visible Spec- lution is equivalent to 7 to 13z of that from 25 mLofthe trophotometry <2.24>, and compare the spectrum with the standard solution. Reference Spectrum: both spectra exhibit similar intensities System performance: When the procedure is run with 25 of absorption at the same wavelengths. mL of the sample solution under the above operating condi- (2) Determine the infrared absorption spectrum of tions, the number of theoretical plates and the symmetry fac- Timolol Maleate as directed in the potassium bromide disk tor of the peak of timolol are not less than 1500 and not more method under Infrared Spectrophotometry <2.25>,andcom- than 2.5, respectively. pare the spectrum with the Reference Spectrum: both spectra System repeatability: When the test is repeated 6 times with exhibit similar intensities of absorption at the same wave 25 mL of the standard solution under the above operating numbers. conditions, the relative standard deviation of the peak area of (3) To 5 mL of a solution of Timolol Maleate (1 in 500) timolol is not more than 2.0z. add 1 drop of potassium permanganate TS: the red color of the TS disappears immediately. Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum, 20 1009C, 3 hours). Optical rotation <2.49> [a]D : －5.7 – －6.29(after drying, 1.25 g, 1 mol/L hydrochloric acid TS, 25 mL, 100 mm). Residue on ignition <2.44> Not more than 0.1z (1 g). pH <2.54> The pH of a solution prepared by dissolving 1.0 Assay Weigh accurately about 0.8 g of Timolol Maleate, g of Timolol Maleate in 20 mL of water is between 3.8 and previously dried, dissolve in 90 mL of acetic acid (100), and 4.3. titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio- metric titration). Perform a blank determination in the same Purity (1) Clarity and color of solution—Dissolve 1.0 g of manner, and make any necessary correction. Timolol Maleate in 20 mL of water: the solution is clear, and its absorbance at 440 nm, determined as directed under Each mL of 0.1 mol/L perchloric acid VS

Ultraviolet-visible Spectrophotometry <2.24>,isnotmore ＝43.25 mg of C13H24N4O3S.C4H4O4 than 0.05. Containers and storage Containers—Tight containers. (2) Heavy metals <1.07>—Proceed with 2.0 g of Timolol Maleate according to Method 4, and perform the test. Pre- pare the control solution with 2.0 mL of Standard Lead Solu- tion (not more than 10 ppm). Tinidazole (3) Related substances—Dissolve 30 mg of Timolol Male- チニダゾール ate 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 100 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol- lowing conditions, and determine each peak area by the auto- C H N O S: 247.27 matic integration method: the area of the peak other than 8 13 3 4 1-[2-(Ethylsulfonyl)ethyl]-2-methyl-5-nitro-1H-imidazole timolol and maleic acid is not larger than 1/5 times the peak [19387-91-8] area of timolol from the standard solution, and the total area of the peaks other than the peak of timolol and maleic acid is Tinidazole, when dried, contains not less than not larger than 1/2 times the peak area of timolol from the 98.5z and not more than 101.0z of C H N O S. standard solution. 8 13 3 4 Operating conditions— Description Tinidazole occurs as a light yellow, crystalline Detector: An ultraviolet absorption photometer powder. (wavelength: 280 nm). It is soluble in acetic anhydride and in acetone, sparingly Column: A stainless steel column 4.6 mm in inside di- soluble in methanol, slightly soluble in ethanol (99.5), and ameter and 25 cm in length, packed with phenylsilanized sili- very slightly soluble in water. ca gel for liquid chromatography (5 m in particle diameter). m Identiˆcation (1) Determine the absorption spectrum of a Column temperature: A constant temperature of about solution of Tinidazole in methanol (1 in 50,000) as directed 409C. under Ultraviolet-visible Spectrophotometry <2.24>,and Mobile phase: Dissolve 1.9 g of sodium 1-hexanesulfonate compare the spectrum with the Reference Spectrum: both in 1800 mL of water, add 6.0 mL of triethylamine and 8.0 spectra exhibit similar intensities of absorption at the same mL of formic acid, adjust to pH 3.0 with formic acid, and wavelengths. add water to make 2000 mL. To 1400 mL of this solution add (2) Determine the infrared absorption spectrum of 500 mL of methanol and 100 mL of acetonitrile. 1176 Tipepidine Hibenzate / O‹cial Monographs JP XV

Tinidazole as directed in the potassium bromide disk method 3-(Dithien-2-ylmethylene)-1-methylpiperidine mono[2-(4- under Infrared Spectrophotometry <2.25>,andcomparethe hydroxybenzoyl)benzoate] [31139-87-4] spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Tipepidine Hibenzate, when dried, contains not less than 98.5z of C H NS .C H O . Melting point <2.60> 125 – 1299C 15 17 2 14 10 4 Description Tipepidine Hibenzate occurs as a white to light Purity (1) Sulfate <1.14>—To 2.0 g of Tinidazole add 100 yellow, crystalline powder. It is odorless and tasteless. mL of water, boil for 5 minutes, cool, add water to make 100 It is freely soluble in acetic acid (100), slightly soluble in mL, and filter. Take 25 mL of the filtrate, and add 1 mL of methanol and in ethanol (95), very slightly soluble in water, dilute hydrochloric acid and water to make 50 mL. Use this and practically insoluble in diethyl ether. solution as the test solution, and perform the test. Prepare the control solution with 0.45 mL of 0.005 molWL sulfuric Identification (1) Dissolve 0.01 g of Tipepidine Hibenzate acid VS (not more than 0.043z). in 5 mL of sulfuric acid: an orange-red color develops. (2) Heavy metals <1.07>—Proceed with 1.0 g of Tinida- (2) Dissolve 0.3 g of Tipepidine Hibenzate in 10 mL of zole according to Method 4, and perform the test. Prepare sodium hydroxide TS and 5 mL of water, and extract with the control solution with 2.0 mL of Standard Lead Solution two 20-mL portions of chloroform. Wash the chloroform ex- (not more than 20 ppm). tracts with 10 mL of water, and filter the chloroform layer. (3) Arsenic <1.11>—Prepare the test solution with 2.0 g Evaporate the filtrate on a water bath to dryness, and dis- of Tinidazole according to Method 3, and perform the test solve the residue in 0.5 mL of 1 molWL hydrochloric acid TS (not more than 1 ppm). and 5 mL of water. To 2 mL of this solution add 5 mL of (4) Related substances—Dissolve 50 mg of Tinidazole in Reinecke salt TS: a light red precipitate is formed. 2 mL of acetone, and use this solution as the sample solution. (3) Determine the absorption spectrum of a solution of Pipet 1 mL of the sample solution, add acetone to make ex- Tipepidine Hibenzate in ethanol (99.5) (1 in 100,000) as actly 200 mL, and use this solution as the standard solution. directed under Ultraviolet-visible Spectrophotometry <2.24>, Perform the test with these solutions as directed under Thin- and compare the spectrum with the Reference Spectrum; layer Chromatography <2.03>.Spot10mLeachofthesample both spectra exhibit similar intensities of absorption at the solution and standard solution on a plate of silica gel with same wavelengths. fluorescent indicator for thin-layer chromatography. De- (4) Determine the infrared absorption spectrum of velop the plate with a mixture of ethyl acetate and diethyla- Tipepidine Hibenzate, previously dried, as directed in the mine (19:1) to a distance of about 10 cm, air-dry the plate, potassium bromide disk method under Infrared Spec- heat at 1009C for 5 minute, and cool. Examine under ultrav- trophotometry <2.25>, and compare the spectrum with the iolet light (main wavelength: 254 nm): the spots other than Reference Spectrum: both spectra exhibit similar intensities the principal spot from the sample solution are not more in- of absorption at the same wave numbers. tense than the spot from the standard solution. Melting point <2.60> 189 – 1939C Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, Purity (1) Clarity of solution—Dissolve 1.0 g of Tipepi- 2 hours). dine Hibenzate in 10 mL of acetic acid (100): the solution is Residue on ignition <2.44> Not more than 0.1z (1 g). clear. Perform the test with this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: its absorbance Assay Weigh accurately about 0.35 g of Tinidazole, previ- at 400 nm is not more than 0.16. ously dried, dissolve in 50 mL of acetic anhydride, and titrate (2) Heavy metals <1.07>—Proceed with 2.0 g of Tipepi- <2.50> with 0.1 molWL perchloric acid VS (potentiometric dine Hibenzate according to Method 2, and perform the test. titration). Perform a blank determination, and make any Prepare the control solution with 2.0 mL of Standard Lead necessary correction. Solution (not more than 10 ppm). Each mL of 0.1 molWL perchloric acid VS (3) Arsenic <1.11>—Prepare the test solution with 1.0 g

＝ 24.73 mg of C8H13N3O4S of Tipepidine Hibenzate according to Method 3, and perform the test (not more than 2 ppm). Containers and storage Containers—Tight containers. (4) Related substances—(i) Dissolve 10 mg of Tipep- Storage—Light-resistant. idine Hibenzate in 20 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so- lution, add the mobile phase to make exactly 100 mL, and use Tipepidine Hibenzate this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu- チペピジンヒベンズ酸塩 tion as directed under Liquid Chromatography <2.01> ac- cording to the following conditions. Determine each peak area of both solutions by the automatic integration method: the total area of all peaks other than the area of the hibenzic acid and tipepidine from the sample solution is not larger than the peak area of the tipepidine from the standard solu- tion. Operating conditions— Detector: An ultraviolet absorption photometer C H NS .C H O : 517.66 15 17 2 14 10 4 (wavelength: 254 nm). JP XV O‹cial Monographs / Tipepidine Hibenzate Tablets 1177

Column: A stainless steel column 4.6 mm in inside di- zate and 4 mg of xanthene in 50 mL of the mobile phase. ameter and 15 cm in length, packed with octadecylsilanized When the procedure is run with 10 mL of this solution under silica gel for liquid chromatography (5 mminparticledi- the above operating conditions, hibenzic acid, tipepidine and ameter). xanthene are eluted in this order with the resolution between Column temperature: A constant temperature of about the peaks of tipepidine and xanthene being not less than 3. 509C. System repeatability: When the test is repeated 6 times with Mobile phase: A mixture of a solution of ammonium 20 mL of the standard solution under the above operating acetate (1 in 100) and tetrahydrofuran (32:13). conditions, the relative standard deviation of the peak area of Flow rate: Adjust the ‰ow rate so that the retention time of tipepidine is not more than 3.0z. tipepidine is about 12 minutes. Loss on drying <2.41> Not more than 0.5z (1 g, 609C, in Time span of measurement: As long as the retention time vacuum, phosphorus (V) oxide, 3 hours). of tipepidine beginning after the solvent peak. System suitability— Residue on ignition <2.44> Not more than 0.1z (1 g). Test for required detection: To exactly 2 mL of the stan- Assay Weigh accurately about 1 g of Tipepidine Hibenzate, dard solution add the mobile phase to make exactly 20 mL. previously dried, dissolve in 40 mL of acetic acid (100), and Conˆrm that the peak area of tipepidine obtained from 20 mL titrate <2.50> with 0.1 molWL perchloric acid VS until the of this solution is equivalent to 7 to 13z of that of tipepidine color of the solution changes from purple through blue to obtained from 20 mL of the standard solution. green (indicator: 3 drops of crystal violet TS). Perform a System performance: Dissolve 10 mg of Tipepidine Hiben- blank determination, and make any necessary correction. zate and 3 mg of propyl parahydroxybenzoate in 100 mL of the mobile phase. When the procedure is run with 20 mLof Each ml of 0.1 molWL perchloric acid VS this solution under the above operating conditions, hibenzic ＝ 51.77 mg of C15H17NS2.C14H10O4 acid, tipepidine and propyl parahydroxybenzoate are eluted Containers and storage Containers— Well-closed contain- in this order with the resolution between the peaks of tipepi- ers. dine and propyl parahydroxybenzoate being not less than 3. Storagle—Light-resistant. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of tipepidine is not more than 1.5z. Tipepidine Hibenzate Tablets (ii) Dissolve 10 mg of Tipepidine Hibenzate in 20 mL of チペピジンヒベンズ酸塩錠 the mobile phase, and use this solution as the sample solu- tion. 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 20 mLeachofthesam- Tipepidine Hibenzate Tablets contain not less than ple solution and standard solution as directed under Liquid 95z and not more than 105z of the labeled amount of Chromatography <2.01> according to the following condi- tipepidine hibenzate (C15H17NS2.C14H10O4:517.66). tions, and determine each peak area by the automatic integra- Method of preparation Prepare as directed under Tablets, tion method: the total area of all peaks other than the area of with Tipepidine Hibenzate. the hibenzic acid and tipepidine from the sample solution is Identification (1) To a quantity of powdered Tipepidine not larger than 1W2 times the peak area of the tipepidine from Hibenzate Tablets, equivalent to 44 mg of Tipepidine Hiben- the standard solution. zate according to the labeled amount, add 5 mL of water, Operating conditions— shake for 1 minute, add 10 mL of sodium hydroxide TS, and Detector: An ultraviolet absorption photometer extract with two 20-mL portions of chloroform. Combine the (wavelength: 254 nm). extracts, wash with 10 mL of water, and filter the chloroform Column: A stainless steel column 4.6 mm in inside di- layer. Evaporate the filtrate on a water bath to dryness, dis- ameter and 15 cm in length, packed with octadecylsilanized solve the residue in 0.2 mL of 1 mol L hydrochloric acid TS silica gel for liquid chromatography (5 mminparticledi- W and 2 mL of water, and add 5 mL of Reinecke salt TS: a light ameter). red precipitate is formed. Column temperature: A constant temperature of about (2) To a quantity of powdered Tipepidine Hibenzate 409C. Tablets, equivalent to 11 mg of Tipepidine Hibenzate accord- Mobile phase: A mixture of methanol and a solution of ingtothelabeledamount,add30mLofethanol(99.5),and ammonium acetate (1 in 500) (13:7). warm for 10 minutes with occasional shaking. After cooling, Flow rate: Adjust the ‰ow rate so that the retention time of add ethanol (99.5) to make 50 mL, and filter. To 1 mL of the tipepidine is about 10 minutes. filtrate add ethanol (99.5) to make 20 mL, and determine the Time span of measurement: As long as the retention time absorption spectrum of this solution as directed under of tipepidine beginning after the solvent peak. Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a System suitability— maximum between 282 nm and 286 nm. Test for required detection: To exactly 2 mL of the stan- dard solution add the mobile phase to make exactly 20 mL. Dissolution <6.10> Perform the test according to the follow- Conˆrm that the peak area of tipepidine obtained from 20 mL ing method: it meets the requirement. of this solution is equivalent to 7 to 13z of that of tipepidine Perform the test with 1 tablet of Tipepidine Hibenzate obtained from 20 mL of the standard solution. Tablets at 50 revolutions per minute according to the Paddle System performance: Dissolve 12 mg of Tipepidine Hiben- method, using 900 mL of water as the dissolution medium. 1178 Titanium Oxide / O‹cial Monographs JP XV

Use the dissolved solution 30 minutes after starting the test as sulfate in diluted phosphoric acid (1 in 1000) (1 in 500), the sample solution. Separately, weigh accurately about 0.11 acetonitrile and 2-propanol (3:2:1). g of tipepidine hibenzate for assay, previously dried in a Flow rate: Adjust the flow rate so that the retention time of desiccator (in vacuum, phosphorus (V) oxide, 609C) for 3 tipepidine is about 7 minutes. hours, and dissolve in 80 mL of diluted ethanol (99.5) (3 in 4) System suitability— by warming occasionally. After cooling, add diluted ethanol System performance: When the procedure is run with 20 (99.5) (3 in 4) to make exactly 100 mL, then pipet 20 mL of mL of the standard solution under the above operating condi- this solution, add water to make exactly 900 mL, and use this tions, tipepidine and the internal standard are eluted in this solution as the standard solution. Determine the absor- order with the resolution between these peaks being not less bances, AT1 and AS1,at286nm,andAT2 and AS2,at360nm than 10. of the sample solution and standard solution as directed un- System repeatability: When the test is repeated 6 times with der Ultraviolet-visible Spectrophotometry <2.24>. The disso- 20 mL of the standard solution under the above operating lution rate of Tipepidine Hibenzate Tablets in 30 minutes is conditions, the relative standard deviation of the ratios of the not less than 80z. peak area of tipepidine to that of the internal standard is not more than 1.0z. Dissolution rate (z) with respect to the labeled amount

of tipepidine hibenzate (C15H17NS2.C14H10O4) Containers and storage Containers—Tight containers. ＝ WS × {(AT － AT2/AS － AS2)} × (20/C) Storage—Light-resistant.

WS: Amount (mg) of tipepidine hibenzate for assay. C: Labeled amount (mg) of tipepidine hibenzate Titanium Oxide (C15H17NS2.C14H10O4)in1tablet. Assay Weigh accurately and powder not less than 20 酸化チタン Tipepidine Hibenzate Tablets. Weigh accurately a portion of the powder, equivalent to about 22 mg of tipepidine hiben- zate (C15H17NS2.C14H10O4), add 10 mL of diluted acetic acid TiO2:79.87 (100) (1 in 2) and 30 mL of methanol, and warm for 10 minutes with occasional shaking. After cooling, add diluted Titanium Oxide, when dried, contains not less than methanol (1 in 2) to make exactly 50 mL, and filter. Discard 98.5z of TiO2. the first 10 mL of the filtrate, pipet the subsequent 5 mL, add Description Titanium Oxide occurs as a white powder. It is exactly 5 mL of the internal standard solution, then add odorless and tasteless. dilutedmethanol(1in2)tomake25mL,andusethissolu- It is practically insoluble in water, in ethanol (99.5) and in tion as the sample solution. Separately, weigh accurately diethyl ether. about 22 mg of tipepidine hibenzate for assay, previously It dissolves in hot sulfuric acid and in hydro‰uoric acid, dried in a desiccator (in vacuum, phosphorus (V) oxide, and does not dissolve in hydrochloric acid, in nitric acid and 609C) for 3 hours, dissolve in 10 mL of diluted acetic acid in dilute sulfuric acid. (100)(1in2)and30mLofmethanol,andadddiluted When fused by heating with potassium hydrogen sulfate, methanol (1 in 2) to make exactly 50 mL. Pipet 5 mL of this with potassium hydroxide, or with potassium carbonate, it solution, add exactly 5 mL of the internal standard solution, changes to soluble salts. then add diluted methanol (1 in 2) to make exactly 25 mL, Shake 1 g of Titanium Oxide with 10 mL of water: the mix- and use this solution as the standard solution. Perform the ture is neutral. test with 20 mL each of the sample solution and standard so- lution as directed under Liquid Chromatography <2.01> ac- Identiˆcation Heat 0.5 g of Titanium Oxide with 5 mL of cording to the following conditions, and calculate the ratios, sulfuric acid until white fumes are evolved, cool, add cauti- Q T and QS, of the peak area of tipepidine to that of the inter- ously water to make 100 mL, and ˆlter. To 5 mL of the nal standard, respectively. ˆltrate add 2 to 3 drops of hydrogen peroxide TS: a yellow- red color develops. Amount (mg) of tipepidine hibenzate

(C15H17NS2.C14H10O4) Purity (1) Lead—Place 1.0 g of Titanium Oxide in a plati- ＝ WS×(QT/QS) num crucible, add 10.0 g of potassium hydrogen sulfate, heat gently with caution at the beginning, then raise the tempera- W : Amount (mg) of tipepidine hibenzate for assay S ture gradually, and heat strongly with occasional shaking un- Internal standard solution—A solution of dibucaine til the contents fuse to yield a clear liquid. Cool, add 30 mL hydrochloride in methanol (1 in 2000). of a solution of diammonium hydrogen citrate (9 in 20) and Operating conditions— 50 mL of water, dissolve by heating on a water bath, cool, Detector: An ultraviolet absorption photometer add water to make 100 mL, and use this solution as the sam- (wavelength: 254 nm). ple stock solution. Take 25 mL of the solution to a separator, Column: A stainless steel column 4.6 mm in inside di- add 10 mL of a solution of ammonium sulfate (2 in 5) and 5 ameter and 15 cm in length, packed with octadecylsilanized drops of thymol blue TS, neutralize with ammonia TS, and silica gel for liquid chromatography (5 mminparticledi- add 2.5 mL of ammonia TS. To this solution add exactly 20 ameter). mL of a solution of dithizone in n-butyl acetate (1 in 500), Column temperature: A constant temperature of about shake for 10 minutes, and use this n-butyl acetate solution as 409C. the sample solution. Separately, place 6.0 mL of Standard Mobile phase: A mixture of a solution of sodium lauryl Lead Solution in a platinum crucible, proceed as directed in JP XV O‹cial Monographs / Tizanidine Hydrochloride 1179 the sample solution, and use this solution as the standard so- smoke stops evolving. Heat strongly between 9009Cand lution. Determine the absorbances of the sample solution and 9509C to constant mass, cool, and weigh as titanium oxide standard solution as directed under Atomic Absorption Spec- (TiO2). trophotometry <2.23> according to the following conditions: Containers and storage Containers—Well-closed contain- the absorbance of the sample solution is smaller than that of ers. the standard solution (not more than 60 ppm). Gas: Combustible gas—Acetylene gas or hydrogen gas Supporting gas—Air Lamp: Lead hollow-cathode lamp Tizanidine Hydrochloride Wavelength: 283.3 nm チザニジン塩酸塩 (2) Arsenic <1.11>—Perform the test with 20 mL of the sample stock solution obtained in (1) as the test solution: the stain is not deeper than the following standard stain. Standard stain: Proceed in the same manner without Titanium Oxide, transfer 20 mL of the obtained solution to a generator bottle, add 2.0 mL of Standard Arsenic Solution, and proceed in the same manner as the test with the test solu- tion (not more than 10 ppm). C H ClN S.HCl: 290.17 (3) Water-soluble substances—Shake thoroughly 4.0 g of 9 8 5 5-Chloro-N-(4,5-dihydro-1H-imidazol-2-yl)- Titanium Oxide with 50 mL of water, and allow to stand 2,1,3-benzothiadiazole-4-amine monohydrochloride overnight. Shake thoroughly with 2 mL of ammonium chlo- [64461-82-1] ride TS, add further 2 mL of ammonium chloride TS if neces- sary, and allow titanium oxide to settle. Add water to make Tizanidine Hydrochloride, when dried, contains 200 mL, shake thoroughly, and ˆlter through double ˆlter not less than 99.0z and not more than 101.0z of paper. Discard the ˆrst 10 mL of the ˆltrate, evaporate 100 C H ClN S.HCl. mL of the clear ˆltrate on a water bath, and heat strongly at 9 8 5 6509C to constant mass: the mass of the residue is not more Description Tizanidine Hydrochloride occurs as a white to than 5.0 mg. light yellowish white crystalline powder. It is soluble in water, slightly soluble in ethanol (99.5), and Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, practically insoluble in acetic anhydride and in acetic acid 3 hours). (100). Assay Weigh accurately about 0.2 g of Titanium Oxide, Melting point: about 2909C (with decomposition). previously dried, transfer to a crucible, and add 3 g of potas- Identiˆcation (1) Determine the absorption spectrum of a sium disulfate. Cover, and heat gently at ˆrst, gradually raise solution of Tizanidine Hydrochloride in diluted 1 mol/L am- the temperature, and then heat the fused contents for 30 monia TS (1 in 10) (1 in 125,000) as directed under Ultra- minutes. Continue heating for 30 minutes at a higher temper- violet-visible Spectrophotometry <2.24>, and compare the ature to make the fused mixture a deep yellow-red, almost spectrum with the Reference Spectrum: both spectra exhibit clear liquid. Cool, transfer the contents of the crucible to a similar intensities of absorption at the same wavelengths. 250-mL beaker, wash the crucible with a mixture of 75 mL of (2) Determine the infrared absorption spectrum of water and 2.5 mL of sulfuric acid into the beaker, and heat Tizanidine Hydrochloride as directed in the potassium chlo- on a water bath until the solution becomes almost clear. Dis- ride disk method under Infrared Spectrophotometry <2.25>, solve 2 g of L-tartaric acid in the solution, add 2 to 3 drops of and compare the spectrum with the Reference Spectrum: bromothymol blue TS, neutralize with ammonia TS, and both spectra exhibit similar intensities of absorption at the acidify with 1 to 2 mL of diluted sulfuric acid (1 in 2). Pass same wave numbers. hydrogen sulˆde su‹ciently through the solution, add 30 mL (3) A solution of Tizanidine Hydrochloride (1 in 50) of ammonia TS, again saturate the solution with hydrogen responds to the Qualitative Tests <1.09> for chloride. sulˆde, allow to stand for 10 minutes, and ˆlter. Wash the precipitate on the ˆlter paper with ten 25-mL portions of a Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of mixture of ammonium L-tartrate solution (1 in 100) and am- Tizanidine Hydrochloride according to Method 3, and per- monium sulˆde TS (9:1). When the precipitate is ˆltered and form the test. Prepare the control solution with 2.0 mL of washed, prevent iron (II) sulˆde from oxidation by ˆlling the Standard Lead Solution (not more than 20 ppm). solution on the ˆlter paper. Combine the ˆltrate and the (2) Related substances—Dissolve 60 mg of Tizanidine washings, add 40 mL of diluted sulfuric acid (1 in 2), and boil Hydrochloride in 10 mL of a mixture of water and acetoni- to expel hydrogen sulˆde. Cool, and dilute with water to trile (17:3), and use this solution as the sample solution. Pipet make400mL.Addgradually40mLofcupferronTStothe 1 mL of the sample solution, add the mixture of water and solution with stirring, and allow to stand. After sedimenta- acetonitrile (17:3) to make exactly 200 mL, and use this solu- tion of a yellow precipitate, add again cupferron TS until a tion as the standard solution. Perform the test with exactly 10 white precipitate is produced. Filter by slight suction using mL of the sample solution and standard solution as directed quantitative ˆlter paper, wash with twenty portions of diluted under Liquid Chromatography <2.01> according to the fol- hydrochloric acid (1 in 10), and remove water by stronger lowing conditions, and determine each peak area by the auto- suction at the last washing. Dry the precipitate together with matic integration method: the area of the peak other than the ˆlter paper at 709C, transfer to a tared crucible, and heat tizanidine is not larger than 1/5 times the peak area of tizani- very gently at ˆrst, and raise the temperature gradually after dine with the standard solution. 1180 Tobramycin / O‹cial Monographs JP XV

Operating conditions— ers. Detector: An ultraviolet absorption photometer (wavelength: 230 nm for about 3 minutes after sample injec- tion and 318 nm subsequently). Tobramycin Column: A stainless steel column 4.6 mm in inside diameter and 12.5 cm in length, packed with octadecyl- トブラマイシン silanized silica gel for liquid chromatography (5 mmin particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water and formic acid (200:1), adjusted to pH 8.5 with ammonia water (28). Mobile phase B: A mixture of acetonitrile and the mobile phase A (4:1). Flowing of the mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table.

Time after injection Mobile phase Mobile phase of sample (min) A(volz) B(volz)

0–10 81ª 68 19 ª 32 C18H37N5O9:467.51 10–13 68 32 3-Amino-3-deoxy-a-D-glucopyranosyl-(1ª6)- 13–26 68ª 10 32 ª 90 [2,6-diamino-2,3,6-trideoxy-a-D-ribo-hexopyranosyl- 26–28 10 90 (1ª4)]-2-deoxy-D-streptamine [32986-56-4]

Flow rate: Adjust the ‰ow rate so that the retention time of Tobramycin is an aminoglycoside substance having tizanidine is about 7 minute. antibacterial activity produced by the growth of Strep- Time span of measurement: About 4 times as long as the tomyces tenebrarius. retention time of tizanidine beginning after the solvent peak. It contains not less than 900 mg (potency) and not System suitability— more than 1060 mg (potency) per mg, calculated on the Test for required detectability: Measure exactly 2 mL of anhydrous basis. The potency of Tobramycin is ex- the standard solution, and add the mixture of water and pressed as mass (potency) of tobramycin (C18H37N5O9). acetonitrile (17:3) to make exactly 10 mL. Conˆrm that the Description Tobramycin occurs as a white to pale yellowish peak area of tizanidine obtained with 10 L of this solution is m white powder. equivalent to 14 to 26z of that with 10 mL of the standard so- It is very soluble in water, freely soluble in formamide, lution. slightly soluble in methanol, and very slightly soluble in System performance: Dissolve 2 mg each of Tizanidine ethanol (95). Hydrochloride and p-toluenesulfonic acid monohydrate in It is hygroscopic. 100 mL of the mixture of water and acetonitrile (17:3). When the procedure is run with 10 mL of this solution under the Identiˆcation (1) Determine the spectrum of a solution of above operating conditions, p-toluenesulfonic acid and Tobramycin in heavy water for nuclear magnetic resonance tizanidine are eluted in this order with the resolution between spectroscopy (1 in 125) as directed under Nuclear Magnetic these peaks being not less than 10. Resonance Spectroscopy <2.21> (1H), using sodium 3- System repeatability: When the test is repeated 6 times with trimethylsilylpropanesulfonate for nuclear magnetic 10 mL of the standard solution under the above operating resonance spectroscopy as an internal reference compound: it conditions, the relative standard deviation of the peak area of exhibits a double signal A at around d5.1 ppm, a multiple tizanidine is not more than 2.0z. signal B between d2.6 ppm and d4.0 ppm, and a multiple sig- nal C between d1.0 ppm and d2.1 ppm. The ratio of the in- Loss on drying <2.41> Not more than 0.2z (1 g, 1059C, 3 tegrated intensity of these signals, A:B:C, is about 1:8:2. hours). (2) Dissolve 10 mg each of Tobramycin and Tobramycin Residue on ignition <2.44> Not more than 0.1z (1 g). Reference Standard in 1 mL of water, and use these solutions as the sample solution and the standard solution. Perform Assay Weigh accurately about 0.2 g of Tizanidine the test with these solutions as directed under Thin-layer Hydrochloride, previously dried, dissolve in 60 mL of a Chromatography <2.03>.Spot4mL of the sample solution mixture of acetic anhydride and acetic acid (100) (7:3) with and standard solution on a plate of silica gel for thin-layer the aid of warming. After cooling, titrate 2.50 with 0.1 < > chromatography. Develop the plate with a mixture of ammo- mol/L perchloric acid VS (potentiometric titration). Perform nia TS, 1-butanol and methanol (5:5:2) to a distance of about a blank determination in the same manner, and make any 10 cm, and air-dry the plate. Spray evenly ninhydrin TS on necessary correction. the plate, and heat at 1009C for 5 minutes: the Rf values of Each mL of 0.1 mol/L perchloric acid VS the principal spots obtained from the sample solution and the

＝29.02 mg of C9H8ClN5S.HCl standard solution are the same. 20 Containers and storage Containers—Well-closed contain- Optical rotation <2.49> [a]D : ＋138 – ＋1489(1 g calculated JP XV O‹cial Monographs / Tocopherol 1181 on the anhydrous basis, water, 25 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 0.10 g of Tobramycin in 10 mL of water is between 9.5 and 11.5. Tocopherol Purity (1) Clarity and color of solution—Dissolve 1.0 g of Vitamin E Tobramycin in 10 mL of water: the solution is clear and dl-a-Tocopherol colorless to pale yellow. (2) Heavy metals <1.07>—Proceed with 1.0 g of トコフェロール Tobramycin according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). (3) Related substances—Dissolve 80 mg of Tobramycin in 10 mL of diluted ammonia solution (28) (1 in 250), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add diluted ammonia solution (28) (1 in 250) to make exactly 100 mL, and use this solution as the standard C29H50O2: 430.71 solution. Perform the test with these solutions as directed un- 2,5,7,8-Tetramethyl-2-(4,8,12-trimethyltridecyl)chroman- der Thin-layer Chromatography <2.03>.Spot5mLofthe 6-ol [10191-41-0] samplesolutionandstandardsolutiononaplateofsilicagel for thin-layer chromatography. Develop the plate with a mix- Tocopherol contains not less than 96.0z and not ture of ammonia solution (28), ethanol (95) and 2-butanone more than 102.0z of C29H50O2. (1:1:1) to a distance of about 10 cm, air-dry the plate, then Description Tocopherol is a clear, yellow to red-brown, vis- further dry at 1109C for 10 minutes. Immediately spray even- cous liquid. It is odorless. ly a mixture of water and sodium hypochlorite TS (4:1) on It is miscible with ethanol (99.5), with acetone, with chlo- the plate, air-dry the plate, then spray potassium iodide- roform, with diethyl ether and with vegetable oils. starch TS on the plate: the spot other than the principal spot It is freely soluble in ethanol (95), and practically insoluble from the sample solution is not more intense than the spot in water. from the standard solution. It is optically inactive. Water <2.48> Not more than 11.0z (0.1 g, volumetric titra- It is oxidized by air and light, and acquires a dark red tion, direct titration). Use a mixture of formamide for water color. determination and methanol for water determination (3:1) in- Identification (1) Dissolve 0.01 g of Tocopherol in 10 mL stead of methanol for water determination. of ethanol (99.5), add 2 mL of nitric acid, and heat at 759C Residue on ignition <2.44> Not more than 1.0z (0.5 g). for 15 minutes: a red to orange color develops. (2) Determine the infrared absorption spectrum of Assay Perform the test according to the Cylinder-plate Tocopherol as directed in the liquid film method under In- method as directed under Microbial Assay for Antibiotics frared Spectrophotometry <2.25>, and compare the spectrum <4.02> according to the following conditions. with the Reference Spectrum or the spectrum of Tocopherol (i) Test organism—Bacillus subtilis ATCC 6633 Reference Standard: both spectra exhibit similar intensities (ii) Culture medium—Use the medium i in 1) under (1) of absorption at the same wave numbers. Agar media for seed and base layer. 1z (iii) Standard solutions—Weigh accurately an amount of Absorbance <2.24> E 1cm (292 nm): 71.0 – 76.0 (10 mg, Tobramycin Reference Standard, equivalent to about 25 mg ethanol (99.5), 200 mL). (potency), dissolve in 0.1 mol/L phosphate buŠer solution, 20 Refractive index <2.45> nD : 1.503 – 1.507 pH 8.0 to make exactly 25 mL, and use this solution as the 20 standard stock solution. Keep the standard stock solution be- Specific gravity <2.56> d20: 0.947 – 0.955 tween 59Cand159C, and use within 30 days. Take exactly a Purity (1) Clarity and color of solution—Dissolve 0.10 g suitable amount of the standard stock solution before use, of Tocopherol in 10 mL of ethanol (99.5): the solution is add 0.1 mol/L phosphate buŠer solution, pH 8.0 to make so- clear and has no more color than Matching Fluid C. lutionssothateachmLcontains8mg (potency) and 2 mg (2) Heavy metals <1.07>—Proceed with 1.0 g of (potency), and use these solutions as the high concentration Tocopherol according to Method 4, and perform the test. standard solution and low concentration standard solution, Prepare the control solution with 2.0 mL of Standard Lead respectively. Solution (not more than 20 ppm). (iv) Sample solutions—Weigh accurately an amount of Tobramycin, equivalent to about 25 mg (potency), and dis- Assay Dissolve about 50 mg each of Tocopherol and To- solve in 0.1 mol/L phosphate buŠer solution, pH 8.0 to make copherol Reference Standard, accurately weighed, in ethanol exactly 25 mL. Take exactly a suitable amount of this solu- (99.5) to make exactly 50 mL, and use these solutions as the tion, add 0.1 mol/L phosphate buŠer solution, pH 8.0 to sample solution and standard solution. Perform the test with make solutions so that each mL contains 8 mg (potency) and 2 exactly 20 mLeachofthesesolutionsasdirectedunderLiquid mg (potency), and use these solutions as the high concentra- Chromatography <2.01> according to the following condi- tion sample solution and low concentration sample solution, tions, and determine the peak heights, HT and HS,of respectively. tocopherol in the sample solution and standard solution.

Containers and storage Containers—Tight containers. Amount (mg) of C29H50O2 ＝ WS × (HT/HS) 1182 Tocopherol Acetate / O‹cial Monographs JP XV

Tocopherol Acetate as directed in the liquid film method un- W : Amount (mg) of Tocopherol Reference Standard S der Infrared Spectrophotometry <2.25>, and compare the Operating conditions— spectrum with the Reference Spectrum or the spectrum of Detector: An ultraviolet absorption photometer (wave- Tocopherol Acetate Reference Standard: both spectra exhibit length: 292 nm). similar intensities of absorption at the same wave numbers. Column: A stainless steel column 4.6 mm in inside di- Absorbance <2.24> E 1z (284 nm): 41.0 – 45.0 (10 mg, ameter and 15 cm in length, packed with octadecylsilanized 1cm ethanol (99.5), 100 mL). silica gel for liquid chromatography (5 mLinparticle 20 diameter). Refractive index <2.45> nD : 1.494 – 1.499 Column temperature: A constant temperature of about Specific gravity <2.56> d20: 0.952 – 0.966 359C. 20 Mobile phase: A mixture of methanol and water (49:1). Purity (1) Clarity and color of solution—Dissolve 0.10 g Flow rate: Adjust the ‰ow rate so that the retention time of of Tocopherol Acetate in 10 mL of ethanol (99.5): the solu- tocopherol is about 10 minutes. tion is clear, and has no more color than the following con- System suitability— trol solution. System performance: Dissolve 0.05 g each of Tocopherol Control solution: To 0.5 mL of Ferric Chloride Colorimet- and tocopherol acetate in 50 mL of ethanol (99.5). When the ric Stock Solution add 0.5 molWL hydrochloric acid TS to procedure is run with 20 mL of this solution under the above make 100 mL. operating conditions, tocopherol and tocopherol acetate are (2) Heavy metals <1.07>—Carbonize 1.0 g of Tocopherol eluted in this order with the resolution between these peaks Acetate by gentle heating. Cool, add 10 mL of a solution of being not less than 2.6. magnesium nitrate hexahydrate in ethanol (95) (1 in 10), and System repeatability: When the test is repeated 5 times with ignite the ethanol to burn. Cool, add 1 mL of sulfuric acid, 20 mL of the standard solution under the above operating proceed according to Method 4, and perform the test. Pre- conditions, the relative standard deviation of the peak height- pare the control solution with 2.0 mL of Standard Lead Solu- s of tocopherol is not more than 0.8z. tion (20 ppm). (3) a-Tocopherol—Dissolve 0.10 g of Tocopherol Containers and storage Containers—Tight containers. Acetate in exactly 10 mL of hexane, and use this solution as Storage—Light-resistant, and well-filled, or under nitro- the sample solution. Separately, dissolve 50 mg of gen atmosphere. Tocopherol Reference Standard in hexane to make exactly 100 mL. Pipet 1 mL of this solution, add hexane to make ex- actly 10 mL, and use this solution as the standard solution. Tocopherol Acetate Perform the test with these solutions as directed under Thin- layer Chromatography <2.03>.Spot10mLeachofthesample Vitamin E Acetate solution and standard solution on a plate of silica gel for dl-a-Tocopherol Acetate thin-layer chromatography. Develop the plate with a mixture of toluene and acetic acid (100) (19:1) to a distance of about トコフェロール酢酸エステル 10 cm, and air-dry the plate. Spray evenly a solution of iron (III) chloride hexahydrate in ethanol (99.5) (1 in 500) on the plate, then spray evenly a solution of a,a?-dipyridyl in ethanol (99.5) (1 in 200) on the same plate, and allow to stand for 2 to 3 minutes: the spot from the sample solution cor- responding to that from the standard solution is not larger than and not more intense than the spot from the standard solution. C31H52O3:472.74 2,5,7,8-Tetramethyl-2-(4,8,12-trimethyltridecyl)chroman- Assay Dissolve 50 mg each of Tocopherol Acetate and 6-yl acetate [7695-91-2] Tocopherol Acetate Reference Standard, accurately weighed, in ethanol (99.5) to make exactly 50 mL, and use these solu- Tocopherol Acetate contains not less than 96.0z tions as the sample solution and standard solution. Perform and not more than 102.0z of C31H52O3. the test with exactly 20 mLeachofthesesolutionsasdirected under Liquid Chromatography <2.01> according to the fol- Description Tocopherol Acetate is a clear, colorless or yel- lowing conditions, and determine the peak heights, HT and low, viscous and odorless liquid. HS, of tocopherol acetate in the sample solution and the stan- It is miscible with ethanol (99.5), with acetone, with chlo- dard solution, respectively. roform, with diethyl ether, with hexane and with fixed oils. It is freely soluble in ethanol (95), and practically insoluble Amount (mg) of C31H52O3 ＝ WS × (HT/HS) in water. WS: Amount (mg) of Tocopherol Acetate Reference It is optically inactive. Standard It is affected by air and light. Operating conditions— Identification (1) Dissolve 0.05 g of Tocopherol Acetate Detector: An ultraviolet absorption photometer (wave- in 10 mL of ethanol (99.5), add 2 mL of nitric acid, and heat length: 284 nm). at 759C for 15 minutes: a red to orange color is produced. Column: A stainless steel column 4.6 mm in inside di- (2) Determine the infrared absorption spectrum of ameter and 15 cm in length, packed with octadecylsilanized JP XV O‹cial Monographs / Tocopherol Calcium Succinate 1183 silica gel (5 mm in particle diameter). Reference Spectrum: both spectra exhibit similar intensities Column temperature: A constant temperature of about of absorption at the same wave numbers. 359C. (3) Dissolve 5 g of Tocopherol Calcium Succinate in 30 Mobile phase: A mixture of methanol and water (49:1). mL of chloroform, add 10 mL of hydrochloric acid, shake Flow rate: Adjust the ‰ow rate so that the retention time of for 10 minutes, then draw off the water layer, and neutralize tocopherol acetate is about 12 minutes. with ammonia TS: the solution responds to the Qualitative System suitability— Tests <1.09> for calcium salt. System performance: Dissolve 0.05 g each of Tocopherol Absorbance <2.24> E 1z (286 nm): 36.0 – 40.0 (10 mg, chlo- Acetate and tocopherol in 50 mL of ethanol (99.5). When the 1cm roform, 100 mL). procedure is run with 20 mL of this solution under the above operating conditions, tocopherol and tocopherol acetate are Purity (1) Clarity and color of solution—Dissolve 0.10 g eluted in this order with the resolution between these peaks of Tocopherol Calcium Succinate in 10 mL of chloroform: being not less than 2.6. the solution is clear, and has no more color than the follow- System repeatability: When the test is repeated 5 times with ing control solution. 20 mL of the standard solution under the above operating Control solution: To 0.5 mL of Ferric Chloride Colorimet- conditions, the relative standard deviation of the peak height- ric Stock Solution add 0.5 molWL hydrochloric acid TS to s of tocopherol acetate is not more than 0.8z. make 100 mL. (2) Alkalinity—To 0.20 g of Tocopherol Calcium Suc- Containers and storage Containers—Tight containers. cinate add 10 mL of diethyl ether, 2 mL of water, 1 drop of Storage—Light-resistant. phenolphthalein TS and 0.10 mL of 0.1 molWL hydrochloric acid VS, and shake: no red color develops in the water layer. (3) Chloride <1.03>—Dissolve 0.10 g of Tocopherol Cal- Tocopherol Calcium Succinate cium Succinate in 4 mL of acetic acid (100), add 20 mL of water and 50 mL of diethyl ether, shake thoroughly, and col- Vitamin E Calcium Succinate lect the water layer. To the diethyl ether layer add 10 mL of トコフェロールコハク酸エステルカルシウム water, shake, and collect the water layer. Combine the water layers, add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solu- tion. Prepare the control solution in the same manner using 0.60 mL of 0.01 molWLhydrochloricacidVSinplaceof Tocopherol Calcium Succinate (not more than 0.212z). (4) Heavy metals <1.07>—Proceed with 1.0 g of Tocopherol Calcium Succinate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of

C66H106CaO10: 1099.62 Standard Lead Solution (not more than 20 ppm). Monocalcium bis｛3-[2,5,7,8-tetramethyl-2-(4,8,12- (5) Arsenic <1.11>—Prepare the test solution with 1.0 g trimethyltridecyl)chroman-6-yloxycarbonyl]propanoate｝ of Tocopherol Calcium Succinate according to Method 3, [14638-18-7] and perform the test (not more than 2 ppm). (6) a-Tocopherol—Dissolve 0.10 g of Tocopherol Calci- Tocopherol Calcium Succinate, when dried, con- um Succinate in exactly 10 mL of chloroform, and use this tains not less than 96.0z and not more than 102.0z of solution as the sample solution. Separately, dissolve 50 mg of C66H106CaO10. Tocopherol Reference Standard in chloroform to make ex- actly 100 mL. Pipet 1 mL of this solution, add chloroform to Description Tocopherol Calcium Succinate occurs as a make exactly 10 mL, and use this solution as the standard so- white to yellowish white powder. It is odorless. lution. Perform the test with these solutions as directed under It is freely soluble in chloroform and in carbon tetrachlo- Thin-layer Chromatography <2.03>.Spot10mLeachofthe ride, and practically insoluble in water, in ethanol (95) and in sample solution and standard solution on a plate of silica gel acetone. for thin-layer chromatography. Develop the plate with a mix- Shake 1 g of Tocopherol Calcium Succinate with 7 mL of ture of toluene and acetic acid (100) (19:1) to a distance of acetic acid (100): it dissolves, and produces a turbidity after about 10 cm, and air-dry the plate. Spray evenly a solution of being allowed to stand for a while. iron (III) chloride hexahydrate in ethanol (99.5) (1 in 500) on It dissolves in acetic acid (100). the plate, then spray evenly a solution of a-a?-dipyridyl in It is optically inactive. ethanol (99.5) (1 in 200) on the same plate, and allow to stand Identification (1) Dissolve 0.05 g of Tocopherol Calcium for 2 to 3 minutes: the spots from the sample solution cor- Succinate in 1 mL of glacial aetic acid, add 9 mL of ethanol responding to the spots from the standard solution is not (99.5), and mix. To this solution add 2 mL of fuming nitric larger than and not more intense than the spots from the acid, and heat at 759C for 15 minutes: a red to orange color standard solution. develops. Loss on drying <2.41> Not more than 2.0z (1 g, in vacuum, (2) Dissolve 0.08 g of Tocopherol Calcium Succinate, phosphorus (V) oxide, 24 hours). previously dried, in 0.2 mL of carbon tetrachloride. Deter- mine the infrared absorption spectrum of the solution as Assay Weigh accurately about 50 mg each of Tocopherol directed in the liquid film method under Infrared Spec- Calcium Succinate and Tocopherol Succinate Reference trophotometry <2.25>, and compare the spectrum with the Standard, previously dried, dissolve in a mixture of ethanol 1184 Tocopherol Nicotinate / O‹cial Monographs JP XV

(99.5) and diluted acetic acid (100) (1 in 5) (9:1) to make ex- It is freely soluble in ethanol (99.5), and practically insolu- actly 50 mL, and use these solutions as the sample solution ble in water. and the standard solution. Pipet exactly 20 mLeachofthe A solution of Tocopherol Nicotinate in ethanol (99.5) sample solution and standard solution, and perform the test (1 in 10) shows no optical rotation. as directed under Liquid Chromatography <2.01> according It is affected by light. to the following operating conditions. Determine the peak Identification (1) Determine the absorption spectrum of a heights, H and H , of tocopherol succinate in these solu- T S solution of Tocopherol Nicotinate in ethanol (99.5) (1 in tions, respectively. 20,000) as directed under Ultraviolet-visible Spectrophoto-

