DEVELOPMENT AND VALIDATION OF A RP-HPLC METHOD FOR THE DETERMINATION OF ZIDOVUDINE AND RELATIVE SUBSTANCES IN BULK

A reversed- phase High Performance Liquid Chromatography (RP-HPLC) method for the rapid and accurate quantification of zidovudine (AZT) and related substances. The detection was carried out by Shimadzu RP-C18 ODS (250×4.6mm, 5µm) column using mixture of water and Methanol (70:30) with a flow rate of 1ml/min. The eluents were detected at 270nm and the Retention time for zidovudine, zidovudine B and C impurity was found to be 9.5, 11.3 and 3.5 min. A linear responses for zidovudine and impurities B and C were 5-25 µg/ml, 1-5 µg/ml with correlation factor of 0.998, 0.998 and 0.997 respectively. The method was validated as per ICH guidelines for specificity, linearity, accuracy and robustness. The validated method was successfully employed for the simultaneous determination zidovudine (AZT) and related substances. Keywords: zidovudine (AZT), impurities B and C, RP-HPLC anti-viral