Cyanocobalamin.

. IDENTIFICATION • A. ULTRAVIOLET ABSORPTION 〈197U〉 Cupric Sulfate Injection Wavelength range: 200–700 nm Sample solution: Prepare as directed for Sample solution in » Cupric Sulfate Injection is a sterile solution of the Assay. Cupric Sulfate in W ater for Injection. It contains Acceptance criteria: The absorption spectrum exhibits maxima at 278 ± 1 nm, 361 ± 1 nm, and 550 ± 2 nm. The abnot less than 90.0 per cent and not more than sorbance ratio A361/A278 is 1.70–1.90, and the absorbance ra110.0 percent of the labeled amount of copper tio A361/A550 is 3.15–3.40. (Cu). • B. Sample solution: Fuse 1 mg of Cyanocobalamin with 50 Packaging and storage—Preserve in single-dose or multiplemg of potassium pyrosulfate in a por celain crucible. Cool, dose containers, preferably of T ype I or T ype II glass. break up the mass with a glass rod, add 3 mL of water, and Labeling—Label the Injection to indicate that it is to be diluted dissolve by boiling. to the appropriate strength with Sterile W ater for Injection or Analysis: Add 1 drop of phenolphthalein TS, and add soother suitable fluid prior to administration. dium hydroxide solution (100 mg/mL), dropwise, until just pink. Add 500 mg of sodium acetate, 0.5 mL of 1 N acetic USP Reference standards 〈11〉— acid, and 0.5 mL of nitroso R salt solution (2 mg/mL). USP Endotoxin RS Acceptance criteria: A red or orange-red color appears at Identification—The Assay preparation, prepared as directed in once. Add 0.5 mL of hydrochloric acid, and boil for 1 min: the Assay, exhibits an absorption maximum at about 325 nm the red color persists. when tested as directed for Procedure in the Assay. • C. Bacterial endotoxins 〈85〉—It contains not more than 250.0 Sample solution: Dissolve 5 mg of Cyanocobalamin in 5 USP Endotoxin Units per mg of copper. mL of water in a 50-mL distilling flask connected with a short, water-cooled condenser. To the flask add 2.5 mL of pH 〈791〉: between 2.0 and 3.5. hypophosphorous acid, close, and boil gently but short of Particulate matter 〈788〉: meets the requirements for distillation for 10 min. Then distill 1 mL into a test tube small-volume injections. containing 1 mL of sodium hydroxide solution (20 mg/mL). Other requirements—It meets the requirements under InjecAnalysis: To the test tube add 4 drops of cold saturated tions 〈1〉. ferrous ammonium sulfate solution, shake gently, then add Assay— 30 mg of sodium fluoride, and bring the contents to a boil. Sodium chloride solution, Copper stock solution, and Standard Immediately add, dropwise, 5 N sulfuric acid until a clear preparations—Prepare as directed in the Assay under Cupric solution results, then add 3–5 drops more of the acid. Chloride Injection. Acceptance criteria: A blue or blue-green color develops within a few min. Assay preparation—Transfer an accurately measured volume of Injection, equivalent to about 2 mg of copper, to a 100-mL ASSAY volumetric flask, dilute with water to volume, and mix. Pipet 15 • PROCEDURE mL of this solution into a 100-mL volumetric flask. From the Standard solution: 30 μg/mL of cyanocobalamin from USP labeled amount of sodium chloride, if any, in the Injection, calCyanocobalamin RS in water culate the amount, in mg, of sodium chloride in the initial diluSample solution: 30 μg/mL of Cyanocobalamin in water tion, and add sufficient Sodium chloride solution to bring the Instrumental conditions total sodium content of this flask to 13.5 mg. Dilute with water (See Spectrophotometry and Light-Scattering 〈851〉.) to volume, and mix. Mode: UV Procedure—Proceed as directed for Procedure in the Assay Analytical wavelength: 361 nm under Cupric Chloride Injection. Cell: 1 cm Blank: Water Analysis Samples: Standard solution and Sample solution