Amount (mg) C66H106CaO10 metry <2.24>, and compare the spectrum with the Reference ＝ WS×(HT/HS)×1.0358 Spectrum or the spectrum of a solution of Tocopherol Nicotinate Reference Standard prepared in the same manner W : Amount (mg) of Tocopherol Succinate Reference S as the sample solution: both spectra exhibit similar intensities Standard of absorption at the same wavelengths. Operating conditions— (2) Determine the infrared spectrum of Tocopherol Detector: An ultraviolet absorption photometer Nicotinate, if necessary melt by warming, as directed in the (wavelength: 284 nm). liquid film method under the Infrared Spectrophotometry Column: A stainless steel column about 4 mm in inside di- <2.25>, and compare the spectrum with the Reference Spec- ameter and 15 to 30 cm in length, packed with octadecyl- trum or the spectrum of Tocopherol Nicotinate Reference silanized silica gel (5 to 10 mL in particle diameter). Standard: both spectra exhibit similar intensities of absorp- Column temperature: Room temperature. tion at the same wave numbers. Mobile phase: A mixture of methanol, water and acetic Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of acid (100) (97:2:1). Tocopherol Nicotinate according to Method 4, and perform Flow rate: Adjust the flow rate so that the retention time of the test. Prepare the control solution with 2.0 mL of Stan- tocopherol succinate is about 8 minutes. dard Lead Solution (not more than 20 ppm). Selection of column: Dissolve 0.05 g each of tocopherol (2) Arsenic <1.11>—Prepare the test solution with 1.0 g succinate and tocopherol in 50 mL of a mixture of ethanol of Tocopherol Nicotinate according to Method 4, and per- (99.5) and diluted acetic acid (100) (1 in 5) (9:1). Proceed with form the test (not more than 2 ppm). 20 mL of this solution under the above operating conditions, (3) Related substances—Dissolve 0.05 g of Tocopherol and calculate the resolution. Use a column giving elution of Nicotinate in 50 mL of ethanol (99.5), and use this solution as tocopherol succinate and tocopherol in this order with the the sample solution. Pipet 7 mL of this solution, add ethanol resolution between these peaks being not less than 2.0. (99.5) to make exactly 200 mL, and use this solution as the System repeatability: Repeat the test five times with the standard solution. Perform the test with exactly 10 mLeach standard solution under the above operating conditions: the of the sample solution and standard solution as directed un- relative standard deviation of the peak height of tocopherol der Liquid Chromatography <2.01> according to the follow- succinate is not more than 0.8z. ing conditions. Determine each peak area of both solutions Containers and storage Containers—Tight containers. by the automatic integration method: the total area of the Storage—Light-resistant. peaks other than tocopherol nicotinate from the sample solu- tion is not larger than the peak area of tocopherol nicotinate from the standard solution, and the area of a peak which has Tocopherol Nicotinate a retention time 0.8 to 0.9 times that of tocopherol nicotinate from the sample solution is not larger than 4W7ofthepeak Vitamin E Nicotinate area of tocopherol nicotinate from the standard solution. dl-a-Tocopherol Nicotinate Operating conditions— Detector, column, and column temperature: Proceed as トコフェロールニコチン酸エステル directed in the operating conditions in the Assay. Mobile phase: A mixture of methanol and water (19:1). Flow rate: Adjust the flow rate so that the retention time of tocopherol nicotinate is about 20 minutes. Time span of measurement: About 1.5 times as long as the retention time of tocopherol nicotinate beginning after the solvent peak. System suitability— Test for required detection: To exactly 1 mL of the sample solution add ethanol (99.5) to make exactly 100 mL, and use C H NO : 535.80 35 53 3 this solution as the test solution for system suitability. Pipet 1 2,5,7,8-Tetramethyl-2-(4,8,12-trimethyltridecyl)chroman- mL of the test solution for system suitability, add ethanol 6-yl nicotinate [51898-34-1] (99.5) to make exactly 10 mL. Confirm that the peak area of Tocopherol Nicotinate contains not less than 96.0z tocopherol nicotinate obtained from 10 mL of this solution is of nicotinic acid dl-a-tocopherol (C H NO ). equivalent to 7 to 13z of that of tocopherol nicotinate ob- 35 53 3 tained from 10 mL of the test solution for system suitability. Description Tocopherol Nicotinate occurs as a yellow to System performance: Dissolve 0.05 g of Tocopherol orange-yellow liquid or solid. Nicotinate and 0.25 g of tocopherol in 100 mL of ethanol JP XV O‹cial Monographs / Todralazine Hydrochloride Hydrate 1185

(99.5). When the procedure is run with 10 mLofthissolution monohydrochloride monohydrate [3778-76-5,anhydride] under the above operating conditions, tocopherol and tocopherol nicotinate are eluted in this order with the resolu- Todralazine Hydrochloride Hydrate contains not tion between these peaks being not less than 8. less than 98.5z of todralazine hydrochloride System repeatability: When the test is repeated 6 times with (C11H12N4O2.HCl: 268.70), calculated on the anhy- 10 mL of the standard solution under the above operating drous basis. conditions, the relative standard deviation of the peak areas Description Todralazine Hydrochloride Hydrate occurs as of tocopherol nicotinate is not more than 2.0z. white crystals or crystalline powder. It has a slight, charac- Assay Weigh accurately about 50 mg each of Tocopherol teristic odor, and has a bitter taste. Nicotinate and Tocopherol Nicotinate Reference Standard, It is very soluble in formic acid, freely soluble in methanol, dissolve each in ethanol (99.5) to make exactly 50 mL, and soluble in water, sparingly soluble in ethanol (95), and practi- use these solutions as the sample solution and standard solu- cally insoluble in diethyl ether. tion, respectively. Perform the test with exactly 5 mLeachof The pH of a solution of Todralazine Hydrochloride Hy- the sample solution and standard solution as directed under drate (1 in 200) is between 3.0 and 4.0. Liquid Chromatography <2.01> according to the following Identification (1) To 2 mL of a solution of Todralazine conditions, and determine peak areas, A and A ,of T S Hydrochloride Hydrate (1 in 200) add 5 mL of silver nitrate- tocopherol nicotinate of these solutions. ammonia TS: the solution becomes turbid, and a black

Amount (mg) of C35H53NO3 precipitate is formed. ＝ WS×(AT/AS) (2) Determine the absorption spectrum of a solution of Todralazine Hydrochloride Hydrate in 0.1 mol/L W : Amount (mg) of Tocopherol Nicotinate S hydrochloric acid TS (3 in 100,000) as directed under Ultrav- Operating conditions— iolet-visible Spectrophotometry <2.24>, and compare the Detector: An ultraviolet absorption photometer spectrum with the Reference Spectrum: both spectra exhibit (wavelength: 264 nm). similar intensities of absorption at the same wavelengths. Column: A stainless steel column 4.6 mm in inside di- (3) Determine the infrared absorption spectrum of ameter and 15 cm in length, packed with octadecylsilanized Todralazine Hydrochloride Hydrate as directed in the potas- silica gel for liquid chromatography (5 mminparticledi- sium chloride disk method under the Infrared Spectrophoto- ameter). metry <2.25>, and compare the spectrum with the Reference Column temperature: A constant temperature of about Spectrum: both spectra exhibit similar intensities of absorp- 359C. tion at the same wave numbers. Mobile phase: Methanol (4) A solution of Todralazine Hydrochloride Hydrate (1 Flow rate: Adjust the flow rate so that the retention time of in 50) responds to the Qualitative Tests <1.09> (1) for chlo- tocopherol nicotinate is about 10 minutes. ride. System suitability— Purity (1) Clarity and color of solution—Dissolve 0.30 g System performance: Dissolve 0.05 g of Tocopherol of Todralazine Hydrochloride Hydrate in 10 mL of water: Nicotinate and 0.25 g of tocopherol in 100 mL of ethanol the solution is clear and colorless to pale yellow. (99.5). When the procedure is run with 5 mL of this solution (2) Sulfate <1.14>—Proceed the test with 2.0 g of under the above operating conditions, tocopherol and Todralazine Hydrochloride Hydrate. Prepare the control so- tocopherol nicotinate are eluted in this order with the resolu- lution with 0.50 mL of 0.005 molWL sulfuric acid VS (not tion between these peaks being not less than 3. more than 0.012z). System repeatability: When the test is repeated 6 times with (3) Heavy metals <1.07>—Proceed with 1.0 g of Todrala- 5 mL of the standard solution under the above operating con- zine Hydrochloride Hydrate according to Method 2, and per- ditions: the relative standard deviation of the peak areas of form the test. Prepare the control solution with 2.0 mL of tocopherol nicotinate is not more than 0.8z Standard Lead Solution (not more than 20 ppm). Containers and storage Containers—Tight containers. (4) Arsenic <1.11>—Prepare the test solution with 1.0 g Storage—Light-resistant. of Todralazine Hydrochloride Hydrate according to Method 1, and perform the test (not more than 2 ppm). (5) Related substances—Dissolve 50 mg of Todralazine Todralazine Hydrochloride Hydrate Hydrochloride Hydrate in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the Ecarazine Hydrochloride sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform トドララジン塩酸塩水和物 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 both solutions by the automatic integration method: the total area of the peaks other than the peak of todralazine from the sample solution is not larger than the peak area of todralazine from the standard solution.

C11H12N4O2.HCl.H2O: 286.71 Operating conditions— Ethyl 2-(phthalazin-1-yl)hydrazinecarboxylate Detector: An ultraviolet absorption photometer 1186 Tofisopam / O‹cial Monographs JP XV

(wavelength: 240 nm). 4-methyl-5H-2,3- [22345-47-7] Column: A stainless steel column 3.9 mm in inside di- ameter and 30 cm in length, packed with octadecylsilanized Tofisopam, when dried, contains not less than silica gel for liquid chromatography (10 mminparticledi- 98.0z of C22H26N2O4. ameter). Description Tofisopam occurs as a pale yellowish white, Column temperature: A constant temperature of about crystalline powder. 259C. It is freely soluble in acetic acid (100), soluble in acetone, Mobile phase: Dissolve 1.10 g of sodium 1-heptane sul- sparingly soluble in ethanol (95), slightly soluble in diethyl fonate in 1000 mL of diluted methanol (2 in 5). Adjust the ether, and practically insoluble in water. pH of the solution to between 3.0 and 3.5 with acetic acid A solution of Tofisopam in ethanol (95) (1 in 100) shows (100). no optical rotation. Flow rate: Adjust the flow rate so that the retention time of todralazine is about 8 minutes. Identification (1) Determine the absorption spectrum of a Time span of measurement: About twice as long as the solution of Tofisopam in ethanol (95) (1 in 100,000) as direct- retention time of todralazine beginning after the solvent ed under Ultraviolet-visible Spectrophotometry <2.24>,and peak. compare the spectrum with the Reference Spectrum: both System suitability— spectra exhibit similar intensities of absorption at the same Test for required detection: To exactly 5 mL of the stan- wavelengths. dard solution add the mobile phase to make exactly 25 mL. (2) Determine the infrared absorption spectrum of Confirm that the peak area of todralazine obtained from 10 Tofisopam, previously dried, as directed in the potassium mL of this solution is equivalent to 15 to 25z of that of bromide disk method under Infrared Spectrophotometry todralazine obtained from 10 mL of the standard solution. <2.25>, and compare the spectrum with the Reference Spec- System performance: Dissolve 5 mg each of Todralazine trum: both spectra exhibit similar intensities of absorption at Hydrochloride Hydrate and potassium biphthalate in 100 mL thesamewavenumbers. of the mobile phase. When the procedure is run with 10 mLof Melting point <2.60> 155 – 1599C this solution under the above operating conditions, phthalic acidandtodralazineareelutedinthisorderwiththeresolu- Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of tion between these peaks being not less than 8. Tofisopam according to Method 2, and perform the test. Pre- System repeatability: When the test is repeated 6 times with pare the control solution with 1.0 mL of Standard Lead Solu- 10 mL of the standard solution under the above operating tion (not more than 10 ppm). conditions, the relative standard deviation of the peak areas (2) Arsenic <1.11>—Prepare the test solution with 1.0 g of todralazine is not more than 2.0z. of Tofisopam according to Method 3, and perform the test (not more than 2 ppm). Water <2.48> 6.0–7.5z (0.5 g, direct titration). (3) Related substances—Dissolve 0.05 g of Tofisopam in Residue on ignition <2.44> Not more than 0.1z (1 g). 10 mL of acetone, and use this solution as the sample solu- tion. Pipet 1 mL of the sample solution, add acetone to make Assay Weigh accurately about 0.4 g of Todralazine exactly 25 mL, pipet 1 mL of this solution, add acetone to Hydrochloride, dissolve in 5 mL of formic acid, add 70 mL make exactly 20 mL, and use this solution as the standard so- of acetic anhydride, and titrate <2.50> with 0.1 molWLper- lution. Perform the test with these solutions as directed under chloric acid VS (potentiometric titration). Perform a blank Thin-layer Chromatography <2.03>.Spot10mLeachofthe determination, and make any necessary correction. sample solution and standard solution on a plate of silica gel Each mL of 0.1 molWL perchloric acid VS with fluorescent indicator for thin-layer chromatography.

＝ 26.87 mg of C11H12N4O2.HCl Develop the plate with a mixture of ethyl acetate, acetone, methanol and formic acid (24:12:2:1) to a distance of about Containers and storage Containers—Tight containers. 10 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 Tofisopam spot from the standard solution. トフィソパム Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum, silica gel, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.2 g of Tofisopam, previ- ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 molWL perchloric acid VS (potentiometric titration). Perform a blank determination, and make any necessary correction. Each mL of 0.1 molWL perchloric acid VS

＝ 38.25 mg of C22H26N2O4

C22H26N2O4:382.45 Containers and storage Containers—Tight containers. (5RS)-1-(3,4-Dimethoxyphenyl)-5-ethyl-7,8-dimethoxy- Storage—Light-resistant. JP XV O‹cial Monographs / Tolazamide 1187

solution (28) (10 in 11) (200:100:60:23) to a distance of about 12 cm, and air-dry the plate. Heat the plate at 1109Cfor10 Tolazamide minutes, and immediately expose to chlorine for 2 minutes. Expose the plate to cold wind until a very pale blue color de- トラザミド velops when 1 drop of potassium iodide-starch TS is placed on a site below the starting line on the plate. Spray evenly potassium iodide-starch TS on the plate: the spot from the sample solution corresponding to the spot from the standard solution (2) is not more intense than the spot from the stan- dard solution (2), and the spots other than the principal and above spots from the sample solution are not more intense than the spot from the standard solution (1). C14H21N3O3S: 311.40 N-(Azepan-1-ylcarbamoyl)-4-methylbenzenesulfonamide (4) N-Aminohexamethyleneimine—To 0.50 g of [1156-19-0] Tolazamide add 2.0 mL of acetone, stopper the flask tightly, shake vigorously for 15 minutes. Add 8.0 mL of disodium Tolazamide, when dried, contains not less than hydrogenphosphate-citric acid buffer solution, pH 5.4, shake, allow to stand for 15 minutes, and filter. To the 97.5z and not more than 102.0z of C14H21N3O3S. filtrate add 1.0 mL of trisodium ferrous pentacyanoamine Description Tolazamide occurs as a white to pale yellow, TS, and shake: the color developing within 30 minutes is not crystalline powder. It is odorless. deeper than that of the following control solution. It is freely soluble in chloroform, soluble in acetone, slight- Control solution: Dissolve 0.125 g of N-aminohex- ly soluble in ethanol (95) and in n-butylamine, and practically amethyleneimine in acetone to make exactly 100 mL. Pipet 1 insoluble in water and in diethyl ether. mL of this solution, and add acetone to make exactly 100 Melting point: about 1689C (with decomposition). mL. To 2.0 mL of this solution add 8.0 mL of disodium Identification (1) Dissolve 0.02 g of Tolazamide in 5 mL hydrogenphosphate-citric acid buffer solution, pH 5.4, of water and 1 mL of n-butylamine, add 2 to 3 drops of cop- shake, and proceed in the same manner. per (II) sulfate TS, and shake well. Shake well this solution Loss on drying <2.41> Not more 0.5z (1 g, in vacuum, with 5 mL of chloroform, and allow to stand: a green color 609C, 3 hours). develops in the chloroform layer. (2) Determine the absorption spectrum of a solution of Residue on ignition <2.44> Not more than 0.2z (1 g). Tolazamide in ethanol (95) (1 in 100,000) as directed under Assay Weigh accurately about 30 mg each of Tolazamide Ultraviolet-visible Spectrophotometry <2.24>,andcompare and Tolazamide Reference Standard, previously dried, dis- the spectrum with the Reference Spectrum or the spectrum of solve each in 10 mL of the internal standard solution, and use a solution of Tolazamide Reference Standard prepared in the these solutions as the sample solution and standard solution, same manner as the sample solution: both spectra exhibit respectively. Perform the test with 10 mLeachofthesample similar intensities of absorption at the same wavelengths. solutionandstandardsolutionasdirectedunderLiquid (3) Determine the infrared absorption spectrum of Chromatography <2.01> according to the following condi- Tolazamide, previously dried, as directed in the potassium tions, and calculate the ratios, Q T and QS, of the peak area of bromide disk method under the Infrared Spectrophotometry tolazamide to that of the internal standard, respectively. <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of previously dried Tolazamide Refer- Amount (mg) of C14H21N3O3S ence Standard: both spectra exhibit similar intensities of ab- ＝ WS×(QT/QS) sorption at the same wave numbers. WS: Amount (mg) of Tolazamide Reference Standard Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Internal standard solution—A solution of in Tolazamide according to Method 2, and perform the test. ethanol-free chloroform (3 in 2000). Prepare the control solution with 2.0 mL of Standard Lead Operating conditions— Solution (not more than 20 ppm). Detector: An ultraviolet absorption photometer (2) Arsenic <1.11>—Prepare the test solution with 1.0 g (wavelength: 254 nm). of Tolazamide according to Method 3, and perform the test Column: A stainless steel column 4.6 mm in inside di- (not more than 2 ppm). ameter and 30 cm in length, packed with silica gel for liquid (3) Related substances—Dissolve 0.20 g of Tolazamide in chromatography (5 mm in particle diameter). acetone to make exactly 10 mL, and use this solution as the Column temperature: A constant temperature of about sample solution. Pipet 1 mL of the sample solution, add ace- 259C. tone to make exactly 200 mL, and use this solution as the Mobile phase: A mixture of hexane, water-saturated standard solution (1). Separately, dissolve 20 mg of p-tol- hexane, tetrahydrofuran, ethanol (95) and acetic acid (100) uenesulfonamide in acetone to make exactly 200 mL, and use (475:475:20:15:9). this solution as the standard solution (2). Perform the test Flow rate: Adjust the flow rate so that the retention time of with these solutions as directed under Thin-layer Chro- tolazamide is about 12 minutes. matography <2.03>.Spot10mL each of the sample solution System suitability— and standard solutions (1) and (2) on a plate of silica gel for System performance: When the procedure is run with 10 thin-layer chromatography. Develop the plate with a mixture mL of the standard solution under the above operating condi- of chloroform, methanol, cyclohexane and diluted ammonia tions, the internal standard and tolazamide are eluted in this 1188 Tolbutamide / O‹cial Monographs JP XV order with the resolution between these peaks being not less 3 hours). than 5. Residue on ignition <2.44> Not more than 0.1z (1 g). System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operating Assay Weigh accurately about 0.5 g of Tolbutamide, previ- conditions, the relative standard deviation of the ratios of the ously dried, and dissolve in 30 mL of neutralized ethanol. peak area of tolazamide to that of the internal standard is not Add 20 mL of water, and titrate <2.50> with 0.1 molWLsodi- more than 1.0z. um hydroxide VS (indicator: 3 drops of phenolphthalein TS). Containers and storage Containers—Well-closed contain- Each mL of 0.1 molWL sodium hydroxide VS ers. ＝ 27.04 mg of C12H18N2O3S Containers and storage Containers—Well-closed contain- Tolbutamide ers. トルブタミド Tolbutamide Tablets

トルブタミド錠

Tolbutamide Tablets contain not less than 95z and

C12H18N2O3S: 270.35 not more than 105z of the labeled amount of tol- N-(Butylcarbamoyl)-4-methylbenzenesulfonamide butamide (C12H18N2O3S: 270.35). [64-77-7] Method of preparation Prepare as directed under Tablets, with Tolbutamide. Tolbutamide, when dried, contains not less than 99.0z of C12H18N2O3S. Identification Shake a quantity of powdered Tolbutamide Tablets, equivalent to 0.5 g of Tolbutamide according to the Description Tolbutamide occurs as white crystals or crystal- labeled amount, with 50 mL of chloroform, filter, and line powder. It is odorless or has a slight, characteristic odor. evaporate the filtrate to dryness. Proceed with the residue as It is tasteless. directed in the Identification under Tolbutamide. It is soluble in ethanol (95), slightly soluble in diethyl ether, and practically insoluble in water. Dissolution <6.10> Perform the test according to the fol- lowing method: it meets the requirement. Identification (1) Boil 0.2 g of Tolbutamide with 8 mL of Take 1 tablet of Tolbutamide Tablets at 100 revolutions diluted sulfuric acid (1 in 3) under a reflux condenser for 30 per minute according to the using 900 mL of phosphate minutes. Cool the solution in ice water, collect the precipitat- buffer solution, pH 7.4, as the dissolution medium. Take 20 ed crystals, recrystallize from water, and dry at 1059Cfor3 mL or more of the dissolved solution 30 minutes after the hours: the crystals melt <2.60> between 1359C and 1399C. start of the test, and filter through a membrane filter (less (2) Render the filtrate obtained in (1) alkaline with about than 0.8 mm in pore size). Discard the first 10 mL of the 20 mL of a solution of sodium hydroxide (1 in 5), and heat: filtrate, pipet the subsequent V mL, add water to make exact- an ammonia-like odor is perceptible. ly V ? mL of a solution containing about 10 mgoftol-

Melting point <2.60> 126 – 1329C butamide (C12H18N2O3S) per mL according to the labeled amount, and use this solution as the sample solution. Purity (1) Acidity—Warm 3.0 g of Tolbutamide with 150 Separately, weigh accurately about 50 mg of Tolbutamide mL of water at 709C for 5 minutes, allow to stand for 1 hour Reference Standard, previously dried at 1059C for 3 hours, in ice water, and filter. To 25 mL of the filtrate add 2 drops dissolve in 10 mL of methanol, and add phosphate buffer so- of methyl red TS and 0.20 mL of 0.1 molWLsodium lution, pH 7.4, to make exactly 100 mL. Pipet 2 mL of this hydroxide VS: a yellow color develops. solution, add water to make exactly 100 mL, and use this so- (2) Chloride <1.03>—To 40 mL of the filtrate obtained in lution as standard solution. Determine the absorbances, A (1) add 6 mL of dilute nitric acid and water to make 50 mL. T and A , of the sample solution and the standard solution at Perform the test using this solution as the test solution. Pre- S 226 nm as directed under Ultraviolet-visible Spectrophoto- pare the control solution with 0.25 mL of 0.01 molWL metry <2.24>, respectively. The dissolution rate of Tol- hydrochloric acid VS (not more than 0.011z). butamide Tablets after 30 minutes should be not less than (3) Sulfate <1.14>—To 40 mL of the filtrate obtained in 80z. (1) add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solu- Dissolution rate (z) to labeled amount tion. Prepare the control solution with 0.35 mL of 0.005 mol of tolbutamide (C12H18N2O3S) WL sulfuric acid VS (not more than 0.021z). ＝ WS × (AT/AS) × (V?/V) × (90/C) × (1/5) (4) Heavy metals <1.07>—Proceed with 2.0 g of Tol- W : Amount (mg) of Tolbutamide Reference Standard butamide according to Method 2, and perform the test. Pre- S C: Labeled amount (mg) of tolbutamide (C H N O S) per pare the control solution with 2.0 mL of Standard Lead Solu- 12 18 2 3 tablet tion (not more than 10 ppm). Assay Weigh accurately and powder not less than 20 Tol- Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, butamide Tablets. Weigh accurately a portion of the powder, JP XV O‹cial Monographs / Solution 1189

equivalent to about 0.5 g of tolbutamide (C12H18N2O3S), dis- residue by igniting between 5009Cand6009C for 1 hour. solve in 50 mL of neutralized ethanol, add 25 mL of water, Proceed according to Method 2, and perform the test with 50 and titrate <2.50> with 0.1 molWL sodium hydroxide VS (indi- mL of the test solution so obtained. Prepare the control solu- cator: 3 drops of phenolphthalein TS). tion as follows: to 11 mL of nitric acid add 1 mL of sulfuric acid, 1 mL of perchloric acid and 2 mL of hydrochloric acid, Each mL of 0.1 molWL sodium hydroxide VS proceed in the same manner as the test solution, and add 2.0 ＝ 27.04 mg of C H N O S 12 18 2 3 mL of Standard Lead Solution and water to make 50 mL (not Containers and storage Containers—Well-closed contain- more than 20 ppm). ers. (2) Related substances—Dissolve 0.50 g of Tolnaftate in 10 mL of chloroform, and use this solution as the sample so- lution. Pipet 2 mL of the sample solution, and add chlo- Tolnaftate roform to make exactly 100 mL. Pipet 5 mL of this solution, add chloroform to make exactly 100 mL, and use this solu- トルナフタート tion as the standard solution. Perform the test with these so- lutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mLeachofthesamplesolutionandstandardsolution on a plate of silica gel with fluorescent indicator for thin-lay- er chromatography. Develop the plate with toluene to a dis- tance of about 10 cm, and air-dry the plate. Allow the plate to stand in iodine vapor for 5 minutes, and examine under ultraviolet light (wavelength: 254 nm): the spots other than C19H17NOS: 307.41 the principal spot from the sample solution are not more in- O-Naphthalen-2-yl N-methyl-N-(3- tense than the spot from the standard solution. methylphenyl)thiocarbamate [2398-96-1] Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum Tolnaftate, when dried, contains not less than at a pressure not exceeding 0.67 kPa, 659C, 3 hours). 98.0z of C19H17NOS. Residue on ignition <2.44> Weigh accurately about 2.0 g of Description Tolnaftate occurs as a white powder. It is odor- Tolnaftate, and carbonize by gradual heating. Moisten the less. substance with 1 mL of sulfuric acid, heat gradually until It is freely soluble in chloroform, sparingly soluble in white fumes are no longer evolved, and ignite between 4509C diethyl ether, slightly soluble in methanol and in ethanol (95), and 5509C for about 2 hours to constant mass: the residue is and practically insoluble in water. not more than 0.1z. Identification (1) To 0.2 g of Tolnaftate add 20 mL of Assay Weigh accurately about 50 mg of Tolnaftate and potassium hydroxide-ethanol TS and 5 mL of water, and Tolnaftate Reference Standard, previously dried, dissolve heat under a reflux condenser for 3 hours. After cooling, to each in 200 mL of methanol by warming in a water bath, 10 mL of this solution add 2 mL of acetic acid (100), and cool, and add methanol to make exactly 250 mL. Pipet 5 mL shake with 1 mL of lead (II) acetate TS: a black precipitate is each of the solutions, to each add methanol to make exactly formed. 100 mL, and use these solutions as the sample solution and (2) Determine the absorption spectrum of a solution of standard solution, respectively. Determine the absorbances, Tolnaftate in methanol (1 in 100,000) as directed under AT and AS, of the sample solution and standard solution at Ultraviolet-visible Spectrophotometry <2.24>,andcompare 257 nm as directed under Ultraviolet-visible Spectrophoto- the spectrum with the Reference Spectrum or the spectrum of metry <2.24>. a solution of Tolnaftate Reference Standard prepared in the Amount (mg) of C19H17NOS same manner as the sample solution: both spectra exhibit ＝ WS×(AT/AS) similar intensities of absorption at the same wavelengths. (3) Determine the infrared absorption spectrum of Tol- WS: Amount (mg) of Tolnaftate Reference Standard naftate, previously dried, as directed in the potassium Containers and storage Containers—Tight containers. bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of previously dried Tolnaftate Refer- ence Standard: both spectra exhibit similar intensities of ab- Tolnaftate Solution sorption at the same wave numbers. トルナフタート液 Melting point <2.60> 111 – 1149C (after drying). Purity (1) Heavy metals <1.07>—Carbonize 1.0 g of Tol- Tolnaftate Solution contains not less than 90z and naftate by gentle heating. After cooling, add 5 mL of nitric not more than 110z of the labeled amount of tol- acid and 1 mL of sulfuric acid, and heat until white fumes are naftate (C H NOS: 307.41). evolved. After cooling, add 2 mL of nitric acid, and heat un- 19 17 til white fumes are evolved. After cooling, add 2 mL of nitric Method of preparation Prepare as directed under Liquids acid and 0.5 mL of perchloric acid, and heat gradually until and Solutions, with Tolnaftate. white fumes are evolved. Repeat this procedure twice, and Identification (1) Spot 1 drop of Tolnaftate Solution on heat until white fumes are no longer evolved. Incinerate the filter paper. Spray hydrogen hexachloroplatinate (IV)-potas- 1190 Tolperisone Hydrochloride / O‹cial Monographs JP XV sium iodide TS on the paper: a light yellow color develops in the spot. (2) To a volume of Tolnaftate Solution, equivalent to Tolperisone Hydrochloride 0.02 g of Tolnaftate according to the labeled amount, add chloroform to make 10 mL, and use this solution as the sam- トルペリゾン塩酸塩 ple solution. Separately, dissolve 0.02 g of Tolnaftate Refer- ence Standard in 10 mL of chloroform, and use this solution as the standard solution. Perform the test with these solu- tions 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 with fluorescent indicator for thin-lay- er chromatography. Develop the plate with toluene to a dis- C16H23NO.HCl: 281.82 tance of about 12 cm, and air-dry the plate. Examine under (2RS)-2-Methyl-1-(4-methylphenyl)-3-piperidin-1- ultraviolet light (main wavelength: 254 nm): the spot from ylpropan-1-one monohydrochloride [3644-61-9] the sample solution and that from the standard solution show the same Rf value. Tolperisone Hydrochloride, when dried, contains not less than 98.5z of C16H23NO.HCl. Assay Pipet a volume of Tolnaftate Solution, equivalent to Description Tolperisone Hydrochloride occurs as a white, about 20 mg of tolnaftate (C19H17NOS), add exactly 4 mL of the internal standard solution, then add chloroform to make crystalline powder. It has a slight, characteristic odor. 50 mL, and use this solution as the sample solution. It is very soluble in acetic acid (100), freely soluble in water Separately, weigh accurately about 0.4 g of Tolnaftate Refer- and in ethanol (95), soluble in acetic anhydride, slightly solu- ence Standard, previously dried in vacuum at a pressure not ble in acetone, and practically insoluble in diethyl ether. exceeding 0.67 kPa at 659C for 3 hours, and dissolve in chlo- It is hygroscopic. roform to make exactly 100 mL. Pipet 5 mL of this solution, The pH of a solution of Tolperisone Hydrochloride (1 in add exactly 4 mL of the internal standard solution, then add 20) is between 4.5 and 5.5. chloroform to make 50 mL, and use this solution as the stan- Melting point: 167 – 1749C dard solution. Perform the test with 10 mLeachofthesample Identification (1) Dissolve 0.2 g of Tolperisone solution and standard solution as directed under Liquid Hydrochloride in 2 mL of ethanol (95), add 2 mL of 1,3- Chromatography <2.01> according to the following condi- dinitrobenzene TS and 2 mL of sodium hydroxide TS, and tions, and calculate the ratios, Q T and QS, of the peak area of heat: a red color develops. tolnaftate to that of the internal standard, respectively. (2) To 5 mL of a solution of Tolperisone Hydrochloride (1 in 20) add 2 to 3 drops of iodine TS: a red-brown Amount (mg) of tolnaftate (C19H17NOS) ＝ WS×(QT/QS)×(1/20) precipitate is produced. (3) Dissolve 0.5 g of Tolperisone Hydrochloride in 5 mL WS: Amount (mg) of Tolnaftate Reference Standard of water, add 2 mL of ammonia TS, and filter. Acidify 5 mL Internal standard solution—A solution of diphenyl phthalate of the filtrate with dilute nitric acid: the solution responds to in chloroform (3 in 200). the Qualitative Tests <1.09> for chloride. Operating conditions— 1z Absorbance <2.24> E 1cm (257 nm): 555 – 585 (after drying, Detector: An ultraviolet absorption photometer 5 mg, ethanol (95), 500 mL). (wavelength: 254 nm). Column: A stainless steel column about 4 mm in inside di- Purity (1) Clarity and color of solution—Dissolve 1.0 g of ameter and 15 to 30 cm in length, packed with octadecyl- Tolperisone Hydrochloride in 10 mL of water: the solution is silanized silica gel for liquid chromatography (5 to 10 mmin clear and colorless. particle diameter). (2) Sulfate <1.14>—Perform the test using 4.0 g of Tol- Column temperature: A constant temperature of about perisone Hydrochloride. Prepare the control solution with 259C. 0.40 mL of 0.005 molWL sulfuric acid VS (not more than Mobile phase: A mixture of methanol and water (7:3). 0.005z). Flow rate: Adjust the flow rate so that the retention time of (3) Heavy metals <1.07>—Proceed with 1.0 g of Tolperi- tolnaftate is about 14 minutes. sone Hydrochloride according to Method 2, and perform the Selection of column: Proceed with 10 mL of the standard test. Prepare the control solution with 2.0 mL of Standard solution under the above operating conditions, and calculate Lead Solution (not more than 20 ppm). the resolution. Use a column giving elution of the internal (4) Piperidine hydrochloride—Dissolve 0.20 g of Tolperi- standard and tolnaftate in this order with the resolution be- sone Hydrochloride in water to make exactly 10 mL, and use tween these peaks being not less than 5. this solution as the sample solution. Separately, dissolve 20 mg of piperidine hydrochloride in water to make exactly 1000 Containers and storage Containers—Tight containers. mL, and use this solution as the standard solution. Transfer 5.0 mL each of the sample solution and standard solution to different separators, add 0.1 mL each of a solution of copper (II) sulfate pentahydrate (1 in 20), then add 0.1 mL each of ammonia solution (28) and exactly 10 mL each of a mixture of isooctane and carbon disulfide (3:1), and shake vigorously for 30 minutes. Immediately after allowing to stand, separate JP XV O‹cial Monographs / Tranexamic Acid 1191 the isooctane-carbon disulfide mixture layer, and dehydrate tion, pH 3.5, mix, add 1.2 mL of thioacetamide TS , mix with anhydrous sodium sulfate. Perform the test with these immediately, and use this solution as the sample solution. solutions as directed under Ultraviolet-visible Spectrophoto- Separately, proceed in the same manner as above with a metry <2.24>: the absorbance of the sample solution at 438 mixture of 1 mL of Standard Lead Solution, 2 mL of the nm is not more than that of the standard solution. sample stock solution and 9 mL of water, and use the solu- tion so obtained as the standard solution. Separately, pro- Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum, ceed in the same manner with a mixture of 10 mL of water silica gel, 3 hours). and 2 mL of the sample stock solution, and use the solution Residue on ignition <2.44> Not more than 0.1z (1 g). so obtained as the control solution. Conform that the color of the standard solution is slightly darker than that of the Assay Weigh accurately about 0.5 g of Tolperisone control solution. Compare the sample solution and the stan- Hydrochloride, previously dried, dissolve in 70 mL of a mix- dard solution 2 minutes after they are prepared: the color of ture of acetic anhydride and acetic acid (100) (7:3), and titrate the sample solution is not more intense than that of the stan- <2.50> with 0.1 molWL perchloric acid VS (potentiometric dard solution (not more than 10 ppm). titration). Perform a blank determination, and make any (4) Arsenic <1.11>—Prepare the test solution by dissolv- necessary correction. ing 1.0 g of Tranexamic Acid in 10 mL of water, and perform Each mL of 0.1 molWL perchloric acid VS the test (not more than 2 ppm).

＝ 28.18 mg C16H23NO.HCl (5) Related substances—Dissolve 0.20 g of Tranexamic Acid in water to make exactly 20 mL, and use this solution as Containers and storage Containers—Well-closed contain- the sample solution. Pipet 5 mL of the sample solution, and ers. add water to make exactly 100 mL. Pipet 1 mL of this solu- tion, add water to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL Tranexamic Acid each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol- トラネキサム酸 lowing conditions, and determine each peak area by the auto- matic integration method: the area multiplied by relative response factor 1.2 of the peak, having the relative retention time of about 1.5 with respect to tranexamic acid, is not more than 2/5 of the peak area of tranexamic acid from the stan- dard solution, and the area of the peak, having the relative

C8H15NO2: 157.21 retention time of about 2.1 with respect to tranexamic acid, is trans-4-(Aminomethyl)cyclohexanecarboxylic acid not more than 1/5 of the peak area of tranexamic acid from [1197-18-8] the standard solution. The area of each peak other than tranexamic acid and other than the peaks mentioned above is Tranexamic Acid, when dried, contains not less than not more than 1/5 of the peak area of tranexamic acid from 98.0z and not more than 101.0z of C8H15NO2. the standard solution. For this comparison, use the area of the peaks, having the relative retention time of about 1.1 and Description Tranexamic Acid occurs as white crystals or about 1.3, after multiplying by their relative response factors crystalline powder. 0.005 and 0.006, respectively. The total area of the peaks It is freely soluble in water, and practically insoluble in other than tranexamic acid is not more than the peak area of ethanol (99.5). tranexamic acid from the standard solution. Identiˆcation Determine the infrared absorption spectrum Operating conditions— of Tranexamic Acid as directed in the potassium bromide Detector, column, column temperature, mobile phase, and disk method under Infrared Spectrophotometry <2.25>,and ‰ow rate: Proceed as directed in the operating conditions in compare the spectrum with the Reference Spectrum or the the Assay. spectrum of Tranexamic Acid Reference Standard: both Time span of measurement: About 3 times as long as the spectra exhibit similar intensities of absorption at the same retention time of tranexamic acid beginning after the solvent wave numbers. peak. System suitability— pH <2.54> The pH of a solution prepared by dissolving 1.0 Test for required detectability: To exactly 5 mL of the stan- g of Tranexamic Acid in 20 mL of water is between 7.0 and dard solution add water to make exactly 25 mL. Conˆrm that 8.0. the peak area of tranexamic acid obtained from 20 mLofthis Purity (1) Clarity and color of solution—Dissolve 1.0 g of solution is equivalent to 14 to 26z of that from 20 mLofthe Tranexamic Acid in 10 mL of water: the solution is clear and standard solution. colorless. System performance: Proceed as directed in the system (2) Chloride <1.03>—Perform the test with 1.0 g of suitability in the Assay. Tranexamic Acid. Prepare the control solution with 0.40 mL System repeatability: When the test is repeated 6 times with of 0.01 mol/L hydrochloric acid VS (not more than 0.014z). 20 mL of the standard solution under the above operating (3) Heavy metals—Dissolve 2.0 g of Tranexamic Acid in conditions, the relative standard deviation of the peak area of water to make 20 mL, and use this solution as the sample tranexamic acid is not more than 7z. stock solution. To 12 mL of the sample stock solution add Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 mL of hydrochloric acid-ammonium acetate buŠer solu- 1192 Tranexamic Acid Capsules / O‹cial Monographs JP XV

2 hours). velops. Residue on ignition <2.44> Not more than 0.1z (1 g). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Assay Weigh accurately about 50 mg each of Tranexamic Acid and Tranexamic Acid Reference Standard, previously Dissolution <6.10> Perform the test according to the follow- dried, dissolve in water to make exactly 25 mL, and use these ing method: it meets the requirement. solutions as the sample solution and standard solution. Per- Perform the test with 1 capsule of Tranexamic Acid Cap- form the test with exactly 20 mL each of the sample solution sules at 50 revolutions per minute according to the Paddle and standard solution as directed under Liquid Chro- methed using a sinker and using 900 mL of water as the disso- matography <2.01> according to the following conditions, lution medium. Withdraw 20 mL or more of the dissolved so- and determine the peak areas, AT and AS,oftranexamicacid. lution 15 minutes after starting the test, and ˆlter through a membrane ˆlter with pore size of not more than 0.45 mm.

Amount (mg) of C8H15NO2＝WS×(AT/AS) Discard the ˆrst 10 mL of the ˆltrate, pipet the subsequent V mL, add water to make exactly V? mL so that each mL con-

WS: Amount (mg) of Tranexamic Acid Reference Standard tains about 0.28 mg of tranexamic acid (C8H15NO2) accord- Operating conditions— ing to the labeled amount, and use this solution as the sample Detector: An ultraviolet absorption photometer solution. Separately, weigh accurately about 28 mg of (wavelength: 220 nm). Tranexamic Acid Reference Standard, previously dried at Column: A stainless steel column 6.0 mm in inside 1059C for 2 hours, dissolve in water to make exactly 100 mL, diameter and 25 cm in length, packed with octadecylsilanized and use this solution as the standard solution. Perform the silica gel for liquid chromatography (5 mminparticle test with exactly 10 mL each of the sample solution and stan- diameter). dard solution as directed under Liquid Chromatography Column temperature: A constant temperature of about <2.01> according to the following conditions, and determine

259C. the peak areas of tranexamic acid, AT and AS. The dissolu- Mobile phase: Dissolve 11.0 g of sodium dihydrogen phos- tion rate in 15 minutes is not less than 80z. phate in 500 mL of water, and add 5 mL of triethylamine and Dissolution rate (z) with respect to the labeled amount of 1.4 g of sodium lauryl sulfate. Adjust the pH to 2.5 with tranexamic acid (C H NO ) phosphoric acid or diluted phosphoric acid (1 in 10), add 8 15 2 ＝W ×(A /A )×(V?/V)×(1/C)×900 water to make 600 mL, and add 400 mL of methanol. S T S

Flow rate: Adjust the ‰ow rate so that the retention time of WS: Amount (mg) of Tranexamic Acid Reference Stan- tranexamic acid is about 20 minutes. dard

System suitability— C: Labeled amount (mg) of tranexamic acid (C8H15NO2)in System performance: To 5 mL of the standard solution 1capsule add 1 mL of a solution of 4-(aminomethyl)benzoic acid (1 in Operating conditions— 10,000) and water to make 50 mL. When the procedure is run Detector: An ultraviolet absorption photometer with 20 mL of this solution under the above operating condi- (wavelength: 220 nm). tions, tranexamic acid and 4-(aminomethyl)benzoic acid are Column: A stainless steel column 4.6 mm in inside di- eluted in this order with the resolution between these peaks ameter and 15 cm in length, packed with octadecylsilanized being not less than 5. silica gel for liquid chromatography (5 mminparticledi- System repeatability: When the test is repeated 6 times with ameter). 20 mL of the standard solution under the above operating Column temperature: A constant temperature of about conditions, the relative standard deviation of the peak area of 259C. tranexamic acid is not more than 0.6z. Mobile phase: Dissolve 11.0 g of anhydrous sodium di- Containers and storage Containers—Well-closed contain- hydrogen phosphate in 500 mL of water, and add 10 mL of ers. triethylamine and 1.4 g of sodium lauryl sulfate. Adjust the pH to 2.5 with phosphoric acid, add water to make 600 mL, and add 400 mL of methanol. Tranexamic Acid Capsules Flow rate: Adjust the ‰ow rate so that the retention time of tranexamic acid is about 8 minutes. トラネキサム酸カプセル System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating condi- Tranexamic Acid Capsules contain not less than tions, the number of theoretical plates and the symmetry fac- 95.0z and not more than 105.0z of the labeled tor of the peak of tranexamic acid are not less than 4000 and amount of tranexamic acid (C8H15NO2:157.21). not more than 2.0, respectively. Method of preparation Prepare as directed under Capsules, System repeatability: When the test is repeated 6 times with with Tranexamic Acid. 10 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of Identiˆcation Take an amount of powdered contents of tranexamic acid is not more than 2.0z. Tranexamic Acid Capsules, equivalent to 0.5 g of Tranexam- ic Acid according to the labeled amount, add 50 mL of water, Assay Weigh accurately the mass of the contents of not less shake well, and ˆlter. To 5 mL of the ˆltrateadd 1 mL ofnin- than 20 Tranexamic Acid Capsules, and powder. Weigh ac- hydrin TS, and heat for 3 minutes: a dark purple color de- curately an amount of the powder, equivalent to about 0.1 g JP XV O‹cial Monographs / Tranexamic Acid Tablets 1193 of tranexamic acid (C H NO ), add 30 mL of water, shake 8 15 2 Foreign insoluble matter <6.06> Perform the test according well, and add water to make exactly 50 mL. Centrifuge, ˆlter to Method 1: it meets the requirement. the supernatant liquid through a membrane ˆlter with pore size of not more than 0.45 mm, discard the ˆrst 10 mL of the Insoluble particulate matter <6.07> Perform the test accord- ˆltrate, and use the subsequent ˆltrate as the sample solution. ing to Method 1: it meets the requirement. Separately, weigh accurately about 50 mg of Tranexamic Sterility <4.06> Perform the test according to the Membrane Acid Reference Standard, previously dried at 1059Cfor2 ˆltration method: it meets the requirement. hours, dissolve in water to make exactly 25 mL, and use this solution as the standard solution. Perform the test with ex- Assay Take accurately a volume of Tranexamic Acid actly 30 mL each of the sample solution and standard solution Injection, equivalent to about 0.1 g of tranexamic acid as directed under Liquid Chromatography <2.01> according (C8H15NO2), add water to make exactly 50 mL, and use this to the following conditions, and determine the peak areas, AT solution as the sample solution. Separately, weigh accurately and AS, of tranexamic acid. about 50 mg of Tranexamic Acid Reference Standard, previously dried at 1059C for 2 hours, dissolve in water to Amount (mg) of tranexamic acid (C H NO ) 8 15 2 make exactly 25 mL, and use this solution as the standard so- ＝W ×(A /A )×2 S T S lution. Perform the test with exactly 30 mLeachofthesample

WS: Amount (mg) of Tranexamic Acid Reference Standard solutionandstandardsolutionasdirectedunderLiquid Chromatography <2.01> according to the following Operating conditions— conditions, and determine the peak areas, A and A ,of Detector, column, and mobile phase: Proceed as directed T S tranexamic acid. in the operating conditions in the Assay under Tranexamic Acid. Amount (mg) of tranexamic acid (C H NO ) Column temperature: A constant temperature of about 8 15 2 ＝W ×(A /A ) 359C. S T S

Flow rate: Adjust the ‰ow rate so that the retention time of WS: Amount (mg) of Tranexamic Acid Reference Standard tranexamic acid is about 16 minutes. Operating conditions— System suitability— Detector, column, and mobile phase: Proceed as directed System performance: To 5 mL of the standard solution in the operating conditions in the Assay under Tranexamic add 1 mL of a solution of 4-(aminomethyl)benzoic acid (1 in Acid. 10,000) and water to make 50 mL. When the procedure is run Column temperature: A constant temperature of about with 30 mL of this solution under the above operating condi- 359C. tions, tranexamic acid and 4-(aminomethyl)benzoic acid are Flow rate: Adjust the ‰ow rate so that the retention time of eluted in this order with the resolution between these peaks tranexamic acid is about 16 minutes. being not less than 3. System suitability— System repeatability: When the test is repeated 6 times with System performance: To 5 mL of the standard solution 30 mL of the standard solution under the above operating add 1 mL of a solution of 4-(aminomethyl)benzoic acid (1 in conditions, the relative standard deviation of the peak area of 10,000) and water to make 50 mL. When the procedure is run tranexamic acid is not more than 1.0z. with 30 mL of this solution under the above operating condi- Containers and storage Containers—Tight containers. tions, tranexamic acid and 4-(aminomethyl)benzoic acid are eluted in this order with the resolution between these peaks being not less than 3. Tranexamic Acid Injection System repeatability: When the test is repeated 6 times with 30 mL of the standard solution under the above operating トラネキサム酸注射液 conditions, the relative standard deviation of the peak area of tranexamic acid is not more than 1.0z. Tranexamic Acid Injection is an aqueous injection. Containers and storage Containers—Hermetic containers. It contains not less than 95.0z and not more than 105.0z of the labeled amount of tranexamic acid (C8H15NO2: 157.21). Tranexamic Acid Tablets Method of preparation Prepare as directed under トラネキサム酸錠 Injections, with Tranexamic Acid. Description Tranexamic Acid Injection is a clear and color- Tranexamic Acid Tablets contain not less than less liquid. 95.0z and not more than 105.0z of the labeled Identiˆcation To a volume of Tranexamic Acid Injection, amount of tranexamic acid (C8H15NO2: 157.21). equivalent to 50 mg of Tranexamic Acid according to the Method of preparation Prepare as directed under Tablets, labeled amount, add water to make 5 mL, add 1 mL of with Tranexamic Acid. ninhydrin TS, and heat: a dark purple color develops. Identiˆcation To an amount of powdered Tranexamic Acid pH <2.54> 7.0 – 8.0 Tablets, equivalent to 0.5 g of Tranexamic Acid according to Bacterial endotoxins <4.01> Not more than 0.12 EU/mg. the labeled amount, add 50 mL of water, shake well, and ˆlter. To 5 mL of the ˆltrate add 1 mL of ninhydrin TS, and Extractable volume <6.05> It meets the requirement. 1194 Trapidil / O‹cial Monographs JP XV heat for 3 minutes: a dark purple color develops. [15421-84-8] Uniformity of dosage units <6.02> It meets the requirement Trapidil, when dried, contains not less than 98.5z of the Mass variation test. of C10H15N5. Dissolution Being specified separately. Description Trapidil occurs as a white to pale yellowish Assay Weigh accurately the mass of not less than 20 white, crystalline powder. Tranexamic Acid Tablets, and powder. Weigh accurately a It is very soluble in water and in methanol, freely soluble in quantity of the powder, equivalent to about 5 g of tranexamic ethanol (95), in acetic anhydride and in acetic acid (100), and acid (C8H15NO2), add 150 mL of water, disintegrate the sparingly soluble in diethyl ether. tablets completely with the aid of ultrasonic waves, and add The pH of a solution of Trapidil (1 in 100) is between 6.5 water to make exactly 200 mL. Centrifuge, pipet 4 mL of the and 7.5. supernatant liquid, and add water to make exactly 50 mL. Identification (1) To 5 mL of a solution of Trapidil (1 in Filter through a membrane ˆlter with pore size of not more 50) add 3 drops of Dragendorff's TS: an orange color de- than 0.45 mm, discard the ˆrst 10 mL of the ˆltrate, and use velops. the subsequent ˆltrate as the sample solution. Separately, (2) Determine the absorption spectrum of a solution of weigh accurately about 50 mg of Tranexamic Acid Reference Trapidil (1 in 125,000) as directed under Ultraviolet-visible Standard, previously dried at 1059C for 2 hours, dissolve in Spectrophotometry <2.24>, and compare the spectrum with water to make exactly 25 mL, and use this solution as the the Reference Spectrum: both spectra exhibit similar intensi- standard solution. Perform the test with exactly 30 mLeach ties of absorption at the same wavelengths. of the sample solution and standard solution as directed un- 1z der Liquid Chromatography <2.01> according to the follow- Absorbance <2.24> E 1cm (307 nm): 860 – 892 (after drying, ing conditions, and determine the peak areas, AT and AS,of 20 mg, water, 2500 mL). tranexamic acid. Melting point <2.60> 101 – 1059C Amount (mg) of tranexamic acid (C H NO ) 8 15 2 Purity (1) Clarity and color of solution—Dissolve 2.5 g of ＝W ×(A /A )×100 S T S Trapidil in 10 mL of water: the solution is clear and colorless

