Development and Validation of Novel RP-HPLC Method for the Simultaneous Estimation of Itraconazole and Secnidazole in Bulk and Pharmaceutical Formulations

Simple, accurate and precise RP-HPLC method for the simultaneous estimation of Itraconazole (ITZ) and Secnidazole (SEC) in bulk and pharmaceutical formulations has been developed and validated. The chromatographic analysis was performed on Shimadzu model equipped with SPD-20 AD UV detector, isocratic pump LC- 20 AD and loop injector. The chromatographic separation was performed using C18 column (250mm x 4.6mm i.d. and 5µm particle size). The analytes were monitored at 257nm using methanol: potassium dihydrogen phosphate buffer (PDP) pH 5.5 in the ratio of 90:10v/v with a flow rate of 1.5ml/min. The retention time was found to be 6.037 min for ITZ and 3.788 min for SEC. The linearity was found to be in the range of 10-50µg/ml for both ITZ and SEC with a regression coefficient of 0.999 and 0.998 for ITZ and SEC respectively. The method was validated in accordance with ICH guidelines for its linearity, precision, accuracy, robustness, ruggedness, LOD and LOQ. The recovery studies were carried out which confirmed the accuracy of the proposed method. LOD values for ITZ & SEC was found to be 0.583µg/ml and 0.241µg/ml, respectively. LOQ values for ITZ and SEC was found to be 1.749µg/ml and 0.732µg/ml, respectively. The developed method was also found to be precise and robust for the simultaneous determination of ITZ and SEC in bulk and pharmaceutical formulations.