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Capsicum Oleoresin and Homocapsaicin

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Capsicum Oleoresin and Homocapsaicin Printed on: Wed Jan 06 2021, 02:44:36 AM Official Status: Currently Official on 06-Jan-2021 DocId: 1_GUID-1560FD9B-BE0F-495E-9994-C5718733DB4C_2_en-US (EST) Printed by: Jinjiang Yang Official Date: Official as of 01-May-2019 Document Type: USP @2021 USPC 1 nordihydrocapsaicin, nonivamide, decanylvanillinamide, Capsicum Oleoresin and homocapsaicin. DEFINITION ASSAY Capsicum Oleoresin is an alcoholic extract of the dried ripe · CONTENT OF TOTAL CAPSAICINOIDS fruits of Capsicum. It contains NLT 6.5% of total Mobile phase: A mixture of acetonitrile and diluted capsaicinoids, calculated as the sum of capsaicin, phosphoric acid (1 in 1000) (2:3) dihydrocapsaicin, nordihydrocapsaicin, nonivamide, Standard solution A: 0.2 mg/mL of USP Capsaicin RS in decanylvanillinamide, and homocapsaicin, all calculated on methanol the anhydrous basis. The nonivamide content is NMT 5% of Standard solution B: 0.1 mg/mL of USP the total capsaicinoids, calculated on the anhydrous basis. Dihydrocapsaicin RS in methanol [CAUTIONÐCapsicum Oleoresin is a powerful irritant, and Sample solution: 5 mg/mL of Capsicum Oleoresin in even in minute quantities produces an intense burning methanol. Pass a portion of this solution through a filter of sensation when it comes in contact with the eyes and 0.2-µm pore size, and use the filtrate as the Sample solution. tender parts of the skin. Care should be taken to protect Chromatographic system the eyes and to prevent contact of the skin with (See Chromatography á621ñ, System Suitability.) Capsicum Oleoresin.] Mode: LC IDENTIFICATION Detector: UV 281 nm Å · A. THIN-LAYER CHROMATOGRAPHY Column: 4.6-mm × 25-cm; end-capped, 5-µm, 150 , Standard solution A: 0.4 mg/mL of USP Capsaicin RS in packing L11 methanol Column temperature: 30° Standard solution B: 0.4 mg/mL of USP Flow rate: 1.0 mL/min Dihydrocapsaicin RS in methanol Injection volume: 20 µL Sample solution: 10 mg/mL of Capsicum Oleoresin in System suitability hexanes Samples: Standard solution A and Sample solution Chromatographic system Suitability requirements Adsorbent: Chromatographic reverse phase octadecyl silyl Resolution: NLT 1.5 between the capsaicin peak and the silica gel with an average particle size of 5 µm (HPTLC nonivamide peak that occurs at a retention time of 0.95 plates) relative to 1.0 for capsaicin, Sample solution Application volume: 2 µL, as 8-mm bands Relative standard deviation: NMT 2.0% for the Developing solvent system: A mixture of methanol and capsaicin peak, Standard solution A water (8:2) Analysis Derivatization reagent A: 0.25 mg/mL of Samples: Standard solution A, Standard solution B, and dichloroquinonechlorimide in ethyl acetate Sample solution Derivatization reagent B: Ammonium hydroxide solution Identify the capsaicin and dihydrocapsaicin peaks in the Analysis Sample solution chromatogram by comparison with the Samples: Standard solution A, Standard solution B, and chromatograms of Standard solution A and Standard Sample solution solution B, respectively. Identify the peaks corresponding Apply the Samples as bands to a suitable to nordihydrocapsaicin, nonivamide, high-performance thin-layer chromatographic plate. decanylvanillinamide, and homocapsaicin using the Use a saturated chamber, and condition the plate to a approximate relative retention times provided in Table 1. relative humidity of about 33% using a suitable device. Develop the chromatograms over a distance of 6 cm. Table 1 Remove the plate from the chamber, dry, derivatize with Approximate Official Relative Derivatization reagent A, dry, expose to vapors of Retention Derivatization reagent B until blue bands develop, and Analyte Time examine under white light. System suitability: Standard solution A shows a blue band Nordihydrocapsaicin 0.89 at about one-third of the chromatogram, and Standard Nonivamide 0.95 solution B shows a blue band at an RF right below that from Standard solution A. Capsaicin 1.00 Acceptance criteria: The Sample solution exhibits a blue Decanylvanillinamide 1.34 band at about one-third of the chromatogram, similar in position and color to the capsaicin band in the Homocapsaicin 