WS: Amount (mg) of Tranexamic Acid Reference Standard to pale yellow. (2) Chloride <1.03>—Perform the test with 0.5 g of Operating conditions— Trapidil. Prepare the control solution with 0.25 mL of 0.01 Detector, column, and mobile phase: Proceed as directed molWL hydrochloric acid VS (not more than 0.018z). in the operating conditions in the Assay under Tranexamic (3) Ammonium—Place 0.05 g of Trapidil in a glass-stop- Acid. pered conical flask, thoroughly moisten with 10 drops of so- Column temperature: A constant temperature of about dium hydroxide TS, and stopper the flask. Allow it to stand 359C. at 379C for 15 minutes: the gas evolved does not change Flow rate: Adjust the ‰ow rate so that the retention time of moistened red litmus paper to blue. tranexamic acid is about 16 minutes. (4) Heavy metals <1.07>—Dissolve 1.0 g of Trapidil in 40 System suitability— mL of water, and add 1.5 mL of dilute hydrochloric acid, 2 System performance: To 5 mL of the standard solution mL of dilute acetic acid and water to make 50 mL. Perform add 1 mL of a solution of 4-(aminomethyl)benzoic acid (1 in the test using this solution as the test solution. Prepare the 10,000) and water to make 50 mL. When the procedure is run control solution as follows: to 1.0 mL of Standard Lead So- with 30 mL of this solution under the above operating condi- lution add 2 mL of dilute acetic acid and water to make 50 tions, tranexamic acid and 4-(aminomethyl)benzoic acid are mL (not more than 10 ppm). eluted in this order with the resolution between these peaks (5) Arsenic <1.11>—Prepare the test solution with 1.0 g being not less than 3. of Trapidil according to Method 1, and perform the test (not System repeatability: When the test is repeated 6 times with more than 2 ppm). 30 mL of the standard solution under the above operating (6) Related substances—Dissolve 0.10 g of Trapidil in 4 conditions, the relative standard deviation of the peak area of mL of methanol, and use this solution as the sample solution. tranexamic acid is not more than 1.0z. Pipet 1 mL of the sample solution, and add methanol to Containers and storage Containers—Tight containers. make exactly 20 mL. Pipet 1 mL of this solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions Trapidil as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a トラピジル plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform, ethanol (95) and acetic acid (100) (85:13:2) to a distance of about 10 cm, and air-dry the plate. Allow the plate to stand in iodine vapor for 60 minutes: the spots other than the principal spot from the sample solution are not more intense than the spot from the standard solution. C H N :205.26 10 15 5 Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum, 7-Diethylamino-5-methyl[1,2,4]triazolo[1,5-a]pyrimidine silica gel, 609C, 3 hours). JP XV O‹cial Monographs / Triamcinolone 1195

(not more than 0.021z). Residue on ignition <2.44> Not more than 0.1z (1 g). (2) Heavy metals <1.07>—Proceed with 1.0 g of Trepibu- Assay Weigh accurately about 0.2 g of Trapidil, previously tone according to Method 2, and perform the test. Prepare dried, dissolve in 20 mL of acetic acid (100), and titrate the control solution with 2.0 mL of Standard Lead Solution <2.50> with 0.1 molWL perchloric acid VS (potentiometric (not more than 20 ppm). titration). Perform a blank determination, and make any (3) Related substances—Dissolve 0.10 g of Trepibutone necessary correction. in 10 mL of acetone, and use this solution as the sample solu- tion. Pipet 2 mL of the sample solution, add acetone to make Each mL of 0.1 molWL perchloric acid VS exactly 100 mL. To exactly 10 mL of this solution add ace- ＝ 20.53 mg of C H N 10 15 5 tone to make exactly 100 mL, and use this solution as the Containers and storage Containers—Tight containers. standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>.Spot10 mL each of the sample solution and standard solution on a Trepibutone plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of トレピブトン isopropylether, acetone, water and formic acid (100:30:3:3) to a distance of about 10 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. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). C16H22O6: 310.34 4-Oxo-4-(2,4,5-triethoxyphenyl)butanoic acid Residue on ignition <2.44> Not more than 0.1z (1 g). [41826-92-0] Assay Weigh accurately about 0.5 g of Trepibutone, previ- Trepibutone, when dried, contains not less than ously dried, dissolve in 50 mL of ethanol (95), add 50 mL of water, and titrate <2.50> with 0.1 molWL sodium hydroxide 98.5z of C16H22O6. VS (indicator: 5 drops of phenolphthalein TS). Perform a Description Trepibutone occurs as white to yellowish white blank determination, and make any necessary correction. crystals or crystalline powder. It is odorless, and is tasteless or has a slight, characteristic aftertaste. Each mL of 0.1 molWL sodium hydroxide VS It is soluble in acetone, sparingly soluble in ethanol (95), ＝ 31.03 mg of C16H22O6 slightly soluble in diethyl ether, and practically insoluble in Containers and storage Containers—Tight containers. water. Storage—Light-resistant. It dissolves in sodium hydroxide TS. Identification (1) Determine the absorption spectrum of a solution of Trepibutone in diluted dilute sodium hydroxide Triamcinolone TS (1 in 10) (1 in 100,000) as directed under Ultraviolet-visi- ble Spectrophotometry <2.24>, and compare the spectrum トリアムシノロン with the Reference Spectrum: both spectra exhibit similar in- tensities of absorption at the same wavelengths. (2) Determine the spectrum of a solution of Trepibutone in deuterated chloroform for the nuclear magnetic resonance spectroscopy (1 in 10), using tetramethylsilane for the nuclear magnetic resonance spectroscopy as an internal reference compound, as directed under Nuclear Magnetic Resonance 2.21 1 Spectroscopy < > ( H): it exhibits a sharp multiple signal A C21H27FO6: 394.43 at around d 1.5 ppm, a triplet signal B at around d 2.7 ppm, a 9-Fluoro-11b,16a,17,21-tetrahydroxypregna-1,4-diene- triplet signal C at around d 3.3 ppm, a multiple signal D at 3,20-dione [124-94-7] around d 4.2 ppm, a sharp single signal E at around d 6.4 ppm, a sharp single signal F at around d 7.4 ppm, and a sin- Triamcinolone, when dried, contains not less than gle signal G at around d 10.5 ppm. The ratio of integrated in- 97.0z and not more than 103.0z of C21H27FO6. tensity of each signal, A:B:C:D:E:F:G, is about 9:2:2:6:1:1:1. Description Triamcinolone occurs as a white, crystalline powder. It is odorless. Melting point <2.60> 146 – 1509C It is freely soluble in N,N-dimethylformamide, slightly Purity (1) Chloride <1.03>—Dissolve 0.5 g of Trepibutone soluble in methanol, in ethanol (95) and in acetone, and prac- in 30 mL of acetone, and add 6 mL of dilute nitric acid and tically insoluble in water, in 2-propanol and in diethyl ether. water to make 50 mL. Perform the test using this solution as Melting point: about 2649C (with decomposition). the test solution. Prepare the control solution as follows: to Identification (1) Dissolve 1 mg of Triamcinolone in 6 mL 0.30 mL of 0.01 molWL hydrochloric acid VS add 30 mL of of ethanol (95), add 5 mL of 2,6-di-tert-butylcresol TS and 5 acetone, 6 mL of dilute nitric acid and water to make 50 mL mL of sodium hydroxide TS, and heat on a water bath for 30 1196 Triamcinolone Acetonide / O‹cial Monographs JP XV minutes under a reflux condenser: a red-purple color de- Mobile phase: A mixture of water and acetonitrile (3:1). velops. Flow rate: Adjust the ‰ow rate so that the retention time of (2) Add5mLofwaterand1mLofFehling'sTSto0.01 triamcinolone is about 10 minutes. g of Triamcinolone, and heat: a red precipitate is produced. System suitability— (3) Proceed with 0.01 g of Triamcinolone as directed un- System performance: When the procedure is run with 10 der Oxygen Flask Combustion Method <1.06>,usingamix- mL of the standard solution under the above operating condi- ture of 0.5 mL of 0.01 molWL sodium hydroxide TS and 20 tions, triamcinolone and the internal standard are eluted in mL of water as the absorbing liquid. When combustion is this order with the resolution between these peaks being not completed, shake vigorously so as to absorb the gas evolved: less than 2.0. the solution responds to the Qualitative Tests <1.09> for fluo- System repeatability: When the test is repeated 6 times with ride. 10 mL of the standard solution under the above operating (4) Determine the infrared absorption spectrum of conditions, the relative standard deviation of the ratios of the Triamcinolone, previously dried, as directed in the potassium peak height of triamcinolone to that of the internal standard bromide disk method under Infrared Spectrophotometry is not more than 1.5z. <2.25>, and compare the spectrum with the Reference Spec- Containers and storage Containers—Tight containers. trum or the spectrum of previously dried Triamcinolone Storage—Light-resistant. Reference Standard: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve 0.1 g each of Triam- cinolone and Triamcinolone Reference Standard in 7 mL of a Triamcinolone Acetonide mixture of 2-propanol and water (2:1), respectively, by トリアムシノロンアセトニド warming. Allow the solutions to cool in ice to effect crystals, filter, then wash the formed crystals with two 10-mL portions of water, and repeat the test on the dried crystals.

20 Optical rotation <2.49> [a] D : ＋65 – ＋719(after drying, 0.1 g, N,N-dimethylformamide, 10 mL, 100 mm). Purity Heavy metals <1.07>—Proceed with 0.5 g of Triam- cinolone according to Method 2, and perform the test. Pre- pare the control solution with 1.5 mL of Standard Lead Solu- tion (not more than 30 ppm). C24H31FO6: 434.50 Loss on drying <2.41> Not more than 2.0z (0.5 g, in vacu- 9-Fluoro-11b,21-dihydroxy-16a,17- um, phosphorus (V) oxide, 609C, 3 hours). (1-methylethylidenedioxy)pregna-1,4-diene-3,20-dione [76-25-5] Residue on ignition <2.44> Not more than 0.3z (0.5 g, platinum crucible). Triamcinolone Acetonide, when dried, contains not Assay Dissolve about 20 mg each of Triamcinolone and less than 97.0z and not more than 103.0z of Triamcinolone Reference Standard, previously dried and ac- C24H31FO6. curately weighed, in a solution of L-ascorbic acid in methanol Description Triamcinolone Acetonide occurs as a white, (1 in 1000) to make exactly 50 mL. Pipet 5 mL each of these crystalline powder. It is odorless. solutions, add exactly 5 mL each of the internal standard so- It is sparingly soluble in ethanol (99.5), in acetone, and in lution, add a solution of L-ascorbic acid in methanol (1 in 1,4-dioxane, slightly soluble in methanol and in ethanol (95), 1000) to make 20 mL, and use these solutions as the sample andpracticallyinsolubleinwaterandindiethylether. solution and standard solution. Perform the test with 10 mL Melting point: about 2909C (with decomposition). each of these solutions as directed under Liquid Chromatog- raphy <2.01> according to the following conditions, and cal- Identification (1) Dissolve 2 mg of Triamcinolone Acetonide in 40 mL of ethanol (95), add 5 mL of 2,6-di-tert- culate the ratios, QT and Q S, of the peak height of tri- amcinolone to that of the internal standard, respectively. butylcresol TS and 5 mL of sodium hydroxide TS, and heat on a water bath under a reflux condenser for 20 minutes: a Amount (mg) of C21H27FO6 ＝ WS × (Q T/Q S) green color develops. (2) Add 5 mL of water and 1 mL of Fehling's TS to 0.01 WS: Amount (mg) of Triamcinolone Reference Standard g of Triamcinolone Acetonide, and heat: a red precipitate is Internal standard solution—Dissolve 15 mg of methyl para- produced. hydroxybenzoate in a solution of L-ascorbicacidinmethanol (3) Proceed with 0.01 g of Triamcinolone Acetonide as (1 in 1000) to make 100 mL. directed under Oxygen Flask Combustion Method <1.06>,us- Operating conditions— ing a mixture of 0.5 mL of 0.01 molWL sodium hydroxide TS Detector: An ultraviolet absorption photometer (wave- and 20 mL of water as the absorbing liquid. When combus- length: 254 nm). tion is completed, shake vigorously so as to absorb the gas Column: A stainless steel column 4.0 mm in inside di- evolved: the solution responds to the Qualitative Tests <1.09> ameter and 15 cm in length, packed with octadecylsilanized for fluoride. silica gel (5 mm in particle diameter). (4) Determine the absorption spectrum of a solution of Column temperature: A constant temperature of about Triamcinolone Acetonide in ethanol (95) (1 in 100,000) as 259C. directed under Ultraviolet-visible Spectrophotometry <2.24>, JP XV O‹cial Monographs / Triamterene 1197 and compare the spectrum with the Reference Spectrum or Operating conditions— the spectrum of a solution of Triamcinolone Acetonide Detector: An ultraviolet absorption photometer Reference Standard prepared in the same manner as the sam- (wavelength: 240 nm). ple solution: both spectra exhibit similar intensities of ab- Column: A stainless steel column 4.6 mm in inside di- sorption at the same wavelengths. ameter and 30 cm in length, packed with octadecylsilanized (5) Determine the infrared absorption spectrum of silica gel (10 mminparticlediameter). Triamcinolone Acetonide, previously dried, as directed in the Column temperature: A constant temperature of about potassium bromide disk method under the Infrared Spec- 259C. trophotometry <2.25>, and compare the spectrum with the Mobile phase: A mixture of water and acetonitrile (3:1). Reference Spectrum or the spectrum of previously dried Flow rate: Adjust the flow rate so that the retention time of Triamcinolone Acetonide Reference Standard: both spectra triamcinolone acetonide is about 13 minutes. exhibit similar intensities of absorption at the same wave System suitability— numbers. If any difference appears between the spectra, dis- System performance: When the procedure is run with 10 solve 0.1 g each of Triamcinolone Acetonide and Triamcino- mL of the standard solution under the above operating condi- lone Acetonide Reference Standard in 20 mL of ethanol (95), tions, the internal standard and triamcinolone acetonide are respectively, then evaporate the ethanol to dryness, and eluted in this order with the resolution between these peaks repeat the test on the dried residue. being not less than 6. System repeatability: When the test is repeated 6 times with Optical rotation <2.49> [a]20: ＋100 – ＋1079(after drying, D 10 mL of the standard solution under the above operating 0.1g,1,4-dioxane,10mL,100mm). conditions, the relative standard deviation of the ratios of the Purity (1) Heavy metals <1.07>—Proceed with 0.5 g of peak height of triamcinolone acetonide to that of the internal Triamcinolone Acetonide according to Method 2, and per- standard is not more than 1.0z. form the test. Prepare the control solution with 1.5 mL of Containers and storage Containers—Tight containers. Standard Lead Solution (not more than 30 ppm). Storage—Light-resistant. (2) Related substances—Dissolve 40 mg of Triamcino- lone Acetonide in 4 mL of acetone, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add acetone to make exactly 100 mL, and use this solution as the Triamterene standard solution. Perform the test with these solutions as トリアムテレン directed under Thin-layer Chromatography <2.03>.Spot20 mLeachofthesamplesolutionandstandardsolutionona plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chlo- roform and methanol (93:7) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot C H N : 253.26 from the sample solution are not more intense than the spot 12 11 7 6-Phenylpteridine-2,4,7-triamine [396-01-0] from the standard solution. Loss on drying <2.41> Not more than 2.0z (0.5 g, in vacu- Triamterene, when dried, contains not less than um, phosphorus (V) oxide, 609C, 3 hours). 98.5z of C12H11N7. Residue on ignition <2.44> Not more than 0.2z (0.5 g, Description Triamterene occurs as a yellow, crystalline platinum crucible). powder. It is odorless, and tasteless. It is sparingly soluble in dimethylsulfoxide, very slightly Assay Dissolve about 20 mg each of Triamcinolone soluble in acetic acid (100), and practically insoluble in water, Acetonide and Triamcinolone Acetonide Reference Stan- in ethanol (95), and in diethyl ether. dard, previously dried and accurately weighed, in methanol It dissolves in nitric acid and in sulfuric acid, but does not to make exactly 50 mL. Pipet 10 mL each of these solutions, dissolve in dilute nitric acid, in dilute sulfuric acid and in di- add exactly 10 mL each of the internal standard solution, lute hydrochloric acid. then add the mobile phase to make 50 mL, and use these solu- tions as the sample solution and standard solution. Perform Identification (1) To 0.01 g of Triamterene add 10 mL of the test with 20 mL each of these solutions as directed under water, heat, and filter after cooling: the filtrate shows a pur- Liquid Chromatography <2.01> according to the following ple fluorescence. To 2 mL of the filtrate add 0.5 mL of conditions, and calculate the ratios, Q T and Q S, of the peak hydrochloric acid: the fluorescence disappears. height of triamcinolone acetonide to that of the internal stan- (2) The filtrate obtained in (1) responds to the Qualitative dard, respectively. Tests <1.09> for primary aromatic amines. (3) Dissolve 0.01 g of Triamterene in 100 mL of acetic Amount (mg) of C H FO 24 31 6 acid (100), and to 10 mL of the solution add water to make ＝ W ×(Q /Q ) S T S 100 mL. Determine the absorption spectrum of the solution

WS: Amount (mg) of Triamcinolone Acetonide Reference as directed under Ultraviolet-visible Spectrophotometry Standard <2.24>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at —A solution of in Internal standard solution thesamewavelengths. methanol (1 in 50,000). 1198 Trichlormethiazide / O‹cial Monographs JP XV

Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Identiˆcation (1) Determine the absorption spectrum of a Triamterene according to Method 2, and perform the test. solution of Trichlormethiazide in ethanol (95) (3 in 250,000) Prepare the control solution with 2.0 mL of Standard Lead as directed under Ultraviolet-visible Spectrophotometry Solution (not more than 20 ppm). <2.24>, and compare the spectrum with the Reference Spec- (2) Arsenic <1.11>—Prepare the test solution with 1.0 g trum or the spectrum of a solution of Trichlormethiazide of Triamterene according to Method 3, and perform the test Reference Standard prepared in the same manner as the sam- (not more than 2 ppm). ple solution: both spectra exhibit similar intensities of ab- (3) Related substances—Dissolve 0.10 g of Triamterene sorption at the same wavelengths. in 20 mL of dimethylsulfoxide. To 2 mL of this solution add (2) Determine the infrared absorption spectrum of methanol to make 50 mL, and use this solution as the sample Trichlormethiazide as directed in the potassium bromide disk solution. Pipet 1 mL of the sample solution, add methanol to method under Infrared Spectrophotometry <2.25>,andcom- make exactly 200 mL, and use this solution as the standard pare the spectrum with the Reference Spectrum or the spec- solution. Perform the test with these solutions as directed un- trum of Trichlormethiazide Reference Standard: both spectra der Thin-layer Chromatography <2.03>.Spot5mLeachof exhibit similar intensities of absorption at the same wave the sample solution and standard solution on a plate of silica numbers. gel for thin-layer chromatography. Develop the plate with a (3) Perform the test with Trichlormethiazide as directed mixture of ethyl acetate, ammonia solution (28) and under Flame Coloration Test <1.04> (2): a green color ap- methanol (9:1:1) to a distance of about 10 cm, and air-dry the pears. plate. Examine the plate under ultraviolet light (main Purity (1) Chloride <1.03>—Dissolve 1.0 g of Trichlor- wavelength: 365 nm): the spots other than the principal spot methiazide in 30 mL of acetone, add 6 mL of dilute nitric from the sample solution are not more intense than the spot acid and water to make 50 mL, and perform the test using from the standard solution. this solution as the test solution. Prepare the control solution Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, as follows: to 1.0 mL of 0.01 molWL hydrochloric acid VS 4 hours). add 30 mL of acetone, 6 mL of dilute nitiric acid and water to make 50 mL (not more than 0.036z). Residue on ignition <2.44> Not more than 0.10z (1 g). (2) Sulfate <1.14>—Dissolve 1.0 g of Trichlormethiazide Assay Weigh accurately about 0.15 g of Triamterene, previ- in 30 mL of acetone, add 1 mL of dilute hydrochloric acid ously dried, and dissolve in 100 mL of acetic acid (100) by and water to make 50 mL, and perform the test using this so- warming. Titrate <2.50> with 0.05 molWL perchloric acid VS lution as the test solution. Prepare the control solution as fol- (indicator: 2 drops of crystal violet TS). Perform a blank de- lows: to 1.0 mL of 0.005 molWL sulfuric acid VS add 30 mL termination, and make any necessary correction. of acetone, 1 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.048z). Each mL of 0.05 molWL perchloric acid VS (3) Heavy metals <1.07>—Proceed with 1.0 g of Trichlor- ＝ 12.663 mg of C H N 12 11 7 methiazide according to Method 2, and perform the test. Pre- Containers and storage Containers—Well-closed contain- pare the control solution with 2.0 mL of Standard Lead Solu- ers. tion (not more than 20 ppm). (4) Arsenic <1.11>—Prepare the test solution with 0.6 g of Trichlormethiazide according to Method 5, using 20 mL Trichlormethiazide of N,N-dimethylformamide, and perform the test (not more than 3.3 ppm). トリクロルメチアジド (5) Related substances―Dissolve 25 mg of Trichlor- methiazide in 50 mL of acetonitrile, and use the solution as the sample solution. Perform the test with 10 mLofthesam- ple solution as directed under Liquid Chromatography <2.01> according to the following conditions, determine each peak area by the automatic integration method, and calculate the amount of related substances by the area percentage method: the amount of 4-amino-6-chlorobenzene-1,3-disulfonamide, C H Cl N O S :380.66 8 8 3 3 4 2 having the relative retention time of about 0.3 with respect to (3RS)-6-Chloro-3-dichloromethyl-3,4-dihydro-2H-1,2,4- trichlormethiazide, is not more than 2.0z, and the total benzothiadiazine-7-sulfonamide 1,1-dioxide [133-67-5] amount of the related substances is not more than 2.5z. Operating conditions— Trichlormethiazide, when dried, contains not less Detector: An ultraviolet absorption photometer than 97.5z and not more than 102.0z of (wavelength: 268 nm). C H Cl N O S . 8 8 3 3 4 2 Column: A stainless steel column 4.6 mm in inside Description Trichlormethiazide occurs as a white powder. diameter and 15 cm in length, packed with phenylsilanized It is freely soluble in N,N-dimethylformamide and in silica gel for liquid chromatography (5 mminparticle acetone, slightly soluble in acetonitrile and in ethanol (95), diameter). and practically insoluble in water. Column temperature: A constant temperature of about A solution of Trichlormethiazide in acetone (1 in 50) shows 259C. no optical rotation. Mobile phase A: A mixture of diluted phosphoric acid Melting point: about 2709C (with decomposition). (1 in 1000) and acetonitrile (3:1). JP XV O‹cial Monographs / Trichlormethiazide Tablets 1199

Mobile phase B: A mixture of acetonitrile and diluted Detector: An ultraviolet absorption photometer phosphoric acid (1 in 1000) (3:1). (wavelength: 268 nm). Flowing of the mobile phase: Control the gradient by Column: A stainless steel column 4.6 mm in inside mixing the mobile phases A and B as directed in the following diameter and 15 cm in length, packed with phenylsilanized table. silica gel for liquid chromatography (5 mminparticle diameter). Time after injection Mobile phase Mobile phase Column temperature: A constant temperature of about of sample (min) A(volz) B(volz) 259C. Mobile phase: A mixture of diluted phosphoric acid (1 in 0–10 100 0 1000) and acetonitrile (3:1). 10–20 100ª 00ª 100 Flow rate: Adjust the ‰ow rate so that the retention time of trichlormethiazide is about 8 minutes. Flow rate: 1.5 mL per minute System suitability— Time span of measurement: About 2.5 times as long as the System performance: When the procedure is run with retention time of trichlormethiazide beginning after the sol- 10 mL of the standard solution under the above operating vent peak. conditions, the internal standard and trichlormethiazide are System suitability— eluted in this order with the resolution between these peaks Test for required detectability: To exactly 1 mL of the sam- being not less than 2.0. ple solution add acetonitrile to make exactly 50 mL, and use System repeatability: When the test is repeated 6 times with this solution as the solution for system suitability test. Pipet 10 mL of the standard solution under the above operating 1 mL of the solution, and add acetonitrile to make exactly conditions, the relative standard deviation of the ratio of the 20 mL. Conˆrm that the peak area of trichlormethiazide peak area of trichlormethiazide to that of the internal stan- obtained from 10 mLofthissolutionisequivalentto3.5to dard is not more than 1.0z. 6.5z of that of trichlormethiazide obtained from 10 mLof Containers and storage Containers—Well-closed contain- the solution for system suitability test. ers. System performance: To 5 mL of the solution for system suitability test add 5 mL of water, and warm in a water bath at 609C for 30 minutes. When the procedure is run with 10 mL of this solution, after cooling, under the above operat- Trichlormethiazide Tablets ing conditions, 4-amino-6-chlorobenzene-1,3-disulfonamide and trichlormethiazide are eluted in this order, the relative トリクロルメチアジド錠 retention time of 4-amino-6-chlorobenzene-1,3-disul- fonamide with respect to trichlormethiazide is about 0.3, and Trichlormethiazide Tablets contain not less than the number of theoretical plates and the symmetry factor of 93.0z and not more than 107.0z of the labeled the peak of trichlormethiazide are not less than 5000 and not amount of trichlormethiazide (C8H8Cl3N3O4S2: more than 1.2, respectively. 380.66). System repeatability: When the test is repeated 3 times with 10 mL of the solution for system suitability test under the Method of preparation Prepare as directed under Tablets, above operating conditions, the relative standard deviation with Trichlormethiazide. of the peak area of trichlormethiazide is not more than 2.0z. Identiˆcation To an amount of pulverized Trichlor- Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 methiazide Tablets, equivalent to 4 mg of Trichlormethiazide hours). according to the labeled amount, add 10 mL of acetone, shake vigorously for 5 minutes, centrifuge, and use the super- Residue on ignition <2.44> Not more than 0.1z (1 g). natant liquid as the sample solution. Separately, dissolve 4 Assay Weigh accurately about 25 mg of Trichlormethiazide mg of Trichlormethiazide Reference Standard in 10 mL of and Trichlormethiazide Reference Standard, previously acetone, and use this solution as the standard solution. Per- dried, and dissolve separately in exactly 20 mL of the internal form the test with these solutions as directed under Thin-lay- standard solution. To 1 mL of these solutions add acetoni- er Chromatography <2.03>.Spot5mL each of the sample so- trile to make 20 mL, and use these solutions as the sample so- lutionandstandardsolutiononaplateofsilicagelwith lution and the standard solution. Perform the test with 10 mL ‰uorescent indicator for thin-layer chromatography. Develop each of the sample solution and standard solution as directed the plate with a mixture of ethyl acetate, hexane and under Liquid Chromatography <2.01> according to the fol- methanol (10:4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: lowing conditions, and determine the ratios, QT and QS,of the peak area of trichlormethiazide to that of the internal 254 nm): the principal spots from the sample solution and the standard. standard solution show the same Rfvalue. Purity Related substances—Pulverize a suitable amount of Amount (mg) of C8H8Cl3N3O4S2＝WS×(QT/QS) Trichlormethiazide Tablets in an agate mortar. Take an WS: Amount (mg) of Trichlormethiazide Reference Stan- amount of the powder, equivalent to 10 mg of Trichlor- dard methiazide according to the labeled amount, add 20 mL of Internal standard solution—A solution of 3-nitrophenol in acetonitrile, shake vigorously for 15 minutes, centrifuge, and acetonitrile (1 in 800). use the supernatant liquid as the sample solution. Perform Operating conditions— the test with 10 mL of the sample solution as directed under 1200 Trichlormethiazide Tablets / O‹cial Monographs JP XV

Liquid Chromatography <2.01> according to the following ing to the following method: it meets the requirement of the conditions, determine each peak area by the automatic in- Content uniformity test. tegration method, and calculate the amount of each related Perform the test according to the following method: it substance by the area percentage method: the amount of 4- meets the requirements of the Content Uniformity Test. To amino-6-chlorobenzene-1,3-disulfoneamide, having the rela- one tablet of Trichlormethiazide Tablets add 5 mL of diluted tive retention time of about 0.3 with respect to trichlor- phosphoric acid (1 in 50) to disintegrate. Add exactly an methiazide, is not more than 4.0z, and the total amount of amount of the internal standard solution, equivalent to 10 the peaks other than trichlormethiazide is not more than mL per 2 mg of trichlormethiazide (C8H8Cl3N3O4S2)accord- 5.0z. ing to the labeled amount, add acetonitrile to make 25 mL, Operating conditions— shake vigorously for 15 minutes, and centrifuge. To an Detector: An ultraviolet absorption photometer amount of the supernatant liquid add the mobile phase to (wavelength: 268 nm). make a solution so that it contains about 40 mg of trichlor-

Column: A stainless steel column 4.6 mm in inside methiazide (C8H8Cl3N3O4S2) in each mL, and use this solu- diameter and 15 cm in length, packed with phenylsilanized tion as the sample solution. Separately, weigh accurately silica gel for liquid chromatography (5 mminparticle about 20 mg of Trichlormethiazide Reference Standard, diameter). previously dried at 1059C for 3 hours, and dissolve in Column temperature: A constant temperature of about acetonitrile to make exactly 100 mL. Pipet 5 mL of this solu- 259C. tion, add exactly 5 mL of the internal standard solution, add Mobile phase A: A mixture of diluted phosphoric acid 10 mL of acetonitrile and 5 mL of diluted phosphoric acid (1 (1 in 1000) and acetonitrile (3:1). in 50), and use this solution as the standard solution. Per- Mobile phase B: A mixture of acetonitrile and diluted form the test with 20 mL each of the sample solution and stan- phosphoric acid (1 in 1000) (3:1). dard solution as directed under Liquid Chromatography Flowing of the mobile phase: Control the gradient by <2.01> according to the conditions described in the Assay, mixing the mobile phases A and B as directed in the following and determine the ratios, QT and QS, of the peak area of table. trichlormethiazide to that of the internal standard.

Amount (mg) of trichlormethiazide (C8H8Cl3N3O4S2) Time after injection Mobile phase Mobile phase ＝WS×(QT/QS)×C×(1/20) of sample (min) A(volz) B(volz) WS: Amount (mg) of Trichlormethiazide Reference 0–10 100 0 Standard 10–20 100ª 00ª 100 C: Labeled amount (mg) of trichlormethiazide

(C8H8Cl3N3O4S2) per tablet Flow rate: 1.5 mL/minute Internal standard solution—A solution of 3-nitrophenol in Time span of measurement: About 2.5 times as long as the acetonitrile (1 in 5000). retention time of trichlormethiazide beginning after the sol- vent peak. Dissolution <6.10> Perform the test according to the follow- System suitability— ing method: it meets the requirement. Test for required detectability: Dissolve 25 mg of Trichlor- Perform the test with 1 tablet of Trichlormethiazide methiazide in 50 mL of acetonitrile. Pipet 1 mL of this solu- Tablets at 50 revolutions per minute according to the Paddle tion, add acetonitrile to make exactly 50 mL, and use this so- method, using 900 mL of water as the dissolution medium. lution as the solution for system suitability test. Pipet 1 mL Withdraw 20 mL or more of the dissolution medium 15 of the solution for system suitability test, and add acetonitrile minutes after starting the test, and ˆlter through a membrane to make exactly 20 mL. Conˆrm that the peak area of ˆlterwithporesizeofnotmorethan0.45mm. Discard the trichlormethiazide obtained from 10 mLofthissolutionis ˆrst 10 mL of the ˆltrate, pipet the subsequent V mL of the equivalent to 3.5 to 6.5z of that obtained from 10 mLofthe ˆltrate, add diluted phosphoric acid (1 in 50) to make exactly solution for system suitability test. V? mL so that each mL contains about 1.1 mgoftrichlor-

System performance: To 5 mL of the solution for system methiazide (C8H8Cl3N3O4S2) according to the labeled suitability test add 5 mL of water, and warm in a water bath amount, and use this solution as the sample solution. of 609C for 30 minutes. When the procedure is run with 10 Separately, weigh accurately about 22 mg of Trichlor- mL of this solution, after cooling, under the above operating methiazide Reference Standard, previously dried at 1059C conditions, 4-amino-6-chlorobenzene-1,3-disulfonamide and for 3 hours, and dissolve in acetonitrile to make exactly trichlormethiazide are eluted in this order, the relative reten- 200 mL. Pipet 2 mL of this solution, add diluted phosphoric tion time of 4-amino-6-chlorobenzene-1,3-disulfonamide acid (1 in 50) to make exactly 200 mL, and use this solution as with respect to trichlormethiazide is about 0.3, and the num- the standard solution. Perform the test with exactly 40 mL ber of theoretical plates and the symmetry factor of the peak each of the sample solution and standard solution as directed of trichlormethiazide are not less than 5000 and not more under Liquid Chromatography <2.01> according to the fol- than 1.2, respectively. lowing conditions, and determine the peak areas, ATa and System repeatability: When the test is repeated 3 times with ASa, of trichlormethiazide obtained with the sample solution 10 mL of the solution for system suitability test under the and the standard solution, and the area, ATb,ofthepeak, above operating conditions, the relative standard deviation having the relative retention time of about 0.3 with respect to of the peak area of trichlormethiazide is not more than 2.0z. trichlormethiazide, obtained with the sample solution. The dissolution rate in 15 minutes is not less than 75z. Uniformity of dosage units <6.02> Perform the test accord- Dissolution rate (z) with respect to the labeled amount of JP XV O‹cial Monographs / Trichomycin 1201

trichlormethiazide (C8H8Cl3N3O4S2) diameter). ＝WS×[(ATa＋ATb×0.95)/ASa]×(V?/V)×(1/C)×(9/2) Column temperature: A constant temperature of about 259C. W : Amount (mg) of Trichlormethiazide Reference Stan- S Mobile phase: A mixture of diluted phosphoric acid (1 in dard 1000) and acetonitrile (3:1). C: Labeled amount (mg) of trichlormethiazide Flow rate: Adjust the ‰ow rate so that the retention time of (C H Cl N O S ) per tablet 8 8 3 3 4 2 trichlormethiazide is about 8 minutes. Operating conditions— System suitability— Proceed as directed in the operating conditions in the System performance: When the procedure is run with Assay. 20 mL of the standard solution under the above operating System suitability— conditions, the internal standard and trichlormethiazide are System performance: Dissolve 25 mg of Trichlor- eluted in this order with the resolution between these peaks methiazide in 50 mL of acetonitrile. To 1 mL of this solution being not less than 2.0. add acetonitrile to make 50 mL. To 5 mL of this solution add System repeatability: When the test is repeated 6 times with 5 mL of water, and heat at 609Cinawaterbathfor30 20 mL of the standard solution under the above operating minutes. After cooling, when the procedure is run with 10 mL conditions, the relative standard deviation of the ratio of the of this solution under the above operating conditions, 4- peak area of trichlormethiazide to that of the internal stan- amino-6-chlorobenzene-1,3-disulfonamide and trichlor- dard is not more than 1.0z. methiazide are eluted in this order, the relative retention time Containers and storage Containers—Tight containers. of 4-amino-6-chlorobenzene-1,3-disulfonamide with respect to trichlormethiazide is about 0.3, and the number of theoret- ical plates and the symmetry factor of the peak of trichlor- methiazide are not less than 5000 and not more than 1.2, re- Trichomycin spectively. トリコマイシン System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of trichlormethiazide is not more than 2.0z. Assay Weigh accurately the mass of not less than 20 Trichlormethiazide Tablets, and pulverize the tablets in an agate mortar. Weigh accurately an amount of the powder, equivalent to about 2 mg of trichlormethiazide (C8H8Cl3N3O4S2)accordingtothelabeledamount,add5mL Trichomycin A ofdilutedphosphoricacid(1in50)andexactly10mLofthe 33-(3-Amino-3,6-dideoxy-b-D-mannopyranosyloxy)-17- internal standard solution, add 10 mL of acetonitrile, shake [6-(4-aminophenyl)-4-hydroxy-1-methyl-6-oxohexyl]- vigorously for 15 minutes, and centrifuge. To 2 mL of the su- 1,3,5,9,11,37-hexahydroxy-18-methyl-13,15-dioxo-16,39- pernatant liquid add 2 mL of the mobile phase, and use this dioxabicyclo[33.3.1]nonatriaconta-19,21,23,25,27,29,31- solution as the sample solution. Separately, weigh accurately heptaene-36-carboxylic acid [12698-99-6] about 40 mg of Trichlormethiazide Reference Standard, Trichomycin B previously dried at 1059C for 3 hours, and dissolve in 33-(3-Amino-3,6-dideoxy-b-D-mannopyranosyloxy)-17- acetonitrile to make exactly 200 mL. Pipet 5 mL of this solu- [6-(4-aminophenyl)-4-hydroxy-1-methyl-6-oxohexyl]- tion, add exactly 5 mL of the internal standard solution, add 1,3,5,7,9,37-hexahydroxy-18-methyl-13,15-dioxo-16,39- 10 mL of acetonitrile and 5 mL of diluted phosphoric acid (1 dioxabicyclo[33.3.1]nonatriaconta-19,21,23,25,27,29,31- in 50), and use this solution as the standard solution. Per- heptaene-36-carboxylic acid [12699-00-2] form the test with 20 mL each of the sample solution and stan- [1394-02-1,Trichomycin] dard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine Trichomycin is a mixture of polyene macrolide sub- the ratios, QT and QS, of the peak area of trichlormethiazide stances having antifungal and antiprotozoal activities to that of the internal standard. produced by the growth of Streptomyces hachijoensis.

Amount (mg) of trichlormethiazide (C8H8Cl3N3O4S2) It contains not less than 7000 Units per mg, calculat- ＝WS×(QT/QS)×(1/20) ed on the dried basis. The potency of Trichomycin is expressed as unit based on the amount of trichomycin. WS: Amount (mg) of Trichlormethiazide Reference One unit of Trichomycin is equivalent to 0.05 mgof Standard trichomycin. Internal standard solution—A solution of 3-nitrophenol in Description Trichomycin occurs as a yellow to yellow- acetonitrile (1 in 5000). brown powder. Operating conditions— It is practically insoluble in water, in ethanol (99.5) and in Detector: An ultraviolet absorption photometer tetrahydrofuran. (wavelength: 268 nm). It dissolves in dilute sodium hydroxide TS. Column: A stainless steel column 4.6 mm in inside It is hygroscopic. diameter and 15 cm in length, packed with phenylsilanized silica gel for liquid chromatography (5 mminparticle Identiˆcation (1) To 2 mg of Trichomycin add 2 mL of 1202 Triclofos Sodium / O‹cial Monographs JP XV sulfuric acid: a blue color appears, and the color is changed light, using light-resistant vessels. Weigh accurately an to a blue-purple after allowing to stand. amount of Trichomycin and Trichomycin Reference Stan- (2) Dissolve 1 mg of Trichomycin in 50 mL of a solution dard, equivalent to about 150,000 units, dissolve them of sodium hydroxide (1 in 200). Determine the absorption separately in a mixture of tetrahydrofuran for liquid chro- spectrum of this solution as directed under Ultraviolet-visible matography and water (3:1) to make exactly 100 mL, and use Spectrophotometry <2.24>: it exhibits maxima between 359 these solutions as the sample solution and the standard solu- nm and 365 nm, between 378 nm and 384 nm, and between tion. Perform the test with exactly 20 mLeachofthesample 400 nm and 406 nm. solutionandstandardsolutionasdirectedunderLiquid Chromatography <2.01> according to the following condi- Content ratio of the active principle Conduct this proce- tions, and determine the peak areas, A and A ,oftrichomy- dure without exposure to daylight, using light-resistant ves- T S cin. sels. Dissolve about 10 mg of Trichomycin in 50 mL of a mix- ture of tetrahydrofuran for liquid chromatography and water Amount (unit) of trichomycin ＝ WS × (AT/AS) (3:1), and use this solution as the sample solution. Perform W : Amount (unit) of Trichomycin Reference Standard the test with 5 mL of the sample solution as directed under S Liquid Chromatography <2.01> according to the following Operating conditions— conditions, determine the peak areas by the automatic in- Detector: An ultraviolet absorption photometer (wave- tegration method, and calculate the amount of trichomycin A length: 360 nm). and trichomycin B by the area percentage method: the Column: A stainless steel column 4.6 mm in inside di- amount of trichomycin A is between 20z and 40z,andthat ameter and 15 cm in length, packed with silica gel for liquid of trichomycin B is between 15z and 25z.Therelative chromatography (10 mm in particle diameter). retention time of trichomycin B with respect to trichomycin Column temperature: A constant temperature of about A is about 1.2. 259C. Operating conditions— Mobile phase: Dissolve 15 g of ammonium acetate in 120 Detector: An ultraviolet absorption photometer (wave- mL of water, and add 1000 mL of acetonitrile for liquid chro- length: 360 nm). matography and 700 mL of methanol. Column: A stainless steel column 4.6 mm in inside di- Flow rate: Adjust the ‰ow rate so that the retention time of ameter and 15 cm in length, packed with octadecylsilanized trichomycin is about 6 minutes. silica gel for liquid chromatography (5 mminparticledi- System suitability— ameter). System performance: Dissolve 5 mg of Trichomycin and 1 Column temperature: A constant temperature of about mg of chloride in 100 mL of a mixture of tetra- 259C. hydrofuran for liquid chromatography and water (3:1). Mobile phase: Dissolve 3.4 g of potassium dihydrogen When the procedure is run with 20 mL of this solution under phosphate and 1.7 g of sodium lauryl sulfate in a mixture of the above operating conditions, berberine and trichomycin 600 mL of water and 400 mL of acetonitrile for liquid chro- are eluted in this order with the resolution between these matography. peaks being not less than 4. Flow rate: Adjust the ‰ow rate so that the retention time of System repeatability: When the test is repeated 6 times with trichomycin A is about 8 minutes. 20 mL of the standard solution under the above operating Time span of measurement: About 4 times as long as the conditions, the relative standard deviation of the peak area of retention time of trichomycin A. trichomycin is not more than 2.0z. System suitability— Containers and storage Containers—Tight containers. Test for required detectability: Measure exactly 5 mL of Storage—Light-resistant, and in a cold place. the sample solution, add a mixture of tetrahydrofuran for liquid chromatography and water (3:1) to make exactly 50 mL, and use this solution as the solution for system suitabil- ity test. Pipet 5 mL of the solution for system suitability test, Triclofos Sodium and add a mixture of tetrahydrofuran for liquid chro- matography and water (3:1) to make exactly 30 mL. Conˆrm Monosodium Trichloroethyl Phosphate that the peak area of trichomycin A obtained from 5 mLof トリクロホスナトリウム this solution is equivalent to 12 to 22z of that from 5 mLof the solution for system suitability test. System performance: When the procedure is run with 5 mL of the solution for system suitability test under the above operating conditions, trichomycin A and trichomycin B are C2H3Cl3NaO4P: 251.37 eluted in this order with the resolution between these peaks Monosodium 2,2,2-trichloroethyl being not less than 2.5. monohydrogenphosphate [7246-20-0] System repeatability: When the test is repeated 6 times with 5 mL of the solution for system suitability test under the Triclofos Sodium, when dried, contains not less than above operating conditions, the relative standard deviation 97.0 z and not more than 102.0 z of of the peak area of trichomycin A is not more than 2.0z. C2H3Cl3NaO4P, and not less than 41.0z and not more Loss on drying <2.41> Not more than 5.0z (1 g, in vacuum, than 43.2z of chlorine (Cl: 35.45). 609C, 3 hours). Description Triclofos Sodium is a white, crystalline pow- Assay Conduct this procedure without exposure to day- JP XV O‹cial Monographs / Triclofos Sodium Syrup 1203 der. g of Triclofos Sodium, previously dried, place in a Kjeldhal It is freely soluble in water, slightly soluble in ethanol (95), flask, add 2 mL of sulfuric acid and 2.5 mL of nitric acid, and practically insoluble in diethyl ether. and heat until brown gas are not evolved. After cooling, add It is hygroscopic. 1 mL of nitric acid, heat until white fumes are produced, and cool. Repeat this procedure until the solution becomes color- Identification (1) Determine the infrared absorption spec- less. Transfer this solution to a flask using 150 mL of water, trum of Triclofos Sodium as directed in the potassium add 50 mL of molybdenum (III) oxide-citric acid TS, heat bromide disk method under Infrared Spectrophotometry gently to boil, add gradually 25 mL of quinoline TS with stir- <2.25>, and compare the spectrum with the Reference Spec- ring, and heat on a water bath for 5 minutes. After cooling, trum: both spectra exhibit similar intensities of absorption at filter the precipitate, and wash repeatedly with water until the thesamewavenumbers. washing does not indicate acidity. Transfer the precipitate to (2) To 0.5 g of Triclofos Sodium add 10 mL of nitric a flask using 100 mL of water, add exactly 50 mL of 0.5 molW acid, evaporate on a water bath to dryness, and ignite further L sodium hydroxide VS, dissolve, and titrate <2.50> with 0.5 over a flame. Dissolve the residue in 5 mL of water, and filter molWL hydrochloric acid VS until the color of the solution it necessary: the filtrate responds to Qualitative Tests <1.09> changes from purple to yellow (indicator: 3 drops of for sodium salt. phenolphthalein-thymol blue TS). Perform a blank determi- (3) To 0.1 g of Triclofos Sodium add 1 g of anhydrous nation. sodium carbonate, and heat for 10 minutes. After cooling, dissolve the residue in 40 mL of water, filter if necessary, and Each mL of 0.5 molWL sodium hydrochloride VS render the filtrate acidic with dilute nitric acid: the solution ＝ 4.834 mg of C2H3Cl3NaO4P responds to the Qualitative Tests <1.09> (2) for chloride. The (2) Chlorine—Weigh accurately about 10 mg of Triclofos remainder of the filtrate responds to the Qualitative Tests Sodium, previously dried, perform the test according to the <1.09> (1) for chloride and to the Qualitative Tests <1.09> for procedure of determination for chlorine as directed under phosphate. Oxygen Flask Combustion Method <1.06>,using1mLof1 pH <2.54> Dissolve 1.0 g of Triclofos Sodium in 50 mL of molWL sodium hydroxide TS and 20 mL of water as the ab- water: the pH of this solution is between 3.0 and 4.5. sorbing liquid. Purity (1) Clarity and color of solution—Dissolve 1.0 g of Containers and storage Containers—Tight containers. Triclofos Sodium in 50 mL of water: the solution is clear and colorless. (2) Chloride <1.03>—Perform the test with 0.20 g of Triclofos Sodium Syrup Triclofos Sodium. Prepare the control solution with 1.0 mL of 0.01 molWLhydrochloricacidVS(notmorethan0.178z). Monosodium Trichloroethyl Phosphate Syrup (3) Heavy metals <1.07>—Proceed with 1.0 g of Triclofos Sodium according to Method 1, and perform the test. Pre- トリクロホスナトリウムシロップ pare the control solution with 2.0 mL of Standard Lead Solu- tion (not more than 20 ppm). (4) Arsenic <1.11>—Prepare the test solution with 1.0 g Triclofos Sodium Syrup contains not less than 90z of Triclofos Sodium according to Method 1, and perform the and not more than 110z of the labeled amount of test (not more than 2 ppm). triclofos sodium (C2H3Cl3NaO4P: 251.37). (5) Free phosphoric acid—Weigh accurately about 0.3 g Method of preparation Prepare as directed under Syrups, of Triclofos Sodium, previously dried, dissolve in water to with Triclofos Sodium. make exactly 100 mL, and use this solution as the sample so- lution. Pipet 5 mL each of the sample solution and Standard Identification (1) Weigh a portion of Triclofos Sodium Phosphoric Acid Solution, add 2.5 mL of hexaammonium Syrup, equivalent to 0.25 g of Triclofos Sodium according to heptamolybdate-sulfuric acid TS and 1 mL of 1-amino-2- the labeled amount, add 40 mL of water, shake well, add 5 naphthol-4-sulfonic acid TS, shake, add water to make exact- mLofdilutedsulfuricacid(3in50),andextractwith25mL ly 25 mL, and allow to stand at 209C for 30 minutes. Perform of 3-methyl-1-butanol. Take 5 mL of the extract, evaporate the test with these solutions, using a solution obtained in the on a water bath to dryness, and add 1 mL of diluted sulfuric same manner with 5 mL of water as the blank, as directed un- acid (1 in 2) and 1 mL of a solution of potassium perman- der Ultraviolet-visible Spectrophotometry <2.24>. Determine ganate (1 in 20) to the residue. Heat in a water bath for 5 minutes, add 7 mL of water, and then add a solution of oxal- the absorbances, AT and AS, of each solution from the sam- ple solution and Standard Phosphoric Acid Solution at 740 ic acid dihydrate (1 in 20) until the color of the solution disap- nm: the content of the free phosphoric acid is not more than pears. To 1 mL of this solution add 1 mL of pyridine and 1 1.0z. mL of a solution of sodium hydroxide (1 in 5), and heat in a water bath, while shaking, for 1 minute: a light red color de- Content (z) of the free phosphoric acid (H3PO4) velops in the pyridine layer. ＝ (AT/AS) × (1/W) × 287.8 (2) Take 10 mL of the extract obtained in (1), evaporate W: Amount (mg) of the sample taken. on a water bath to dryness, add 1 g of anhydrous sodium car- bonate to the residue, and heat for 10 minutes. After cooling, Loss on drying <2.41> Not more than 5.0z (1 g, in vacuum, dissolve the residue in 40 mL of water, filter if necessary, and 1009C, 3 hours). render the filtrate acidic with dilute nitric acid: the solution Assay (1) Triclofos sodium—Weigh accurately about 0.2 responds to the Qualitative Tests <1.09> (2)forchloride.The 1204 Trihexyphenidyl Hydrochloride / O‹cial Monographs JP XV remainder of the filtrate responds to the Qualitative Tests Collect the precipitate, wash with a small amount of water, <1.09> (1) for chloride and to the Qualitative Tests <1.09> for recrystallize from methanol, and dry in a desiccator (in vacu- phosphate. um, silica gel) for 2 hours: the crystals so obtained melt <2.60> between 1139Cand1179C. pH <2.54> 6.0 – 6.5 (3) The sample solution obtained in (1) responds to the Assay Weigh accurately a portion of Triclofos Sodium Qualitative Tests <1.09> (2) for chloride. Syrup, equivalent to 0.13 g of Triclofos Sodium according to pH <2.54> Dissolve 1.0 g of Trihexyphenidyl Hydrochloride the labeled amount, add 15 mL of water, 1 mL of sodium in 100 mL of water by warming, and cool: the pH of this so- hydroxide TS and 15 mL of diethyl ether, shake for 1 minute, lution is between 5.0 and 6.0. and separate the water layer. Wash the diethyl ether layer with 1 mL of water, and combine the washing with above Purity (1) Clarity and color of solution—Dissolve 1.0 g of water layer. To this solution add 2.5 mL of diluted sulfuric Trihexyphenidyl Hydrochloride in 100 mL of water by warm- acid (3 in 50), and extract with four 10-mL portions of 3- ing: the solution is clear and colorless. methyl-1-butanol. Combine the 3-methyl-1-butanol extracts, (2) Heavy metals <1.07>—Dissolve 1.5 g of Trihex- and add 3-methyl-1-butanol to make exactly 50 mL. Measure yphenidyl Hydrochloride in 60 mL of water by warming on a exactly 10 mL each of this solution, and dilute with potassi- water bath at 809C, cool, and filter. To 40 mL of the filtrate um hydroxide-ethanol TS. Place in a glass ampule, fire-seal, add 2 mL of dilute acetic acid and water to make 50 mL, and mix, and heat at 1209C for 2 hours in an autoclave. After perform the test using this solution as the test solution. Pre- cooling, transfer the contents to a flask, add 20 mL of diluted pare the control solution with 2.0 mL of Standard Lead Solu- nitric acid (63 in 500) and exactly 25 mL of 0.02 molWLsilver tion, 2 mL of dilute acetic acid and water to make 50 mL (not nitrate VS, shake well, and titrate <2.50> the excess silver ni- more than 20 ppm). trate with 0.02 molWL ammonium thiocyanate VS (indicator: (3) Piperidylpropiophenone—Dissolve 0.10 g of Trihex- 2 to 3 drops of ammonium iron (III) sulfate TS). Perform a yphenidyl Hydrochloride in 40 mL of water and 1 mL of 1 blank determination. molWL hydrochloric acid VS by warming, cool, and add water to make 100 mL. Determine the absorbance of this so- Each mL of 0.02 molWLsilvernitrateVS lution at 247 nm as directed under Ultraviolet-visible Spec- ＝ 1.676 mg of C H Cl NaO P 2 3 3 4 trophotometry <2.24>: the absorbance is not more than 0.50. Containers and storage Containers—Tight containers. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, Storage—In a cold place. 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Trihexyphenidyl Hydrochloride Assay Weigh accurately about 0.5 g of Trihexyphenidyl Hydrochloride, previously dried, dissolve in 50 mL of a mix- トリヘキシフェニジル塩酸塩 ture of acetic anhydride and acetic acid (100) (1:1), and titrate <2.50> with 0.1 molWL perchloric acid-dioxane VS (potentio- metric titration). Perform a blank determination, and make any necessary correction. Each mL of 0.1 molWL perchloric acid-dioxane VS

