Development and validation of RP-HPLC method for the determination of diphenamid in its formulations

Abstract

A simple, selective, precise and accurate High Performance Liquid Chromatographic method for the analysis of Diphenamid in its formulations was developed and validated in the present study. The mobile phase consist a mixture of 20mM Ammonium Phosphate (adjusted pH 3.0 with OPA) and Methanol in the proportion 60: 40 (v/v). This was found to give sharp peak of Diphenamid at a run time of 10 min. HPLC analysis of Diphenamid was carried out at a wave length of 250 nm with a flow rate of 1.0mL/ min. The linear regression analysis data for the calibration curve showed a good linear relationship with a regression coefficient 0.999 in the concentration range of 50% to 150%. The linear regression equation was y = 2159.x - 136.1. The developed method was employed with a high degree of precision and accuracy for the analysis of Diphenamid. The method was validated for accuracy, precision, robustness, ruggedness and specificity. The Precision, accuracy, sensitivity, short retention time and composition of the mobile phase indicated that this method is useful for the quantification of Diphenamid.