Thiamine Mononitrate (thye' a min mon'' oh nye' trate). C12H17N5O4S327.36 Thiazolium, 3-[(4-amino-2-methyl-5-pyrimidinyl)methyl]-5-(2-hydroxyethyl)-4-methyl-, nitrate (salt); Thiamine nitrate (salt) [532-43-4]. DEFINITION Thiamine Mononitrate contains NLT 98.0% and NMT 102.0% of thiamine mononitrate (C12H17N5O4S), calculated on the dried basis. IDENTIFICATION • A. Sample solution: 20 mg/mL of Thiamine Mononitrate Analysis: To 2 mL of the Sample solution add 2 mL of sulfuric acid. Cool, and superimpose 2 mL of ferrous sulfate TS. Acceptance criteria: A brown ring is produced at the junction of the two liquids. • B. Sample: 5 mg Analysis: Dissolve the Sample in a mixture of 1 mL of lead acetate TS and 1 mL of 2.5 N sodium hydroxide. Heat for several min on a steam bath. Acceptance criteria: After dissolution of the Sample, a yellow color is produced. After heating the solution, the color changes to brown and on standing, a precipitate of lead sulfide separates. • C. A solution of Thiamine Mononitrate yields a white precipitate with mercuric chloride TS, and a red-brown precipitate with iodine TS. It also yields a precipitate with mercuricpotassium iodide TS, and with trinitrophenol TS. • D. Sample solution: Dissolve 5 mg in 5 mL of 0.5 N sodium hydroxide. Analysis: To the Sample solution add 0.5 mL of potassium ferricyanide TS and 5 mL of isobutyl alcohol. Shake the mixture vigorously for 2 min, and allow the liquid layers to separate. Acceptance criteria: When illuminated from above by a vertical beam of UV light and viewed at a right angle to this beam, the airliquid meniscus shows a vivid blue fluorescence, which disappears when the mixture is slightly acidified, but reappears when it is again made alkaline. ASSAY • Procedure Solution A: 0.005 M sodium 1-octanesulfonate in dilute glacial acetic acid (1 in 100) Solution B: Methanol and acetonitrile (3:2) Mobile phase: Solution A and Solution B (60:40) Internal standard solution: 2.0% (v/v) of methylbenzoate in methanol Standard solution: Prepare a 1-mg/mL solution of USP Thiamine Hydrochloride RS in Mobile phase. Transfer 20.0 mL of this solution and 5.0 mL of Internal standard solution to a 50-mL volumetric flask, and dilute with Mobile phase to volume. The Standard solution contains 400 µg/mL of thiamine hydrochloride. Sample solution: Prepare a 2-mg/mL solution of Thiamine Mononitrate in Mobile phase. Transfer 10.0 mL of this solution and 5.0 mL of Internal standard solution to a 50-mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system Mode: LC Detector: UV 254 nm Column: 4-mm × 30-cm; packing L1 Flow rate: 1 mL/min. [NoteThe flow rate may be adjusted as needed to obtain a retention time of about 12 min for thiamine hydrochloride. ] Injection size: 10 µL System suitability Sample: Standard solution Suitability requirements Resolution: NLT 4.0 between the thiamine and methylbenzoate peaks Tailing factor: NMT 2.0 for the thiamine peak Column efficiency: NLT 1500 theoretical plates for the thiamine peak Relative standard deviation: NMT 2.0% for the ratios of the thiamine peak area to the internal standard peak area Analysis Samples: Standard solution and Sample solution Calculate the percentage of thiamine mononitrate (C12H17N5O4S) in the portion of Thiamine Mononitrate taken: Result = (RU/RS) × (CS/CU) × (Mr1/Mr2) × 100
Acceptance criteria: 98.0%102.0% on the dried basis IMPURITIES • Residue on Ignition 281: NMT 0.2% • Chloride and Sulfate, Chloride 221 Standard: 0.40 mL of 0.020 N hydrochloric acid Sample: 500 mg of Thiamine Mononitrate Acceptance criteria: NMT 0.06% • Related Compounds Solution A, Solution B, and Mobile phase: Proceed as directed in the Assay. Sample solution: 1.0 mg/mL of Thiamine Mononitrate in Mobile phase Chromatographic system Mode: LC Detector: UV 254 nm Column: 4.0-mm × 15-cm; packing L1 Flow rate: 0.75 mL/min Injection size: 10 µL Analysis Sample: Sample solution Allow the Sample solution to elute for NLT three times the retention time of the main peak. Calculate the percentage of total secondary peaks in the portion of Thiamine Mononitrate taken: Result = (rU/rT) × 100
Acceptance criteria: NMT 1.0% SPECIFIC TESTS • Loss on Drying 731: Dry 500 mg at 105 for 2 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS • Packaging and Storage: Preserve in tight, light-resistant containers. Auxiliary Information Please check for your question in the FAQs before contacting USP.
USP35NF30 Page 4837 Chromatographic Column Chromatographic columns text is not derived from, and not part of, USP 35 or NF 30. |