＝ 33.793 mg of C20H31NO.HCl Containers and storage Containers—Tight containers. C20H31NO.HCl: 337.93 (1RS)-1-Cyclohexyl-1-phenyl-3-(piperidin-1-yl)propan-1-ol monohydrochloride [52-49-3] Trihexyphenidyl Hydrochloride Trihexyphenidyl Hydrochloride, when dried, con- Tablets tains not less than 98.5z of C20H31NO.HCl. トリヘキシフェニジル塩酸塩錠 Description Trihexyphenidyl Hydrochloride occurs as a white, crystalline powder. It is odorless, and has a bitter taste. Trihexyphenidyl Hydrochloride Tablets contain not It is soluble in ethanol (95), sparingly soluble in acetic acid less than 93z and not more than 107z of the labeled (100), slightly soluble in water, very slightly soluble in acetic amount of trihexyphenidyl hydrochloride anhydride, and practically insoluble in diethyl ether. (C H NO.HCl: 337.93). Melting point: about 2509C (with decomposition). 20 31 Method of preparation Prepare as directed under Tablets, Identification (1) Dissolve 1 g of Trihexyphenidyl with Trihexyphenidyl Hydrochloride. Hydrochloride in 100 mL of water by warming, and cool. Use this solution as the sample solution. To 5 mL of the sam- Identification (1) Weigh a quantity of powdered Trihex- ple solution add 1 mL of a solution of 2,4,6-trinitrophenol in yphenidyl Hydrochloride Tablets, equivalent to 0.1 g of Tri- chloroform (1 in 50), and shake vigorously: a yellow hexyphenidyl Hydrochloride according to the labeled precipitate is formed. amount, add 30 mL of chloroform, shake, and filter. (2) To 20 mL of the sample solution obtained in (1) add 2 Evaporate the filtrate on a water bath to dryness. Dissolve mL of sodium hydroxide TS: a white precipitate is formed. the residue in 10 mL of water by warming, cool, and use this JP XV O‹cial Monographs / Trihexyphenidyl Hydrochloride Tablets 1205 solution as the sample solution. With 5 mL of the sample so- more than 0.8 mm. Discard the first 10 mL of the filtrate, and lution, proceed as directed in the Identification (1) under Tri- use the subsequent as the sample solution. Separately, weigh hexyphenidyl Hydrochloride. accurately about 10 mg of Trihexyphenidyl Hydrochloride (2) Shake a quantity of powdered Trihexyphenidyl Reference Standard, previously dried at 1059C for 3 hours, Hydrochloride Tablets, equivalent to 0.01 g of Trihex- and dissolve in 2nd fluid for dissolution test to make exactly yphenidyl Hydrochloride according to the labeled amount, 100 mL. Pipet 2 mL of this solution, add 2nd fluid for disso- with 5 mL of chloroform, filter, and use the filtrate as the lution test to make exactly 100 mL, and use this solution as sample solution. Dissolve 0.02 g of Trihexyphenidyl the standard solution. Pipet 20 mL each of the sample solu- Hydrochloride Reference Standard in 10 mL of chloroform, tion, the standard solution and 2nd fluid for dissolution test and use this solution as the standard solution. Perform the add exactly 1 mL of diluted acetic acid (31) (1 in 10), and im- test with these solutions as directed under Thin-layer Chro- mediately add 5 mL of bromocresol green-sodium hydroxide- matography <2.03>.Spot10mL each of the sample solution acetic acid-sodium acetate TS, and shake. Then, add exactly and standard solution on a plate of silica gel for thin-layer 10 mL each of dichloromethane, shake well, centrifuge, and chromatography. Develop the plate with a mixture of chlo- take the dichloromethane layer. Determine the absorbances, roform and methanol (9:1) to a distance of about 10 cm, and AT, AS and AB, of these dichloromethane layers at 415 nm as air-dry the plate. Spray evenly hydrogen hexachloroplatinate directed under Ultraviolet-visible Spectrophotometry <2.24>, (IV)-potassium iodide TS on the plate: the spots from the using dichloromethane as a blank. The dissolution rate of sample solution and the standard solution show a blue-purple Trihexyphenidyl Hydrochloride Tablets in 30 minutes should color and the same Rfvalue. be not less than 70z. (3) The sample solution obtained in (1) responds to the Dissolution rate (z) with respect to the labeled amount Qualitative Tests <1.09> (2) for chloride. of trihexyphenidyl hydrochloride (C20H31NO.HCl) Uniformity of dosage units <6.02> Perform the test accord- ＝ WS × [(AT － AB)/(AS － AB)] × (1/ C ) × 18 ing to the following method: it meets the requirement of the W : Amount (mg) of Trihexyphenidyl Hydrochloride Content uniformity test. S Reference Standard To one tablet of Trihexyphenidyl Hydrochloride Tablets C: Labeled amount (mg) of trihexyphenidyl hydrochloride add 2 mL of dilute hydrochloric acid and 60 mL of water, (C H NO.HCl) in 1 tablet disintegrate by vigorous shaking for 10 minutes, and warm 20 31 on a water bath with occasional shaking for 10 minutes. Assay Weigh accurately and powder not less than 20 Tri- Cool, add 2 mL of methannol, and add water to make exactly hexyphenidyl Hydrochloride Tablets. Weigh accurately a V mL of the solution contains about 20 mgoftrihexyphenidyl portion of the powder, equivalent to about 5 mg of trihex- hydrochloride (C20H31NO.HCl) per ml. Centrifuge, if neces- yphenidyl hydrochloride (C20H31NO.HCl), dissolve in 2 mL sary, and use the supernatant liquid as the sample solution. of dilute hydrochloric acid and 60 mL of water by warming Separately, dissolve about 20 mg of Trihexyphenidyl on a water bath for 10 minutes with occasional shaking. Af- Hydrochloride Reference Standard (determine previously its ter cooling, add 2 mL of methanol and water to make exactly loss on drying <2.41> in the same manner as Trihexyphenidyl 100 mL, and use this solution as the sample solution. Dis- Hydrochloride) in methanol to make exactly 20 mL. Pipet 2 solve about 50 mg of Trihexyphenidyl Hydrochloride Refer- mL of this solution, and add 2 mL of dilute hydrochloric acid ence Standard (determine previously its loss on drying <2.41> and water to make exactly 100 mL, and use this solution as in the same manner as Trihexyphenidyl Hydrochloride), the standard solution. Pipet 10 mL each of the sample solu- weighed accurately, in methanol, add methanol to make ex- tion and the standard solution, transfer to glass-stoppered actly 20 mL. Pipet 2 mL of this solution, add 2 mL of dilute centrifuge tubes, add exactly 10 mL of bromocresol purple- hydrochloric acid and water to make exactly 100 mL, and use dipotassium hydrogenphosphate-citric acid TS and 15 mL of this solution as the standard solution. Pipet 10 mL each of chloroform, stopper tightly, shake well, and centrifuge. Pipet the sample solution and the standard solution into glass-stop- 10 mL each of the chloroform layers, add chloroform to pered centrifuge tubes, add exactly 10 mL each of make exactly 50 mL. Determine the absorbances, AT and AS, bromocresol purple-dipotassium hydrogenphosphate-citric of the subsequent solutions of the sample solution and stan- acid TS and 15 mL each of chloroform, stopper tightly, dard solution at 408 nm as directed under Ultraviolet-visible shake thoroughly, and centrifuge. Pipet 10 mL each of the Spectrophotometry <2.24>, respectively. chloroform layers, and add chloroform to make exactly 50 mL. Determine the absorbances, A and A , of the subse- Amount (mg) of trihexyphenidyl hydrochloride T S quent solutions of the sample solution and standard solution (C H NO.HCl) 20 31 at 408 nm as directed under Ultraviolet-visible Spectrophoto- ＝ W ×(A /A )×(V/1000) S T S metry <2.24>, respectively. W : Amount (mg) of Trihexyphenidyl Hydrochlochloride S Amount (mg) of trihexyphenidyl hydrochloride Reference Standard, calculated on the dried basis (C20H31NO.HCl) Dissolution <6.10> Perform the test according to the follow- ＝ WS×(AT/AS)×(1/10) ing method: it meets the requirement. W : Amount (mg) of Trihexyphenidyl Hydrochloride Perform the test with 1 tablet of Trihexyphenidyl S Reference Standard, calculated on the dried basis Hydrochloride Tablets at 50 revolutions per minute accord- ing to the Paddle method, using 900 mL of 2nd fluid for dis- Containers and storage Containers—Tight containers. solution test as the dissolution medium. Take 30 mL or more of the dissolved solution 30 minutes after starting the test, and filter through a membrane filter with pore size of not 1206 Trimebutine Maleate / O‹cial Monographs JP XV

dard solution, and the total area of the peaks other than maleic acid and trimebutine is not more than the peak area of Trimebutine Maleate trimebutine from the standard solution. Operating conditions— トリメブチンマレイン酸塩 Detector: An ultraviolet absorption photometer (wave- length: 254 nm). Column: A stainless steel column 4.6 mm in inside di- ameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticledi- ameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 650 mL of diluted perchloric acid (17 in

C22H29NO5.C4H4O4: 503.54 20,000), previously adjusted the pH to 3.0 with a solution of (2RS)-2-Dimethylamino-2-phenylbutyl 3,4,5- ammonium acetate (1 in 1000), add 1 g of sodium 1-pen- trimethoxybenzoate monomaleate [34140-59-5] tanesulfonate to dissolve. To 650 mL of this solution add 350 mL of acetonitrile. Trimebutine Maleate, when dried, contains not less Flow rate: Adjust the ‰ow rate so that the retention time of than 98.5z and not more than 101.0z of trimebutine trimebutine is about 9 minutes. Time span of measurement: About 2 times as long as the maleate (C22H29NO5.C4H4O4). retention time of trimebutine beginning after the peak of Description Trimebutine Maleate occurs as white, crystals maleic acid. or crystalline powder. System suitability— It is freely soluble in N,N-dimethylformamide and in acetic Test for required detectability: Measure exactly 5 mL of acid (100), soluble in acetonitrile, and slightly soluble in the standard solution, and add a mixture of 0.01 mol/L water and in ethanol (99.5). hydrochloric acid TS and acetonitrile (13:7) to make exactly It dissolves in 0.01 mol/L hydrochloric acid TS. 20 mL. Conˆrm that the peak area of trimebutine obtained A solution of it in N,N-dimethylformamide (1 in 20) shows from 20 mLofthissolutionisequivalentto20to30z of that no optical rotation. from 20 mL of the standard solution. Identiˆcation (1) Determine the absorption spectrum of a System performance: Dissolve 40 mg of Trimebutine Male- solution of Trimebutine Maleate in 0.01 mol/L hydrochloric ate and 20 mg of hydrochloride in 100 mL of a acid TS (1 in 50,000) as directed under Ultraviolet-visible mixture of 0.01 mol/L hydrochloric acid TS and acetonitrile Spectrophotometry <2.24>, and compare the spectrum with (13:7). When the procedure is run with 20mL of this solution the Reference Spectrum: both spectra exhibit similar intensi- under the above operating conditions, trimebutine and im- ties of absorption at the same wavelengths. ipramine are eluted in this order with the resolution between (2) Determine the infrared absorption spectrum of these peaks being not less than 2.5. Trimebutine Maleate as directed in the potassium bromide System repeatability: When the test is repeated 6 times with disk method under Infrared Spectrophotometry <2.25>,and 20 mL of the standard solution under the above operating compare the spectrum with the Reference Spectrum: both conditions, the relative standard deviation of the peak area of spectra exhibit similar intensities of absorption at the same trimebutine is not more than 5z. wave numbers. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 Melting point <2.60> 131 – 1359C hours). Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Residue on ignition <2.44> Not more than 0.1z (1 g). Trimebutine Maleate according to Method 2, and perform Assay Weigh accurately about 0.8 g of Trimebutine Male- the test. Prepare the control solution with 2.0 mL of Stan- ate, previously dried, dissolve in 70 mL of acetic acid (100), dard Lead Solution (not more than 10 ppm). and titrate <2.50> with 0.1 mol/L perchloric acid VS until the (2) Arsenic <1.11>—Prepare the test solution with 2.0 g color of the solution changes from purple through blue to of Trimebutine Maleate according to Method 3, and perform blue-green (indicator: 3 drops of crystal violet TS). Perform a the test (not more than 1 ppm). blank determination, and make any necessary correction. (3) Related substances—Dissolve 0.10 g of Trimebutine Maleate in 100 mL of a mixture of 0.01 mol/L hydrochloric Each mL of 0.1 mol/L perchloric acid VS acid TS and acetonitrile (13:7), and use this solution as the ＝ 50.35 mg of C22H29NO5.C4H4O4 sample solution. Pipet 1 mL of the sample solution, add a Containers and storage Containers—Well-closed contain- mixture of 0.01 mol/L hydrochloric acid TS and acetonitrile ers. (13:7) to make exactly 250 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 un- der Liquid Chromatography <2.01> according to the follow- ing conditions, and determine each peak area by the automat- ic integration method: the area of the peak other than maleic acid and trimebutine from the sample solution is not more than 1/2 times the peak area of trimebutine from the stan- JP XV O‹cial Monographs / Trimetazidine Hydrochloride Tablets 1207

Assay Weigh accurately about 0.12 g of Trimetazidine Hydrochloride, dissolve in 5 mL of formic acid, add exactly Trimetazidine Hydrochloride 15 mL of 0.1 mol/L perchloric acid VS, and heat at 90 – 1009C for 30 minutes. After cooling, add 45 mL of acetic トリメタジジン塩酸塩 acid (100), and titrate <2.50> the excess perchloric acid with 0.1 mol/L sodium acetate VS (potentiometric titration). Per- form a blank determination in the same manner. Each mL of 0.1 mol/L perchloric acid VS

＝16.96 mg of C14H22N2O3.2HCl Containers and storage Containers—Tight containers. C14H22N2O3.2HCl: 339.26 1-(2,3,4-Trimethoxybenzyl)piperazine dihydrochloride [13171-25-0] Trimetazidine Hydrochloride Trimetazidine Hydrochloride contains not less than Tablets 98.0 z and not more than 101.0 z of C14H22N2O3.2HCl, calculated on the dried basis. トリメタジジン塩酸塩錠 Description Trimetazidine Hydrochloride occurs as a white, crystalline powder. Trimetazidine Hydrochloride Tablets contain not It is very soluble in water and in formic acid, sparingly less than 94.0z and not more than 106.0z of the soluble in methanol, and slightly soluble in ethanol (99.5). labeled amount of trimetazidine hydrochloride The pH of a solution of Trimetazidine Hydrochloride (1 in (C14H22N2O3.2HCl: 339.26). 20) is between 2.3 and 3.3. Method of preparation Prepare as directed under Tablets, Melting point: about 2279C (with decomposition). with Trimetazidine Hydrochloride. Identiˆcation (1) Determine the absorption spectrum of a Identiˆcation Shake a quantity of powdered Trimetazidine solution of Trimetazidine Hydrochloride in 0.1 mol/L Hydrochloride Tablets, equivalent to 10 mg of Trimetazidine hydrochloric acid TS (1 in 6250) as directed under Ultra- Hydrochloride according to the labeled amount, with 10 mL violet-visible Spectrophotometry <2.24>, and compare the of a mixture of ethanol (95) and water (3:1), and ˆlter. spectrum with the Reference Spectrum: both spectra exhibit Evaporate the ˆltrate on a water bath, add 2 mL of water to similar intensities of absorption at the same wavelengths. the residue, and shake. To 1 mL of this solution add 1 mL of (2) Determine the infrared absorption spectrum of p-benzoquinone TS, boil gently for 2 to 3 minutes, and cool: Trimetazidine Hydrochloride as directed in the potassium a red color develops. chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- Uniformity of dosage units <6.02> Perform the test accord- trum: both spectra exhibit similar intensities of absorption at ing to the following method: it meets the requirement of the thesamewavenumbers. Content uniformity test. (3) A solution of Trimetazidine Hydrochloride (1 in 50) To 1 tablet of Trimetazidine Hydrochloride Tablets add 15 responds to the Qualitative Tests <1.09> for chloride. mL of a mixture of 0.1 mol/L hydrochloric acid TS and ethanol(99.5)(1:1),andtreatwithultrasonicwavesfor10 Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of minutes. Shake the solution for 10 minutes, and add the mix- Trimetazidine Hydrochloride according to Method 2, and ture of 0.1 mol/L hydrochloric acid TS and ethanol (99.5) perform the test. Prepare the control solution with 2.0 mL of (1:1) to make exactly 20 mL. Centrifuge, pipet V mL of the Standard Lead Solution (not more than 10 ppm). supernatant liquid, equivalent to about 0.75 mg of trimetazi- (2) Related substances—Dissolve 0.5 g of Trimetazidine dine hydrochloride (C H N O .2HCl), add exactly 2.5 mL Hydrochloride in 10 mL of water, and use this solution as the 14 22 2 3 of the internal standard solution, add 0.1 mol/L hydrochlor- sample solution. Pipet 1 mL of the sample solution, add ic acid TS to make 50 mL, and use this solution as the sample water to make exactly 100 mL, and use this solution as the solution. Separately, weigh accurately about 30 mg of standard solution. Perform the test with these solutions as trimetazidine hydrochloride for assay, separately determined directed under Thin-layer Chromatography <2.03>.Spot10 the water content <2.48> in the same manner as Trimetazidine mLeachofthesamplesolutionandstandardsolutionona Hydrochloride, and dissolve in the mixture of 0.1 mol/L plate of silica gel with ‰uorescent indicator for thin-layer hydrochloric acid TS and ethanol (99.5) (1:1) to make exactly chromatography. Develop the plate with a mixture of 200mL.Pipet5mLofthissolution,addexactly5mLofthe diethylamine and cyclohexane (1:1) to a distance of about 10 internal standard solution and 0.1 mol/L hydrochloric acid cm, air-dry the plate, and then dry at 1109C for 1 hour. After TS to make 50 mL, and use this solution as the standard solu- cooling, examine under ultraviolet light (main wavelength: tion. Proceed as directed in the Assay. 254 nm): the spot other than the principal spot and the spot at the starting point are not more intense than the spots from Amount (mg) of trimetazidine hydrochloride the standard solution. (C14H22N2O3.2HCl)＝WS×(QT/QS)×(1/2V)

Water <2.48> Not more than 1.5z (2 g, direct titration). WS: Amount (mg) of trimetazidine hydrochloride for as- say, calculated on the anhydrous basis Residue on ignition <2.44> Not more than 0.1z (1 g). Internal standard solution—A solution of parahydroxyben- 1208 Trimethadione / O‹cial Monographs JP XV zoic acid in the mixture of 0.1 mol/L hydrochloric acid TS and use this solution as the sample solution. Separately, and ethanol (99.5) (1:1) (7 in 40,000). weigh accurately about 30 mg of trimetazidine hydrochloride for assay, separately determined the water content <2.48> in Dissolution <6.10> Perform the test according to the follow- the same manner as Trimetazidine Hydrochloride, and dis- ing method: it meets the requirement. solve in the mixture of 0.1 mol/L hydrochloric acid TS and Perform the test with 1 tablet of Trimetazidine Hydrochlo- ethanol (99.5) (1:1) to make exactly 200 mL. To exactly 5 mL ride Tablets at 50 revolutions per minute according to the of this solution add exactly 5 mL of the internal standard so- Paddle method using 900 mL of water as the dissolution lutionand0.1mol/LhydrochloricacidTStomake50mL, medium. Withdraw not less than 20 mL of the dissolved solu- and use this solution as the standard solution. Perform the tion 45 minutes after starting the test, and ˆlter through a test with 10 mL each of the sample solution and standard so- membrane ˆlter with a pore size of not more than 0.45 mm. lution as directed under Liquid Chromatography <2.01> ac- Discard the ˆrst 10 mL of the ˆltrate, pipet V mL of the sub- cording to the following conditions, and determine the ratios, sequent ˆltrate, and add water to make exactly V? mL so that Q and Q , of the peak area of trimetazidine to that of the in- each mL contains about 3.3 mg of trimetazidine hydrochlo- T S ternal standard. ride (C14H22N2O3.2HCl) according to the labeled amount. Pipet 3 mL of this solution, add exactly 3 mL of 0.1 mol/L Amount (mg) of trimetazidine hydrochloride hydrochloric acid TS, and use this solution as the sample so- (C14H22N2O3.2HCl)＝WS×(QT/QS)×1/10 lution. Separately, weigh accurately about 17 mg of W : Amount (mg) of trimetazidine hydrochloride for as- trimetazidine hydrochloride for assay, separately determined S say, calculated on the anhydrous basis the water content in the same manner as Trimetazidine Hydrochloride, and dissolve in water to make exactly 100 Internal standard solution—A solution of parahydroxyben- mL. Pipet 5 mL of this solution, and add water to make ex- zoic acid in the mixture of 0.1 mol/L hydrochloric acid TS actly 50 mL. Pipet 5 mL of this solution, and add water to and ethanol (99.5) (1:1) (7 in 40,000). make exactly 25 mL. Pipet 3 mL of this solution, add exactly Operating conditions— 3 mL of 0.1 mol/L hydrochloric acid TS, and use this solu- Detector: An ultraviolet absorption photometer tion as the standard solution. Perform the test with exactly 50 (wavelength: 230 nm). mLeachofthesamplesolutionandstandardsolutionas Column: A stainless steel column 4.6 mm in inside di- directed under Liquid Chromatography <2.01> according to ameter and 15 cm in length, packed with octadecylsilanized the following conditions, and determine the peak areas, AT silica gel for liquid chromatography (5 mminparticledi- and AS, of trimetazidine. The dissolution rate in 45 minutes is ameter). not less than 80z. Column temperature: A constant temperature of about 409C. Dissolution rate (z) with respect to the labeled amount of Mobile phase: A mixture of 0.05 mol/L potassium di- trimetazidine hydrochloride (C H N O .2HCl) 14 22 2 3 hydrogen phosphate TS, pH 3.0 and methanol (17:3). ＝W ×(A /A )×(V?/V)×(1/C)×18 S T S Flow rate: Adjust the ‰ow rate so that the retention time of

WS: Amount (mg) of trimetazidine hydrochloride for as- trimetazidine is about 7 minutes. say, calculated on the anhydrous basis System suitability— C: Labeled amount (mg) of trimetazidine hydrochloride System performance: When the procedure is run with 10

(C14H22N2O3.2HCl) in 1 tablet mL of the standard solution under the above operating condi- tions, trimetazidine and the internal standard are eluted in Operating conditions— this order with the resolution between these peaks being not Proceed as directed in the Assay. less than 3. System suitability— System repeatability: When the test is repeated 6 times with System performance: When the procedure is run with 50 10 mL of the standard solution under the above operating mL of the standard solution under the above operating condi- conditions, the relative standard deviation of the ratio of the tions, the number of theoretical plates and the symmetry fac- peak area of trimetazidine to that of the internal standard is tor of the peak of trimetazidine are not less than 5000 and not not more than 1.0z. more than 1.5, respectively. System repeatability: When the test is repeated 6 times with Containers and storage Containers—Tight containers. 50 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of trimetazidine is not more than 1.5z. Trimethadione Assay Weigh accurately not less than 20 tablets of トリメタジオン Trimetazidine Hydrochloride Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 3 mg of trimetazidine hydrochloride (C14H22N2O3.2HCl), add about 15 mL of a mixture of 0.1 mol/L hydrochloric acid TS and ethanol (99.5) (1:1), and treat with ultrasonic waves for 10 minutes. Then shake for 10 minutes, add the mixture of C H NO : 143.14 0.1 mol/L hydrochloric acid TS and ethanol (99.5) (1:1) to 6 9 3 3,5,5-Trimethyl-1,3-oxazolidine-2,4-dione [127-48-0] make exactly 20 mL, and centrifuge. To exactly 5 mL of the supernatant liquid add exactly 5 mL of the internal standard solutionand0.1mol/LhydrochloricacidTStomake50mL, Trimethadione, when dried, contains not less than 98.0z of C6H9NO3. JP XV O‹cial Monographs / Trimetoquinol Hydrochloride Hydrate 1209

tion (1) under Trimethadione. Description Trimethadione occurs as white crystals or crys- (2) Determine the infrared absorption spectrum of a solu- talline powder. It has a camphor-like odor. tion of the residue obtained in (1) in chloroform (1 in 50) in a It is very soluble in ethanol (95) and in chloroform, freely 0.1-mm fixed sodium chloride cell, as directed in the solution soluble in diethyl ether, and soluble in water. method under Infrared Spectrophotometry <2.25>:itexhibits Identification (1) To 5 mL of a solution of Trimethadione absorption at the wave numbers of about 2960 cm－1, 1814 (1 in 50) add 2 mL of barium hydroxide TS: a precipitate is cm－1,1735cm－1, 1445 cm－1,1394cm－1, 1290 cm－1,1100 formed immediately. cm－1 and 1055 cm－1. (2) Determine the infrared absorption spectrum of a solu- Assay Weigh accurately and powder not less than 20 tion of Trimethadione in chloroform (1 in 50) as directed in Trimethadione Tablets. Weigh accurately a portion of the the solution method under Infrared Spectrophotometry powder, equivalent to about 1 g of trimethadione <2.25>, using a 0.1-mm fixed sodium chloride cell, and com- (C H NO ), add 50 mL of ethanol (95), and boil gently for 15 pare the spectrum with the Reference Spectrum: both spectra 6 9 3 minutes under a reflux condenser. Filter the warm ethanol exhibit similar intensities of absorption at the same wave (95) solution into a 100-mL volumetric flask through a glass numbers. filter (G4), and wash the residue with three 10-mL portions of Meting point <2.60> 45–479C warm ethanol (95). Combine the washings with the filtrate in the flask, cool, and add ethanol (95) to make exactly 100 mL. Purity Heavy metals <1.07>—Proceed with 2.0 g of Pipet 25 mL of the solution into a glass-stoppered conical Trimethadione according to Method 1, and perform the test. flask, add 25 mL of water and exactly 30 mL of 0.1 molWL Prepare the control solution with 2.0 mL of Standard Lead sodium hydroxide VS, stopper, allow to stand for 15 minutes Solution (not more than 10 ppm). with occasional shaking, and titrate <2.50> the excess sodium Loss on drying <2.41> Not more than 0.5z (1 g, silica gel, hydroxide with 0.1 molWL hydrochloric acid VS (indicator: 4 6 hours). drops of cresol red TS). Perform a blank determination. Residue on ignition <2.44> Not more than 0.1z (1 g). Each mL of 0.1 molWL sodium hydroxide VS ＝ 14.31 mg of C H NO Assay Weigh accurately about 0.4 g of Trimethadione, 6 9 3 previously dried, in a glass-stoppered conical flask, dissolve Containers and storage Containers—Tight containers. in 5 mL of ethanol (95), add exactly measured 50 mL of 0.1 Storage—Not exceeding 309C. molWL sodium hydroxide VS, stopper, and allow to stand for 15 minutes with occasional shaking. Titrate <2.50> the excess sodium hydroxide with 0.1 molWL hydrochloric acid VS (in- Trimetoquinol Hydrochloride dicator: 4 drops of cresol red TS). Perform a blank determi- nation. Hydrate Each mL of 0.1 molWL sodium hydroxide VS Tretoquinol Hydrochloride ＝ 14.31 mg of C6H9NO3 トリメトキノール塩酸塩水和物 Containers and storage Containers—Tight containers. Storage—Not exceeding 309C.

Trimethadione Tablets

トリメタジオン錠

Trimethadione Tabelts contain not less than 94z C19H23NO5.HCl.H2O: 399.87 and not more than 106z of the labeled amount of (1S)-1-(3,4,5-Trimethoxybenzyl)-1,2,3,4-tetrahydro- trimethadione (C6H9NO3:143.14). isoquinoline-6,7-diol monohydrochloride monohydrate Method of preparation Prepare as directed under Tablets, [18559-59-6,anhydride] with Trimethadione. Trimetoquinol Hydrochloride Hydrate contains not Identification (1) Weighaportionofpowdered less than 98.5z and not more than 101.0z of trimeto- Trimethadione Tablets, equivalent to 1 g of Trimethadione quinol hydrochloride (C19H23NO5.HCl: 381.85), calcu- according to the labeled amount, add 10 mL of petroleum lated on the anhydrous basis. benzin, and shake frequently for 15 minutes. Decant, remove the petroleum benzin, add another 10 mL of petroleum ben- Description Trimetoquinol Hydrochloride Hydrate occurs zin, and repeat the extraction in the same manner. To the as white, crystals or crystalline powder. residue add 25 mL of diethyl ether, allow to stand for 20 It is freely soluble in methanol, and sparingly soluble in minutes with occasional shaking, filter, evaporate the filtrate water and in ethanol (99.5). at room temperature, and dry the residue in a desiccator (sili- Melting point: about 1519C (with decomposition, after ca gel) for 6 hours: the residue melts <2.60> between 449Cand drying in vacuum, 1059C, 4 hours). 479C. Proceed with this residue as directed in the Identifica- Identiˆcation (1) Determine the absorption spectrum of a 1210 Dental Triozinc Paste / O‹cial Monographs JP XV solution of Trimetoquinol Hydrochloride Hydrate in 0.01 retention time of trimetoquinol beginning after the solvent mol/L hydrochloric acid TS (1 in 20,000) as directed under peak. Ultraviolet-visible Spectrophotometry <2.24>,andcompare System suitability— the spectrum with the Reference Spectrum: both spectra ex- Test for required detection: To exactly 2 mL of the stan- hibit similar intensities of absorption at the same dard solution add the mobile phase to make exactly 20 mL. wavelengths. Conˆrm that the peak area of trimetoquinol obtained from (2) Determine the infrared absorption spectrum of 20 mLofthissolutionisequivalentto7to13z of that of Trimetoquinol Hydrochloride Hydrate as directed in the trimetoquinol obtained from 20 mL of the standard solution. potassium chloride disk method under Infrared Spec- System performance: Dissolve 5 mg of Trimetoquinol trophotometry <2.25>, and compare the spectrum with the Hydrochloride Hydrate and 1 mg of procaine hydrochloride Reference Spectrum: both spectra exhibit similar intensities in 50 mL of the mobile phase. When the procedure is run of absorption at the same wave numbers. with 20 mL of this solution under the above operating condi- (3) A solution of Trimetoquinol Hydrochloride Hydrate tions, procaine and trimetoquinol are eluted in this order (1 in 50) responds to the Qualitative Tests <1.09> (1) for chlo- with the resolution between these peaks being not less than 4. ride. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operating Optical rotation <2.49> [a]20: －16 – －199(0.25 g, calculat- D conditions, the relative standard deviation of the peak area of ed on the anhydrous basis, water, after warming and cooling, trimetoquinol is not more than 2.0z. 25 mL, 100 mm). Water <2.48> 3.5–5.5z (0.3 g, volumetric titration, direct pH <2.54> Dissolve 1.0 g of Trimetoquinol Hydrochloride titration). Hydrate in 100 mL of water by warming, and cool: the pH of this solution is between 4.5 and 5.5. Residue on ignition <2.44> Not more than 0.1z (1 g). Purity (1) Clarity and color of solution—Dissolve 0.10 g Assay Weigh accurately about 0.5 g of Trimetoquinol of Trimetoquinol Hydrochloride Hydrate in 10 mL of water Hydrochloride Hydrate, dissolve in 2 mL of 0.1 molWL by warming: the solution is clear and colorless. hydrochloric acid VS and 70 mL of ethanol (99.5) with (2) Sulfate <1.14>—Perform the test with 0.5 g of thorough shaking, and titrate <2.50> with 0.1 molWL potassi- Trimetoquinol Hydrochloride Hydrate. Prepare the control um hydroxide-ethanol VS (potentiometric titration). Calcu- solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not late the consumed volume of 0.1 molWL potassium hydrox- more than 0.038z). ide-ethanol VS between the first inflection point and of the (3) Heavy metals <1.07>—Proceed with 1.0 g of Trimeto- second inflection point. quinol Hydrochloride Hydrate according to Method 2, and Each mL of 0.1 molWL potassium hydroxide-ethanol VS perform the test. Prepare the control solution with 2.0 mL of ＝ 38.19 mg of C H NO .HCl Standard Lead Solution (not more than 20 ppm). 19 23 5 (4) Related substances—Dissolve 50 mg of Trimeto- Containers and storage Containers—Well-closed contain- quinol Hydrochloride Hydrate in 50 mL of the mobile phase, ers. and use this solution as the sample solution. Pipet 1 mL of Storage—Light-resistant. this solution, add the mobile phase 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 stan- Dental Triozinc Paste dard solution, as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each 歯科用トリオジンクパスタ peak area of both solutions by the automatic integration method: the total area of the peaks other than that of trimetoquinol from the sample solution is not larger than the Dental Triozinc Paste consists of a powder contain- peak area of trimetoquinol from the standard solution. ing Paraformaldehyde, Thymol, anhydrous zinc sul- Operating conditions— fate and Zinc Oxide, and a solution containing Cresol, Detector: An ultraviolet absorption photometer (wave- Potash Soap and Glycerin. Suitable amounts of the length: 283 nm). two components are triturated before use. Column: A stainless steel column 4.6 mm in inside di- Method of preparation ameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mminparticledi- (1) The powder ameter). Paraformaldehyde, ˆnely powdered 10 g Column temperature: A constant temperature of about Thymol, ˆnely powdered 3 g 409C. Zinc Sulfate Hydrate 9 g Mobile phase: Dissolve 2 g of potassium dihydrogen phos- Zinc Oxide 82 g phate and 2 g of sodium 1-pentane sulfonate in 1000 mL of To make about 100 g water. Adjust with phosphoric acid to a pH between 2.8 and 3.2,andˆlterthroughamembraneˆlterwithporesizeof0.4 Heat Zinc Sulfate Hydrate at about 2509C to obtain anhy- mm. Add 200 mL of acetonitrile to 800 mL of the ˆltrate. drous zinc sulfate, cool, and pulverize to a ˆne powder. Mix Flow rate: Adjust the ‰ow rate so that the retention time of homogeneously this powder with Thymol, Paraformalde- trimetoquinol is about 7 minutes. hyde, and Zinc Oxide. Time span of measurement: About twice as long as the (2) The solution JP XV O‹cial Monographs / L-Tryptophan 1211

Cresol 40 g dilute acetic acid and water to make 50 mL (not more than 20 Potash Soap 40 g ppm). Glycerin 20 g (3) N-Ethyl-g-picolylamine—Dissolve 0.10 g of To make 100 g Tropicamide in 5 mL of water by heating, add 1 mL of a so- lution of acetaldehyde (1 in 20), and shake well. Add 1 to 2 Dissolve Potash Soap in a mixture of Cresol and Glycerin. drops of sodium pentacyanonitrosylferrate (III) TS and 1 to 2 Description The powder occurs as a ˆne, white powder, drops of sodium hydrogen carbonate TS, and shake: no blue having a characteristic odor. The solution is a clear, yellow- color develops. brown to red-brown, viscous liquid, having the odor of (4) Tropic acid—To 10 mg of Tropicamide add 5 mg of cresol. sodium borate and 7 drops of 4-dimethylaminobenzaldehyde TS, and heat in a water bath for 3 minutes. Cool in ice water, Containers and storage Containers—Tight containers. and add 5 mL of acetic anhydride: no red-purple color de- velops. Loss on drying <2.41> Not more than 0.30z (1 g, in vacu- Tropicamide um, silica gel, 24 hours). トロピカミド Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.5 g of Tropicamide, previ- ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 molWL perchloric acid VS (indicator: 3 drops of crystal violet TS). Perform a blank determination, and make any necessary correction.

C17H20N2O2: 284.35 Each mL of 0.1 molWL perchloric acid VS (2RS)-N-Ethyl-3-hydroxy-2-phenyl-N-(pyridin-4- ＝ 28.44 mg of C17H20N2O2 ylmethyl)propanamide [1508-75-4] Containers and storage Containers—Tight containers. Storage—Light-resistant. Tropicamide, when dried, contains not less than 98.5z of C17H20N2O2. Description Tropicamide occurs as a white, crystalline pow- L-Tryptophan der. It is odorless, and has a bitter taste. It is freely soluble in ethanol (95) and in chloroform, slight- L-トリプトファン ly soluble in water and in diethyl ether, and practically insolu- ble in petroleum ether. It dissolves in dilute hydrochloric acid. The pH of a solution of Tropicamide (1 in 500) is between 6.5 and 8.0. Identification (1) To 5 mg of Tropicamide add 0.5 mL of a solution of ammonium vanadate (V) in sulfuric acid, (1 in C H N O :204.23 200), and heat: a blue-purple color develops. 11 12 2 2 (2S)-2-Amino-3-(indol-3-yl)propanoic acid [73-22-3] (2) Dissolve 5 mg of Tropicamide in 1 mL of ethanol (95) and 1 mL of water, add 0.1 g of 1-chloro-2,4-dinitrobenzene, L-Tryptophan, when dried, contains not less than and heat on a water bath for 5 minutes. Cool, and add 2 to 3 98.5z of C H N O . drops of a solution of sodium hydroxide (1 in 10) and 3 mL 11 12 2 2 of ethanol (95): a red-purple color develops. Description L-Tryptophan occurs as white to yellowish white crystals or crystalline powder. It is odorless, and has a Absorbance <2.24> E 1z (255 nm): 166 – 180 (after drying, 1cm slightly bitter taste. 5mg,2molWL hydrochloric acid TS, 200 mL). It is freely soluble in formic acid, slightly soluble in water, Melting point <2.60> 96–999C and very slightly soluble in ethanol (95). It dissolves in dilute hydrochloric acid. Purity (1) Chloride <1.03>—Dissolve 1.0 g of Tropica- mide in 30 mL of ethanol (95), add 6 mL of dilute nitric acid Identification Determine the infrared absorption spectrum and water to make 50 mL, and perform the test using this so- of L-Tryptophan, previously dried, as directed in the potassi- lution as the test solution. Prepare the control solution with um bromide disk method under Infrared Spectrophotometry 0.45 mL of 0.01 molWL hydrochloric acid VS, 30 mL of <2.25>, and compare the spectrum with the Reference Spec- ethanol (95), 6 mL of dilute nitric acid, and add water to trum: both spectra exhibit similar intensities of absorption at make 50 mL (not more than 0.016z). thesamewavenumbers. (2) Heavy metals <1.07>—Dissolve 1.0 g of Tropicamide Optical rotation <2.49> [a]20: －30.0 – －33.09Weigh ac- in 30 mL of ethanol (95), add 2 mL of dilute acetic acid and D curately about 0.25 g of L-Tryptophan, previously dried, and water to make 50 mL, and perform the test using this solution dissolve in 20 mL of water by warming. After cooling, add as the test solution. Prepare the control solution with 2.0 mL water to make exactly 25 mL, and determine the optical rota- of Standard Lead Solution, 30 mL of ethanol (95), 2 mL of tion of the solution in a 100-mm cell. 1212 Tubocurarine Chloride Hydrochloride Hydrate / O‹cial Monographs JP XV pH <2.54> Dissolve 1.0 g in 100 mL of water by warming, and cool: the pH of this solution is between 5.4 and 6.4. Tubocurarine Chloride Purity (1) Clarity of solution—Dissolve 0.20 g of L-Tryp- tophan in 10 mL of 2 molWL hydrochloric acid TS: the solu- Hydrochloride Hydrate tion is clear. (2) Chloride <1.03>—Dissolve 0.5 g of L-Tryptophan in 6 Tubocurarine Hydrochloride mL of dilute nitric acid, and add water to make 50 mL. Per- ツボクラリン塩化物塩酸塩水和物 form the test using this solution as the test solution. Prepare the control solution with 0.30 mL of 0.01 molWL hydrochloric acid VS (not more than 0.021z). (3) Sulfate <1.14>—Dissolve 0.6 g of L-Tryptophan in 40 mL of water and 1 mL of dilute hydrochloric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.35 mL of 0.005 molWL sulfuric acid VS (not more than 0.028z). (4) Ammonium <1.02>—Perform the test with 0.25 g of L-Tryptophan. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). (5) Heavy metals <1.07>—Proceed with 1.0 g of L-Tryp- C H ClN O .HCl.5H O: 771.72 tophan according to Method 4, and perform the test. Prepare 37 41 2 6 2 7 ,12 -Dihydroxy-6,6 -dimethoxy-2,2 ,2 - the control solution with 2.0 mL of Standard Lead Solution ? ? ? ? ? trimethyltubocuraranium chloride monohydrochloride (not more than 20 ppm). pentahydrate [41354-45-4] (6) Arsenic <1.11>—Dissolve 1.0 g of L-Tryptophan in 3 mL of 1 molWL hydrochloric acid TS and 2 mL of water by heating, and perform the test with this solution as the test so- Tubocurarine Chloride Hydrochloride Hydrate con- lution (not more than 2 ppm). tains not less than 98.0z of tubocurarine chloride hydrochloride (C H ClN O .HCl: 681.65), calculated (7) Related substances—Dissolve 0.30 g of L-Tryptophan 37 41 2 6 in 1 mL of 1 molWL hydrochloric acid TS, add water to make on the dried basis. 50 mL, and use this solution as the sample solution. Pipet 1 Description Tubocurarine Chloride Hydrochloride Hydrate mL of the sample solution, and add water to make exactly 50 occurs as white crystals or crystalline powder. It is odorless. mL. Pipet 5 mL of this solution, add water to make exactly It is sparingly soluble in water and in ethanol (95), slightly 20 mL, and use this solution as the standard solution. Per- soluble in acetic acid (100), and practically insoluble in form the test with these solutions as directed under Thin-lay- diethyl ether and in chloroform. er Chromatography <2.03>.Spot5mLeachofthesampleso- The pH of a solution of Tubocurarine Chloride lution and standard solution on a plate of silica gel for thin- Hydrochloride Hydrate (1 in 100) is between 4.0 and 6.0. layer chromatography. Develop the plate with a mixture of 1- Melting point: about 2709C (with decomposition). butanol, water and acetic acid (100) (3:1:1) to a distance of Identification (1) To 20 mL of a solution of Tubocurarine about 10 cm, and dry the plate at 809C for 30 minutes. Spray Chloride Hydrochloride Hydrate (1 in 2000) add 0.2 mL of evenly a solution of ninhydrin in acetone (1 in 50) on the sulfuric acid and 2 mL of a solution of potassium iodate (1 in plate, and heat at 809C for 5 minutes: the spots other than 100), shake, and heat on a water bath for 30 minutes: a yel- the principal spot from the sample solution are not more in- low color is produced. tense than the spot from the standard solution. (2) To 1 mL of a solution of Tubocurarine Chloride Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, Hydrochloride Hydrate (1 in 100) add 1 mL of a solution of 3 hours). Reinecke salt (1 in 25): a red precipitate is formed. (3) Determine the absorption spectrum of a solution of Residue on ignition <2.44> Not more than 0.1z (1 g). Tubocurarine Chloride Hydrochloride Hydrate (3 in 100,000) Assay Weigh accurately about 0.2 g of L-Tryptophan, as directed under Ultraviolet-visible Spectrophotometry previously dried, dissolve in 3 mL of formic acid, add 50 mL <2.24>, and compare the spectrum with the Reference Spec- of acetic acid (100), and titrate <2.50> with 0.1 molWLper- trum or the spectrum of a solution of Tubocurarine Chloride chloric acid VS (potentiometric titration). Perform a blank Reference Standard prepared in the same manner as the sam- determination, and make any necessary correction. ple solution: both spectra exhibit similar intensities of ab- sorption at the same wavelengths. Each mL of 0.1 molWL perchloric acid VS (4) A solution of Tubocurarine Chloride Hydrochloride ＝ 20.42 mg of C H N O 11 12 2 2 Hydrate (1 in 50) responds to Qualitative Tests <1.09> (2) for Containers and storage Containers—Tight containers. chloride.

