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 inpharmaceutical formulation. The chromatographic separation was accomplished on C8 column by using acetonitrileand potassium dihydrogen phosphate as the mobile phase (60:40 v/v) having a flow rate of 0.8 ml/minute. Theeluent was detected at 254 nm, simultaneously for both the drugs. The retention time for pyrimethamine andsulfamethoxypyrazine was found to be 3.33 and 4.21 minutes, respectively. According to the International Conferenceon Harmonisation guidelines, the develop method was validated in terms of accuracy, precision, linearity, limit ofdetection, limit of quantitation, robustness, and stress degradation studies. This validated method can be suggested forthe routine simultaneous laboratory analysis of pyrimethamine and sulfamethoxypyrazine.