Ascorbic Acid Injection
»Ascorbic Acid Injection is a sterile solution,in Water for Injection,of Ascorbic Acid prepared with the aid of Sodium Hydroxide,Sodium Carbonate,or Sodium Bicarbonate.It contains not less than 90.0percent and not more than 110.0percent of the labeled amount of ascorbic acid (C6H8O6).
Packaging and storage— Preserve in light-resistant,single-dose containers,preferably of Type Ior Type IIglass.
Labeling— In addition to meeting the requirements for Labelingunder Injections á1ñ,fused-seal containers of the Injection in concentrations of 250mg per mLand greater are labeled to indicate that since pressure may develop on long storage,precautions should be taken to wrap the container in a protective covering while it is being opened.
Identification—
A: To a volume of Injection,equivalent to 40mg of ascorbic acid,add 4mLof 0.1Nhydrochloric acid,then add 4drops of methylene blue TS,and warm to 40:the deep blue color becomes appreciably lighter or is completely discharged within 3minutes.
B: The retention time of the major peak in the chromatogram of the Assay preparationcorresponds to that of the Standard preparation,obtained as directed in the Assay.
C: It responds to the flame test for Sodium á191ñ.
Bacterial endotoxins á85ñ— It contains not more than 1.2USP Endotoxin Units per mg of ascorbic acid.
pHá791ñ: between 5.5and 7.0.
Limit of oxalate— Dilute a volume of Injection,equivalent to 50mg of ascorbic acid,with water to 5mL.Add 0.2mLof acetic acid and 0.5mLof calcium chloride TS:no turbidity is produced in 1minute.
Other requirements— It meets the requirements under Injections á1ñ.
Assay—
Mobile phase— 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±0.05.Make adjustments if necessary (see System Suitabilityunder Chromatography á621ñ).
Standard preparation— 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—Refrigerate 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— Dilute the Injection,quantitatively,and stepwise if necessary,with Mobile phaseto obtain a solution having a concentration of about 0.5mg per mL.[NOTE—Refrigerate 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 á621ñ)—The liquid chromatograph is equipped with a 245-nm detector and a 6-mm ×150-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— 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),
in which Cis the concentration,in mg per mL,of USP Ascorbic Acid RSin the Standard preparation;Dis the dilution factor;and rUand rSare the peak responses obtained from the Assay preparationand the Standard preparation,respectively.
Auxiliary Information— Staff Liaison:Lawrence Evans,III,Ph.D.,Scientist
Expert Committee:(DSN)Dietary Supplements:Non-Botanicals
USP28–NF23Page 178
Phone Number:1-301-816-8389