A Validated RP-HPLC Method for Estimation of Hepatitis B Surface Antigen (Hbsag) in Bulk Preparation
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A Simple Selective, rapid, precise and accurate reverse phase high pressure liquid chromatographic method has been developed for the estimation of Hepatitis B surface antigen (HBsAg) in pharmaceutical formulation. The method was carried out on a Gracesmart C18 5 micron (250 mm x 4.6mm i.d.) column with mobile phase consisting of 5 mmol potassium dihydrogen phosphate: Methanol (50: 50 v/v) and pH was unadjusted. The flow rate was kept 1.0 mL/min. Detection was carried out at 280 nm. The retention time of HBsAg was found to be 7.80 min. The developed method was validated for linearity, accuracy, precision (inter day and intra day), limit of detection and limit of quantitation.
KEYWORDS:Hepatitis B; RP-HPLC; HBsAg; Surface antigen