Storage—Light-resistant. 20 Optical rotation <2.49> [a]D : ＋210 – ＋2209(0.1 g, calcu- lated on the dried basis, water, 10 mL, after allowing to stand for 3 hours, 100 mm). Purity (1) Clarity and color of solution—Dissolve 0.10 g of Tubocurarine Chloride Hydrochloride Hydrate in 10 mL JP XV O‹cial Monographs / Tulobuterol Hydrochloride 1213 of ethanol (95): the solution is clear and colorless. rine Chloride Hydrochloride Hydrate. (2) Chloroform-soluble substances—Weigh accurately (3) To a volume of Tubocurarine Chloride Hydrochlo- about 0.2 g of Tubocurarine Chloride Hydrochloride Hy- ride Injection, equivalent to 3 mg of Tubocurarine Chloride drate, calculated on the dried basis, add 200 mL of water and Hydrochloride Hydrate according to the labeled amount, add 1 mL of a saturated solution of sodium hydrogen carbonate, water to make 100 mL, and determine the absorption spec- and extract with three 20-mL portions of chloroform. Com- trum of the solution as directed under Ultraviolet-visible bine the chloroform extracts, wash with 10 mL of water, Spectrophotometry <2.24>: it exhibits a maximum between filter the chloroform solution through absorbent cotton into 279 nm and 281 nm, and a minimum between 253 nm and a tared beaker, wash the absorbent cotton with two 5-mL 257 nm. portions of chloroform, and combine the filtrate and the Optical rotation <2.49> a20: ＋0.35 – ＋0.429(200 mm), cal- washings. Evaporate the chloroform on a water bath, and D culated with reference to the value of solution containing 1 dry the residue at 1059C for 1 hour: the mass of the residue is mg of Tubocurarine Chloride Hydrochloride Hydrate per not more than 2.0z of the mass of Tubocurarine Chloride mL, according to the labeled amount of Tubocurarine Chlo- Hydrochloride Hydrate taken. Add 10 mL of water to the ride Hydrochloride Injection. residue: the residue does not dissolve. Add 1 mL of hydrochloric acid, and stir: the residue dissolves. pH <2.54> 3.0–6.0 Loss on drying <2.41> 9–12z (0.5 g, in vacuum, phospho- Extractable volume <6.05> It meets the requirement. rus (V) oxide, 1059C, 4 hours). Assay Measure exactly a volume of Tubocurarine Chloride Residue on ignition <2.44> Not more than 0.2z (0.5 g). Hydrochloride Injection, equivalent to about 15 mg of tubocurarine chloride hydrochloride hydrate (C H ClN O Assay Weigh accurately about 0.5 g of Tubocurarine Chlo- 37 41 2 6 .HCl.5H O), add water to make exactly 500 mL, and use this ride Hydrochloride Hydrate, add 20 mL of acetic acid (100), 2 solution as the sample solution. Separately, weigh accurately and dissolve by warming on a water bath. After cooling, add about 15 mg of Tubocurarine Chloride Hydrochloride Refer- 60 mL of acetic anhydride, and titrate <2.50> with 0.1 molWL ence Standard (separately determine the loss on drying <2.41> perchloric acid VS (potentiometric titration). Perform a in the same manner as Tubocurarine Chloride Hydrochloride blank determination, and make any necessary correction. Hydrate), dissolve in water to make exactly 500 mL, and use Each mL of 0.1 molWL perchloric acid VS this solution as the standard solution. Determine the absor-

＝ 34.08 mg of C37H41ClN2O6.HCl bances, AT and AS, of the sample solution and the standard solution at 280 nm as directed under Ultraviolet-visible Spec- Containers and storage Containers—Tight containers. trophotometry <2.24>, respectively. Storage—Light-resistant. Amount (mg) of tubocurarine chloride hydrochloride

hydrate (C37H41ClN2O6.HCl.5H2O) Tubocurarine Chloride ＝ WS×(AT/AS)×1.1321

Hydrochloride Injection WS: Amount (mg) of Tubocurarine Chloride Hydrochlo- ride Reference Standard, calculated on the dried basis Tubocurarine Hydrochloride Injection Containers and storage Containers—Hermetic containers. Storage—Light-resistant, and under Nitrogen atmosphere. ツボクラリン塩化物塩酸塩注射液

Tubocurarine Chloride Hydrochloride Injection is Tulobuterol Hydrochloride an aqueous solution for injection. It contains not less than 93z and not more than 107z of the labeled ツロブテロール塩酸塩 amount of tubocurarine chloride hydrochloride hy- drate (C37H41ClN2O6.HCl.5H2O: 771.72). Method of preparation Prepare as directed under Injec- tions, with Tubocurarine Chloride Hydrochloride Hydrate. Description Tubocurarine Chloride Hydrochloride Injec- C12H18ClNO.HCl: 264.19 tion is a clear, colorless liquid. (1RS)-1-(2-Chlorophenyl)-2-(1,1- Identification (1) To a volume of Tubocurarine Chloride dimethylethyl)aminoethanol monohydrochloride Hydrochloride Injection, equivalent to 0.01 g of Tubocura- [56776-01-3] rine Chloride Hydrochloride Hydrate according to the la- beled amount, add water to make 20 mL, and proceed as Tulobuterol Hydrochloride, when dried, contains directed in the Identification (1) under Tubocurarine Chlo- not less than 98.5z of C12H18ClNO.HCl. ride Hydrochloride Hydrate. Description Tulobuterol Hydrochloride occurs as white (2) Proceed with a volume of Tubocurarine Chloride crystals or crystalline powder. Hydrochloride Injection, equivalent to 3 mg of Tubocurarine It is very soluble in methanol, freely soluble in water, in Chloride Hydrochloride Hydrate according to the labeled ethanol (95) and in acetic acid (100), sparingly soluble in acet- amount, as directed in the Identification (2) under Tubocura- ic anhydride, and very slightly soluble in diethyl ether. 1214 Turpentine Oil / O‹cial Monographs JP XV

A solution of Tulobuterol Hydrochloride (1 in 20) shows no optical rotation. Melting point: about 1639C Turpentine Oil Identification (1) Determine the absorption spectrum of a solution of Tulobuterol Hydrochloride (1 in 2500) as directed Oleum Terebinthinae under Ultraviolet-visible Spectrophotometry 2.24 ,and < > テレビン油 compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Turpentine Oil is the essential oil distilled with steam (2) Determine the infrared absorption spectrum of from the wood or balsam of Pinus species (Pinaceae). Tulobuterol Hydrochloride, previously dried, as directed in Description Turpentine Oil is a clear, colorless to pale yel- the potassium bromide disk method under Infrared Spec- low liquid. It has a characteristic odor and a pungent, bitter trophotometry <2.25>, and compare the spectrum with the taste. Reference Spectrum: both spctra exhibit similar intensities of Turpentine Oil (1 mL) is miscible with 5 mL of ethanol (95) absorption at the same wave numbers. and this solution is neutral. (3) A solution of Tulobuterol Hydrochloride (1 in 20) 20 responds to the Qualitative Tests <1.09> for chloride. Refractive index <2.45> nD : 1.465 – 1.478 20 Purity (1) Clarity and color of solution—Dissolve Speciˆc gravity <1.13> d20: 0.860 – 0.875 1.0 g of Tulobuterol Hydrochloride in 10 mL of water: the Purity (1) Foreign matter—Turpentine Oil has no oŠen- solution is clear and colorless. sive odor. Shake 5 mL of Turpentine Oil with 5 mL of a solu- (2) Heavy metals <1.07>—Proceed with 2.0 g of tion of potassium hydroxide (1 in 6): the aqueous layer does Tulobuterol Hydrochloride according to Method 1, and per- not show a yellow-brown to dark brown color. form the test. Prepare the control solution with 2.0 mL of (2) Hydrochloric acid-coloring substances—Shake 5 mL Standard Lead Solution (not more than 10 ppm). of Turpentine Oil with 5 mL of hydrochloric acid, and allow (3) Related substances—Dissolve 0.30 g of Tulobuterol to stand for 5 minutes: the hydrochloric acid layer is light yel- Hydrochloride in 5 mL of methanol, and use this solution as low and not brown in color. the sample solution. Pipet 1 mL of the sample solution, add (3) Mineral oil—Place 5 mL of Turpentine Oil in a Cassia methanol to make exactly 50 mL, and use this solution as the ‰ask, cool to a temperature not exceeding 159C, add drop- standard solution. Perform the test with these solutions as wise 25 mL of fuming sulfuric acid while shaking, warm be- directed under Thin-layer Chromatography <2.03>.Usea tween 609Cand659C for 10 minutes, and add sulfuric acid to plate previously developed with the upper-layer of a mixture raise the lower level of the oily layer to the graduated portion of ethyl acetate and ammonia solution (28) (200:9) to the top of the neck: not more than 0.1 mL of oil separates. of the plate and air-dried. Spot 5 mL each of the sample solu- tion and standard solution on a plate of silica gel with Distilling range <2.57> 150–1709C, not less than 90 volz. fluorescent indicator for thin-layer chromatography. De- Containers and storage Containers—Tight containers. velop the plate with the upper layer of a mixture of ethyl Storage—Light-resistant. acetate and ammonia solution (28) (200:9) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the Ubidecarenone principal spot and the spot of starting point from the sample solution are not more intense than the spot from the standard ユビデカレノン solution. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu- um, 609C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.5 g of Tulobuterol Hydrochloride, previously dried, dissolve in 80 mL of a mix- ture of acetic anhydride and acetic acid (100) (7:3), and titrate C59H90O4:863.34 <2.50> with 0.1 molWL perchloric acid VS (potentiometric (2 E,6E,10E,14E,18E,22E,26E,30E,34E,38E)-2- titration). Perform a blank determination, and make any (3,7,11,15,19,23,27,31,35,39-Decamethyltetraconta- necessary correction. 2,6,10,14,18,22,26,30,34,38-decaen-1-yl)-5,6-dimethoxy-3- methyl-1,4-benzoquinone [303-98-0] Each mL of 0.1 molWL perchloric acid VS ＝ 26.42 mg of C H ClNO.HCl 12 18 Ubidecarenone contains not less than 98.0z of Containers and storage Containers—Tight containers. C59H90O4, calculated on the anhydrous basis. Description Ubidecarenone occurs as a yellow to orange crystalline powder. It is odorless and has no taste. It is soluble in diethyl ether, very slightly soluble in ethanol (99.5), and practically insoluble in water. JP XV O‹cial Monographs / Ulinastatin 1215

It is gradually decomposed and colored by light. (wavelength: 275 nm). Melting point: about 489C Column: A stainless steel column about 5 mm in inside di- ameter and about 15 cm in length, packed with octadecyl- Identification (1) Dissolve 0.05 g of Ubidecarenone in 1 silanized silica gel for liquid chromatography (5 mminparti- mL of diethyl ether, and add 10 mL of ethanol (99.5). To 2 cle diameter). mL of this solution add 3 mL of ethanol (99.5) and 2 mL of Column temperature: A constant temperature of about dimethyl malonate, then add dropwise 1 mL of a solution of 359C. potassium hydroxide (1 in 5), and mix: a blue color appears. Mobile phase: A mixture of methanol and ethanol (99.5) (2) Determine the infrared absorption spectrum of (13:7). Ubidecarenone as directed in the potassium bromide disk Flow rate: Adjust the flow rate so that the retention time of method under Infrared Spectrophotometry <2.25>,andcom- ubidecarenone is about 10 minutes. pare the spectrum with the Reference Spectrum or the spec- Selection of column: Dissolve 0.01 g each of Ubidecare- trum of Ubidecarenone Reference Standard: both spectra ex- none and ubiquinone–9 in 20 mL of ethanol (99.5) by warm- hibit similar intensities of absorption at the same wave num- ing at about 509C for 2 minutes. After cooling, proceed with bers. 5 mL of this solution under the above operating conditions, Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of and calculate the resolution. Use a column giving elution of Ubidecarenone according to Method 4, and perform the test. ubiquinone-9 and ubidecarenone in this order with the reso- Prepare the control solution with 2.0 mL of Standard Lead lution between these peaks being not less than 4. Solution (not more than 20 ppm). System repeatability: Repeat the test five times with the (2) Related substances—Dissolve 0.05 g of Ubidecare- standard solution under the above operating conditions: the none in 50 mL of ethanol (99.5) by warming at about 509C relative standard deviation of the peak areas of ubidecare- for 2 minutes, and after cooling use this solution as the sam- none is not more than 0.8z. ple solution. To exactly 1 mL of the sample solution add Containers and storage Containers—Tight containers. ethanol (99.5) to make exactly 100 mL, and use this solution Storage—Light-resistant. as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol- lowing conditions. Determine each peak area of both solu- Ulinastatin tions by the automatic integration method: the total area of ウリナスタチン the peaks other than the peak of ubidecarenone from the sample solution is not larger than the peak area of ubidecare- none from the standard solution. Operating conditions— Ulinastatin is a solution of a glycoprotein having Detector, column, column temperature, mobile phase, trypsin inhibiting activity, which is separated and puri- flow rate, and selection of column: Proceed as directed in the fied from human . operating conditions in the Assay. It contains ulinastatin of not less than 45,000 Units Detection sensitivity: Adjust the detection sensitivity so per mL and not less than 2500 Units per mg protein. that the peak height of ubidecarenone obtained from 5 mLof Description Ulinastatin occurs as a light brown to brown, thestandardsolutionisbetween20mmand40mm. clear liquid. Time span of measurement: About 2 times of the retention Identification (1) Dilute a suitable amount of Ulinastatin time of ubidecarenone beginning after the solvent peak. with water to make a solution containing 4000 Units of Water <2.48> Not more than 0.20z (1 g, direct titration). ulinastatin per mL. To 1 mL of this solution add 1 mL of a solution of phenol (1 in 20), then carefully add 5 mL of sul- Residue on ignition <2.44> Not more than 0.1z (1 g). furic acid, and mix: an orange to red-orange color develops. Assay Weigh accurately about 50 mg each of Ubidecare- (2) Dilute a suitable quantity of Ulinastatin with water to none and Ubidecarenone Reference Standard (separately de- make a solution containing 2000 units per mL. Determine the termined the water content <2.48> inthesamemanneras absorption spectrum of the solution as directed under Ultrav- Ubidecarenone) dissolve each in 40 mL of ethanol (99.5) by iolet-visible Spectrophotometry <2.24>, and compare the warming at about 509C for 2 minutes, and after cooling add spectrum with the Reference Spectrum: both spectra exhibit ethanol (99.5) to make exactly 50 mL each, and use these so- similar intensities of absorption at the same wavelengths. lutions as the sample solution and the standard solution. Per- (3) Dilute a suitable amount of Ulinastatin with pH 7.8 form the test with exact 5 mL each of the sample solution and 2,2?,2!-nitrilotrisethanol buffer solution to make a solution standard solution as directed under Liquid Chromatography containing 500 Units of ulinastatin per mL, and use this solu- <2.01> according to the following conditions, and determine tion as the sample solution. Use the same buffer solution as peak areas, AT and AS, of ubidecarenone of these solution. the control solution. To 0.1 mL each of the sample solution and the control solution add 1.6 mL of the buffer solution Amount (mg) of C H O 59 90 4 and 0.2 mL of trypsin TS for test of ulinastatin, mix, and al- ＝ W ×(A /A ) S T S low them to stand in a water bath at 259C for 1 minute. Then WS: Amount (mg) of Ubidecarenone Reference Standard, add 1 mL of N-a-benzoyl-L-arginine-4-nitroanilide TS, mix, calculated on the anhydrous basis and allow them to stand at 259C for 2 minutes: the solution obtained with the sample solution develops no color while Operating conditions— that obtained with the control solution develops a yellow Detector: An ultraviolet absorption photometer 1216 Ulinastatin / O‹cial Monographs JP XV color. (i) Tris buffer solution for polyacrylamide gel elec- (4) To 1.5 g of Powdered Agar add 100 mL of pH 8.4 trophoresis A Dissolve 18.2 g of 2-amino-2-hydroxymethyl- boric acid-sodium hydroxide buffer solution, dissolve by 1,3-propanediol in 80 mL of water, adjust to pH 8.8 with 6 warming in a water bath, then pour immediately into a Petri molWLhydrochloricacidTS,andaddwatertomake100mL. dish placed horizontally so that the agar layer is about 2 mm (ii) Tris buffer solution for polyacrylamide gel elec- in thickness. After the agar becomes hard, bore two wells trophoresis B Dissolve 6.0 g of 2-amino-2-hydroxymethyl- about 2.5 mm in diameter with a separation of 6 mm from 1,3-propanediol in 80 mL of water, adjust to pH 8.8 with 6 each other. In one of the wells place 10 mL of a solution of molWLhydrochloricacidTS,andaddwatertomake100mL. Ulinastatin containing 500 Units per mL in pH 8.4 boric acid- (iii) Tris buffer solution for polyacrylamide gel elec- sodium hydroxide buffer solution, and in the other well place trophoresis C Dissolve 3.0 g of 2-amino-2-hydroxymethyl- 10 mL of anti-ulinastatin rabbit serum, cover the dish to 1,3-propanediol and 14.4 g of glycine in water to make 1000 avoid drying of the agar, and allow to stand for overnight at mL. a room temperature: a clear precipitin line appears between (iv) Acrylamide solution for polyacrylamide gel elec- the wells. trophoresis Dissolve 30 g of acrylamide and 0.8 g of N,N?- methylenebisacrylamide in water to make 100 mL. pH <2.54> 6.0 – 8.0 (v) Gel for separation Mix gently 15 mL of tris buffer Specific activity When calculated from the results obtained solution for polyacrylamide gel electrophoresis A, 20 mL of by the Assay and the following method, the specific activity is acrylamide solution for polyacrylamide gel electrophoresis, not less than 2500 Units per 1 mg protein. 24.5 mL of water, 0.022 mL of N,N,N?,N?- (i) Sample solution—To an exactly measured volume of tetramethylethylenediamine, 0.32 mL of 10z ammonium Ulinastatin, equivalent to about 10,000 Units according to peroxodisulfate TS and 0.3 mL of 1 molWL sodium sulfite TS, the labeled amount, add water to make exactly 20 mL. pour into a plate for slab gel preparation, then cover the gel (ii) Standard solutions—Weigh accurately about 10 mg of mixture with a layer of water, and allow to set for 1 hour. bovine serum albumin for test of ulinastatin, and dissolve in (vi) Gel for concentration Remove the water layer on water to make exactly 20 mL. To a suitable volume of this so- the gel for separation, and pour a mixture of 2.5 mL of tris lution add water to make four solutions containing exactly buffer solution for polyacrylamide gel electrophoresis B, 2.66 300, 200, 100 and 50 mg of the albumin per mL, respectively. mL of acrylamide solution for polyacrylamide gel elec- (iii) Procedure—Pipet 0.5 mL each of the sample solution trophoresis, 14.6 mL of water, 0.01 mL of N,N,N?,N?- and standard solutions, put them in glass test tubes about 18 tetramethylethylenediamine, 0.2 mL of 10z ammonium mm in internal diameter and about 130 mm in length, add ex- peroxodisulfate TS and 0.04 mL of 1 molWLsodiumsulfite actly 5 mL of alkaline copper TS, mix, and allow the tubes to TS on the gel. Then position a plastic sample well former so standinawaterbathat309C for 10 minutes. Then add exact- that the height of the gel for concentration is about 15 mm, ly 0.5 mL of diluted Folin’sTS(1in2),mix,andwarminthe and allow to set for 2 hours. water bath for 20 minutes. Determine the absorbances of (vii) Procedure these solutions at 750 nm as directed under Ultraviolet-visible Electrophoresis—Set the gel in an apparatus for slab gel Spectrophotometry <2.24> using a solution obtained in the electrophoresis, and fill the upper and lower reservoirs with same manner with 0.5 mL of water as the blank. tris buffer solution for polyacrylamide gel electrophoresis C. Plot the absorbances of the standard solutions on the verti- Introduce carefully 10 mL each of the sample solution and cal axis and their protein concentrations on the horizontal standard solution into the wells using a different well for each axis to prepare a calibration curve, and determine the protein solution, and allow electrophoresis to proceed using the elec- content of the sample solution from its absorbance by using trodeofthelowerreservoirastheanode.Switchoffthepow- this curve. Then calculate the amount of protein per mL of er supply when the bromophenol blue band has migrated to Ulinastatin. about 10 mm from the bottom of the gel. Staining—Dissolve 2.0 g of Coomassie brilliant blue R-250 Purity (1) Heavy metals <1.07>—Proceed with 10 mL of in 400 mL of a mixture of methanol and 100 mL of acetic Ulinastatin according to Method 2, and perform the test. acid (100), add water to make 1000 mL, and use this solution Prepare the control solution with 1.0 mL of Standard Lead as the staining solution. Stain the gel for 2 hours in the stain- Solution (not more than 1 ppm). ingsolutionwarmedto409C. (2) Related substances—To a suitable volume of Decolorization—To 100 mL of methanol and 75 mL of Ulinastatin add water to make a solution containing exactly acetic acid (100) add water to make 1000 mL, and use this so- 12,500 Units per mL, and use this solution as the sample lution as the rinsing solution. Immerse the gel removed from stock solution. To exactly 0.25 mL of the sample stock solu- the staining solution in the rinsing solution to decolorise. tion add exactly 0.2 mL of glycerin and exactly 0.05 mL of (3) Kallidinogenase—Dilute a suitable volume of 0.05z bromophenol blue TS, mix, and use this solution as Ulinastatin with water so that each mL of the solution con- the sample solution. Separately, to exactly 1 mL of the sam- tains about 50,000 Units, and use this solution as the sample ple stock solution add water to make exactly 100 mL. To ex- solution. Take exactly 0.4 mL of the sample solution into a actly 0.25 mL of this solution add exactly 0.2 mL of glycerin test tube, add exactly 0.5 mL of pH 8.2 tris buffer solution, and exactly 0.05 mL of 0.05z bromophenol blue TS, mix, mix, and allow the tube to stand in a water bath at 37 ± and use this solution as the standard solution. Perform the 0.29C for 5 minutes. Add exactly 0.1 mL of substrate TS for following test with the sample solution and the standard solu- kallidinogenase assay (4), mix, allow the tube to stand in the tion: the bands other than the principal band obtained from waterbathof37± 0.29C for exactly 30 minutes, then add the sample solution are not more intense than the band from exactly 0.1 mL of diluted acetic acid (100) (1 in 2), mix, and the standard solution in the electrophoretogram. use this solution as the test solution. Separately, take exactly JP XV O‹cial Monographs / Urapidil 1217

0.4 mL of the sample solution in a test tube, add exactly 0.5 guinea pig of the second group. Observe the signs of respira- mL of pH 8.2 tris buffer solution, mix, and allow the tube to tory distress, collapse or death of the animals for 30 minutes stand in the water bath of 37 ± 0.29C for 35 minutes. Then after each intravenous injection and 24 hours later: the add exactly 0.1 mL of diluted acetic acid (100) (1 in 2), mix, animals of the first group exhibit none of the signs mentioned add exactly 0.1 mL of substrate TS for kallidinogenase assay above, and all the animals of the second group exhibit sym- (4), mix, and use this solution as the control solution. Deter- ptoms of respiratory distress or collapse and not less than 3 mine the absorbances of the test solution and the control so- animals are killed. lution at 405 nm as directed under Ultraviolet-visible Spec- Toxicity Inject intravenously 0.50 mL of Ulinastatin into trophotometry <2.24> using water as the blank, and calculate each of five well-fed, healthy albino mice weighing 18 to 25 g: the difference between them: the difference is not more than no mouse dies within 48 hours after injection. If any mouse 0.050. dies within 48 hours, repeat the test using 5 albino mice Molecular mass Dilute a suitable volume of Ulinastatin weighing 19 to 21 g: all the animals survive for 48 hours. with the mobile phase so that each mL of the solution con- Assay Measure exactly a suitable volume of Ulinastatin, di- tains about 6500 Units, and use this solution as the sample so- lute with 2,2?,2!-nitrilotrisethanol buffer solution, pH 7.8 so lution. Separately, dissolve 1.0 mg each of g-globulin (mol. that each mL of the solution contains about 150 Units ac- mass: 160,000), bovine serum albumin for test of ulinastatin cording to the labeled amount, and use this solution as the (mol. mass: 67,000), and myoglobin (mol. mass: 17,000) in sample solution. Separately, dilute a suitable volume of about 1 mL of the mobile phase, and use this solution as the Ulinastatin Reference Standard with 2,2?,2!- molecular mass reference solution. Perform the test with 50 nitrilotrisethanol buffer solution, pH 7.8 so that each mL of mL each of the sample solution and molecular mass reference the solution contains exactly 300, 200, 100, 50 or 0 Units, and solution as directed under Liquid Chromatography <2.01> ac- use these solutions as the standard solutions. 2,2?,2!- cording to the following conditions. Prepare a calibration Nitrilotrisethanol buffer solution, pH 7.8 and N-a-benzoyl- curve by plotting the logarithm of molecular masses on the L-arginine-4-nitroanilide TS are warmed in a water bath of 25 vertical axis and the retention times (min) of the molecular ± 19C for use as described below. Take exactly 0.1 mL each mass reference substances on the horizontal axis, and deter- of the sample solution and the standard solutions in test mine the molecular mass of the sample using the calibration tubes, add exactly 1.6 mL of 2,2?,2!-nitrilotrisethanol buffer curve and the retention time obtained with the sample solu- solution, pH 7.8 mix, and put the tubes in the water bath of tion:themolecularmassis67,000± 5000. 25 ± 19C. One minute after addition of the buffer solution Operating conditions— add exactly 0.2 mL of ice-cooled trypsin TS for test of Detector: An ultraviolet absorption photometer ulinastatin, mix, and put the tubes again in the water bath. (wavelength: 280 nm). One minute later add exactly 1 mL of N-a-benzoyl-L-ar- Column: A stainless steel column about 7 mm in inside di- ginine-4-nitroanilide TS, mix, and then put the tubes in the ameter and about 60 cm in length, packed with porous silica water bath. Exactly 2 minutes later add exactly 0.1 mL of gel for liquid chromatography (10 – 12 mminparticledi- diluted acetic acid (100) (1 in 2) to stop the enzyme reaction, ameter). and determine the absorbances of the solutions so obtained at Column temperature: A constant temperature of about 405 nm as directed under Ultraviolet-visible Spectrophoto- 259C. metry <2.24> using water as the blank. Prepare a calibration Mobile phase: Dissolve 16.33 g of potassium dihydrogen- curve using the absorbances obtained with the standard solu- phosphate and 124.15 g of ethylene glycol in water to make tions, and calculate ulinastatin Units in the sample solution 1000 mL. If necessary, adjust to pH 4.0 with phosphoric from its absorbance by using this curve. acid. Flow rate: Adjust the flow rate so that the retention time of Containers and storage Containers—Tight containers. bovine serum albumin is about 36 minutes. Storage—Preserve at －209Corlower. Selection of column: Proceed with 50 mLofthemolecular mass reference solution according to the above operating conditions, and calculate the resolution. Use a column from Urapidil which g-globulin, bovine serum albumin and myoglobin are eluted in this order with the resolution between their peaks ウラピジル being not less than 1.5, respectively. Antigenicity Dilute a suitable volume of Ulinastatin with isotonic sodium chloride solution so that each mL of the so- lution contains 15,000 Units, and use this solution as the sam- ple solution. Inject 0.10 mL of the sample solution on 3 occa- sions at intervals of 2 days into the peritoneal cavity of each of 4 well-nourished, healthy guinea pigs weighing 250 to 300 g. Inject 0.10 mL of horse serum into the peritoneal cavity of each of 4 guinea pigs of another group as a control. Inject C20H29N5O3:387.48 0.20 mL of the sample solution intravenously into each of 2 6-｛3-[4-(2-Methoxyphenyl)piperazin-1-yl]propylamino｝- guinea pigs of the first group 14 days after the first in- 1,3-dimethyluracil [34661-75-1] traperitoneal injection and into each of the remaining 2 guinea pigs 21 days after the injection, and inject 0.20 mL of Urapidil, when dried, contains not less than 98.0z horse serum intravenously in the same manner into each and not more than 101.0z of C20H29N5O3. 1218 Urea / O‹cial Monographs JP XV

Description Urapidil occurs as white to pale yellowish, white, crystals or crystalline powder. It has a bitter taste. It is freely soluble in acetic acid (100), sparingly soluble in Urea ethanol (95) and in acetone, and very slightly soluble in 尿素 water. Identiˆcation (1) Determine the absorption spectrum of a solution of Urapidil in ethanol (95) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>,and compare the spectrum with the Reference Spectrum: both CH4N2O: 60.06 spectra exhibit similar intensities of absorption at the same Urea [57-13-6] wavelengths. (2) Determine the infrared absorption spectrum of Ura- Urea contains not less than 99.0z of CH4N2O. pidil as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spec- Description Urea occurs as colorless to white crystals or trum with the Reference Spectrum: both spectra exhibit simi- crystalline powder. It is odorless, and has a cooling, saline lar intensities of absorption at the same wave numbers. taste. It is very soluble in water, freely soluble in boiling ethanol Melting point <2.60> 156 – 1619C (95), soluble in ethanol (95), and very slightly soluble in Purity (1) Chloride <1.03>—Dissolve 3.0 g of Urapidil in diethyl ether. 40 mL of acetone and 6 mL of dilute nitric acid, add water to A solution of Urea (1 in 100) is neutral. make 50 mL, and perform the test using this solution as the Identification (1) Heat 0.5 g of Urea: it liquefies and the test solution. Prepare the control solution as follows. To 0.25 odor of ammonia is perceptible. Continue heating until the mL of 0.01 mol/L hydrochloric acid VS add 40 mL of ace- liquid becomes turbid, then cool. Dissolve the resulting lump tone, 6 mL of dilute nitric acid and water to make 50 mL (not in a mixture of 10 mL of water and 2 mL of sodium more than 0.003z). hydroxide TS, and add 1 drop of copper (II) sulfate TS: a (2) Heavy metals <1.07>—Proceed with 1.0 g of Urapidil reddish purple color develops. according to Method 4, and perform the test. Prepare the (2) Dissolve 0.1 g of Urea in 1 mL of water, and add 1 control solution with 2.0 mL of Standard Lead Solution (not mL of nitric acid: a white, crystalline precipitate is formed. more than 20 ppm). (3) Related substances—Dissolve 40 mg of Urapidil in 5 Melting point <2.60> 132.5 – 134.59C mL of ethanol (95), and use this solution as the sample solu- Purity (1) Chloride <1.03>—Perform the test with 2.0 g of tion. Pipet 1 mL of the sample solution, add ethanol (95) to Urea. Prepare the control solution with 0.40 mL of 0.01 molW make exactly 200 mL, and use this solution as the standard LhydrochloricacidVS(notmorethan0.007z). solution. Perform the test with these solutions as directed un- (2) Sulfate <1.14>—Perform the test with 2.0 g of Urea. der Thin-layer Chromatography <2.03>.Spot5mLeachof Prepare the control solution with 0.40 mL of 0.005 molWL the sample solution and standard solution on a plate of silica sulfuric acid VS (not more than 0.010z). gel with ‰uorescent indicator for thin-layer chromatography, (3) Heavy metals <1.07>—Proceed with 1.0 g of Urea ac- develop with a mixture of ethyl acetate, ethanol (95) and am- cording to Method 1, and perform the test. Prepare the con- monia water (28) (22:13:1) to a distance of about 15 cm, and trol solution with 2.0 mL of Standard Lead Solution (not air-dry the plate. Examine under ultraviolet light (main more than 20 ppm). wavelength: 254 nm): the spot other than the principal spot (4) Ethanol-insoluble substances—Dissolve 5.0 g of Urea appears not more than one and it is not more intense than the in 50 mL of warm ethanol (95), filter through a tared glass spot from the standard solution. filter (G4), wash the residue with 20 mL of warm ethanol Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 (95), and dry at 1059C for 1 hour: the mass of the residue is hours). not more than 2.0 mg. Residue on ignition <2.44> Not more than 0.2z (1 g). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 70 mg of Urapidil, previous- Assay Weigh accurately about 0.2 g of Urea, dissolve in ly dried, dissolve in 80 mL of acetic acid (100), and titrate water, and make exactly 200 mL. Measure exactly 5 mL of <2.50> with 0.1 mol/L perchloric acid VS (potentiometric this solution into a Kjeldahl flask, and proceed as directed titration). Perform a blank determination in the same man- under Nitrogen Determination <1.08>. ner, and make any necessary correction. Each mL of 0.005 molWL sulfuric acid VS Each mL of 0.1 mol/L perchloric acid VS ＝ 0.3003 mg of CH4N2O ＝12.92 mg of C20H29N5O3 Containers and storage Containers—Well-closed contain- Containers and storage Containers—Tight containers. ers. JP XV O‹cial Monographs / Urokinase 1219

of each of 2 or more of well-nourished, healthy guinea pigs weighing about 350 g, and inject 0.5 mL of the sample solu- Urokinase tion into the peritoneal cavity of each of 2 or more of well- nourished, healthy mice aged about 5 weeks. Observe the ウロキナーゼ conditions of the animals for more than 7 days: all the animals exhibit no abnormalities. High molecular mass urokinase Dilute Urokinase with [ ] 9010-53-1 gelatin-phosphate buŠer solution so that each mL of the solu- tion contains 10,000 Units, and use this solution as the sam- Urokinase is an enzyme, obtained from human u- ple solution. Perform the test with 100 mL of the sample solu- rine, that activates plasminogen, and has the molecular tion as directed under Liquid Chromatography <2.01> ac- mass of about 54,000. cording to the following conditions. Determine the peak It is a solution using a suitable buŠer solution as the areas of two peaks eluted closely at about 35 minutes having solvent. smaller retention time, A , and larger retention time, A ,by It contains not less than 60,000 Units per mL, and a b the automatic integration method: the value, AaW(Aa＋Ab), is not less than 120,000 Units per mg of protein. not less than 0.85. Description Urokinase is a clear and colorless liquid. Operating conditions— The pH is between 5.5 and 7.5. Apparatus: Use a pumping system for the mobile phase, a sample injection port, a column, a pumping system for the Identiˆcation (1) Dissolve 0.07 g of ˆbrinogen in 10 mL reaction reagent, a reaction coil, a reaction chamber, a spec- of phosphate buŠer solution, pH 7.4. To this solution add 1 tro‰uorometer and a recorder. Attach a 3-way tube to the mL of a solution of thrombin containing 10 Units per mL in outlet for the mobile phase of the column, connect the pump- isotonic sodium chloride solution, mix, place in a Petri dish ing system for the reaction reagent and the reaction coil, and about 90 mm in inside diameter, and keep horizontally until join outlet of the reaction coil to the spectro‰uorometer. the solution is coagulated. On the surface drop 10 Lofaso- m Detector: Spectro‰uorometer (excitation wavelength: 365 lution of Urokinase containing 100 Units per mL in gelatin- nm, ‰uorescence wavelength: 460 nm). tris buŠer solution, and stand for overnight: lysis circle is ap- Column: A stainless steel column 7.5 mm in inside di- peared. ameter and 60 cm in length, packed with porous silica gel for (2) Dissolve 1.0 g of Powdered Agar in 100 mL of boric liquid chromatography (10 to 12 mm in particle diameter). acid-sodium hydroxide buŠer solution, pH 8.4, by warming, Column temperature: A constant temperature of about and pour the solution into a Petri dish until the height come 209C. to about 2 mm. After cooling, make two wells of 2.5 mm in Reaction coil: A stainless steel column 0.25 mm in inside diameter with the space of 6 mm. To each well place separate- diameter and 150 cm in length. ly 10 mL of a solution of Urokinase containing 30,000 Units Reaction coil temperature: 379C per mL in isotonic sodium chloride solution and 10 Lofan- m Mobile phase: Gelatin-phosphate buŠer solution. ti-urokinase serum, and stand for overnight: a clear precipi- Flow rate of mobile phase: 0.5 mL per minute. tin line is appeared. Reaction reagent: 7-(Glutarylglycyl-L-arginylamino)-4- Purity (1) Heavy metals <1.07>—Proceed with 2.0 mL of methylcoumarin TS. Urokinase according to Method 2, and perform the test. Pre- Flow rate of reaction reagent: 0.75 mL per minute. pare the control solution with 2.0 mL of Standard Lead Solu- Selection of column: Adjust the pH of Urokinase to 7.5 tion (not more than 10 ppm). with sodium hydroxide TS, allow to stand at 379C for over 24 (2) Blood group substances—Dilute Urokinase with iso- hours, and add gelatin-phosphate buŠer solution to make the tonic sodium chloride solution so that each mL of the solu- solution containing 20,000 Units per mL. Proceed with 100 tion contains 12,000 Units, and use this solution as the sam- mL of this solution under the above operating conditions, and ple solution. To anti-A type antibody for blood typing add calculate the resolution. Use a column giving elution of high isotonic sodium chloride solution to dilute each 32, 64, 128, molecular mass urokinase (mol. wt.: 54,000) and low molecu- 256, 512 and 1024 times, place separately 25 mLeachofthese lar mass urokinase (mol. wt.: 33,000) in this order with the solutions in six wells on the ˆrst and second lane of a V- resolution between these peaks being not less than 1.0. shaped 96-wells microplate. Next, add 25 Lofthesample m Assay (1) Urokinase—Pipet 1 mL of Urokinase, dilute solution into the six wells on the ˆrst lane and 25 Lofiso- m exactly with gelatin-tris buŠer solution so that each mL of the tonic sodium chloride solution into the six wells of the second solution contains about 30 Units, and use this solution as the lane, mix, and allow to stand for 30 min. To each well add 50 sample solution. Add exactly 2 mL of gelatin-tris buŠer solu- L of A-type erythrocyte suspension, mix, allow to stand for m tion to contents of one ampoule of High Molecular Mass 2 hours, and compare the agglutination of erythrocyte in Urokinase Reference Standard to dissolve, pipet 1 mL of this both lanes: dilution factor of anti-A type antibody of the solution, dilute exactly with gelatin-tris buŠer solution so wells which show the agglutination is equal in both lanes. that each mL of the solution contains about 30 Units, and use Perform the same test by using anti-B type antibody for this solution as the standard solution. Place 1.0 mL of L- blood typing and B-type erythrocyte suspension. pyroglutamylglycyl-L-arginine-p-nitroaniline hydrochloride Abnormal toxicity Dilute Urokinase with isotonic sodium TS in two silicon-coated test tubes about 10 mm in inside di- chloride solution so that each mL of the solution contains ameter, warm them in a water bath at 35 ± 0.29Cfor5 12,000 Units, and use this solution as the sample solution. In- minutes, add separately 0.50 mL each of the sample solution ject 5.0 mL of the sample solution into the peritoneal cavity and the standard solution, warm in a water bath at 35 ± 1220 Ursodeoxycholic Acid / O‹cial Monographs JP XV

0.29C for exactly 30 minutes, then add 0.50 mL of diluted (2) Chloride <1.03>—Dissolve 2.0 g of Ursodeoxycholic acetic acid (100) (2 in 5). Determine the absorbances, AT and Acid in 20 mL of acetic acid (100) with shaking, add water to AS, of these solutions at 405 nm as directed under Ultrav- make 200 mL, shake thoroughly, and allow to stand for 10 iolet-visible Spectrophotometry <2.24>,usingwaterasthe minutes. Filter this solution, discard the first 10 mL of the blank. Separately place 1.0 mL of L-pyroglutamylglycyl-L-ar- filtrate, and use the subsequent filtrate as the sample solu- ginine-p-nitroaniline hydrochloride TS in two test tubes, add tion. To 40 mL of the sample solution add 6 mL of dilute 0.50 mL of diluted acetic acid (100) (2 in 5), and 0.50 mL nitric acid and water to make 50 mL. Perform the test using each of the sample solution and the standard solution. Deter- this solution as the test solution. Prepare the control solution mine the absorbances, ATO and ASO, of these solutions at 405 as follows: to 0.25 mL of 0.01 molWL hydrochloric acid VS nm as the same manner, using water as the blank. add 4 mL of acetic acid (100), 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.022z). Amount (Units) of Urokinase (3) Sulfate <1.14>—To 40 mL of the sample solution ob- ＝｛(A －A )/(A －A )｝×a×b T TO S SO tained in (2) add 1 mL of dilute hydrochloric acid and water a: Amount (Units) of urokinase in 1 mL of the standard to make 50 mL, and perform the test using this solution as solution the test solution. Prepare the control solution as follows: to b: Total volume (mL) of the sample solution 0.40 mL of 0.005 molWL sulfuric acid VS add 4 mL of acetic acid (100), 1 mL of dilute hydrochloric acid and water to (2) Protein—Measure exactly a volume of Urokinase, make 50 mL (not more than 0.048z). equivalent to about 15 mg of protein, and perform the test as (4) Heavy metals <1.07>—Proceed with 1.0 g of Ur- directed under Nitrogen Determination <1.08>. sodeoxycholic Acid according to Method 2, and perform the Each mL of 0.005 molWL sulfuric acid VS test. Prepare the control solution with 2.0 mL of Standard ＝0.8754 mg of protein Lead Solution (not more than 20 ppm). (5) Barium—To the solution obtained in (1) add 2 mL of Containers and storage Containers—Tight containers. hydrochloric acid, boil for 2 minutes, cool, filter, and wash Storage—Not exceeding －209C. with water until the last washing makes 100 mL. To 10 mL of the solution add 1 mL of dilute sulfuric acid: no turbidity is produced. Ursodeoxycholic Acid (6) Arsenic <1.11>—Prepare the test solution with 1.0 g of Ursodeoxycholic Acid according to Method 3, and per- Ursodesoxycholic Acid form the test (not more than 2 ppm). (7) Related substances—Dissolve 50 mg of Ursodeoxy- ウルソデオキシコール酸 cholic Acid in a mixture of chloform and ethanol (95) (9:1) to make exactly 25 mL, and use this solution as the sample solu- tion. Separately, dissolve 75 mg of chenodeoxycholic acid for thin-layer chromatography in a mixture of chloroform and ethanol (95) (9:1) to make exactly 100 mL. To exactly 2 mL of this solution add a mixture of chloroform and ethanol (95) (9:1) to make exactly 50 mL, and use this solution as the stan- dard solution (1). Dissolve 25 mg of lithocholic acid for thin- layer chromatography in a mixture of chloroform and C H O :392.57 24 40 4 ethanol (95) (9:1) to make exactly 50 mL. To exactly 1 mL of 3a,7b-Dihydroxy-5b-cholan-24-oic acid [128-13-2] this solution add a mixture of chloroform and ethanol (95) (9:1) to make exactly 50 mL. To exactly 2 mL of this solution Ursodeoxycholic Acid, when dried, contains not less add a mixture of chloroform and ethanol (95) (9:1) to make than 98.5 of C H O . z 24 40 4 exactly 10 mL, and use this solution as the standard solution Description Ursodeoxycholic Acid occurs as white crystals (2). Perform the test with these solutions as directed under or powder. It is odorless, and has a bitter taste. Thin-layer Chromatography <2.03>.Spot10mLeachofthe It is freely soluble in ethanol (95), in ethanol (99.5) and in sample solution and standard solutions (1) and (2) on a plate acetic acid (100), slightly soluble in chloroform, very slightly of silica gel for thin-layer chromatography. Develop the plate soluble in diethyl ether, and practically insoluble in water. with a mixture of chloroform, acetone and acetic acid (100) It dissolves in sodium hydroxide TS. (7:2:1) to a distance of about 10 cm, and air-dry the plate. Dry the plate at 1209C for 30 minutes, spray evenly a solution Identification Dissolve 0.01 g of Ursodeoxycholic Acid in 1 of n-hydrate in ethanol (95) (1 in 5) mL of sulfuric acid, add 1 drop of formaldehyde solution, immediately, and heat at 1209C for 2 to 3 minutes: the spot and allow to stand for 5 minutes. To the solution add 5 mL from the sample solution, corresponding to that from the of water: a blue-green suspended substance is produced. standard solution (1), is not more intense than the spot from 20 Optical rotation <2.49> [a]D : ＋59.0 – ＋62.09 (after the standard solution (1), and the spot other than the prin- drying, 1.0 g, ethanol (99.5), 25 mL, 100 mm). cipal spot and the above spots from the sample solution are not more intense than the spot from the standard solution Melting point <2.60> 200 – 2049C (2). Purity (1) Odor—To 2.0 g of Ursodeoxycholic Acid add Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 100 mL of water, and boil for 2 minutes: no odor is percepti- 2 hours). ble. JP XV O‹cial Monographs / Vancomycin Hydrochloride 1221

(6) Arsenic <1.11>—Proceed with 1.0 g of L-Valine, pre- Residue on ignition <2.44> Not more than 0.2z (1 g). pare the test solution according to Method 2, and perform Assay Weigh accurately about 0.5 g of Ursodeoxycholic the test (not more than 2 ppm). Acid, previously dried, and dissolve in 40 mL of neutralized (7) Related substances—Dissolve 0.10 g of L-Valine in 25 ethanol and 20 mL of water. Add 2 drops of phenolphthalein mL of water, and use this solution as the sample solution. TS, titrate with 0.1 molWL sodium hydroxide VS, and titrate Pipet 1 mL of the sample solution, and add water to make ex- <2.50> again after adding 100 mL of freshly boiled and cooled actly 50 mL. Pipet 5 mL of this solution, add water to make water near the end point. exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin- Each mL of 0.1 molWL sodium hydroxide VS layer Chromatography <2.03>.Spot5mLeachofthesample ＝ 39.26 mg of C H O 24 40 4 solution and standard solution on a plate of silica gel for Containers and storage Containers—Well-closed contain- thin-layer chromatography. Develop the plate with a mixture ers. of 1-butanol, water and acetic acid (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 L-Valine the plate, and heat at 809C for 5 minutes: the spots other than the principal spot from the sample solution are not more L-バリン intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).

Assay Weigh accurately about 0.12 g of L-Valine, previous- C H NO :117.15 5 11 2 ly dried, and dissolve in 3 mL of formic acid, add 50 mL of a- (2S)-2-Amino-3-methylbutanoic acid [72-18-4] cetic acid (100), and titrate <2.50> with 0.1 molWL perchloric acid VS (potentiometric titration). Perform a blank determi- L-Valine, when dried, contains not less than 98.5z nation, and make any necessary correction. of C5H11NO2. Each mL of 0.1 molWL perchloric acid VS Description L-Valine occurs as white crystals or crystalline ＝ 11.72 mg of C H NO powder. It is odorless or has a faint characteristic odor, and 5 11 2 has a slightly sweet taste, which becomes bitter. Containers and storage Containers—Tight containers. It is freely soluble in formic acid, soluble in water, and practically insoluble in ethanol (95). It dissolves in dilute hydrochloric acid. Identification Determine the infrared absorption spectrum Vancomycin Hydrochloride of L-Valine, previously dried, as directed in the potassium バンコマイシン塩酸塩 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 thesamewavenumbers.

