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Ascorbic Acid Injection
Assay¡ª
Mobile phase¡ª Dissolve 15.6g of dibasic sodium phosphate and 12.2g of monobasic potassium phosphate in 2000mLof water,adjust with phosphoric acid to a pHof 2.5¡À0.05.Make adjustments if necessary (see System Suitabilityunder Chromatography ¨¢621ñ.
Standard preparation¡ª Dissolve an accurately weighed quantity of USP Ascorbic Acid RSin Mobile phase,and mix to obtain a solution having a known concentration of about 0.5mg per mL.[NOTE¡ªRefrigerate and store protected from light until use.The solution is stable for at least 24hours.Inject within 3hours after removal from the refrigerator.]
Assay preparation¡ª Dilute the Injection,quantitatively,and stepwise if necessary,with Mobile phaseto obtain a solution having a concentration of about 0.5mg per mL.[NOTE¡ªRefrigerate and store protected from light until use.The solution is stable for at least 24hours.Inject within 3hours after removal from the refrigerator.]
Chromatographic system (see Chromatography ¨¢621ñ¡ªThe liquid chromatograph is equipped with a 245-nm detector and a 6-mm ¡Á150-mm column that contains packing L39.The flow rate is about 0.6mLper minute.Chromatograph the Standard preparation,and record the peak responses as directed for Procedure:the column efficiency is not less than 3500theoretical plates,the tailing factor is not more than 1.6,and the relative standard deviation for replicate injections is not more than 1.5%.
Procedure¡ª Separately inject equal volumes (about 4µL)of the Standard preparationand the Assay preparationinto the chromatograph,record the chromatograms,and measure the responses for the major peak.Calculate the quantity,in mg,of ascorbic acid (C6H8O6)in each mLof the Injection taken by the formula:
CD(rU/rS),

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Make adjustments if necessary (see System Suitability under Chromatography)
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Separately inject equal volumes (about 4µL)of the Standard preparation and the Assay preparationinto the chromatograph
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