A Novel RP-HPLC Method Development and Validation for the Estimation of Pyrimethamine in Bulk and its Pharmaceutical Formulation

U. Srinivasulu*¹, N. Devanna², A. Sandhya ³, B. Siva Sai Kiran³, S. Muneer^3
1Associate Professor of Chemistry, SBSYM Degree College, Kurnool-518004, A.P., India
²Professor and Director, Department of Chemistry, JNTUA-Oil Technological and Pharmaceutical Research Institute, Ananthapuramu-515002, A.P., India.
³Department of Pharmaceutical Sciences, JNTUA-Oil Technological and Pharmaceutical Research Institute, Ananthapuramu-515002, A.P., India

A B S T R A C T
A simple, specific, accurate, and precise reverse phase high performance liquid chromatographic (RP-HPLC) method was developed and validated for the estimation of Pyrimethamine in bulk and its pharmaceutical dosage form. The chromatographic separation was done on Phenomenex Luna reversed phase C₁₈ (250 mm x 4.6 mm, 5μm) column in isocratic mode, with mobile phase containing HPLC grade Acetonitrile: methanol: water (0.1 % Orthophosphoric acid) (10:30:60, v/v) was used. The flow rate was 1mL/min and effluents were monitored at 272 nm. Chromatogram of Pyrimethamine was at 2.85 min. The concentration range was 5-30 μg/ml with correlation coefficient 0.999. The method was validated as per ICH guidelines. The limit of detection and Quantitation was found to be 0.87 and 2.65 μg/ml respectively. All the validation parameters were found to be within the predetermined limits. Proposed method was successfully applied for the quantitative determination of Pyrimethamine in its pharmaceutical dosage form.
Keywords: ICH guidelines, Quantitation, Pyrimethamine and predetermined