20 Optical rotation <2.49> [a]D : ＋26.5 – ＋29.09 (after drying, 2 g, 6 molWL hydrochloric acid TS, 25 mL, 100 mm). pH <2.54> Dissolve 0.5 g of L-Valine in 20 mL of water: the pH of this solution is between 5.5 and 6.5. Purity (1) Clarity and color of solution—Dissolve 0.5 g of L-Valine in 20 mL of water: the solution is clear and color- less. (2) Chloride <1.03>—Perform the test with 0.5 g of L-Va- line. Prepare the control solution with 0.30 mL of 0.01 molWL hydrochloric acid VS (not more than 0.021z). (3) Sulfate <1.14>—Perform the test with 0.6 g of L-Va- line. Prepare the control solution with 0.35 mL of 0.005 molW L sulfuric acid VS (not more than 0.028z). (4) Ammonium <1.02>—Perform the test with 0.25 g of L-Valine. Prepare the control solution with 5.0 mL of Stan- dard Ammonium Solution (not more than 0.02 ). z C66H75Cl2N9O24.HCl: 1485.71 (5) Heavy metals <1.07>—Proceed with 1.0 g of L-Valine (1S,2R,18R,19R,22S,25R,28R,40S )-50-[3-Amino- according to Method 1, and perform the test. Prepare the 2,3,6-trideoxy-3-C-methyl-a-L-lyxo-hexopyranosyl- control solution with 2.0 mL of Standard Lead Solution (not (1ª2)-b-D-glucopyranosyloxy]-22-carbamoylmethyl- more than 20 ppm). 5,15-dichloro-2,18,32,35,37-pentahydroxy-19-[(2R)- 1222 Vancomycin Hydrochloride / O‹cial Monographs JP XV

4-methyl-2-(methylamino)pentanoylamino]- vancomycin from the sample solution is not more than the 20,23,26,42,44-pentaoxo-7,13-dioxa-21,24,27,41,43- peak area of vancomycin from the standard solution, and the pentaazaoctacyclo[26.14.2.23,6.214,17.18,12.129,33.010,25.034,39 total area of the peaks other than vancomycin is not more ]pentaconta- than 3 times of the peak area of vancomycin from the stan- 3,5,8,10,12(50),14,16,29,31,33(49),34,36,38,45,47- dard solution. pentadecaene-40-carboxylic acid monohydrochloride Operating conditions— [1404-93-9] Detector: An ultraviolet absorption photometer (wave- length: 280 nm). Vancomycin Hydrochloride is the hydrochloride of a Column: A stainless steel column 4.6 mm in inside di- glycopeptide substance having antibacterial activity ameter and 25 cm in length, packed with octadecylsilanized produced by the growth of Streptomyces orientalis. silica gel for liquid chromatography (5 mminparticledi- It contains not less than 1000 mg (potency) and not ameter). more than 1200 mg (potency) per mg, calculated on the Column temperature: A constant temperature of about anhydrous basis. The potency of Vancomycin 259C. Hydrochloride is expressed as mass (potency) of van- Mobile phase A: A mixture of triethylamine buŠer solu- comycin (C66H75Cl2N9O24: 1449.25). tion, pH 3.2, acetonitrile and tetrahydrofuran (92:7:1). Ad- just the amount of acetonitrile so that the retention time of Description Vancomycin Hydrochloride occurs as a white vancomycin is 7.5 to 10.5 minutes. powder. Mobile phase B: A mixture of triethylamine buŠer solu- It is freely soluble in water, soluble in formamide, slightly tion, pH 3.2, acetonitrile and tetrahydrofuran (70:29:1). soluble in methanol, very slightly soluble in ethanol (95), and Flowing of the mobile phase: Control the gradient by mix- practically insoluble in acetonitrile. ing the mobile phases A and B as directed in the following It is hygroscopic. table. Identiˆcation (1) Determine the absorption spectrum of a solution of Vancomycin Hydrochloride (1 in 10,000) as Time after injection Mobile phase Mobile phase directed under Ultraviolet-visible Spectrophotometry <2.24>, of sample (min) A(volz) B(volz) and compare the spectrum with the Reference Spectrum or 0–12 100 0 the spectrum of a solution of Vancomycin Hydrochloride 12–20 100→ 00→ 100 Reference Standard prepared in the same manner as the sam- 20–22 0 100 ple solution: both spectra exhibit similar intensities of ab- sorption at the same wavelengths. Flow rate: 1.5 mL per minute. (2) Determine the infrared absorption spectrum of Van- Time span of measurement: As long as about 2.5 times of comycin Hydrochloride as directed in the potassium bromide the retention time of vancomycin beginning after the solvent disk method under Infrared Spectrophotometry <2.25>,and peak. compare the spectrum with the Reference Spectrum or the System suitability— spectrum of Vancomycin Hydrochloride Reference Stan- Test for required detectability: Conˆrm that the peak area dard: both spectra exhibit similar intensities of absorption at of vancomycin obtained from 20 mL of the standard solution thesamewavenumbers. is equivalent to 3 to 5z of that from 20 mL of the sample so- (3) Dissolve 20 mg of Vancomycin Hydrochloride in 10 lution. mL of water, and add 1 drop of silver nitrate TS: a white tur- System performance: Dissolve 5 mg of Vancomycin bidity is produced. Hydrochloride in 10 mL of water, heat at 659C for 48 hours, and cool to the ordinal temperature. When the procedure is Optical rotation <2.49> [a]20: －30 – －409(0.2 g calculated D runwith20mL of this solution under the above operating on the anhydrous basis, water, 20 mL, 100 mm). conditions, related substance 1, vancomycin and related sub- pH <2.54> The pH of a solution obtained by dissolving 0.25 stance 2 are eluted in this order, the resolution between the g of Vancomycin Hydrochloride in 5 mL of water is between peaks of the related substance 1 and vancomycin is not less 2.5 and 4.5. than 3, the number of theoretical plates of the peak of van- comycin is not less than 1500, and the related substance 2 is Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of eluted between 15 and 18 minutes. Vancomycin Hydrochloride according to Method 2, and per- System repeatability: When the test is repeated 5 times with form the test. Prepare the control solution with 2.0 mL of 20 mL of the standard solution under the above operating Standard Lead Solution (not more than 20 ppm). conditions, the relative standard deviation of the peak area of (2) Related substances—Dissolve 0.10 g of Vancomycin vancomycin is not more than 2.0z. Hydrochloride in 10 mL of the mobile phase A, and use this solution as the sample solution. Pipet 1 mL of the sample so- Water <2.48> Not more than 5.0z (0.1 g, volumetric titra- lution, add the mobile phase A to make exactly 25 mL, and tion, direct titration. Use a mixture of formamide for water use this solution as the standard solution. Perform the test determination and methanol for water determination (3:1)). with exactly 20 mL each of the sample solution and standard Residue on ignition <2.44> Not more than 1.0z (1 g). solution as directed under the Liquid Chromatography <2.01> according to the following conditions. If necessary, proceed Assay Perform the test according to the Cylinder-plate with 20 mL of the mobile phase A in the same manner to com- method as directed under the Microbial Assay for Antibiotics pensate for the base line. Determine each peak area by the au- <4.02> according to the following conditions. tomatic integration method: the area of each peak other than (i) Test organism—Bacillus subtilis ATCC 6633 JP XV O‹cial Monographs / Vasopressin Injection 1223

(ii) Culture medium—Use the medium i in 1) Medium for clear and colorless to pale yellow, and the absorbance of the test organism [5] under (1) Agar media for seed and base lay- solution, determined at 465 nm as directed under Ultraviolet- er. Adjust the pH of the medium so that it will be 6.2 to 6.4 visible Spectrophotometry <2.24>, is not more than 0.05. after sterilization. (2) Related substances—Dissolve an amount of Van- (iii) Standard solutions—Weigh accurately an amount of comycin Hydrochloride for Injection, equivalent to 0.1 g Vancomycin Hydrochloride Reference Standard, equivalent (potency) of Vancomycin Hydrochloride according to the la- to about 25 mg (potency), dissolve in water to make exactly beled amount, in 10 mL of the mobile phase A, and use this 25 mL, and use this solution as the standard stock solution. solution as the sample solution. Keep the standard stock solution at 59C or below, and use wi- Proceed as directed in the Purity (2) under Vancomycin thin 7 days. Take exactly a suitable amount of the standard Hydrochloride. stock solution before use, add 0.1 mol/L phosphate buŠer Water <2.48> Not more than 5.0z (0.1 g, volumetric titra- solution, pH 4.5 to make solutions so that each mL contains tion, direct titration. Use a mixture of formamide for Karl 100 mg (potency) and 25 mg (potency), and use these solutions Fisher method and methanol for Karl Fisher method (3:1)). asthehighconcentrationstandardsolutionandlowconcen- tration standard solution, respectively. Bacterial endotoxins <4.01> Less than 0.25 EU/mg (poten- (iv) Sample solutions—Weigh accurately an amount of cy). Vancomycin Hydrochloride, equivalent to about 25 mg Uniformity of dosage unit <6.02> It meets the requirement (potency), and dissolve in water to make exactly 25 mL. Take of the Mass Variation Test. exactly a suitable amount of this solution, add 0.1 mol/L phosphate buŠer solution, pH 4.5 to make solutions so that Foreign insoluble matter <6.06> Perform the test according each mL contains 100 mg (potency) and 25 mg (potency), and to the Method 2: it meets the requirement. use these solutions as the high concentration sample solution Insoluble particulate matter <6.07> Perform the test accord- and low concentration sample solution, respectively. ing to the Method 1: it meets the requirement. Containers and storage Containers—Tight containers. Sterility <4.06> Perform the test according to the Membrane ˆltration method: it meets the requirement. Vancomycin Hydrochloride for Assay Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics Injection <4.02> according to the following conditions. (i) Test organism, culture medium, and standard 注射用バンコマイシン塩酸塩 solutions—Proceed as directed in the Assay under Vancomy- cin Hydrochloride. Vancomycin Hydrochloride for Injection is a prepa- (ii) Sample solutions—Weigh accurately the contents of ration for injection which is dissolved before use. not less than 10 Vancomycin Hydrochloride for Injection. It contains not less than 90.0z and not more than Weigh accurately an amount of the content, equivalent to 115.0z of the labeled amount of vancomycin about 25 mg (potency) of Vancomycin Hydrochloride ac- (C66H75Cl2N9O24: 1449.25). cordingtothelabeledamount,anddissolveinwatertomake exactly 25 mL. Take exactly a suitable amount of this solu- Method of preparation Prepare as directed under Injec- tion, add 0.1 mol/L phosphate buŠer solution, pH 4.5 to tions, with Vancomycin Hydrochloride. make solutions so that each mL contains 100 mg (potency) Description Vancomycin Hydrochloride for Injection oc- and 25 mg (potency), and use these solutions as the high con- curs as white masses or a white powder. centration sample solution and low concentration sample so- lution, respectively. Identiˆcation (1) Dissolve an amount of Vancomycin Hydrochloride for Injection, equivalent to 5 mg (potency) of Containers and storage Containers—Hermetic containers. Vancomycin Hydrochloride, in 50 mL of water, and deter- mine the absorption spectrum of this solution as directed un- der Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a Vasopressin Injection maximum between 279 and 283 nm. (2) Dissolve an amount of Vancomycin Hydrochloride バソプレシン注射液 for Injection, equivalent to 20 mg (potency) of Vancomycin Hydrochloride, in 10 mL of water, and add 1 drop of silver nitrate TS: a white turbidity is produced. Vasopressin Injection is an aqueous solution for in- jection. pH <2.54> The pH of a solution prepared by dissolving an It contains synthetic vasopressin or the pressor prin- amount of Vancomycin Hydrochloride for Injection, equiva- ciple, vasopressin, obtained from the posterior lobe of lent to 0.5 g (potency) of Vancomycin Hydrochloride accord- the pituitary of healthy cattles and pigs, from which ing to the labeled amount, in 10 mL of water is between 2.5 the majority of the oxytocic principle, , has and 4.5. been removed. Purity (1) Clarity and color of solution—Dissolve an It contains not less than 85z and not more than amount of Vancomycin Hydrochloride for Injection, equiva- 120z of the labeled vasopressin Units. lent to 0.5 g (potency) of Vancomycin Hydrochloride accord- Method of preparation Prepare as directed under Injec- ing to the labeled amount, in 10 mL of water: the solution is 1224 Vasopressin Injection / O‹cial Monographs JP XV tions, with vasopressin prepared by synthesis or obtained preparation. from the posterior lobe of the pituitary. (iii) Standard solution: Dilute the standard stock solution with isotonic sodium chloride solution so that 0.2 mL of the Description Vasopressin Injection is a clear and colorless obtained solution causes blood pressure increases of between liquid. It is odorless or has a slight, characteristic odor. 35 mmHg and 60 mmHg in test animals when injected ac- pH:3.0–4.0 cording to (vi), and designate this solution as the high-dose

Purity Oxytocic principle—When tested by the following standard solution (SH). Then dilute this solution with isotonic procedure, Vasopressin Injection contains not more than 0.6 sodium chloride solution 1.5 to 2.0 times by volume, and oxytocin Units for each determined 10 vasopressin Units. designate it as the low-dose standard solution (SL). (i) Standard stock solution: Dissolve 200 Units of Oxyto- (iv) Sample solution: Dilute an accurately measured cin Reference Standard, according to the labeled Units, in ex- volume of Vasopressin Injection with isotonic sodium chlo- actly 10 mL of diluted acetic acid (100) (1 in 400). Pipet 1 mL ride solution so that the obtained solution contains the same of this solution, and add diluted acetic acid (100) (1 in 400) to concentration in Units as the high-dose standard solution make exactly 10 mL. Store in a cold place, avoiding freezing. based on the labeled Units, and designate it as the high-dose

Use within 6 months from the date of preparation. sample solution (TH). Then dilute this solution with isotonic (ii) Standard solution: Dilute the standard stock solution sodium chloride solution 1.5 to 2.0 times by volume, and with isotonic sodium chloride solution so that each mL of the designate it as the low-dose sample solution (TL). Make the solution contains 0.020 oxytocin Units. concentration ratio of SH to SL equal to the ratio of TH to TL. (iii) Sample solution: Assume oxytocin Units as equiva- When the sensitivity of an animal is changed, adjust the con- lent to 6W100 of the determined vasopressin Units. Dilute centration of SH and TH before the next set of assay is started. Vasopressin Injection with isotonic sodium chloride solution However,keepthesameratioofSH to SL and TH to TL as in so that each mL of the resulting solution is expected to con- the primary set. tain 0.020 oxytocin Unit. (v) Dose of injection: Although 0.2 mL of each solution (iv) Apparatus: Use the apparatus for the uterus contrac- is usually injected, the dose of injection can be determined tion test, equipped with a thermostatic bath. Maintain a tem- based from preliminary tests or experiences. Inject the same perature of the bath at 379Cto389C with a variation of not volume throughout a set of tests. more than 0.19C during the course of the test. Use a 100-mL (vi) Procedure: Inject subcutaneously 0.7 mL of a solu- Magnus' chamber for suspending the uterus. tion of ethyl (1 in 4) per 100 g of body mass to (v) Test animal: Use healthy, virgin and metestrus guinea anesthetize the test animals and cannulate the trachea. Under pigs weighing between 175 g and 350 g. They have been bred artificial respiration (about 60 strokes per minute), remove a under conditions where they have been completely isolated part of the second cervical vertebra, cut off the spinal cord from the sight and smell of males since the time of weaning. and destroy the brain through the foramen magnum. Insert a (vi) Procedure: Immerse the Magnus' chamber in the cannula filled with isotonic sodium chloride solution into a bath maintained at a constant temperature, add Locke-Rin- femoral vein. Through this cannula, inject the solution pre- ger's solution to the chamber, and introduce oxygen into the pared by dissolving 200 Units of heparin sodium in solution at a moderate rate. Sacrifice a guinea pig by means 0.1 mL of isotonic sodium chloride solution, and then im- of a blow on the head, immediately remove the uterus from mediately inject 0.3 mL of isotonic sodium chloride solution. the body, suspend it in the chamber, and connect one horn of Insert a cannula into a carotid artery, and connect the cannu- the uterus to the lever with a thread. If necessary, weigh the la to a manometer for blood pressure measurement with a lever provided that the mass is not changed throughout the vinyl tube. The cannula and the vinyl tube have previously assay. Start the assay after 15 to 30 minutes when the uterus been filled with isotonic sodium chloride solution. Inject the is completely relaxed. Administer the same quantities, 0.1 to standard and the sample solutions at regular intervals of 10 to 0.5 mL, of the standard solution and the sample solution to 15 minutes into the femoral vein through the cannula fol- the Magnus' chamber alternately twice with regular intervals lowed by 0.3 mL of the isotonic solution when the blood of between 10 and 20 minutes to contract the uterus, finally pressure increases caused by each solution returns to the administer the standard solution in a quantity which is 25z original level. Measure the height of blood pressure increases larger than the preceding doses, and measure the height of within 1 mmHg on the kymogram. Maintain a constant tem- every contraction. The mean height of uterus contraction perature between 209Cand259C during the assay. In ad- caused by the standard solution is equal to or higher than that vance, make four pairs from SH,SL,TH,TL as follows. Ran- caused by the sample solution. The height of contraction domize the order of injection for pairs, but keep the order of caused by the increased dose of the standard solution is dis- injection within pairs as indicated. tinctly higher than those caused by the preceding doses of the Pair 1: SH,TL Pair 2: SL,TH Pair 3: TH,SL Pair 4: standard solution. TL,SH Carry out this assay using the same animals throughout a Extractable volume <6.05> It meets the requirement. set of four pairs of observations. Perform this assay with two Assay (i) Test animals: Use healthy male rats weighing be- sets. If necessary, however, use the different animals for both tween 200 g and 300 g. sets of tests. (ii) Standard stock solution: Dissolve 2000 Units of (vii) Calculation: Subtract increases of blood pressure Vasopressin Reference Standard, according to the labeled caused by the low dose from those caused by the high dose in

Units, in exactly 100 mL of diluted acetic acid (100) (1 in the Pair 1, 2, 3 and 4 of each set, and obtain the responses y1, 400). Pipet 1 mL of this solution, and add diluted acetic acid y2, y3 and y4, respectively. Sum up y1, for each set to obtain (100) (1 in 400) to make exactly 10 mL. Store in a cold place, Y1,andobtainY2, Y3 and Y4 inthesameway. avoiding freezing. Use within 6 months from the date of Units in each mL of Vasopressin Injection JP XV O‹cial Monographs / Verapamil Hydrochloride 1225

＝ antilog M × (Units in each mL of the SH) × (b/a) (2RS)-5-[(3,4-Dimethoxyphenethyl)methylamino]-2- (3,4-dimethoxyphenyl)-2-(1-methylethyl)pentanenitrile M ＝ (IY /Y ) a b monohydrochloride [152-11-4] I ＝ log (SH/SL) ＝ log (TH/TL) Y ＝－Y ＋ Y ＋ Y － Y a 1 2 3 4 Verapamil Hydrochloride, when dried, contains not Yb ＝ Y1 ＋ Y2 ＋ Y3 ＋ Y4 less than 98.5z of C27H38N2O4.HCl. a: Volume (mL) of Vasopressin Injection sampled. Description Verapamil Hydrochloride occurs as a white, b: Total volume (mL) of the high-dose sample solution pre- crystalline powder. It is odorless. pared by diluting with isotonic sodium chloride solution. It is freely soluble in methanol, in acetic acid (100) and in chloroform, soluble in ethanol (95) and in acetic anhydride, Compute L (P ＝ 0.95) by the following equation, and con- sparingly soluble in water, and practically insoluble in diethyl firm L to be 0.15 or less. If L exceeds 0.15, repeat the test, ether. improving the conditions of the assay or increasing the num- berofsetsuntilL reaches 0.15 or less. Identification (1) To 2 mL of a solution of Verapamil Hydrochloride (1 in 50) add 5 drops of Reinecke salt TS: a L ＝ 2 (C － 1)(CM2 ＋ I2) light red precipitate is produced. 2 2 2 2 C ＝[Yb /(Yb － 4fs t )] (2) Determine the absorption spectrum of a solution of Verapamil Hydrochloride in 0.01 mol/L hydrochloric acid f:Numberofsets TS (1 in 50,000) as directed under Ultraviolet-visible Spec- 2 2 2 s ＝[Sy － (Y/ f ) － (Y?/4)＋ (Yb /4f)]/n trophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities Sy2: The sum of the squares of y , y , y and y . 1 2 3 4 of absorption at the same wavelengths. Y ＝ Y 2 ＋ Y 2 ＋ Y 2 ＋ Y 2 1 2 3 4 (3) Determine the infrared absorption spectrum of Ver- Y?: The sum of the squares of the sum of y1, y2, y3 apamil Hydrochloride, previously dried, as directed in the and y in each set. 4 potassium chloride disk method under Infrared Spec- n ＝ 3(f － 1) trophotometry <2.25>, and compare the spectrum with the t2: Value shown in the following table against n for Reference Spectrum: both spectra exhibit similar intensities which s2 is calculated. of absorption at the same wave numbers. 2 2 2 (4) A solution of Verapamil Hydrochloride (1 in 50) nt＝F1 nt＝F1 nt＝F1 responds to the Qualitative Tests <1.09> for chloride. 1 161.45 13 4.667 25 4.242 2 18.51 14 4.600 26 4.225 Melting point <2.60> 141 – 1459C 3 10.129 15 4.543 27 4.210 pH <2.54> Dissolve 1.0 g of Verapamil Hydrochloride in 20 4 7.709 16 4.494 28 4.196 mL of freshly boiled and cooled water by warming, and cool: 5 6.608 17 4.451 29 4.183 the pH of this solution is between 4.5 and 6.5. 6 5.987 18 4.414 30 4.171 7 5.591 19 4.381 40 4.085 Purity (1) Clarity and color of solution—Dissolve 1.0 g of 8 5.318 20 4.351 60 4.001 Verapamil Hydrochloride in 20 mL of water by warming: the 9 5.117 21 4.325 120 3.920 solution is clear and colorless. 10 4.965 22 4.301 ∞ 3.841 (2) Heavy metals <1.07>—Proceed with 1.0 g of Ver- 11 4.844 23 4.279 apamil Hydrochloride according to Method 2, and perform 12 4.747 24 4.260 the test. Prepare the control solution with 2.0 mL of Stan- dard Lead Solution (not more than 20 ppm). (3) Arsenic <1.11>—Prepare the test solution with 1.0 g Containers and storage Containers—Hermetic containers. of Verapamil Hydrochloride according to Method 3, and per- Storage—In a cold place, and avoid freezing. form the test (not more than 2 ppm). (4) Related substances—Dissolve 0.50 g of Verapamil Expiration date 36 months after preparation. Hydrochloride in exactly 10 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample so- lution, add chloroform to make exactly 100 mL, and use this Verapamil Hydrochloride solution as the standard stock solution. Pipet 5 mL of the Iproveratril Hydrochloride standard stock solution, add chloroform to make exactly 100 mL, and use this solution as the standard solution (1). ベラパミル塩酸塩 Separately, pipet 5 mL of standard stock solution, add chlo- roform to make exactly 50 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>.Spot10 mL each of the sample solution and standard solutions (1) and (2) on two plates of silica gel for thin-layer chromatography. With the one plate, develop the plate with a mixture of cyclo- hexane and diethylamine (17:3) to a distance of about 15 cm, air-dry the plate, heat at 1109C for 1 hour, and cool. Exa- C27H38N2O4.HCl: 491.06 mine immediately after spraying evenly iron (III) chloride-io- 1226 Verapamil Hydrochloride Tablets / O‹cial Monographs JP XV dine TS on the plate: the three spots, having more intense take exactly V mL of the subsequent filtrate, add 0.02 mol/L color in the spots other than the principal spot and the origi- hydrochloric acid TS to make exactly V? so that each mL nal point from the sample solution, are not more intense than contains about 40 mg of verapamil hydrochloride (C27H38N2 the spot from the standard solution (2) in color. The remain- O4.HCl), and use this solution as the sample solution. Here- ing spots from the sample solution are not more intense than after, proceed as directed in the Assay. the spot from the standard solution (1) in color. With another Amount (mg) of verapamil hydrochloride (C H N O .HCl) plate, develop the plate with a mixture of toluene, methanol, 27 38 2 4 ＝W ×(A /A )×(V?/V)×(1/25) acetone and acetic acid (100) (14:4:1:1), and perform the test S T S inthesamemanner. WS: Amount (mg) of verapamil hydrochloride for assay Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, Assay To 10 tablets of Verapamil Hydrochloride Tablets 2 hours). add 140 mL of 0.02 mol/L hydrochloric acid TS, disintegrate the tablets by occasional shaking in a water bath at 609Cfor Residue on ignition <2.44> Not more than 0.1z (1 g). about 30 minutes, and then shake for 5 minutes. After cool- Assay Weigh accurately about 0.7 g of Verapamil ing, add 0.02 mol/L hydrochloric acid TS to make exactly Hydrochloride, previously dried, dissolve in 50 mL of a mix- 200 mL, and ˆlter. Discard the ˆrst 20 mL of the ˆltrate, take ture of acetic anhydride and acetic acid (100) (7:3), and titrate a volume of the subsequent filtrate, equivalent to about 4 mg

<2.50> with 0.1 molWL perchloric acid VS (potentiometric of verapamil hydrochloride (C27H38N2O4.HCl), add 0.02 titration). Perform a blank determination, and make any mol/L hydrochloric acid TS to make exactly 100 mL, and use necessary correction. this solution as the sample solution. Separately, weigh ac- curately about 0.1 g of verapamil hydrochloride for assay, Each mL of 0.1 molWL perchloric acid VS previously dried at 1059C for 2 hours, and dissolve in 70 mL ＝ 49.11 mg of C H N O .HCl 27 38 2 4 of 0.02 mol/L hydrochloric acid TS by occasional shaking in Containers and storage Containers—Well-closed contain- a water bath at 609C. After cooling, add 0.02 mol/L ers. hydrochloric acid TS to make exactly 100 mL. Pipet 4 mL of Storage—Light-resistant. this solution, add 0.02 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solu-

tion. Determine the absorbances, AT and AS,ofthesample Verapamil Hydrochloride Tablets solution and the standard solution at 278 nm as directed un- der Ultraviolet-visible Spectrophotometry <2.24>. ベラパミル塩酸塩錠 Amount (mg) of verapamil hydrochloride (C27H38N2O4.HCl) ＝WS×(AT/AS)×(V?/V)×(1/25) Verapamil Hydrochloride Tablets contain not less W : Amount (mg) of verapamil hydrochloride for assay than 95.0z and not more than 105.0z of the labeled S amount of verapamil hydrochloride (C27H38N2O4.HCl: Containers and storage Containers—Tight containers. 491.06). Method of preparation Prepare as directed under Tablets, with Verapamil Hydrochloride. Vinblastine Sulfate Identiˆcation (1) To a quantity of pulverized Verapamil ビンブラスチン硫酸塩 Hydrochloride Tablets, equivalent to 0.2 g of Verapamil Hydrochloride according to the labeled amount, add 70 mL of 0.02 mol/L hydrochloric acid TS, and shake occasionally in a water bath at 609C. After cooling, add 0.02 mol/L hydrochloric acid TS to make 100 mL, and ˆlter. To 3 mL of the ˆltrate add several drops of Reinecke's salt TS: a light red precipitate is formed. (2) To 2 mL of the ˆltrate obtained in (1) add 0.02 mol/L hydrochloric acid TS to make 100 mL, and determine the ab- sorption spectrum of this solution as directed under Ultrav- iolet-visible Spectrophotometry <2.24>: it exhibits maxima between 227 nm and 231 nm, and between 276 nm and 280 C46H58N4O9.H2SO4: 909.05 nm. Methyl (3aR,4R,5S,5aR,10bR,13aR )-4-acetoxy-3a-ethyl- 9-[(5S,7S,9S)-5-ethyl-5-hydroxy-9-methoxycarbonyl- Uniformity of dosage unit <6.02> Perform the test accord- 1,4,5,6,7,8,9,10-octahydro-3,7-methano-3- ing to the following method: it meets the requirement of the azacycloundecino[5,4-b]indol-9-yl]-5-hydroxy-8-methoxy- Content uniformity test. 6-methyl-3a,4,5,5a,6,11,12,13a-octahydro-1H- To 1 tablet of Verapamil Hydrochloride Tablets add 70 indolizino[8,1-cd ]carbazole-5-carboxylate monosulfate mL of 0.02 mol/L hydrochloric acid TS, disintegrate the [143-67-9] tablet by occasional shaking in a water bath at 609Cfor about 30 minutes, and then shake for 5 minutes. After cool- Vinblastine Sulfate contains not less than 96.0z and ing, add 0.02 mol/L hydrochloric acid TS to make exactly not more than 102.0z of C46H58N4O9.H2SO4,calculat- 100 mL, and filter. Discard the first 20 mL of the filtrate, ed on the dried basis. JP XV O‹cial Monographs / Vinblastine Sulfate for Injection 1227

Description Vinblastine Sulfate occurs as a white to pale Loss on drying Perform the test with about 10 mg of yellow powder. Vinblastine Sulfate as directed in Method 2 under the Ther- It is soluble in water, sparingly soluble in methanol, and mal Analysis <2.52> according to the following conditions: practically insoluble in ethanol (99.5). not more than 15.0z. It is hygroscopic. Operating conditions— 20 Optical rotation [a]D : －28 – －359(0.20 g calculated on Heating rate: 59C/minute the dried basis, methanol, 10 mL, 100 mm). Temperature range: room temperature to 2009C Atmospheric gas: dried Nitrogen Identiˆcation (1) Determine the absorption spectrum of a Flow rate of atmospheric gas: 40 mL/minute solution of Vinblastine Sulfate (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>,andcompare Assay Weigh accurately about 10 mg each of Vinblastine the spectrum with the Reference Spectrum or the spectrum of Sulfate and Vinblastine Sulfate Reference Standard (previ- a solution of Vinblastine Sulfate Reference Standard pre- ously determine the loss on drying in the same manner as Vin- pared in the same manner as the sample solution: both spec- blastine Sulfate), dissolve in water to make exactly 25 mL, tra exhibit similar intensities of absorption at the same and use these solutions as the sample solution and the stan- wavelengths. dard solution, respectively. Perform the test with exactly 20 (2) Determine the infrared absorption spectrum of mL each of the sample solution and standard solution as Vinblastine Sulfate as directed in the potassium bromide disk directed under Liquid Chromatography <2.01> according to method under Infrared Spectrophotometry <2.25>,andcom- the following conditions, and determine the peak areas, AT pare the spectrum with the Reference Spectrum or the spec- and AS, of vinblastine. trum of Vinblastine Sulfate Reference Standard: both spectra Amount (mg) of C H N O .H SO ＝W ×(A /A ) exhibit similar intensities of absorption at the same wave 46 58 4 9 2 4 S T S numbers. WS: Amount (mg) of Vinblastine Sulfate Reference (3) A solution of Vinblastine Sulfate (1 in 100) responds Standard, calculated on the dried basis to the Qualitative Tests <1.09> for sulfate. Operating conditions— pH <2.54> Dissolve 15 mg of Vinblastine Sulfate in 10 mL Detector: An ultraviolet absorption photometer of water: the pH of this solution is between 3.5 and 5.0. (wavelength: 262 nm). Column: A stainless steel column 4.6 mm in inside Purity (1) Clarity and color of solution—Dissolve 50 mg diameter and 15 cm in length, packed with octadecylsilanized of Vinblastine Sulfate in 10 mL of water: the solution is clear silica gel for liquid chromatography (5 mminparticle and colorless. diameter). (2) Related substances—Dissolve about 4 mg of Vin- Column temperature: A constant temperature of about blastine Sulfate in 10 mL of water, and use this solution as 259C. the sample solution. Pipet 1 mL of the sample solution, add Mobile phase: To 7 mL of diethylamine add water to make water to make exactly 25 mL, and use this solution as the 500 mL, and adjust the pH to 7.5 with phosphoric acid. To standard solution. Perform the test with exactly 200 mLeach 380 mL of this solution add 620 mL of a mixture of methanol of the sample solution and standard solution as directed un- and acetonitrile (4:1). der Liquid Chromatography <2.01> according to the follow- Flow rate: Adjust the ‰ow rate so that the retention time of ing conditions, and determine each peak area of these solu- vinblastine is about 8 minutes. tions by the automatic integration method: the area of peak System suitability— other than the main peak is not larger than 1/4 of the peak System performance: Dissolve 10 mg each of Vinblastine area of vinblastine from the standard solution, and the total Sulfate and vincristine sulfate in 25 mL of water. When the area of the peaks other than the main peak is not larger than procedure is run with 20 mL of this solution under the above 3/4 of the peak area of vinblastine from the standard operating conditions, vincristine and vinblastine are eluted in solution. this order with the resolution between these peaks being not Operating conditions— less than 4. Detector, column, column temperature, mobile phase, and System repeatability: When the test is repeated 5 times with ‰ow rate: Proceed as directed in the operating conditions in 20 mL of the standard solution under the above operating the Assay. conditions, the relative standard deviation of the peak area of Time span of measurement: About 4 times as long as the vinblastine is not more than 1.0z. retention time of vinblastine beginning after the solvent peak. System suitability— Containers and storage Containers—Tight containers. Test for required detectability: To exactly 2.5 mL of the Storage—Light-resistant, at not exceeding －209C. standard solution add water to make exactly 100 mL. Conˆrm that the peak area of vinblastine obtained from 200 mL of this solution is equivalent to 1.7 to 3.3z of that Vinblastine Sulfate for Injection from 200 mL of the standard solution. System performance: Proceed as directed in the system 注射用ビンブラスチン硫酸塩 suitability in the Assay. System repeatability: When the test is repeated 5 times with 200 mL of the standard solution under the above operating Vinblastine Sulfate for Injection is a preparation for conditions, the relative standard deviation of the peak area of injection, which is dissolved before use. vinblastine is not more than 1.5z. It contains not less than 90.0z and not more than 1228 Vincristine Sulfate / O‹cial Monographs JP XV

110.0z of the labeled amount of vinblastine sulfate amount of water, transfer into a 100-mL volumetric ‰ask, (C46H58N4O9.H2SO4: 909.05). wash each container with water, transfer the washings into the volumetric ‰ask, and add water to make exactly 100 mL. Method of preparation Prepare as directed under Injec- Pipet 10 mL of this solution, add water to make exactly 25 tions, with Vinblastine Sulfate. mL, and use this solution as the sample solution. Separately, Description Vinblastine Sulfate for Injection occurs as weigh accurately about 10 mg of Vinblastine Sulfate Refer- white to pale yellow, light masses or powder. ence Standard (previously determine the loss on drying in the It is freely soluble in water. same manner as Vinblastine Sulfate), dissolve in water to The pH of a solution (1 in 1000) is 3.5 – 5.0. make exactly 25 mL, and use this solution as the standard so- lution. Proceed as directed in the Assay under Vinblastine Identiˆcation Proceed as directed in the Identiˆcation (1) Sulfate. under Vinblastine Sulfate. Amount (mg) of vinblastine sulfate (C H N O .H SO ) Purity Related substances—Dissolve 4 mg of Vinblastine 46 58 4 9 2 4 ＝W ×(A /A )×10 Sulfate for Injection in 10 mL of water, and use this solution S T S as the sample solution. Pipet 1 mL of the sample solution, WS: Amount (mg) of Vinblastine Sulfate Reference add water to make exactly 25 mL, and use this solution as the Standard, calculated on the dried basis standard solution. Perform the test with exactly 200 mLeach Containers and storage Containers—Hermetic containers, of the sample solution and standard solution as directed and colored containers may be used. under Liquid Chromatography <2.01> according to the fol- Storage—Light-resistant, at 2 to 89C. lowing conditions, and determine each peak area by the auto- matic integration method: the area of the peak other than the main peak from the sample solution is not larger than 1/2 of the peak area of vinblastine from the standard solution, and Vincristine Sulfate the total area of the peaks other than the main peak is not ビンクリスチン硫酸塩 larger than 2 times the peak area of vinblastine from the stan- dard solution. Operating conditions— Perform as directed in the operating conditions in Purity (2) under Vinblastine Sulfate. System suitability— Perform as directed in the system suitability in Purity (2) under Vinblastine Sulfate. Bacterial endotoxins <4.01> Less than 10 EU/mg. Uniformity of dosage units <6.02> Perform the test accord- ing to the following method: it meets the requirement of the C46H56N4O10.H2SO4: 923.04 Content uniformity test. Methyl (3aR,4R,5S,5aR,10bR,13aR )-4-acetoxy-3a-ethyl- Dissolve 1 Vinblastine Sulfate for Injection in water to 9-[(5S,7S,9S)-5-ethyl-5-hydroxy-9-methoxycarbonyl- make exactly V mL so that each mL contains about 0.4 mg of 1,4,5,6,7,8,9,10-octahydro-3,7-methano- vinblastine sulfate (C46H58N4O9.H2SO4) according to the la- 3-azacycloundecino[5,4-b]indol-9-yl]-6-formyl-5-hydroxy- beled amount, and use this solution as the sample solution. 8-methoxy-3a,4,5,5a,6,11,12,13a-octahydro-1H- Separately, weigh accurately about 10 mg of Vinblastine Sul- indolizino[8,1-cd ]carbazole-5-carboxylate monosulfate fate Reference Standard (previously determine the loss on [2068-78-2] drying in the same manner as Vinblastine Sulfate), dissolve in water to make exactly 25 mL, and use this solution as the Vincristine Sulfate contains not less than 95.0z and standard solution. Proceed as directed in the Assay under not more than 105.0z of C46H56N4O10.H2SO4,calcu- Vinblastine Sulfate. lated on the dried basis. Amount (mg) of vinblastine sulfate (C46H58N4O9.H2SO4) Description Vincristine Sulfate occurs as a white to light ＝WS×(AT/AS)×(25/V) yellowish white powder.

WS: Amount (mg) of Vinblastine Sulfate Reference It is very soluble in water, and practically insoluble in Standard, calculated on the dried basis ethanol (95) and in diethyl ether. It is hygroscopic. Foreign insoluble matter <6.06> Perform the test according 20 Optical rotation [a]D : ＋28.5 – ＋35.59(0.2 g, calculated to Method 2: it meets the requirement. on the dried basis, water, 10 mL, 100 mm). Insoluble particulate matter <6.07> Perform the test accord- Identification (1) Dissolve 5 mg of Vincristine Sulfate in 2 ing to Method 1: it meets the requirement. mL of cerium (IV) tetraammonium sulfate-phosphoric acid Sterility <4.06> Perform the test according to the Membrane TS: a blue-purple color develops. ˆltration method: it meets the requirement. (2) Determine the absorption spectrum of a solution of Vincristine Sulfate (1 in 50,000) as directed under Ultraviolet- Assay Take an amount of Vinblastine Sulfate for Injection, visible Spectrophotometry <2.24>, and compare the spectrum equivalent to 0.10 g of vinblastine sulfate with the Reference Spectrum: both spectra exhibit similar in- (C46H58N4O9.H2SO4), dissolve each content with a suitable JP XV O‹cial Monographs / Vitamin A Oil 1229 tensities of absorption at the same wavelengths. tion of vincristine and vinblastine in this order with the reso- (3) Dissolve 0.02 g of Vincristine Sulfate in 10 mL of so- lution between these peaks being not less than 4. dium chloride TS, adjust the pH to between 9 and 10 with Detection sensitivity: Adjust the detection sensitivity so ammonia TS, and extract with two 5-mL portions of chlo- that the peak height of vincristine from 20 mL of the standard roform. Wash the combined chloroform extracts with a small solution is between 5 mm and 15 mm. quantity of sodium chloride TS, add a small quantity of an- Time span of measurement: About 3 times as long as the hydrous sodium sulfate, and allow to stand for several retention time of vincristine beginning after the solvent peak. minutes. Filter through a pledget of absorbent cotton, Loss on drying <2.41> Not more than 12.0z (50 mg, in evaporate the filtrate to dryness under reduced pressure, and vacuum, 1059C, 2 hours). dissolve the residue in a small quantity of chloroform. Deter- mine the infrared absorption spectrum of the solution as Assay Weigh accurately about 10 mg of Vincristine Sulfate, directed in the solution method under Infrared Spectrophoto- dissolve in acetic acid-sodium acetate buffer solution, pH metry <2.25>, and compare the spectrum with the Reference 5.0, to make exactly 50 mL. Pipet 5 mL of this solution, add Spectrum: both spectra exhibit similar intensities of absorp- acetic acid-sodium acetate buffer solution, pH 5.0, to make tionatthesamewavenumbers. exactly 50 mL. Determine the absorbance A of this solution (4) A solution of Vincristine Sulfate (1 in 100) responds at the maximum wavelength at about 296 nm as directed un- to the Qualitative Tests <1.09> for sulfate. der Ultraviolet-visible Spectrophotometry <2.24>. pH <2.54> Dissolve 10 mg of Vincristine Sulfate in 10 mL of Amount (mg) of C46H56N4O10.H2SO4 water: the pH of this solution is between 3.5 and 4.5. ＝ (A/177) × 5000 Purity (1) Clarity and color of solution—Dissolve 25 mg Containers and storage Containers—Hermetic containers. of Vincristine Sulfate in 10 mL of water: the solution is clear Storage—Light-resistant, and in a cold place. and colorless. (2) Related substances—Dissolve 25 mg of Vincristine Sulfate in 10 mL of water, and use this solution as the sample Vitamin A Oil solution. Pipet 1 mL of the sample solution, add water to make exactly 20 mL, and use this solution as the standard so- ビタミンA油 lution. Perform the test with exactly 20 mLeachofthesample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi- Vitamin A Oil is synthetic vitamin A esters diluted tions. Determine each peak area of these solutions by the au- with ˆxed oils. tomatic integration method: the total area of the peaks other It contains not less than 30,000 vitamin A Units per than the principal peak of the sample solution is not larger g. than the peak area of vincristine from the standard solution, It may contain suitable antioxidants. and the area of any peak other than the principal peak of the It contains not less than 90.0z and not more than sample solution is not larger than 2W5 of the peak area of vin- 120.0z of the labeled amount of vitamin A. cristine from the standard solution. Description Vitamin A Oil is a yellow to yellow-brown, Operating conditions— clear or slightly turbid oil. It is odorless or has a faint, char- Detector: An ultraviolet absorption photometer acteristic odor. (wavelength: 297 nm). It is decomposed upon exposure to air or light. Column: A stainless steel column about 4 mm in inside di- Identiˆcation Dissolve Vitamin A Oil, Retinol Acetate ameter and about 25 cm in length, packed with octylsilanized Reference Standard and Retinol Palmitate Reference Stan- silica gel for liquid chromatography (5 mminparticledi- dard, equivalent to 15,000 Units, in 5 mL of petroleum ether, ameter). and use these solutions as the sample solution, the standard Column temperature: Room temperature. solution (1) and the standard solution (2), respectively. Per- Mobile phase: Use methanol as the mobile phase A, and a form the test with these solutions as directed under Thin-lay- mixture of water and diethylamine (197:3) adjusted with er Chromatography <2.03>.Spot5 L each of the sample so- phosphoric acid to a pH of 7.5 as the mobile phase B. Run a m lution and standard solutions (1) and (2) on a plate of silica mixture of the mobile phase A and the mobile phase B (31:19) gel for thin-layer chromatography. Develop with a mixture of for24minutesafterinjectionofthesample,andrunamix- cyclohexane and diethyl ether (12:1) to a distance of about 10 ture of the mobile phase A and the mobile phase B for subse- cm, and air-dry the plate. Spray evenly antimony (III) chlo- quent 30 minutes, increasing the composition ratio of the mo- ride TS: the principal spot obtained from the sample solution bile phase A by 1z per minute. For subsequent 4 minutes, has the same color tone and the same f value with the blue run a mixture of the mobile phase A and the mobile phase B, R spot obtained from the standard solution (1) or the standard decreasing the composition ratio of the mobile phase A by solution (2). 7.5z per minute, then continue running a mixture of the mo- bile phase A and the mobile phase B (31:19). Purity (1) Acidity—Dissolve 1.2 g of Vitamin A Oil in 30 Flow rate: Adjust the flow rate so that the retention time of mL of a mixture of neutralized ethanol and diethyl ether vincristine is about 19 minutes. (1:1), boil gently for 10 minutes under a re‰ux condenser, Selection of column: Dissolve 10 mg each of Vincristine cool, and add 5 drops of phenolphthalein TS and 0.60 mL of Sulfate and vinblastine sulfate in 100 mL of water. Proceed 0.1 molWL sodium hydroxide VS: a red color develops. with 20 mL of this solution under the above operating condi- (2) Rancidity—No unpleasant odor of rancid oil is per- tions, and calculate the resolution. Use a column giving elu- ceptible by warming Vitamin A Oil. 1230 Vitamin A Oil Capsules / O‹cial Monographs JP XV

