Stability indicating RP-HPLC method for simultaneous determination of pyrimethamine and sulfamethoxypyrazine in pharmaceutical formulation: Application to method validation

Abstract

The present work takes into account the development of Reverse Phase High Performance Liquid Chromatography (HPLC) for simultaneous method estimation and validation of pyrimethamine and sulfamethoxypyrazine in pharmaceutical formulation. The chromatographic separation was accomplished on C8 column by using acetonitrile and potassium dihydrogen phosphate as the mobile phase (60:40 v/v) having a flow rate of 0.8 ml/minute. The eluent was detected at 254 nm, simultaneously for both the drugs. The retention time for pyrimethamine and sulfamethoxypyrazine was found to be 3.33 and 4.21 minutes, respectively. According to the International Conference on Harmonisation guidelines, the develop method was validated in terms of accuracy, precision, linearity, limit of detection, limit of quantitation, robustness, and stress degradation studies. This validated method can be suggested for the routine simultaneous laboratory analysis of pyrimethamine and sulfamethoxypyrazine.