Development and Validation of RP-HPLC Method for the Simultaneous Estimation of Cefadroxil and Probencid in Bulk and Tablet

A reverse phase high performance liquid chromatographic method was developed for the simultaneous estimation of Cefadroxil (CEF) and Probenecid (PROB) in bulk and tablet dosage form. The separation was achieved by Princeton SPHER-100 C18 column of configuration was selected for further study. (Princeton SPHER-100 C₁₈, 250×4.6 mm, particle size 5 μm.). CEF is freely soluble in methanol and slightly soluble in acetonitrile, water and insoluble in ethanol. Probencid is freely soluble in methanol, slightly soluble in acetonitile, water. Initially methanol and acetronitrile in different ratios were tried and then acetonitril and phosphate buffer (PH 2.5 adjusted with orthophosphoric acid). It was found that acetonitril and phosphate buffer (PH 2.5 adjusted with orthophosphoric acid). In the ratio 60:40 v/v with the flow rate of 1mL/min gives good resolution of peaks with minimum tailing as compared to other mobile phases. Detection was carried out at 242 nm. Retention time of Cefadroxil and Probenecid was found to be 2.213 and 6.62 min, respectively. The method has been validated for linearity, accuracy and precision. Linearity for Cefadroxil was found in the range of 10- 60μg/ml and Probenecid was in the range of 10-60 μg/ml. The percentage recoveries obtained for Cefadroxil and Probenecid were found to be in range of 99.52-99.81 % and 99.50-99.53% respectively.