Powder with 100 mL of water, ˆlter, and to 5 mL of the Assay Proceed as directed in Method 1-1 under Vitamin A ˆltrate add 2.5 mL of sodium hydroxide TS and 0.5 mL of Assay <2.55>. potassium hexacyanoferrate (III) TS. Then add 5 mL of 2- Containers and storage Containers—Tight containers. methyl-1-propanol, shake the mixture vigorously for 2 Storage—Light-resistant, and almost well-ˆlled, or under minutes, allow to stand, and observe under ultraviolet light: Nitrogen atmosphere. the 2-methyl-1-propanol layer shows a blue-purple ‰uores- cence. This ‰uorescence disappears when the mixture is acidi- ˆed, but reappears when it is again made alkaline (thiamine). Vitamin A Oil Capsules (2) Shake 0.1 g of Compound Vitamin B Powder with 100 mL of water, and ˆlter. Perform the following tests with Vitamin A Capsules the ˆltrate (ribo‰avin). (i) The ˆltrate is light yellow-green in color and has an in- ビタミンA油カプセル tense yellow-green ‰uorescence. This color and ‰uorescence of the solution disappears upon the addition of 0.02 g of so- dium hydrosulˆte to 5 mL of the ˆltrate, and again appears Vitamin A Oil Capsules contain not less than 90.0z by shaking the mixture in air. This ‰uorescence disappears and not more than 130.0z of the labeled Units of vita- upon the addition of dilute hydrochloric acid or sodium min A. hydroxide TS. Method of preparation Prepare as directed under Capsules, (ii) To 10 mL of the ˆltrate placed in a glass-stoppered with Vitamin A Oil. test tube add 1 mL of sodium hydroxide TS, after illuminat- ing with a ‰uorescence lamp of 10 to 30 watts at 20-cm dis- Description The content of Vitamin A Oil Capsules con- tance for 30 minutes between 209Cand409C, acidify with forms to the requirements of Description under Vitamin A 0.5 mL of acetic acid (31), and shake thoroughly with 5 mL Oil. of chloroform: the chloroform layer shows yellow-green Identiˆcation Proceed the test with the content of Vitamin ‰uorescence. A Oil Capsules as directed in the Identiˆcation under Vitamin (3) Shake 1 g of Compound Vitamin B Powder with 100 AOil. mL of diluted ethanol (7 in 10), ˆlter, and to 5 mL of the ˆltrate add 2 mL of sodium hydroxide TS and 40 mg of man- Assay Weigh accurately 20 Vitamin A Oil Capsules, and ganese dioxide. Heat on a water bath for 30 minutes, cool, open the capsules to take out the content. Wash the capsules and ˆlter. Add 5 mL of 2-propanol to 1 mL of the ˆltrate, well with a small amount of diethyl ether, allow the capsules and use the solution as the sample solution. To 3 mL of the to stand at ordinal temperature to vaporize the diethyl ether, sample solution add 2 mL of bartibal buŠer solution, 4 mL and weigh accurately. Perform the test with the content as of 2-propanol and 2 mL of a freshly prepared solution of 2,6- directed under Vitamin A Assay <2.55>, and calculate the u- dibromo-N-chloro-1,4-benzoquinone monoimine in ethanol nits of vitamin A per capsule. Before applying Method 1-1, it (95) (1 in 4000) prepared when required for use: a blue color is necessary to know which the sample is, retinol acetate or develops. To 1 mL of the sample solution add 1 mL of a satu- retinol palmitate. rated boric acid solution, and proceed as directed in the same Containers and storage Containers—Well-closed contain- manner as above: no blue color develops (pyridoxine). ers. (4) Shake 0.5 g of Compound Vitamin B Powder with 10 Storage—Light-resistant. mL of ethanol (95), ˆlter, and evaporate 1 mL of the ˆltrate on a water bath to dryness. Add 0.01 g of 2,4- dinitrochlorobenzen to the residue, heat gently for 5–6 se- Compound Vitamin B Powder conds to fuse, and after cooling, add 4 mL of potassium hydroxide-ethanol TS: a red color develops (). 複方ビタミン B 散 (5) Shake 1 g of Compound Vitamin B Powder with 5 mL of diluted ethanol (7 in 10), ˆlter, and use the ˆltrate as the sample solution. Separately, dissolve 0.01 g each of thia- Method of preparation mine mononitrate, ribo‰avin, pyridoxine hydrochloride and Thiamine Nitrate 10 g nicotinamide in 1 mL, 50 mL, 1 mL and 1 mL of water, Ribo‰avin 10 g respectively, and use these solutions as standard solutions (1), Pyridoxine Hydrochloride 10 g (2), (3) and (4). Perform the test with these solutions as Nicotinamide 100 g directed under Thin-layer Chromatography <2.03>.Spot2mL Starch, Lactose Hydrate or each of the sample solution and standard solutions (1), (2), their mixture a su‹cient quantity (3) and (4) on a plate of silica gel with ‰uorescent indicator for thin-layer chromatography. Develop the plate with a mix- To make 1000 g ture of chloroform, ethanol (95) and acetic acid (100) Prepare as directed under Powders, with the above in- (100:50:1) to a distance of about 10 cm, and air-dry the plate. gredients. Examine under ultraviolet light (broad spectrum wavelength): four spots from the sample solution show the Description Compound Vitamin B Powder is orange-yellow same color tone and the same f value as the corresponding in color. It has a slighly bitter taste. R spots from standard solutions (1), (2), (3) and (4). It is slowly aŠected by light. Containers and storage Containers—Well-closed contain- Identiˆcation (1) Shake 2 g of Compound Vitamin B JP XV O‹cial Monographs / Voglibose 1231 ers. bile phase to make exactly 100 mL, and use this solution as Storage—Light-resistant. 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> according to the fol- Voglibose lowing operating conditions, and determine each peak area by the automatic integration method: the total area of the ボグリボース peaks other than voglibose is not larger than 1/5 times the peak area of voglibose from the standard solution. For the calculate of the total area, use the area of the peaks, having the relative retention time of about 1.7, about 2.0 and about 2.3 after multiplying by their relative response factors, 2, 2 and 2.5, respectively. Operating conditions— Apparatus: Use an apparatus consisting of 2 pumps for the mobile phase and reaction reagent transportation, sample in- C H NO : 267.28 10 21 7 jection port, column, reaction coil, cooling coil, detector and 3,4-Dideoxy-4-[2-hydroxy-1-(hydroxymethyl)ethylamino]- recording device, and the reaction coil and cooling coil main- 2-C-(hydroxymethyl)-D-epi-inositol tained at a constant temperature. [83480-29-9] Detector: Fluorophotometer (excitation wavelength: 350 nm, ‰uorescence wavelength: 430 nm) Voglibose contains not less than 99.5z and not Column: A stainless steel column 4.6 mm in inside di- more than 101.0 of C H NO ,calculatedonthean- z 10 21 7 ameter and 25 cm in length, packed with pentaethylenehex- hydrous basis. aaminated polyvinyl alcohol polymer bead for liquid chro- Description Voglibose occurs as white crystals or crystalline matography. powder. Column temperature: A constant temperature of about It is very slightly soluble in water, freely soluble in acetic 259C. acid (100), slightly soluble in methanol, and very slightly Reaction coil: A polytetrafluoroethylene tube 0.5 mm in soluble in ethanol (99.5). inside diameter and 20 m in length It dissolves in 0.1 mol/L hydrochloric acid TS. Cooling coil: A polytetrafluoroethylene tube 0.3 mm in in- side diameter and 2 m in length Identiˆcation (1) Determine the infrared absorption spec- Mobile phase: To 1.56 g of sodium dihydrogen phosphate trum of Voglibose as directed in the potassium bromide disk dihydrate add water to make 500 mL. To this solution add a method under Infrared Spectrophotometry <2.25>,andcom- solution, prepared by dissolving 3.58 g of disodium hydrogen pare the spectrum with the Reference Spectrum: both spectra phosphate dodecahydrate in water to make 500 mL, to adjust exhibit similar intensities of absorption at the same wave to pH 6.5. To 370 mL of this solution add 630 mL of acetoni- numbers. trile. (2) Determine the spectrum of a solution of Voglibose in Reaction reagent: Dissolve 6.25 g of taurine and 2.56 g of heavy water for nuclear magnetic resonance spectroscopy (3 sodium periodate in water to make 1000 mL. in 70) as directed under Nuclear Magnetic Resonance Spec- Reaction temperature: A constant temperature of about troscopy <2.21> (1H), using sodium 3-trimethylsilyl- 1009C propionate-d for nuclear magnetic resonance spectroscopy 4 Cooling temperature: A constant temperature of about as an internal reference compound: it exhibits 2 double sig- 159C nals A at about d 1.5 ppm, 2 double signals B at about d 2.1 Flow rate of the mobile phase: Adjust the ‰ow rate so that ppm, a multiple signal C at about d 2.9 ppm, and a multiple the retention time of voglibose is about 20 minutes. signal D between d 3.4 ppm and d 3.9 ppm. The area intensity Flow rate of the reaction reagent: Same as the ‰ow rate of ratio of each signal, A:B:C:D, is about 1:1:1:10. the mobile phase 20 Optical rotation <2.49> [a]D : ＋45 – ＋489(0.2 g calculated Time span of measurement: About 2.5 times as long as the on the anhydrous basis, 0.1 mol/L hydrochloric acid TS, 20 retention time of voglibose, beginning after the solvent peak mL, 100 mm). System suitability— Test for required detectability: Pipet 10 mL of the standard pH <2.54> Dissolve 1.0 g of Voglibose in 10 mL of water: solution, and add the mobile phase to make 100 mL. Conˆrm the pH of the solution is between 9.8 and 10.4. that the peak area of voglibose obtained from 50 mLofthis Melting point <2.60> 163 – 1689C solution is equivalent to 7 to 13z of that of voglibose ob- tained from 50 mL of the standard solution. Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of System performance: When the procedure is run with 50 Voglibose according to Method 1, and perform the test. Ad- mL of the standard solution under the above operating condi- just the pH of the test solution between 3.0 and 3.5 with di- tions, the number of theoretical plates and the symmetry fac- lute hydrochloric acid instead of dilute acetic acid. Prepare tor of the peak of voglibose are not less than 7000 and be- the control solution with 1.0 mL of Standard Lead Solution tween 0.8 and 1.2, respectively. (not more than 10 ppm). System repeatability: When the test is repeated 6 times with (2) Related substances—Dissolve 50 mg of Voglibose in 50 mL of the standard solution under the above operating 50 mL of the mobile phase, and use this solution as the sam- conditions, the relative standard deviation of the peak area of ple solution. Pipet 1 mL of the sample solution, add the mo- voglibose is not more than 3.0z. 1232 Voglibose Tablets / O‹cial Monographs JP XV

ˆrst 1 mL of the ˆltrate, and use the subsequent ˆltrate as the Water <2.48> Not more than 0.2z (0.5 g, coulometric titra- sample solution. Hereinafter, proceed as directed in the As- tion). say. Residue on ignition <2.44> Not more than 0.1z (1 g). Amount (g) of voglibose (C10H21NO7) Assay Weigh accurately about 0.4 g of Voglibose, dissolve ＝WS×(AT/AS)×(V/500) in 80 mL of acetic acid (100), and titrate <2.50> with 0.1 W : Amount (mg) of voglibose for assay, calculated on the mol/L perchloric acid VS (potentiometric titration). Perform S anhydrous basis a blank determination in the same manner, and make any necessary correction. Assay To 20 tables of Voglibose Tablets add 80 mL of the mobile phase, and completely disintegrate by shaking. To an Each mL of 0.1 mol/L perchloric acid VS exact volume of the solution, equivalent to about 4 mg of ＝26.73 mg of C10H21NO7 voglibose (C10H21NO7) according to the labeled amount, add Containers and storage Containers—Tight containers. the mobile phase to make exactly 100 mL, and centrifuge. Filter the supernatant liquid through a membrane ˆlter with pore size of not more than 0.45 mm. Discard the ˆrst 1 mL of Voglibose Tablets the ˆltrate, and use the subsequent ˆltrate as the sample solu- tion. Separately, weigh accurately about 20 mg of voglibose ボグリボース錠 for assay (previously determine the water <2.48> in the same manner as Voglibose), and dissolve in the mobile phase to make exactly 25 mL. Pipet 5 mL of this solution, add the mo- Voglibose Tablets contain not less than 95.0z and bile phase to make exactly 100 mL, and use this solution as not more than 105.0z of the labeled amount of vogli- the standard solution. Perform the test with exactly 50 mL bose (C10H21NO7:267.28). each of the sample solution and standard solution as directed Method of preparation Prepare as directed under Tablets, under Liquid Chromatography <2.01> according to the fol- with Voglibose. lowing conditions, and determine the peak areas, AT and AS, of voglibose. Identiˆcation Shake vigorously an amount of pulverized Voglibose Tablets, equivalent to 5 mg of Voglibose according Amount (mg) of voglibose (C10H21NO7) to the labeled amount, with 40 mL of water, and centrifuge. ＝WS×(AT/AS)×(1/500) Transfer the supernatant liquid to a chromatographic column W : Amount of voglibose for assay, calculated on the [prepared by pouring 1.0 mL of strongly acidic ion-exchange S dried basis resin (H type) for column chromatography (100 to 200 mmin particle diameter) into a chromatographic column 8 mm in Operating conditions— inside diameter and 130 mm in height], and allow to ‰ow at a Apparatus: Use an apparatus consisting of 2 pumps for the rate of about 5 mL per minute. Then wash the column with mobile phase and reaction reagent transportation, sample in- 200 mL of water, and allow to ‰ow with 10 mL of diluted jection port, column, reaction coil, cooling coil, detector and ammoniaTS(1in4)atarateofabout5mLperminute. recording device, and the reaction coil and cooling coil main- Filter the eŒuent solution two times through a membrane tained at a constant temperature. ˆlter with pore size of not more than 0.22 mm. Evaporate the Detector: Fluorophotometer (excitation wavelength: 350 ˆltrate to dryness at 509C under reduced pressure, dissolve nm, ‰uorescence wavelength: 430 nm) the residue with 0.5 mL of a mixture of water and methanol Column: A stainless steel column 4 mm in inside diameter (1:1), and use this solution as the sample solution. Separate- and 15 cm in length, packed with aminopropylsilanized silica ly, dissolve 20 mg of voglibose for assay in 2 mL of the mix- gel for liquid chromatography (5 mm in particle diameter). ture of water and methanol (1:1), and use this solution as the Column temperature: A constant temperature of about standard solution. Perform the test with these solutions as 259C directed under Thin-layer Chromatography <2.03>.Spot20 Reaction coil: A polytetrafluoroethylene tube 0.5 mm in mLeachofthesamplesolutionandstandardsolutionona inside diameter and 20 m in length plate of silica gel for thin-layer chromatography. Develop the Cooling coil: A polytetrafluoroethylene tube 0.3 mm in in- plate with a mixture of acetone, ammonia water (28) and side diameter and 2 m in length water (5:3:1) to a distance of about 12 cm, air-dry the plate, Mobile phase: To 1.56 g of sodium dihydrogen phosphate and allow to stand in iodine vapors: the principal spot from dihydrate add water to make 500 mL. To this solution add a the sample solution and the spot from the standard solution solution, prepared by dissolving 3.58 g of disodium hydrogen show a yellow-brown color, and the same Rfvalue. phosphate dodecahydrate in water to make 500 mL, to adjust to pH 6.5. To 300 mL of this solution add 600 mL of acetoni- Uniformity of dosage unit 6.02 Perform the test accord- < > trile. ing to the following method: it meets the requirement of the Reaction reagent: Dissolve 6.25 g of taurine and 2.56 g of Content uniformity test. sodium periodate in water to make 1000 mL. To 1 tablet of Voglibose Tablets add exactly mL of the V Reaction temperature: A constant temperature of about mobile phase so that the solution contains about 40 mgof 1009C voglibose (C H NO ) per mL according to the labeled 10 21 7 Cooling temperature: A constant temperature of about amount, disintegrate the tablet completely by shaking, and 159C centrifuge. Filter the supernatant liquid through a membrane Flow rate of mobile phase: Adjust the ‰ow rate so that the ˆlter with pore size of not more than 0.45 m. Discard the m retention time of voglibose is about 20 minutes. JP XV O‹cial Monographs / Warfarin Potassium 1233

Flow rate of reaction reagent: Same as the ‰ow rate of the 20) to make exactly 10 mL, and determine the absorbance at mobile phase 385 nm within 15 minutes as directed under Ultraviolet-visi- System suitability— ble Spectrophotometry <2.24>, using a solution of sodium System performance: Dissolve 2 mg of voglibose for assay hydroxide (1 in 20) as a blank: it does not exceed 0.20. and 0.2 g of lactose monohydrate in 5 mL of water, and add (2) Heavy metals <1.07>—Dissolve 2.0 g of Warfarin the mobile phase to make 50 mL. When the procedure is run Potassium in 30 mL of ethanol (95), add 2 mL of dilute acetic with 50 mL of this solution under the above operating condi- acid and ethanol (95) to make 50 mL. Perform the test using tions, lactose and voglibose are eluted in this order with the this solution as the test solution. Prepare the control solution resolution between these peaks being not less than 4. with 2.0 mL of Standard Lead Solution, 2 mL of dilute acetic System repeatability: When the test is repeated 6 times with acid and ethanol (95) to make 50 mL (not more than 10 50 mL of the standard solution under the above operating ppm). conditions, the relative standard deviation of the peak area of (3) Related substances—Dissolve 0.10 g of Warfarin voglibose is not more than 2.0z. Potassium in 100 mL of a mixture of water and methanol (3:1), and use this solution as the sample solution. Pipet 1 mL Containers and storage Containers—Tight containers. of the sample solution, add the mixture of water and methanol (3:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL Warfarin Potassium each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol- ワルファリンカリウム lowing conditions, and determine each peak area by the auto- matic integration method: each peak area other than warfa- rin is not larger than 1/10 times the peak area of warfarin ob- tained with the standard solution, and the total area of the peaks other than warfarin is not larger than 1/2 times the peak area of warfarin with the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and C H KO : 346.42 19 15 4 ‰ow rate: Proceed as directed in the operating conditions in Monopotassium (1RS)-2-oxo-3-(3-oxo- the Assay. 1-phenylbutyl)chromen-4-olate [ ] 2610-86-8 Timespanofmeasurement:About2timesaslongasthe retention time of warfarin beginning after the solvent peak. Warfarin Potassium, when dried, contains not less System suitability— than 98.0z and not more than 102.0z of C19H15KO4. Test for required detectability: To exactly 1 mL of the stan- Description Warfarin Potassium occurs as a white, crystal- dard solution add the mixture of water and methanol (3:1) to line powder. make exactly 20 mL. Conˆrm that the peak area of warfarin It is very soluble in water, and freely soluble in ethanol obtained with 20 mLofthissolutionisequivalentto3.5to (95). 6.5z of that with 20 mL of the standard solution. It dissolves in sodium hydroxide TS. System performance: Dissolve 20 mg of propyl para- The pH of a solution prepared by dissolving 1.0 g of hydroxybenzoate in 50 mL of methanol, and add water to Warfain Potassium in 100 mL of water is 7.2 – 8.3. make 200 mL. To 5 mL of this solution add 4 mL of a solu- It is colored to light yellow by light. tion of Warfarin Potassium in the mixture of water and A solution of Warfarin Potassium (1 in 10) shows no opti- methanol (3:1) (1 in 2000), and add the mixture of water and cal rotation. methanol (3:1) to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating condi- Identiˆcation (1) Determine the absorption spectrum of a tions, propyl parahydroxybenzoate and warfarin are eluted solution of Warfarin Potassium in 0.02 mol/L potassium in this order with the resolution between these peaks being hydroxide TS (1 in 100,000) as directed under Ultraviolet- not less than 7. visible Spectrophotometry <2.24>, and compare the spectrum System repeatability: When the test is repeated 6 times with with the Reference Spectrum or the spectrum of a slolution of 20 mL of the standard solution under the above operating Warfarin Potassium Reference Standard prepared in the conditions, the relative standard deviation of the peak area of same manner as the sample solution: both spectra exhibit warfarin is not more than 2.0z. similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Loss on drying <2.41> Not more than 4.5z (1 g, 1059C, 3 Warfarin Potassium, previously dried, as directed in the hours). potassium bromide disk method under Infrared Spec- Assay Weigh accurately about 25 mg each of Warfarin trophotometry <2.25>, and compare the spectrum with the Potassium and Warfarin Potassium Reference Standard, Reference Spectrum or the spectrum of Warfarin Potassium previously dried, and dissolve separately in the mixture of Reference Standard: both spectra exhibit similar intensities water and methanol (3:1) to make exactly 50 mL. Pipet 10 of absorption at the same wave numbers. mL each of these solutions, add the mixture of water and (3) A solution of Walfarin Potassium (1 in 250) responds methanol (3:1) to make exactly 50 mL, and use these solu- to the Qualitative Tests <1.09> (1) for potassium salt. tions as the sample solution and the standard solution. Per- Purity (1) Alkaline colored substances—Dissolve 1.0 g of form the test with exactly 20 mL each of the sample solution Warfarin Potassium in a solution of sodium hydroxide (1 in and standard solution as directed under Liquid Chro- 1234 Warfarin Potassium Tablets / O‹cial Monographs JP XV matography <2.01> according to the following conditions, Uniformity of dosage units <6.02> Perform the test accord- and determine the peak areas, A and A , of warfarin. T S ing to the following method: it meets the requirement of the

Amount (mg) of C19H15KO4＝WS×(AT/AS) Content uniformity test. Powder 1 tablet of Warfarin Potassium Tablets, add 40 W : Amount (mg) of Warfarin Potassium Reference Stan- S mL of water, and shake vigorouly for 30 minutes. Add water dard to make exactly V mL of this solution containing about 20 mg

Operating conditions— of warfarin potassium (C19H15KO4) per ml. Filter this solu- Detector: An ultraviolet absorption photometer tion, discard the first 5 mL of the filtrate, and use the subse- (wavelength: 260 nm). quent filtrate as the sample solution. Separately, weigh ac- Column: A stainless steel column 4.6 mm in inside di- curately about 40 mg of Warfarin Potassium Reference Stan- ameter and 25 cm in length, packed with cyanopropyl- dard, previously dried at 1059C for 3 hours, and dissolve in silanized silica gel for liquid chromatography (5 mminparti- water to make exactly 100 mL. Pipet 5 mL of this solution, cle diameter). add water to make exactly 100 mL, and use this solution as Column temperature: A constant temperature of about the standard solution. Pipet 20 mL each of the sample solu- 409C. tion and the standard solution, add 0.05 molWL hydrochloric Mobile phase: A mixture of water, acetonitrile and acetic acid TS to make exactly 25 mL, and use these solutions as the acid (100) (68:32:1). solution T1 and the solution S1, respectively. Separately, Flow rate: Adjust the ‰ow rate so that the retention time of pipet 20 mL each of the sample solution and the standard so- warfarin is about 10 minutes. lution, add 0.05 molWL potassium hydroxide TS to make ex-

System suitability— actly 25 mL, and use these solutions as the solution T2 and System performance: When the procedure is run with 20 the solution S2, respectively. Determine the absorbances, AT mL of the standard solution under the above operating condi- and AS, of the solution T1 and the solution S1 at 272 nm as tions, the number of theoretical plates and the symmetry fac- directed under Ultraviolet-visible Spectrophotometry <2.24>, tor of the peak of warfarin are not less than 8000 and not using the solution T2 and the solution S2 as the blank, respec- more than 1.5, respectively. tively. System repeatability: When the test is repeated 6 times with Amount (mg) of warfarin potassium (C H KO ) 20 mL of the standard solution under the above operating 19 15 4 ＝ W ×(A /A )×(V/2000) conditions, the relative standard deviation of the peak area of S T S warfarin is not more than 1.0z. WS: Amount (mg) of Warfarin Potassium Reference Standard Containers and storage Containers—Tight containers. Storage—Light-resistant. Assay Weigh accurately and powder not less than 20 Warfarin Potassium Tablets. Weigh accurately a portion of the powder, equivalent to about 4 mg of warfarin potassium Warfarin Potassium Tablets (C19H15KO4), add 80 mL of water, shake vigorously for 15 minutes, and add water to make exactly 100 mL. Filter this ワルファリンカリウム錠 solution, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 80 mg of Warfarin Potassium Reference Warfarin Potassium Tablets contain not less than Standard, previously dried at 1059C for 3 hours, and dissolve 95.0z and not more than 105.0z of the labeled in water to make exactly 100 mL. Pipet 5 mL of this solution, amount of warfarin potassium (C19H15KO4:346.42). add water to make exactly 100 mL, and use this solution as the standard solution. Pipet 10 mL each of the sample solu- Method of preparation Prepare as directed under Tablets, tion and the standard solution, add 0.02 molWL hydrochloric with Warfarin Potassium. acid TS to make exactly 20 mL, and use these solutions as the

Identification (1) Determine the absorption spectrum of solution T1 and the solution S1, respectively. Separately, the solution T2 obtained in the Assay, using 0.02 molWL pipet 10 mL each of the sample solution and the standard so- potassium hydroxide TS as the blank, as directed under lution, add 0.02 molWL potassium hydroxide TS to make ex- Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a actly 20 mL, and use these solutions as the solution T2 and maximum between 306 nm and 310 nm, and a minimum be- the solution S2, respectively. Determine the absorbances, AT tween 258 nm and 262 nm. Separately, determine the absorp- and AS, of the solution T1 and the solution S1 at 272 nm as tion spectrum of the solution T1 obtained in the Assay, using directed under Ultraviolet-visible Spectrophotometry <2.24>, 0.02 molWL hydrochloric acid TS as the blank, as directed un- using the solution T2 and the solution S2 as the blank, respec- der Ultraviolet-visible Spectrophotometry <2.24>:itexhibits tively. maxima between 281 nm and 285 nm and between 303 nm Amount (mg) of warfarin potassium (C H KO ) and 307 nm, and a minimum between 243 nm and 247 nm. 19 15 4 ＝ W ×(A /A )×(1/20) (2) Weigh a quantity of Warfarin Potassium Tablets, S T S equivalent to 0.01 g of Warfarin Potassium according to the WS: Amount (mg) of Warfarin Potassium Reference Stan- labeled amount, add 10 mL of acetone, shake, and filter. dard Heat the filtrate on a water bath to evaporate the acetone. To Containers and storage Containers—Tight containers. the residue add 10 mL of diethyl ether and 2 mL of dilute Storage—Light-resistant. hydrochloric acid, and shake: the aqueous layer responds to the Qualitative Tests <1.09> (1) for potassium salt. JP XV O‹cial Monographs / Water for Injection 1235

(4) Nitrogen from nitrate—Transfer 2.0 mL of Water for Injection to a 50-mL beaker, add 1 mL of sodium salicylate- Water sodium hydroxide TS, 1 mL of a solution of sodium chloride (1 in 500) and 1 mL of a solution of ammonium amidosulfate 常水 (1 in 1000), and evaporate on a water bath to dryness. Cool, dissolve in 2 mL of sulfuric acid, allow to stand for 10 minutes with occasional shaking, add 10 mL of water, and

H2O: 18.02 transfer to a Nessler tube. Cool, add 10 mL of a solution of sodium hydroxide (2 in 5) slowly, and add water to make 25 Water meets the quality standards of water supplies mL: no yellow color develops. under Article 4 of the Water Supply Law (the Ministry (5) Nitrogen from nitrite—Transfer 10 mL of Water for of Health, Labour and Welfare Ministerial Ordinance Injection to a Nessler tube, and add 1 mL of a solution of sul- No. 101, May 30, 2003), and also meets the following fanilamide in dilute hydrochloric acid (1 in 100) and 1 mL of requirement: N,N-diethyl-N′-1-naphthylethylenediamine oxalate TS: no pale red color develops. Purity Ammonium <1.02>—Perform the test with 30 mL of (6) Ammonium <1.02>—Perform the test with 30 mL of Water as directed under Ammonium Limit Test. Prepare the Water for Injection as the test solution. Prepare the control control solution as follows: to 0.15 mL of Standard Ammo- solution as follows: to 0.15 mL of Standard Ammonium So- nium Solution add puriˆed water for ammonium limit test to lution add puriˆed water for ammonium limit test to make 30 make 30 mL (not more than 0.05 mg/L). mL (not more than 0.05 mgWL). (7) Heavy metals—To 40 mL of Water for Injection add 2 mL of dilute acetic acid and 1 drop of sodium sulˆde TS: no Water for Injection change occurs. (8) Potassium permanganate-reducing substances—To 注射用水 100 mL of Water for Injection add 10 mL of dilute sulfuric acid, boil, add 0.10 mL of 0.02 mol/L potassium perman- ganate VS, and boil again for 10 minutes: the red color does Water for Injection is water either prepared by distil- not disappear. lation of Water or Puriˆed Water, or by the Reverse (9) Residue on evaporation—Evaporate 100 mL of Osmosis-Ultraˆltration (a reverse osmosis membrane, Water for Injection on a water bath to dryness, and dry the an ultraˆltration membrane or a combined puriˆcation residue at 1059C for 1 hour: the mass of the residue is not system using these membranes) of Puriˆed Water, and more than 1.0 mg. used for the preparation of injections, or for an alter- native usage as a packed Water for Injection, which is For Water for Injection prepared by the Reverse Osmosis- preserved in suitable containers and sterilized. Ultraˆltration for the preparation of injections, perform the When Water for Injection is prepared by the Reverse test for (8) Total organic carbon described below, instead of Osmosis-Ultraˆltration, take precaution against (8) Potassium permanganate-reducing substances. For Water microbial contamination of the purifying system to get for Injection preserved in containers and sterilized, perform comfortable quality being equivalent to that of water the test for (1) Acid or alkali, (2) Chloride, (6) Ammonium prepared by distillation. and (9) Residue on evaporation according to the following Water for Injection for the preparation of injections methods: must be used immediately after preparation. However, (1) Acidity or alkalinity—Shake gently 20 mL of Water it may be stored for a certain period of time, if the for Injection with 0.05 mL of phenol red TS and 0.13 mL of purifying system of water is established for avoiding 0.01 molWL sodium hydroxide VS, and allow to stand for 30 microbial contamination and its growth in preserved seconds: a red color develops. Separately, shake gently 20 mL period. of Water for Injection with 0.05 mL of bromothymol blue Water for Injection preserved in containers and TS and 0.13 mL of 0.01 molWL hydrochloric acid VS, and al- sterilized is used mainly as solvent for injections to be low to stand for 30 seconds: a yellow color develops. dissolved or suspended before use. (2) Chloride—For Water for Injection in containers Water for Injection prepared by distillation and holding a volume not more than 10 mL, add 2.0 mL of dilute packed in containers as sterilized products may be la- nitric acid to 15 mL of Water for Injection, and use this solu- beled ``Distilled Water for Injection'' as a commonly tion as the test solution. Separately, to 0.20 mL of 0.001 mol used name. WL hydrochloric acid VS add water to make 15 mL, then add 2.0 mL of dilute nitric acid, and use this solution as the con- Purity (1) Acidity or alkalinity—To 20 mL of Water for trol solution. Mix the test solution and the control solution Injection add 0.1 mL of methyl red TS for acid or alkali test: separately with 0.30 mL each of silver nitrate TS, allow to a yellow to orange color develops. Separately, to 20 mL of stand for 5 minutes under the protection from sunlight, and Water for Injection add 0.05 mL of bromothymol blue TS: compare the turbidity of the solutions on a black back- no blue color develops. ground: the turbidity of the test solution is not thicker than (2) Chloride—To 50 mL of Water for Injection add 3 that of the control solution (not more than 0.00005z). For dropsofnitricacidand0.5mLofsilvernitrateTS:no Water for Injection in containers holding a volume exceeding change occurs. 10 mL, add 3 drops of nitric acid and 0.5 mL of silver nitrate (3) Sulfate—To 50 mL of Water for Injection add 0.5 TS to 50 mL of Water for Injection: the solution remains un- mL of barium chloride TS: no change occurs. 1236 Puriˆed Water / O‹cial Monographs JP XV changed. dium hydroxide TS, 1 mL of a solution of sodium chloride (1 (6) Ammonium <1.02>—Perform the test with 30 mL of in 500) and 1 mL of a solution of ammonium amidosulfate (1 Water for Injection as the test solution. Prepare the control in 1000), and evaporate on a water bath to dryness. Cool, dis- solution as follows: To 0.6 mL of Standard Ammonium So- solve in 2 mL of sulfuric acid, allow to stand for 10 minutes lution for Water for Injection in containers holding a volume with occasional shaking, add 10 mL of water, and transfer to not more than 10 mL, and 0.3 mL of Standard Ammonium a Nessler tube. Cool, add 10 mL of a solution of sodium Solution for Water for Injection in a volume exceeding 10 hydroxide(2in5)slowly,andaddwatertomake25mL:no mL, add puriˆed water for ammonium limit test to make 30 yellow color develops. mL (not more than 0.2 mgWL for Water for Injection in a (5) Nitrogen from nitrite—Transfer 10 mL of Puriˆed volume not more than 10 mL, and not more than 0.1 mgWL Water to a Nessler tube, and add 1 mL of a solution of sul- for that exceeding 10 mL). fanilamide in dilute hydrochloric acid (1 in 100) and 1 mL of (8) Total organic carbon <2.59>—Apply the test to Water N-(1-naphthyl)-N?-diethylethylenediamine oxalate TS: no for Injection prepared by the Reverse Osmosis-Ultraˆltration pale red color develops. for the preparation of injections: it contains not more than (6) Ammonium <1.02>—Perform the test with 30 mL of 0.50 mg/L of total organic carbon. Puriˆed Water as the test solution. Prepare the control solu- (9) Residue on evaporation—Evaporate 100 mL of tion as follows: to 0.15 mL of Standard Ammonium Solution Water for Injection, and dry the residue at 1059Cfor1hour: add puriˆed water for ammonium limit test to make 30 mL the residue weighs not more than 4.0 mg for Water for Injec- (not more than 0.05 mg/L). tion in a volume not more than 10 mL, and not more than 3.0 (7) Heavy metals—The 40 mL of Puriˆed Water add 2 mg for that exceeding 10 mL. mL of dilute acetic acid and 1 drop of sodium sulˆde TS: no change occurs. Bacterial endotoxins <4.01> Less than 0.25 EUWmL. (8) Potassium permanganate-reducing substances—To Extractable volume <6.05> It meets the requirement. 100 mL of Puriˆed Water add 10 mL of dilute sulfuric acid, boil, add 0.10 mL of 0.02 molWL potassium permanganate Sterility <4.06> Apply the test to Water for Injection VS, and boil again for 10 minutes: the red color does not dis- preserved in containers as sterilized products: it meets the re- appear. quirement. (9) Residue on evaporation—Evaporate 100 mL of Puri- Containers and storage Containers—(1) For the prepara- ˆed Water on a water bath to dryness, and dry the residue at tion of injections, suitable containers, protected from 1059C for 1 hour: the amount of the residue is not more than microbial contamination. 1.0 mg. (2) Hermetic containers for Water for Injection Containers and storage Containers—Tight containers. preserved in the containers as sterilized products. Plastic con- tainers for aqueous infusions may be used. Sterile Puriˆed Water

Puriˆed Water 滅菌精製水 精製水 Sterile Puriˆed Water is sterilized Puriˆed Water. Puriˆed Water is puriˆed Water by hyperˆltration Description Sterile Puriˆed Water is a clear, colorless liq- (reverse osmosis, ultraˆltration), ion-exchange treat- uid. It is odorless and tasteless. ment, distillation or combination of these methods. When prepare Puriˆed Water, be careful to prevent Purity (1) Acidity or alkalinity—To 20 mL of Sterile microbial contamination. Puriˆed Water add 0.1 mL of methyl red TS for acid or alkali Use immediately after puriˆcation. It may be stored test: a yellow to orange color develops. To 20 mL of Sterile for a certain period, if it is in suitable containers Puriˆed Water add 0.05 mL of bromothymol blue TS: no preventing microbial growth. blue color develops. (2) Chloride—To50mLofSterilePuriˆedWateradd3 Description Puriˆed Water is a clear, colorless liquid. It is drops of nitric acid and 0.5 mL of silver nitrate TS: no odorless and tasteless. change occurs. Purity (1) Acidity or alkalinity—To 20 mL of Puriˆed (3) Sulfate—To 50 mL of Sterile Puriˆed Water add 0.5 Wateradd0.1mLofmethylredTSforacidoralkalitest:a mL of barium chloride TS: no change occurs. yellow to orange color develops. To 20 mL of Puriˆed Water (4) Nitrogen from nitrate—Transfer 2.0 mL of Sterile add 0.05 mL of bromothymol blue TS: no blue color de- Puriˆed Water to a 50-mL beaker, add 1 mL of sodium velops. salicylate-sodium hydroxide TS, 1 mL of a solution of sodi- (2) Chloride—To 50 mL of Puriˆed Water add 3 drops um chloride (1 in 500) and 1 mL of a solution of ammonium of nitric acid and 0.5 mL of silver nitrate TS: no change oc- amidosulfate (1 in 1000), and evaporate on a water bath to curs. dryness. Cool, dissolve in 2 mL of sulfuric acid, allow to (3) Sulfate—To 50 mL of Puriˆed Water add 0.5 mL of stand for 10 minutes, with occasional shaking, add 10 mL of barium chloride TS: no change occurs. water, and transfer to a Nessler tube. Cool, add 10 mL of a (4) Nitrogen from nitrate—Transfer 2.0 mL of Puriˆed solution of sodium hydroxide (2 in 5) slowly, and add water Water to a 50-mL beaker, add 1 mL of sodium salicylate-so- to make 25 mL: no yellow color develops. JP XV O‹cial Monographs / Wheat Starch 1237

(5) Nitrogen from nitrite—Transfer 10 mL of Sterile (Gramineae). Puriˆed Water to a Nessler tube, and add 1 mL of a solution Description WheatStarchoccursaswhitemassesor of sulfanilamide in dilute hydrochloric acid (1 in 100) and 1 powder. mL of N-(1-Naphthyl)-N′-diethylethylenediamine oxalate It is practically insoluble in water and in ethanol (99.5). TS: no pale red color develops.  (6) Ammonium <1.02>—Perform the test with 30 mL of Identiˆcation (1) Under a microscope <5.01>,Wheat Sterile Puriˆed Water as the test solution. Prepare the control Starch, preserved in a mixture of water and glycerin (1:1), ap- solution as follows: to 0.15 mL of Standard Ammonium So- pears as large and small sized simple grains, or quite rarely lution add puriˆed water for ammonium limit test to make 30 median sized simple grains; usually, large sized grains about mL (not more than 0.05 mg/L). 10–60mm in diameter, from upper view, disc like or quite (7) Heavy metals—To 40 mL of Sterile Puriˆed Water rarely reniform, centric hilum and striation indistinct or add 2 mL of dilute acetic acid and 1 drop of sodium sulˆde hardly distinct, often cleft on marginal portion visible; from TS: no change occurs. lateral view, narrowly ellipsoid or fusiform, hilum recognized (8) Potassium permanganate-reducing substances—To as a long and slender cleft along with long axis; small sized 100 mL of Sterile Puriˆed Water add 10 mL of dilute sulfuric grains 2 – 10 mm in diameter, spherical or polygonal; a black acid, boil, add 0.10 mL of 0.02 molWL potassium perman- cross, its intersection point on hilum, is observed when grains ganate VS, and boil again for 10 minutes: the red color does are put between two nicol prisms ˆxed at right angle to each not disappear. other. (9) Residue on evaporation—Evaporate 100 mL of (2) To 1 g of Wheat Starch add 50 mL of water, boil for 1 Sterile Puriˆed Water on a water bath to dryness, and dry the minute, and allow to cool: a subtle white-turbid, pasty liquid residue at 1059C for 1 hour: the mass of the residue is not is formed. more than 1.0 mg. (3) To 1 mL of the pasty liquid obtained in (2) add 0.05 mL of diluted iodine TS (1 in 10): a deep blue color is Sterility <4.06> Take 500 mL of Sterile Puriˆed Water, and formed, and the color disappears by heating. perform the test by the Membrane ˆltration method: it meets the requirements. pH <2.54> Put 5.0 g of Wheat Starch in a non-metal vessel, add 25.0 mL of freshly boiled and cooled water, mix gently Containers and storage Containers—Hermetic containers. for 1 minute, and allow to stand for 15 minutes: the pH of Prastic containers for aqueous injections may be used. the solution is between 4.5 and 7.0. Purity (1) Iron—To 1.5 g of Wheat Starch add 15 mL of 2 Weil's Disease and Akiyami mol/L hydrochloric acid TS, mix, ˆlter, and use the ˆltrate as the test solution. To 2.0 mL of Standard Iron Solution add Combined Vaccine water to make 20 mL, and use as the control solution. Put 10 mL each of the test solution and the control solution in test ワイル病秋やみ混合ワクチン tubes, add 2 mL of a solution of citric acid (2 in 10) and 0.1 mL of mercapto acetic acid, and mix. Alkalize with ammonia solution (28) to litmus paper, add water to make 20 mL, and Weil's Disease and Akiyami Combined Vaccine is a mix. Transfer 10 mL each of these solutions into test tubes, liquid for injection containing inactivated Weil's dis- allow to stand for 5 minutes, and compare the color of these ease leptospira, Akiyami A leptospira, Akiyami B lep- solutions against a white background: the color of the test so- tospira and Akiyami C leptospira. The product lacking lution is not more intense than that of the control solution more than a kind of Akiyami leptospira may be pre- (not more than 10 ppm). pared, if necessary. (2) Oxidizing substances—To 4.0 g of Wheat Starch add It conforms to the requirements of Weil's Disease 50.0 mL of water, shake for 5 minutes, and centrifuge. To and Akiyami Combined Vaccine in the Minimum Re- 30.0 mL of the supernatant liquid add 1 mL of acetic acid quirements for Biological Products. (100) and 0.5 to 1.0 g of potassium iodide, shake, and allow Description Weil's Disease and Akiyami Combined Vac- to stand for 25 to 30 minutes at a dark place. Add 1 mL of cine is a white-turbid liquid. 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 correc- Wheat Starch tion: the volume of 0.002 mol/L sodium thiosulfate VS con- sumed is not more than 1.4 mL (not more than 20 ppm, cal- Amylum Tritici culated as hydrogen peroxide). (3) Sulfur dioxide— コムギデンプン (i) Apparatus Use as shown in the ˆgure. (ii) Procedure Introduce 150 mL of water into the boil- This monograph is harmonized with the European ing ‰ask, close the tap of the funnel, and pass carbon dioxide Pharmacopoeia and the U.S. Pharmacopeia. The parts through the whole system at a rate of 100±5mLperminute. of the text that are not harmonized are marked with Pass cooling water through the condenser, and place 10 mL  of hydrogen peroxide-sodium hydroxide TS in the test-tube. symbols ( ) Wheat Starch consists of the starch granules After 15 minutes, remove the funnel without interrupting the obtained from the seeds of Triticum aestivum Linnáe stream of carbon dioxide, and introduce through the opening into the ‰ask about 25 g of Wheat Starch, accurately 1238 White Ointment / O‹cial Monographs JP XV

slight, characteristic odor. Containers and storage Containers—Tight containers.

Whole Human Blood

人全血液

Whole Human Blood is a liquid for injection which is prepared by mixing human blood cells and an an- ticoagulant solution for storage. It conforms to the requirements of Whole Human Blood in the Minimum Requirements for Biological Products. Description Whole Human Blood is a deep red liquid from which the erythrocytes settle upon standing, leaving a yellow supernatant layer. A gray layer which mainly consists of leu- cocytes may appear on the surface of the settled erythrocyte weighed, with the aid of 100 mL of water. Apply tap grease layer. The supernatant layer may become turbid in the to the outside of the connection part of the funnel, and load presence of fat, or may show the faint color of . the funnel. Close the tap of the funnel, pour 80 mL of 2 mol/LhydrochloricacidTSintothefunnel,openthetapto introduce the hydrochloric acid into the ‰ask, and close the tap while several mL of the hydrochloric acid remains, in Wine order to avoid losing sulfur dioxide. Place the ‰ask in a water ブドウ酒 bath, and heat the mixture for 1 hour. Transfer the contents of the test-tube with the aid of a little water to a wide-necked conical ‰ask. Heat in a water bath for 15 minutes, and cool. Wine is an alcoholic liquid obtained by fermenting Add 0.1 mL of bromophenol blue TS, and titrate <2.50> with the juice of the fruits of Linnáe ( ) 0.1 mol/L sodium hydroxide VS until the color changes from Vitis vinifera Vitaceae or allied plants. yellow to violet-blue lasting for at least 20 seconds. Perform a It contains not less than 11 volz and not more than blank determination and make any necessary correction. Cal- 14 volz of ethanol (C H O: 46.07) (by speciˆc gravi- culate the amount of sulfur dioxide by applying the following 2 6 ty), and not less than 0.10 w vz and not more than formula: it is not more than 50 ppm. W 0.40 wWvz of L-tartaric acid (C4H6O6: 150.09). Amount (ppm) of sulfur dioxide＝(V/W)×1000×3.203 It contains no artiˆcial sweetener and no artiˆcial coloring agent. W: Amount (g) of the sample V: Amount (mL) of 0.1 mol/L sodium hydroxide VS Description Wine is a light yellow or reddish purple to red- consumed purple liquid. It has a characteristic and aromatic odor. It has a slightly astringent and faintly irritating taste. Loss on drying <2.41> Not more than 15.0z (1 g, 1309C, 90 20 minutes). Speciˆc gravity <2.56> d20: 0.990 – 1.010 Residue on ignition <2.44> Not more than 0.6z (1 g). Optical rotation <2.49> Boil 160 mL of Wine, neutralize with potassium hydroxide TS, and concentrate to 80 mL on a Containers and storage Containers—Well-closed contain- water bath. Cool, dilute with water to 160 mL, add 16 mL of ers.  lead subacetate TS, shake well, and ˆlter. To 100 mL of the ˆltrate add 10 mL of a saturated solution of sodium sulfate decahydrate, shake well, ˆlter, and use the ˆltrate as the sam- White Ointment ple solution. Allow 20 mL of the sample solution to stand for 24 hours, add 0.5 g of activated charcoal, shake, stopper, 白色軟膏 and allow to stand for 10 minutes. Filter, and observe the op- tical rotation of the ˆltrate in a 200-mm cell. Multiply the op- Method of preparation tical rotation observed by 1.21, and designate as the optical rotation of Wine: it is between －0.39and ＋0.39. White Beeswax 50 g Sorbitan Sesquioleate 20 g Purity (1) Total acid [as L-tartaric acid (C4H6O6)]—To ex- White Petrolatum a su‹cient quantity actly 10 mL of Wine add 250 mL of freshly boiled and cooled water, and titrate <2.50> with 0.1 mol L sodium hydroxide To make 1000 g W VS (indicator: 1 mL of phenolphthalein TS). Prepare as directed under Ointments, with the above Each mL of 0.1 mol L sodium hydroxide VS materials. W ＝7.504 mg of C4H6O6 Description White Ointment is white in color. It has a JP XV O‹cial Monographs / Wine 1239

Total acid is not less than 0.40 wWvz and not more than 0.80 w Wvz.

