Development and Validation of High Performance Liquid Chromatography Assay Method of Phenytoin in Tablet Dosage Form

Abstract

A new, precise and accurate reversed-phase high performance liquid chromatography (RP-HPLC) method was developed for determination of phenytoin. Method was carried out by isocratic technique on a reverse-phase HPLC using a column of C8 inertstil (4.6 x 250) mm, 5µm column with a mobile phase mixture of 500cm3 water, 400cm3 absolute methanol and 100cm3 acetonitrile. The flow rate was adjusted at 2cm3/min, detection wavelength at 235 nm, temperature at 40°C. The retention time was found to be 4.8min. The results of analysis have been validated statistically and recovery studies confirmed the accuracy of the developed method according to ICH guidelines (ICH Q2 R1, 2005).The assay percentage of phenytoin in tablet dosage form was found to be (100.79± 1.067) % .The % of recovery was found to be (99.6 – 102.5) %. The limit of detection (LOD) and limit of quantization (LOQ) were found to be 0.00040755mg/cm3 and 0.001235mg/cm3 respectively.