1.40 chromatogram of Standard solution A, and exhibits a blue band at an RF right below that of capsaicin, similar in Calculate the percentage of capsaicin in the portion of position and color to the dihydrocapsaicin band in the Capsicum Oleoresin taken: chromatogram of Standard solution B. Other bands may be observed in the Sample solution chromatogram. Result = (rU/rS) × (CS/CU)× 100 · B. HPLC Analysis: Proceed as directed in the test for Content of Total rU = peak area of capsaicin from the Sample solution Capsaicinoids. rS = peak area of capsaicin from Standard solution A Acceptance criteria: The Sample solution chromatogram CS = concentration of capsaicin in Standard solution A exhibits the main capsaicinoid peak at the retention time (mg/mL) corresponding to capsaicin in the chromatogram of CU = concentration of Capsicum Oleoresin in the Standard solution A and a peak of lower intensity Sample solution (mg/mL) corresponding to dihydrocapsaicin in the chromatogram of Standard solution B. The Sample solution chromatogram Calculate the percentage of dihydrocapsaicin in the portion shows additional minor peaks corresponding to of Capsicum Oleoresin taken: https://online.uspnf.com/uspnf/document/1_GUID-1560FD9B-BE0F-495E-9994-C5718733DB4C_2_en-US 1/2 Printed on: Wed Jan 06 2021, 02:44:36 AM Official Status: Currently Official on 06-Jan-2021 DocId: 1_GUID-1560FD9B-BE0F-495E-9994-C5718733DB4C_2_en-US (EST) Printed by: Jinjiang Yang Official Date: Official as of 01-May-2019 Document Type: USP @2021 USPC 2 Result = (rU/rS) × (CS/CU) × 100 Acceptance criteria: NLT 6.5% on the anhydrous basis CONTAMINANTS rU = peak area of dihydrocapsaicin from the Sample solution · ELEMENTAL IMPURITIESÐPROCEDURES á233ñ Acceptance criteria rS = peak area of dihydrocapsaicin from Standard solution B Arsenic: NMT 0.5 µg/g Cadmium: NMT 1.0 µg/g CS = concentration of dihydrocapsaicin in Standard solution B (mg/mL) Lead: NMT 5.0 µg/g Mercury: NMT 0.1 µg/g CU = concentration of Capsicum Oleoresin in the Sample solution (mg/mL) Change to read: Calculate the percentage of nonivamide, expressed as ▲ capsaicin, in the portion of Capsicum Oleoresin taken: · ARTICLES OF BOTANICAL ORIGIN, Pesticide Residue Analysis á561ñ▲ (CN 1-May-2019): Meets the requirements Result = (r /r ) × (C /C ) × 100 · ARTICLES OF BOTANICAL ORIGIN, Test for Aflatoxins á561ñ: U S S U Meets the requirements rU = peak area of nonivamide from the Sample solution SPECIFIC TESTS rS = peak area of capsaicin from Standard solution A · LIMIT OF NONIVAMIDE CS = concentration of capsaicin in Standard solution A Analysis: Use the chromatograms and calculations obtained (mg/mL) in the test for Content of Total Capsaicinoids. CU = concentration of Capsaicin Oleoresin in the Calculate the content of nonivamide as percentage of total Sample solution (mg/mL) capsaicinoids: Calculate the sum of the percentages of Result = (PN/PTC) × 100 nordihydrocapsaicin, decanylvanillinamide, and homocapsaicin, expressed as capsaicin, in the portion of PN = percentage of nonivamide as calculated in the Capsicum Oleoresin taken: test for Content of Total Capsaicinoids PTC = percentage of total capsaicinoids as calculated in Result = (ΣrU/rS) × (CS/CU) × 100 the test for Content of Total Capsaicinoids ΣrU = sum of the peak areas of nordihydrocapsaicin, Acceptance criteria: NMT 5% on the anhydrous basis decanylvanillinamide, and homocapsaicin from · WATER DETERMINATION, Method Ia á921ñ the Sample solution Sample: 5.0 g of Capsicum Oleoresin rS = peak area of capsaicin from Standard solution A Acceptance criteria: NMT 8% CS = concentration of capsaicin in Standard solution A (mg/mL) ADDITIONAL REQUIREMENTS · PACKAGING AND STORAGE: Preserve in tight containers. CU = concentration of Capsicum Oleoresin in the Sample solution (mg/mL) · LABELING: Label it to indicate that if separation occurs, it should be warmed and mixed before use. Calculate the content of total capsaicinoids as the sum of · USP REFERENCE STANDARDS á11ñ the percentages of capsaicin, dihydrocapsaicin, USP Capsaicin RS nordihydrocapsaicin, nonivamide, decanylvanillinamide, USP Dihydrocapsaicin RS and homocapsaicin. Official https://online.uspnf.com/uspnf/document/1_GUID-1560FD9B-BE0F-495E-9994-C5718733DB4C_2_en-US 2/2.
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