(2) Volatile acid [as acetic acid (C2H4O2: 60.05)]—Trans- fer 100 mL of Wine to a beaker, add 1 mL of 1 molWLsodi- um hydroxide VS and the same volume of 1 molWLsodium hydroxide VS as that of 0.1 molWLsodiumhydroxideVS titrated in (1) to make the solution alkaline, and concentrate to 50 mL on a water bath. Cool, add water to make 100 mL, transfer to a 1000-mL distillation ‰ask, containing previously added 100 g of sodium chloride. Wash the beaker with 100 mL of water, and combine the washings in the distillation ‰ask. Add 5 mL of a solution of L-tartaric acid (3 in 20), and water lost by distillation. Cool, centrifuge, and decant the su- distil with steam cautiously to maintain the volume of the so- pernatant liquid in another beaker. To this solution add 1 to lution in the ‰ask until 450 mL of the distillate is obtained for 2 drops of dilute sulfuric acid, and allow to stand for 1 hour: 45 minutes. Dilute the distillate to exactly 500 mL with water, a white precipitate is formed. and use this solution as the sample solution. Titrate <2.50> a (5) Arsenic <1.11>—Evaporate 10 mL of Wine on a water 250-mL portion of the sample solution with 0.1 molWLsodi- bath to dryness. Prepare the test solution with the residue ac- um hydroxide VS (indicator: 5 drops of phenolphthalein TS). cording to Method 3, and perform the test (not more than 0.2 Perform a blank determination, and make any necessary cor- ppm). rection. (6) Glycerin—Pipet 100 mL of Wine into a 150-mL por- Each mL of 0.1 molWL sodium hydroxide VS celain dish, and concentrate on a water bath to 10 mL. Add 1

＝6.005 mg of C2H4O2 g of sea sand (No. 1), and make the solution strongly alkaline by adding a solution prepared by dissolving 4 g of calcium The volatile acid is not more than 0.15 wWvz. hydroxide in 6 mL of water. Heat on a water bath with con- (3) Sulfur dioxide—Stopper a 750-mL round-bottomed stant stirring and pushing down any material adhering to the ‰ask with a stopper having two holes. Through one hole, in- wall of the dish until the contents of the dish become soft sert a glass tube A extending nearly to the bottom of the masses. Cool, add 5 mL of ethanol (99.5), and grind to a ‰ask. Through the other hole, insert a glass tube B ending to grue-like substance. Heat on a water bath, add 10 to 20 mL the neck of the ‰ask. Connect the tube B to a Liebig's con- of ethanol (99.5) while agitating, boil, and transfer to a denser, and the end of the condenser to a joint of which inner 100-mL volumetric ‰ask. Wash the dish with seven 10-mL diameter is 5 mm at the lower end. Connect the other end of portions of hot ethanol (99.5), combine the washings with the the joint with a holed rubber stopper to a U tube having three contents of the ‰ask, cool, and add ethanol (99.5) to make bulbs as shown in the Figure. Pass carbon dioxide washed exactly 100 mL. Filter through a dry ˆlter paper, evaporate with a solution of potassium permanganate (3 in 100) 90 mL of the ˆltrate on a water bath, taking care not to boil through the tube A. Displace the air in the apparatus by car- the solution during the evaporation. Dissolve the residue in a bon dioxide, and place 50 mL of a freshly prepared and dilut- small amount of ethanol (99.5), transfer to a 50-mL glass- ed starch TS (1 in 5) and 1 g of potassium iodide in the U stoppered volumetric cylinder, wash with several portions of tube. From the other end of the U tube, add 1 to 2 drops of ethanol (99.5), and add the washings to the solution in the 0.01 molWL iodine VS from a burette. While passing carbon cylinder to make 15 mL. Add three 7.5-mL portions of de- dioxide, remove the stopper of the ‰ask a little, add exactly hydrated diethyl ether, shake vigorously each time, and allow 25 mL of Wine, 180 mL of freshly boiled and cooled water, to stand. When the solution becomes quite clear, transfer to a 0.2 g of tannic acid, and 30 mL of phosphoric acid, and stop- tared, ‰at weighing bottle. Wash the volumetric cylinder with per again. Pass carbon dioxide for further 15 minutes, heat 5 mL of a mixture of dehydrated diethyl ether and ethanol the distillation ‰ask with caution so that 40 to 50 drops of the (99.5) (3:2). Transfer the washings to the weighing bottle, distillate may be obtained in 1 minute. When the color of and evaporate carefully on a water bath. When the liquid starch TS in the U tube is discharged, add 0.01 molWLiodine becomes sticky, dry at 1059C for 1 hour, and cool in a desic- VS dropwise from a burette so that the color of the starch TS cator (silica gel), and weigh: the mass of the residue is not less remains light blue to blue during the distillation. Read the than 0.45 g and not more than 0.90 g. volume of 0.01 molWL iodine VS consumed when exactly 60 (7) Reducing sugars—To a 25-mL portion of the sample minutes have passed after the beginning of distillation. In this solution obtained in the Optical rotation add 50 mL of boil- case, however, the coloration of starch TS produced by 1 ing Fehling's TS, and heat for exactly 2 minutes. Filter the drop of 0.01 molWL iodine VS should persist at least for 1 separated precipitates by a tared glass ˆlter by suction, wash minute. successively with hot water, with ethanol (95) and with

Each mL of 0.01 molWLiodineVS＝0.6406 mg of SO2 diethyl ether, and continue to dry the precipitates by suction. Heat the ˆlter gently at ˆrst, and then strongly until the The amount of sulfur dioxide (SO : 64.06) does not exceed 2 precipitates become completely black. Cool the precipitates 7.5 mg. in a desiccator (silica gel), and weigh as copper (II) oxide: the (4) Total sulfuric acid—Transfer 10 mL of Wine to a mass of cupric oxide does not exceed 0.325 g. beaker, boil, and add 50 mL of a solution prepared by dis- (8) Sucrose—Transfer a 50-mL portion of the sample so- solving 5.608 g of barium chloride dihydrate in 50 mL of lution obtained in the Optical rotation to a 100-mL ‰ask, hydrochloric acid and water to make 1000 mL. Cover the neutralize with diluted hydrochloric acid (1 in 30), followed beaker, and heat on a water bath for 2 hours, supplying the by further addition of 5 mL of diluted hydrochloric acid (1 in 1240 Xylitol / O‹cial Monographs JP XV

30).Heatinawaterbathfor30minutes,cool,neutralizewith ‰ask as a receiver. When about 80 mL of the distillate is ob- a solution of potassium hydroxide (1 in 100), add 4 drops of tained (it takes about 20 minutes), stop the distillation, allow sodium carbonate TS, ˆlter into a 100-mL volumetric ‰ask, to stand in water at 159C for 30 minutes, and add water to wash with water, combine the washings with the ˆltrate, and make exactly 100 mL. Shake well, and determine the speciˆc add water to make 100 mL. To 25 mL of this solution add 50 gravity at 159C according to Method 3 under Speciˆc Gravity 15 mL of boiling Fehling's TS, and proceed as directed in (7), <2.56>: the speciˆc gravity d15 is between 0.982 and 0.985. and weigh as copper (II) oxide. From the number obtained by (2) L-Tartaric acid—Pipet 100 mL of Wine, add 2 mL of multiplying the mass (g) of copper (II) oxide by 2, deduct the acetic acid (100), 0.5 mL of a solution of potassium acetate (1 amount (g) of copper (II) oxide determined in (7), and mul- in 5) and 15 g of powdered potassium chloride, and shake tiply again the number so obtained by 1.2: the number ob- vigorously to dissolve as much as possible. Add 10 mL of tained does not exceed 0.104 (g). ethanol (95), rub the inner wall of the beaker strongly for 1 (9) Benzoic acid, cinnamic acid and salicylic acid—Tran- minute to induce the crystallization, and allow to stand be- sfer exactly 50 mL of the sample solution obtained in (2) to a tween 09Cand59C for more than 15 hours. Filter the crystals separator, add 10 g of sodium chloride and 2 mL of dilute by suction, wash successively the beaker and the crystals with hydrochloric acid, and extract with three 10-mL portions of 3-mL portions of a solution prepared by dissolving 15 g of diethyl ether. Combine the diethyl ether extracts, wash with powdered potassium chloride in 120 mL of diluted ethanol (1 two 5-mL portions of water, and extract with three 10-mL in 6), and repeat the washings ˆve times. Transfer the crystals portions of 0.1 molWL sodium hydroxide VS. Combine the together with the ˆlter paper to a beaker, wash the ˆlter with alkaline extracts, evaporate the diethyl ether by warming on a 50 mL of hot water, combine the washings in the beaker, and water bath, cool, neutralize with 1 molWL hydrochloric acid dissolve the crystals by heating. Titrate <2.50> the solution VS, and add 5 mL of potassium chloride-hydrochloric acid with 0.2 molWL sodium hydroxide VS immediately (indica- buŠer solution and water to make exactly 50 mL. Perform tor: 1 mL of phenolphthalein TS). The number obtained by the test as directed under Ultraviolet-visible Spectrophoto- adding 0.75 to the amount (mL) of 0.2 molWLsodium metry <2.24> with this solution, using a solution prepared in hydroxide VS consumed represents the amount (mL) of 0.2 the same manner instead of the sample solution as the blank: molWL sodium hydroxide VS consumed. the absorbance does not exceed 0.15 at a wavelength between Each mL of 0.2 molWL sodium hydroxide VS 220 nm and 340 nm. ＝30.02 mg of C H O (10) Boric acid—Transfer 50 mL of Wine to a porcelain 4 6 6 dish, add 5 mL of sodium carbonate TS, evaporate on a Containers and storage Containers—Tight containers. water bath to dryness, and ignite: a half portion of the residue does not respond to Qualitative Tests <1.09> (1) for borate. Dissolve another half portion of the residue in 5 mL Xylitol of hydrochloric acid: it does not respond to Qualitative Tests <1.09> (2) for borate. キシリトール (11) Methanol—Wine meets the requirements of the Methanol Test <1.12>, when proceeding with exactly 1 mL of ethanol layer obtained by Method 1 of the Alcohol Number Determination <1.01> and distilling without adding water af- ter shaking with 0.5 g of calcium carbonate. (12) Formaldehyde—To 25 mL of Wine add 5 g of sodi- C5H12O5: 152.15 um chloride and 0.2 g of L-tartaric acid, distil, and obtain 15 meso-Xylitol [87-99-0] mL of the distillate. To 5 mL of the distillate add 5 mL of acetyl acetone TS, mix, and heat on a water bath for 10 Xylitol, when dried, contains not less than 98.0z of minutes: the solution has no more color than that of the fol- C5H12O5. lowing control solution. Description Xylitol occurs as white crystals or powder. It is Control solution: Using 5 mL of water instead of the distil- odorless and has a sweet taste. late, perform the test in the same manner. It is very soluble in water, slightly soluble in ethanol (95). Extract content 1.9–3.5wWvz Pipet 25 mL of Wine It is hygroscopic. to a 200-mL tared beaker containing 10 g of sea sand (No. 1), Identification (1) To 1 mL of a solution of Xylitol (1 in 2) previously dried at 1059C for 2.5 hours, and evaporate to add 2 mL of iron (II) sulfate TS and 1 mL of a solution of so- dryness on a water bath. Dry the residue at 1059Cfor2 dium hydroxide (1 in 5): blue-green color is produced without hours, cool in a desiccator (silica gel), and weigh. turbidity. Total ash 0.13–0.40wWvz Pipet 50 mL of Wine to a (2) Determine the infrared absorption spectrum of tared porcelain dish, and evaporate to dryness on a water Xylitol, previously dried, as directed in the potassium bath. Ignite the residue to the constant mass, cool, and bromide disk method under Infrared Spectrophotometry weigh. <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at Assay (1) Ethanol—Pipet Wine into a 100-mL volumetric thesamewavenumbers. ‰ask at 159C, transfer to a 300- to 500-mL ‰ask, and wash this volumetric ‰ask with two 15-mL portions of water. Add pH <2.54> Dissolve 5.0 g of Xylitol in 10 mL of freshly the washings to the sample in the ‰ask, connect the ‰ask to a boiled and cooled water: the pH of this solution is between distillation tube having a trap, and distil using the volumetric 5.0 and 7.0. JP XV O‹cial Monographs / Dried 1241

tion. Melting point <2.60> 93.0 – 95.09C It contains not less than 95z and not more than Purity (1) Clarity and color of solution—Dissolve 5 g of 105z of the labeled amount of xylitol (C5H12O5: Xylitol in 10 mL of water: the solution is clear and colorless. 152.15). (2) Chloride <1.03>—Perform the test with 2.0 g of Method of preparation Prepare as directed under Injec- Xylitol. Prepare the control solution with 0.30 mL of 0.01 tions, with Xylitol. molWL hydrochloric acid VS (not more than 0.005z). No preservative may be added. (3) Sulfate <1.14>—Perform the test with 4.0 g of Xylitol. Prepare the control solution with 0.50 mL of 0.005 molWL Description Xylitol Injection is a clear, colorless liquid. It sulfuric acid VS (not more than 0.006z). has a sweet taste. (4) Heavy metals <1.07>—Proceed with 4.0 g of Xylitol Identification Measure a volume of Xylitol Injection, according to Method 1, and perform the test. Prepare the equivalent to 0.1 g of Xylitol according to the labeled control solution with 2.0 mL of Standard Lead Solution (not amount, add water to make 10 mL, and use this solution as more than 5 ppm). the sample solution. Separately, dissolve 0.1 g of xylitol in 10 (5) Nickel—Dissolve 0.5 g of Xylitol in 5 mL of water, mL of water, and use this solution as the standard solution. add 3 drops of dimethylglyoxime TS and 3 drops of ammonia Perform the test with these solutions as directed under Thin- TS, and allow to stand for 5 minutes: no red color is layer Chromatography <2.03>.Spot2mLeachofthesample produced. solution and standard solution on a plate of silica gel for (6) Arsenic <1.11>—Prepare the test solution with 1.5 g thin-layer chromatography. Develop the plate with a mixture of Xylitol according to Method 1, and perform the test (not of ethanol (95), ammonia solution (28) and water (25:4:3) to more than 1.3 ppm). a distance of about 10 cm, and air-dry the plate. Spray evenly (7) Sugars—Dissolve 5.0 g of Xylitol in 15 mL of water, silver nitrate-ammonia TS, and dry at 1059Cfor15minutes: add 4.0 mL of dilute hydrochloric acid, and heat in a water the spots from the sample solution and the standard solution bath for 3 hours under a reflux condenser. After cooling, show a blackish brown color and the same Rfvalue. neutralize with sodium hydroxide TS (indicator: 2 drops of methyl orange TS). Then add water to make 50 mL, transfer pH <2.54> 4.5–7.5 10 mL of this solution to a flask, add 10 mL of water and 40 Bacterial endotoxins <4.01> Less than 0.50 EUWmL. mL of Fehling's TS, boil gently for 3 minutes, and allow to stand to precipitate copper (I) oxide. Remove the supernatant Extractable volume <6.05> It meets the requirement. liquid through a glass filter (G4), and wash the precipitate Assay Measure exactly a volume of Xylitol Injection, with warm water until the last washing does not show alkalin- equivalent to about 5 g of xylitol (C H O ) according to the ity. Filter these washings through the glass filter mentioned 5 12 5 labeled amount, and add water to make exactly 250 mL. above. Dissolve the precipitate in the flask in 20 mL of iron Measure exactly 10 mL of this solution, and add water to (III) sulfate TS, filter the solution through the glass filter make exactly 100 mL. Then, pipet 10 mL of this solution into mentioned above, wash with water, combine the washings an iodine flask, and proceed as directed in the Assay under with the filtrate, heat at 809C, and titrate <2.50> with 0.02 Xylitol. molWL potassium permanganate VS: not more than 1.0 mL of 0.02 molWL potassium permanganate VS is consumed. Each mL of 0.1 molWL sodium thiosulfate VS ＝ 1.902 mg of C H O Loss on drying <2.41> Not more than 1.0z (1 g, in vacuum, 5 12 5 phosphorus (V) oxide, 24 hours). Containers and storage Containers—Hermetic containers. Plastic containers for aqueous injections may be used. Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.2 g of Xylitol, previously dried, dissolve in water to make exactly 100 mL. Pipet 10 mL Dried Yeast of this solution into an iodine flask, add 50 mL of potassium periodate TS exactly, and heat in a water bath for 15 minutes. 乾燥酵母 After cooling, add 2.5 g of potassium iodide, stopper, shake well, allow to stand for 5 minutes in a dark place, and titrate <2.50> with 0.1 molWL sodium thiosulfate VS (indicator: 3 Dried Yeast is dried and powdered cells of yeast be- mL of starch TS). Perform a blank determination. longing to Saccharomyces. Each mL of 0.1 molWL sodium thiosulfate VS It contains not less than 400 mg of protein and not less than 100 mg of thiamine compounds [as thiamine ＝ 1.902 mg of C5H12O5 chloride hydrochloride (C12H17ClN4OS.HCl: 337.27)] Containers and storage Containers—Tight containers. in each 1 g. Description Dried Yeast occurs as a light yellowish white to Xylitol Injection brown powder. It has a characteristic odor and taste. Identiˆcation Dried Yeast, when examined under a micro- キシリトール注射液 scope <5.01>, shows isolated cells, spheroidal or oval in shape, and 6 to 12 mminlength. Purity (1) Rancidity—Dried Yeast is free from any un- Xylitol Injection is an aqueous solution for injec- pleasant or rancid odor or taste. 1242 Zaltoprofen / O‹cial Monographs JP XV

(2) Starch—Add iodine TS to Dried Yeast, and examine Detector: An ultraviolet absorption photometer microscopically <5.01>: no or only a few granules are tinted (wavelength: 254 nm). blackish purple. Column: A stainless steel column about 4 mm in inside di- ameter and 15 to 30 cm in length, packed with octadecyl- Loss on drying <2.41> Not more than 8.0z (1 g, 1009C, silanized silica gel for liquid chromatography (5 to 10 mmin 8 hours). particle diameter). Total ash <5.01> Not more than 9.0z (1 g). Column temperature: A constant temperature of about 409C. Assay (1) Protein—Weigh accurately about 50 mg of Mobile phase: Dissolve 2.7 g of potassium dihydrogen- Dried Yeast and perform the test as directed under Nitrogen phosphate in 1000 mL of water, and adjust the pH to 3.5 with Determination <1.08>. diluted phosphoric acid (1 in 10). Dissolve 1.6 g of sodium 1- Amount (mg) of protein in 1 g of Dried Yeast octanesulfonate in 800 mL of this solution, and add 200 mL ＝N×6.25×(1/W) of acetonitrile. Flow rate: Adjust the ‰ow rate so that the retention time of N:Amount(mg)ofnitrogen(N) thiamine is about 8 minutes. W: Amount (g) of sample Selection of column: Proceed with 200 mL of the standard (2) Thiamine—Weigh accurately about 1 g of Dried solution under the above operating conditions, and calculate Yeast, add 1 mL of dilute hydrochloric acid and 80 mL of the resolution. Use a column giving elution of thiamine and water, and heat in a water bath at 809Cto859Cfor30 the internal standard in this order with the resolution between minutes with occasional shaking. After cooling, add water to these peaks being not less than 8. make exactly 100 mL, and centrifuge for 10 minutes. Pipet 4 Containers and storage Containers—Tight containers. mL of the supernatant liquid, add exactly 5 mL of acetic acid-sodium acetate TS and exactly 1 mL of enzyme TS, and allow to stand at 459Cto509C for 3 hours. Place exactly 2 mL of this solution onto a chromatographic column prepared Zaltoprofen by pouring 2.5 mL of a weakly acidic CM-bridged cellulose ザルトプロフェン cation exchanger (H type) (40 to 110 mm in particle diameter) into a chromatographic tube about 1 cm in inside diameter and about 17 cm in length, and elute at the ‰ow rate of about 0.5 mL per minute. Wash the upper part of the column with a small amount of water, and wash the column with two 10-mL portions of water at the ‰ow rate of about 1 mL per minute. Elute the column with two 2.5-mL portions of diluted phos- phoric acid (1 in 50) at the ‰ow rate of about 0.5 mL per C17H14O3S: 298.36 minute, and combine the eluate. To the eluate add exactly 1 (2RS)-2-(10-Oxo-10,11-dihydrodibenzo[b,f]thiepin- mL of the internal standard solution and 0.01 g of sodium 1- 2-yl)propanoic acid octanesulfonate, and after dissolving, use this solution as the [74711-43-6] sample solution. Separately, weigh accurately about 15 mg of Zaltoprofen, when dried, contains not less than 99.0z and

Thiamine Chloride Hydrochloride Reference Standard not more than 101.0z of C17H14O3S. (previously determine the water <2.48> inthesamemanneras Thiamine Chloride Hydrochloride), dissolve in 0.001 molWL Description Zaltoprofenoccursaswhitetolightyellow, hydrochloric acid TS to make exactly 100 mL. Pipet 1 mL of crystals or crystalline powder. this solution, and add the mobile phase to make exactly 100 It is freely soluble in acetone, soluble in methanol and in mL. Pipet 1 mL of this solution, add exactly 1 mL of the in- ethanol (99.5), and practically insoluble in water. ternal standard solution and 3 mL of the mobile phase, and It is gradually decomposed by light. use this solution as the standard solution. Perform the test A solution of Zaltoprofen in acetone (1 in 10) shows no op- with 200 mL each of the sample solution and standard solu- tical rotation. tion as directed under Liquid Chromatography <2.01> ac- Identiˆcation (1) To 0.2 g of Zaltoprofen add 0.5 g of so- cording to the following conditions, and calculate the ratios, dium hydroxide, heat gradually to melt, and then carbonize. Q T and QS, of the peak area of thiamine to that of the inter- After cooling, add 5 mL of diluted hydrochloric acid (1 in 2): nal standard. the gas evolved darkens moisten lead (II) acetate paper. Amount (mg) of thiamine in 1 g of Dried Yeast (2) Determine the absorption spectrum of a solution of Zaltoprofen in ethanol (99.5) (1 in 100,000) as directed under ＝(WS×WT)×(QT/QS)×12.5 Ultraviolet-visible Spectrophotometry <2.24>,andcompare WS: Amount (mg) of Thiamine Chloride Hydrochloride the spectrum with the Reference Spectrum: both spectra ex- Reference Standard, calculated on the anhydrous ba- hibit similar intensities of absorption at the same sis wavelengths. WT: Amount (g) of the sample (3) Determine the infrared absorption spectrum of Zal- Internal standard solution—Dissolve 0.01 g of phenacetin in toprofen as directed in the potassium bromide disk method acetonitrile to make 100 mL, and to 1 mL of this solution add under Infrared Spectrophotometry <2.25>,andcomparethe diluted acetonitrile (1 in 5) to make 100 mL. spectrum with the Reference Spectrum: both spectra exhibit Operating conditions— similar intensities of absorption at the same wave numbers. Melting point <2.60> 135 – 1399C JP XV O‹cial Monographs / Zaltoprofen Tablets 1243

blank determination in the same manner, and make any Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of necessary correction. Zaltoprofen according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Each mL of 0.1 mol/L sodium hydroxide VS

Solution (not more than 10 ppm). ＝29.84 mg of C17H14O3S (2) Arsenic <1.11>—Prepare the test solution with 1.0 g Containers and storage Containers—Tight containers. of Zaltoprofen according to Method 3, using 10 mL of a so- Storage—Light-resistant. lution of magnesium nitrate hexahydrate in ethanol (95) (2 in 25), and perform the test (not more than 2 ppm). (3) Related substances—Dissolve 50 mg of Zaltoprofen in 50 mL of the mobile phase, and use this solution as the Zaltoprofen Tablets sample solution. Pipet 1 mL of the sample solution, and add ザルトプロフェン錠 themobilephasetomakeexactly50mL.Pipet1mLofthis solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test Zaltoprofen Tablets contain not less than 95.0z and with exactly 20 mL each of the sample solution and standard not more than 105.0z of the labeled amount of zal- solution as directed under Liquid Chromatography <2.01> ac- toprofen (C17H14O3S: 298.36). cording to the following conditions, and determine each peak Method of preparation Prepare as directed under Tablets, area by the automatic integration method: the area of the with Zaltoprofen. peak other than zaltoprofen and other than the peak having the relative retention time of about 0.7 with respect to zal- Identiˆcation Powder a suitable amount of Zaltoprofen toprofen from the sample solution is not larger than the peak Tablets. To a portion of the powder, equivalent to 80 mg of area of zaltoprofen from the standard solution. Zaltoprofen, add 30 mL of ethanol (99.5), shake well, and Operating conditions— centrifuge. To 1 mL of the supernatant liquid add ethanol Detector: An ultraviolet absorption photometer (99.5) to make 20 mL. To 2 mL of this solution add ethanol (wavelength: 240 nm). (99.5) to make 25 mL, and determine the absorption spec- Column: A stainless steel column 4.6 mm in inside di- trum of this solution as directed under Ultraviolet-visible ameter and 15 cm in length, packed with octadecylsilanized Spectrophotometry <2.24>: it exhibits maxima between 227 silica gel for liquid chromatography (5 mminparticledi- nm and 231 nm and between 329 nm and 333 nm, and a ameter). shoulder between 241 nm and 245 nm. Column temperature: A constant temperature of about Uniformity of dosage units <6.02> Perform the test accord- 259C. ing to the following method: it meets the requirement of the Mobile phase: A mixture of acetonitrile, water and acetic Content uniformity test. acid (100) (300:200:1) To 1 tablet of Zaltoprofen Tablets add 4 mL of water, and Flow rate: Adjust the ‰ow rate so that the retention time of shake to disintegrate. Add a suitable amount of ethanol (95), zaltoprofen is about 4 minutes. shake, then add ethanol (95) to make exactly mL so that Time span of measurement: About 15 times as long as the V each mL contains about 4 mg of zaltoprofen (C H O S), retention time of zaltoprofen beginning after the solvent 17 14 3 and centrifuge. Pipet 2 mL of the supernatant liquid, add ex- peak. actly 10 mL of the internal standard solution and ethanol (95) System suitability— to make 50 mL, and use this solution as the sample solution. Test for required detectability: To exactly 2 mL of the stan- Proceed as directed in the Assay. dard solution add the mobile phase to make exactly 20 mL.

Conˆrm that the peak area of zaltoprofen obtained with 20 Amount (mg) of zaltoprofen (C17H14O3S) mL of this solution is equivalent to 8 to 12z of that with 20 ＝WS×(QT/QS)×(V/20) mL of the standard solution. : Amount (mg) of zaltoprofen for assay System performance: Dissolve 25 mg of zaltoprofen and 50 WS mg of isopropyl benzoate in 100 mL of ethanol (99.5). Pipet 1 Internal standard solution—A solution of benzyl benzoate in mL of this solution, and add the mobile phase to make ex- acetonitrile (1 in 1000). actly 50 mL. When the procedure is run with 20 mLofthisso- Dissolution <6.10> Perform the test according to the follow- lution under the above operating conditions, zaltoprofen and ing method: it meets the requirement. isopropyl benzoate are eluted in this order with the resolution Perform the test with 1 tablet of Zaltoprofen Tablets at 50 between these peaks being not less than 6. revolutions per minute according to the Paddle method, us- System repeatability: When the test is repeated 6 times with ing 900 mL of 2nd fluid for dissolution test as the dissolution 20 mL of the standard solution under the above operating medium. Withdraw not less than 20 mL of the dissolved solu- conditions, the relative standard deviation of the peak area of tion 30 minutes after start of the test, and ˆlter through a zaltoprofen is not more than 2.0z. membrane ˆlter with pore size of not more than 0.45 mm. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 Discard the ˆrst 10 mL of the ˆltrate, pipet V mL of the sub- hours). sequent ˆltrate, add 2nd fluid for dissolution test to make ex- actly V? mL so that each mL contains about 44 mgofzal- Residue on ignition <2.44> Not more than 0.1z (1 g). toprofen (C17H14O3S) per mL according to the labeled Assay Weigh accurately about 0.5 g of Zaltoprofen, dis- amount, and use this solution as the sample solution. solve in 50 mL of methanol, and titrate <2.50> with 0.1 mol/L Separately, weigh accurately about 22 mg of zaltoprofen for sodium hydroxide VS (potentiometric titration). Perform a assay, previously dried at 1059C for 4 hours, dissolve in 20 1244 Zinc Chloride / O‹cial Monographs JP XV mL of ethanol (99.5), and add 2nd fluid for dissolution test peak area of zaltoprofen to that of the internal standard is to make exactly 100 mL. Pipet 4 mL of this solution, add 2nd not more than 1.0z. fluid for dissolution test to make exactly 20 mL, and use this Containers and storage Containers—Tight containers. solution as the standard solution. Determine the absor- bances, AT and AS, of the sample solution and standard solu- tion at 340 nm as directed under Ultraviolet-visible Spec- trophotometry <2.24>, using 2nd fluid for dissolution test as Zinc Chloride the control. The dissolution rate in 30 minutes is not less than 塩化亜鉛 75z. Dissolution rate (z) with respect to the labeled amount of zaltoprofen (C17H14O3S) ZnCl2: 136.32 ＝WS×(AT/AS)×(V?/V)×(1/C)×180 Zinc Chloride contains not less than 97.0z of WS: Amount (mg) of zaltoprofen for assay C: Labeled amount (mg) of zaltoprofen for assay in 1 ZnCl2. tablet Description Zinc Chloride occurs as white, crystalline pow- der, rods, or masses. It is odorless. Assay To 10 tablets of Zaltoprofen Tablets add 40 mL of It is very soluble in water, and freely soluble in ethanol water, shake to disintegrate, then add a suitable amount of (95), and its solution may sometimes be slightly turbid. The ethanol (95), shake, add ethanol (95) to make exactly 200 solution becomes clear on addition of a small amount of mL, and centrifuge. Pipet an amount of the supernatant liq- hydrochloric acid. uid, equivalent to about 8 mg of zaltoprofen (C H O S), 17 14 3 The pH of an aqueous solution of Zinc Chloride (1 in 2) is add exactly 10 mL of the internal standard solution and between 3.3 and 5.3. ethanol (95) to make 50 mL, and use this solution as the sam- It is deliquescent. ple solution. Separately, weigh accurately about 80 mg of zal- toprofen for assay, previously dried at 1059C for 4 hours, Identiˆcation An aqueous solution of Zinc Chloride (1 in add 4 mL of water and ethanol (95) to make exactly 20 mL. 30) responds to the Qualitative Tests <1.09> for zinc salt and Pipet 2 mL of this solution, add exactly 10 mL of the internal chloride. standard solution and ethanol (95) to make 50 mL, and use Purity (1) Clarity and color of solution—Dissolve 1.0 g of this solution as the standard solution. Perform the test with 5 Zinc Chloride in 10 mL of water and 2 drops of hydrochloric mLeachofthesamplesolutionandstandardsolutionas acid: the solution has no color, and is clear. directed under Liquid Chromatography <2.01> according to (2) Sulfate <1.14>—Perform the test with 2.0 g of Zinc the following conditions, and determine the ratios, Q and T Chloride. Prepare the control solution with 0.40 mL of 0.005 Q , of the peak area of zaltoprofen to that of the internal S mol L sulfuric acid VS (not more than 0.010z). standard. W (3) Ammonium—Dissolve 0.5 g of Zinc Chloride in 5 mL Amount (mg) of zaltoprofen (C17H14O3S) of water, and warm with 10 mL of a solution of sodium ＝WS×(QT/QS)×(1/10) hydroxide (1 in 6): the evolving gas does not change moistened red litmus paper to blue. W : Amount (mg) of zaltoprofen for assay S (4) Heavy metals—Dissolve 0.5 g of Zinc Chloride in 5 Internal standard solution—A solution of benzyl benzoate in mL of water in a Nessler tube, shake thoroughly with 15 mL acetonitrile (1 in 1000) of TS, add 1 drop of sodium sulˆde TS, Operating conditions— allow to stand for 5 minutes, and immediately observe from Detector: An ultraviolet absorption photometer the top downward against a white background: the solution (wavelength: 240 nm). has no more color than the following control solution. Column: A stainless steel column 4.6 mm in inside di- Control solution: To 2.5 mL of Standard Lead Solution ameter and 15 cm in length, packed with octadecylsilanized add 3 mL of water and 15 mL of potassium cyanide TS, silica gel for liquid chromatography (5 mminparticledi- shake thoroughly, and add 1 drop of sodium sulˆde TS (not ameter). more than 50 ppm). Column temperature: A constant temperature of about (5) Alkali earth metals and alkali metals—Dissolve 2.0 g 259C. of Zinc Chloride in 120 mL of water, add ammonium sulˆde Mobile phase: A mixture of acetonitrile, water and acetic TS to complete precipitation, add water to make 200 mL, acid (100) (300:200:1). shake thoroughly, and ˆlter through dry ˆlter paper. Discard Flow rate: Adjust the ‰ow rate so that the retention time of the ˆrst 20 mL of the ˆltrate, take the following 100 mL of zaltoprofen is about 4 minutes. the ˆltrate, evaporate with 3 drops of sulfuric acid to dryness, System suitability— and heat the residue strongly at 6009C to constant mass: the System performance: When the procedure is run with 5 mL mass is not more than 10.0 mg. of the standard solution under the above operating condi- (6) Arsenic <1.11>—Prepare the test solution with 0.40 g tions, zaltoprofen and the internal standard are eluted in this of Zinc Chloride according to Method 1, and perform the test order with the resolution between these peaks being not less (not more than 5 ppm). than 10. (7) Oxychloride—Shake gently 0.25 g of Zinc Chloride System repeatability: When the test is repeated 6 times with with 5 mL of water and 5 mL of ethanol (95), and add 0.3 mL 5 mL of the standard solution under the above operating con- of 1 molWL hydrochloric acid VS: the solution is clear. ditions, the relative standard deviation of the ratio of the Assay Weigh accurately about 0.3 g of Zinc Chloride, add JP XV O‹cial Monographs / Zinc Oxide Oil 1245

0.4 mL of dilute hydrochloric acid and water to make exactly an ethanol (95) solution of a,a?-dipyridyl (1 in 200) and 200 mL. Measure exactly 20 mL of the solution, add 80 mL water to make 50 mL. After allowing to stand for 30 minutes, of water, 2 mL of ammonia-ammonium chloride buŠer solu- compare the color of the both liquids against a white back: tion, pH 10.7, and titrate <2.50> with 0.01 molWL disodium the color of the liquid from the test solution is not stronger dihydrogen ethylenediamine tetraacetate VS (indicator: 0.04 than that from the control solution (not more than 10 ppm). g of eriochrome black T-sodium chloride indicator). (5) Lead—To 2.0 g of Zinc Oxide add 20 mL of water, then add 5 mL of acetic acid (100) with stirring, and heat on a Each mL of 0.01 molWL disodium dihydrogen water bath until solution is complete. Cool, and add 5 drops ethylenediamine tetraacetate VS of potassium chromate TS: no turbidity is produced. ＝1.363 mg of ZnCl 2 (6) Arsenic <1.11>—Dissolve 0.5 g of Zinc Oxide in 5 mL Containers and storage Containers—Tight containers. of dilute hydrochloric acid, use this solution as the test solu- tion, and perform the test (not more than 4 ppm). Loss on ignition <2.43> Not more than 1.0z (1 g, 8509C, Zinc Oxide 1hour). 酸化亜鉛 Assay Weigh accurately about 0.8 g of Zinc Oxide, previ- ously ignited at 8509C for 1 hour, dissolve in 2 mL of water and 3 mL of hydrochloric acid, and add water to exactly 100 ZnO: 81.41 mL. Pipet 10 mL of this solution, add 80 mL of water, then add a solution of sodium hydroxide (1 in 50) until a slight Zinc Oxide, when ignited, contains not less than precipitate is produced. Add 5 mL of ammonia-ammonium 99.0z of ZnO. chloride buŠer solution, pH 10.7, and titrate <2.50> with 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS Description Zinc Oxide occurs as a white, amorphous pow- (indicator: 0.04 g of eriochrome black T-sodium chloride in- der. It is odorless and tasteless. dicator). It is practically insoluble in water, in ethanol (95), in acetic acid (100) and in diethyl ether. Each mL of 0.05 mol/L disodium dihydrogen It dissolves in diute hydrochloric acid and in sodium ethylenediamine tetraacetate VS hydroxide TS. ＝4.071 mg of ZnO It gradually absorbs carbon dioxide from air. Containers and storage Containers—Tight containers. Identification (1) Heat Zinc Oxide strongly: a yellow color develops on strong heating, and disappears on cooling. (2) A solution of Zinc Oxide in dilute hydrochloric acid Zinc Oxide Oil (1 in 10) responds to the Qualitative Tests <1.09> for zinc salt. チンク油 Purity (1) Carbonate, and clarity and color of solution—Mix 2.0 g of Zinc Oxide with 10 mL of water, add 30 mL of dilute sulfuric acid, and heat on a water bath with Zinc Oxide Oil contains not less than 45.0z and not stirring: no effervescence occurs, and the solution obtained is more than 55.0z of zinc oxide (ZnO: 81.41). clear and colorless. (2) Alkalinity—To 1.0 g of Zinc Oxide add 10 mL of Method of preparation water, and boil for 2 minutes. Cool, filter through a glass Zinc Oxide 500 g filter (G3), and to the filtrate add 2 drops of phenolphthalein Fixed oil a su‹cient quantity TS and 0.20 mL of 0.1 molWL hydrochloric acid VS: no color develops. To make 1000 g (3) Sulfate <1.14>—Shake 0.5 g of Zinc Oxide with 40 mL Mix the above ingredients. An appropriate quantity of of water, and filter. Take 20 mL of the filtrate, add 1 mL of Castor Oil or polysorbate 20 may be used partially in place of dilute hydrochloric acid and water to make 50 mL, and per- ˆxed oil. form the test using this solution as the test solution. Prepare the control solution with 0.50 mL of 0.005 molWL sulfuric Description Zinc Oxide Oil is a white to whitish, slimy sub- acid VS (not more than 0.096z). stance, separating a part of its ingredients when stored for a (4) Iron—Dissolve 1.0 g of Zinc Oxide in 50 mL of dilut- prolonged period. ed hydrochloric acid (1 in 2), dissolve 0.1 g of ammonium Identiˆcation Mix thoroughly, and place 0.5 g of Zinc peroxodisulfate in this solution, and extract with 20 mL of 4- Oxide Oil in a crucible, heat gradually raising the tempera- methyl-2-pentanone. Add 30 mL of acetic acid-sodium ture until the mass is thoroughly charred, and then ignite it acetate buffer solution for Iron Limit Test, pH 4.5, to the 4- strongly: a yellow color is produced, and disappears on cool- methyl-2-pentanone layer, extract again, and use the layer of ing. To the residue add 10 mL of water and 5 mL of dilute the buffer solution as the test solution. Separately, perform hydrochloric acid, shake well, and ˆlter. To the ˆltrate add 2 the test in the same manner with 1.0 mL of Standard Iron So- to 3 drops of potassium hexacyanoferrate (II) TS: a white lution, and use the layer so obtained as the control solution. precipitate is formed (zinc oxide). Add 2 mL each of L-ascorbic acid solution for Iron Limit Test (1 in 100) to the test solution and the control solution, Assay Weigh accurately about 0.8 g of Zinc Oxide Oil, mix- respectively, mix, allow to stand for 30 minutes, add 5 mL of ed well, place in a crucible, heat gradually raising the temper- 1246 Zinc Oxide Ointment / O‹cial Monographs JP XV ature until the mass is thoroughly charred, and then ignite Add 80 mL of water to exactly 20 mL of this solution, and until the residue becomes yellow, and cool. Dissolve the add a solution of sodium hydroxide (1 in 50) until a small residue in 1 mL of water and 1.5 mL of hydrochloric acid, amount of precipitates begins to form in the solution. Add 5 and add water to make exactly 100 mL. Pipet 20 mL of this mL of ammonia-ammonium chloride buŠer solution, pH solution, add 80 mL of water, and add a solution of sodium 10.7, and titrate <2.50> with 0.05 mol/L disodium dihydro- hydroxide (1 in 50) until a small amount of precipitates be- gen ethylenediamine tetraacetate VS (indicator: 0.04 g of gins to form in the solution. Add 5 mL of ammonia-ammoni- eriochrome black T-sodium chloride indicator). um chloride buŠer solution, pH 10.7, and titrate <2.50> with Each mL of 0.05 mol/L disodium dihydrogen 0.05 mol/L disodium dihydrogen ethylenediamine tetraa- ethylenediamine tetraacetate VS cetate VS (indicator: 0.04 g of eriochrome black T-sodium ＝4.071 mg of ZnO chloride indicator). Each mL of 0.05 mol/L disodium dihydrogen Containers and storage Containers—Tight containers. ethylenediamine tetraacetate VS ＝4.071 mg of ZnO Containers and storage Containers—Tight containers. Zinc Oxide Starch Powder 亜鉛華デンプン Zinc Oxide Ointment Method of preparation 亜鉛華軟膏 Zinc Oxide 500 g Starch a su‹cient quantity Zinc Oxide Ointment contains not less than 18.5z To make 1000 g and not more than 21.5z of zinc oxide (ZnO: 81.41). Prepare as directed under Powders, with the above in- Method of preparation gredients. Zinc Oxide 200 g Description Zinc Oxide Starch Powder occurs as a white Liquid Para‹n 30 g powder. White Ointment a su‹cient quantity Identiˆcation (1) Place 1 g of Zinc Oxide Starch Powder To make 1000 g in a crucible, heat gradually, raising the temperature until it is charred, and then ignite strongly: a yellow color develops, Prepare as directed under Ointments, with the above in- and disappears on cooling. To the residue add 10 mL of gredients. water and 5 mL of dilute hydrochloric acid, shake well, and Description Zinc Oxide Ointment is white in color. ˆlter. To the ˆltrate add 2 to 3 drops of potassium hexac- yanoferrate (II) TS: a white precipitate is formed (zinc Identiˆcation Place1gofZincOxideOintmentinacruci- oxide). ble, melt by warming, heat gradually raising the temperature (2) Shake well 1 g of Oxide Starch Powder with 10 mL of until the mass is thoroughly charred, and then ignite it water and 5 mL of dilute hydrochloric acid, and ˆlter. Boil strongly: a yellow color is produced, and disappears on cool- the residue on a ˆlter paper with 10 mL of water, cool, and ing. To the residue add 10 mL of water and 5 mL of dilute add 1 drop of iodine TS: a dark blue-purple color is produced hydrochloric acid, shake well, and ˆlter. To the ˆltrate add 2 (starch). to 3 drops of potassium hexacyanoferrate (II) TS: a white precipitate is formed (zinc oxide). Containers and storage Containers—Tight containers. Purity Calcium, magnesium and other foreign inorganic matters—Place 2.0 g of Zinc Oxide Ointment in a crucible, melt by warming, and heat gradually raising the temperature, Zinc Sulfate Hydrate until the mass is thoroughly charred. Ignite the mass strongly 硫酸亜鉛水和物 until the residue becomes uniformly yellow, and cool. Add 6 mL of dilute hydrochloric acid, and heat on a water bath for 5 to 10 minutes: the solution is colorless and clear. Filter the ZnSO .7H O: 287.58 solution, add 10 mL of water to the ˆltrate, and add ammo- 4 2 nia TS until the precipitate ˆrst formed redissolves. Add 2 mL each of ammonium oxalate TS and disodium hydrogen- Zinc Sulfate Hydrate contains not less than 99.0z phosphate TS to this solution: the solution remains un- and not more than 102.0z of ZnSO4.7H2O. changed or becomes very slightly turbid within 5 minutes. Description Zinc Sulfate Hydrate occurs as colorless crys- tals or a white, crystalline powder. It is odorless, and has an Assay Weigh accurately about 2 g of Zinc Oxide Ointment, astringent, characteristic taste. place in a crucible, melt by warming, heat gradually raising It is very soluble in water, and practically insoluble in the temperature until the mass is thoroughly charred, and ethanol (95) and in diethyl ether. then ignite until the residue becomes uniformly yellow, and The pH of a solution of Zinc Sulfate Hydrate (1 in 20) is cool. Dissolve the residue in 1 mL of water and 1.5 mL of between 3.5 and 6.0. hydrochloric acid, and add water to make exactly 100 mL. It effloresces in dry air. JP XV O‹cial Monographs / Zinostatin Stimalamer 1247

Prepare as directed under Ophthalmic Solution, with the Identification A solution of Zinc Sulfate Hydrate (1 in 20) above ingredients. responds to the Qualitative Tests <1.09> for zinc salt and for sulfate. Description Zinc Sulfate Ophthalmic Solution is a clear, colorless liquid. Purity (1) Acidity—Dissolve 0.25 g of Zinc Sulfate Hy- drate in 5 mL of water, and add 1 drop of methyl orange TS: Identiˆcation (1) Zinc Sulfate Ophthalmic Solution no red color develops. responds to the Qualitative Tests <1.09> for zinc salt. (2) Heavy metals <1.07>—Dissolve 1.0 g of Zinc Sulfate (2) Zinc Sulfate Ophthalmic Solution responds to the Hydrate in 10 mL of water contained in a Nessler tube. Add Qualitative Tests <1.09> for borate. 20 mL of potassium cyanide TS, and mix well. Add 2 drops (3) Zinc Sulfate Ophthalmic Solution responds to the of sodium sulfide TS, and allow the mixture to stand for 5 Qualitative Tests <1.09> for chloride. minutes. Observe vertically against a white background, the Assay Pipet accurately 25 mL of Zinc Sulfate Ophthalmic color of the solution is not more intense than the following Solution, add 100 mL of water and 2 mL of ammonia-ammo- control solution. nium chloride buŠer solution, pH 10.7, and titrate <2.50> Control solution: To 1.0 mL of Standard Lead Solution with 0.01 molWL disodium dihydrogen ethylenediamine add 10 mL of water and 20 mL of potassium cyanide TS, and tetraacetate VS (indicator: 0.04 g of eriochrome black T-sodi- mix well. Add 2 drops of sodium sulfide TS (not more than um chloride indicator). 10 ppm). (3) Alkali earth metals and alkali metals—Dissolve 2.0 g Each mL of 0.01 molWL disodium dihydrogen of Zinc Sulfate Hydrate in 150 mL of water, add a suitable ethylenediamine tetraacetate VS amount of ammonium sulfide TS to complete the precipita- ＝2.876 mg of ZnSO4.7H2O tion, and add water to make exactly 200 mL. Shake well, and Containers and storage Containers—Tight containers. filter through a dry filter paper. Discard the first 20 mL of the filtrate, take exactly 100 mL of the subsequent filtrate, evaporate to dryness, and ignite as directed under Residue on Ignition <2.44>: the mass of the residue is not more than 5.0 Zinostatin Stimalamer mg. ジノスタチン スチマラマー (4) Arsenic <1.11>—Prepare the test solution with 1.0 g of Zinc Sulfate Hydrate according to Method 1, and perform the test (not more than 2 ppm). Assay Weigh accurately about 0.3 g of Zinc Sulfate Hy- drate, and dissolve in water to make exactly 100 mL. Measure exactly 25 mL of this solution, add 100 mL of water and 2 mL of ammonia-ammonium chloride buffer solution, pH 10.7, and titrate <2.50> with 0.01 molWL disodium dihydoro- gen ethylenediamine tetraacetate VS (indicator: 0.04 g of eriochrome black T-sodium chloride indicator). Each mL of 0.01 molWL disodium dihydrogen ethylenediamine tetraacetate VS

＝ 2.876 mg of ZnSO4.7H2O Containers and storage Containers—Tight containers. (4S,6R,11R,12R)-11-[a-D-2,6-Dideoxy-2-(methylamino)- galactopyranosyloxy]-4-[(4R)-2-oxo-1,3-dioxolan-4-yl]- Zinc Sulfate Ophthalmic Solution 5-oxatricyclo[8.3.0.04,6]trideca-1(13),9-diene-2,7-diyn-12-yl 2-hydroxy-7-methoxy-5-methylnaphthalene-1- 硫酸亜鉛点眼液 carboxylate [123760-07-6]

Zinc Sulfate Ophthalmic Solution contains not less than 0.27 w/vz and not more than 0.33 w/vz of zinc sulfate hydrate (ZnSO4.7H2O: 287.58). Method of preparation Zinc Sulfate Hydrate 3 g Boric Acid 20 g Sodium Chloride 5 g Fennel Oil 2 mL Puriˆed Water a su‹cient quantity

To make 1000 mL 1248 Zinostatin Stimalamer / O‹cial Monographs JP XV

solution, pH 7.0, 10 mL).

20 Optical rotation <2.49> [a]D : －30.0 – －38.09(20 mg cal- culated on the anhydrous basis, 0.05 molWL phosphate buffer solution, pH 7.0, 5 mL, 100 mm). pH <2.54> Dissolve 10 mg of Zinostatin Stimalamer in 1 mL of water: the pH of the solution is between 4.5 and 5.5. Purity (1) Clarity and color of solution—Being specified separately. (2) Heavy metals <1.07>—Weighaccurately40mgof Zinostatin Stimalamer, place in a crucible, carbonize and in- cinerate according to Method 2, add 2 mL of hydrochloric acid, and evaporate on a water bath to dryness. After cool-

ing, weigh the residue WT g. Then, moisten the residue with 0.1 mL of diluted hydrochloric acid (1 in 5), add 1 mL of water, 85 mL of diluted ammonia TS (1 in 2) and 0.1 mL of

dilute acetic acid, and add water so that the mass is WT ＋ 2.0 g. Adjust the pH of this solution to 3.2 to 3.4 with diluted ammoniaTS(1in20)ordilutedhydrochloricacid(1in50),

add water so that the mass is WT ＋ 2.5 g, and use this solu- tion as the test solution. Separately, prepare the blank solu- tion in the same manner without the sample. Separately, take Zinostatin Stimalamer consists 1 molecule of 2 mL of nitric acid, 5 drops of sulfuric acid and 2 mL of zinostatin, consisting of chromophore and apoprotein hydrochloric acid, and evaporate to dryness according to

(polypeptide consisting of 113 amino acid residues) and Method 2. After cooling, weigh the residue WS g. Then, 2 molecules of partially butyl-esteriˆed styrene-maleic moisten the residue with 0.1 mL of diluted hydrochloric acid acid alternate copolymer, and has average molecular (1 in 5), and proceed in the same manner as directed in the mass of about 15,000. The alternate copolymer is preparation of the test solution. After adjusting the pH of the bound an amido bond to a-amino group of alanine of solution so obtained to 3.2 to 3.4, add 80 mLofStandard

N-terminal and to e-amino group of lysine 20 of the Lead Solution, and add water so that the mass is WS ＋ 2.5 g, apoprotein. and use this solution as the control solution. Add 10 mLeach It contains not less than 900 mg (potency) and not of diluted sodium sulfide TS (1 in 6) to the test solution, the more than 1080 mg (potency) per mg, calculated on the blank solution and the control solution, mix, and allow to anhydrous basis. The potency of Zinostatin stand for 5 minutes. Determine the absorbances, AT, AO and Stimalamer is expressed as mass (potency) of zinostatin AS of the test solution, the blank solution and the control so- stimalamer. lution at 400 nm as directed under Ultraviolet-visible Spec- Description Zinostatin Stimalamer occurs as a pale yellow trophotometry <2.24>: AT － AO is not larger than AS － AO powder. (not more than 20 ppm). It is freely soluble in water, and practically insoluble in (3) Related substances—Being specified separately. ethanol (95) and in diethyl ether. (4) Inorganic salts of manufacturing process origin—Being specified separately. Identification (1) Dissolve 10 mg of Zinostatin Stimalamer in 1 mL of sodium hydroxide TS, and add a drop Water <2.48> Not more than 12.0z (10 mg, coulometric of copper (II) sulfate TS: a purple color develops. titration). (2) Dissolve 1 mg of Zinostatin Stimalamer in 1 mL of Assay Perform the test according to the Cylinder-plate 0.05 molWL phosphate buffer solution, pH 7.0, add 0.5 mL method as directed under Microbial Assay for Antibiotics of a solution of trichloroacetic acid (1 in 5), and shake: a <4.02> according to the following conditions. Perform the white precipitate is formed. procedures of (iii), (iv) and (v) without exposure to direct or (3) Determine the absorption spectra of solutions of indirect sunlight. Zinostatin Stimalamer and Zinostatin Stimalamer Reference (i) Test organism—Micrococcus luteus ATCC 9341 Standard in 0.05 molWL phosphate buffer solution, pH 7.0 (1 (ii) Culture medium—Use the medium i in 3) Medium for in 2500) as directed under Ultraviolet-visible Spectrophoto- other organisms under (1) Agar media for seed and base lay- metry <2.24>, and compare these spectra: both spectra exhibit er. Adjust the pH of the medium so that it will be 7.9 to 8.1 similar intensities of absorption at the same wavelengths. after sterilization. (4) Determine the infrared absorption spectra of Zinosta- (iii) Standard solutions—Weigh accurately an amount of tin Stimalamer and Zinostatin Stimalamer Reference Stan- Zinostatin Stimalamer Reference Standard equivalent to dard as directed in the potassium bromide disk method under about 20 mg (potency), dissolve in 0.1 molWL phosphate Infrared Spectrophotometry <2.25>, and compare these spec- buffer solution, pH 8.0 to make exactly 50 mL, and use this tra: both spectra exhibit similar intensities of absorption at solution as the high concentration standard solution. Pipet 5 thesamewavenumbers. mLofthehighconcentrationstandardsolution,add0.1mol 1z WL phosphate buffer solution, pH 8.0, to make exactly 20 Absorbance <2.24> E1cm (268 nm): 15.5 – 18.5 (4 mg calcu- lated on the anhydrous basis, 0.05 molWL phosphate buffer mL, and use this solution as the low concentration standard JP XV O‹cial Monographs / Zinostatin Stimalamer 1249 solution. (iv) Sample solutions—Weigh accurately an amount of Zinostatin Stimalamer equivalent to about 20 mg (potency), dissolve in 0.1 molWL phosphate buffer solution, pH 8.0 to make exactly 50 mL, and use this solution as the high concen- tration sample solution. Pipet 5 mL of the high concentration sample solution, add 0.1 molWL phosphate buffer solution, pH 8.0, to make exactly 20 mL, and use this solution as the low concentration sample solution. (v) Procedure—Allow to stand at 3 to 59C for 2 hours be- fore incubation. Containers and storage Containers—Tight containers. Storage—Light-resistant, and not exceeding －209C.