Development and validation of RP-HPLC method for simultaneous estimation of metformin hydrochloride and glipizide in bulk and pharmaceutical dosage form

Abstract

A simple, accurate, economical and precise reverse phase high performance liquid chromatographic (RP-HPLC) method has been developed for simultaneous estimation of Metformin hydrochloride and Glipizide in bulk and pharmaceutical dosage form. The separation of Metformin hydrochloride and Glipizide was achieved by using Cosmosil C 18 (250 mm × 4.6 ID, particle size-5 micron) column as stationary phase with the mobile phase comprising of methanol and water (60:40 ,pH 3 adjusted with ortho-phosphoric acid) in an isocratic mode and flow rate of 0.8 ml/min with UV detection at 226 nm. The retention time of Metformin hydrochloride and Glipizide were found to be 4.159 min and 5.571 min respectively. The proposed method was validated according to ICH guidelines for the parameters like linearity, accuracy, precision, percent recovery, robustness, ruggedness, limit of detection and limit of quantitation. The % RSD is found to be less than 2 % and the tailing factor for both the drugs are found to be less than 2. The number of therotical plates for Metformin hydrochloride and Glipizide were found to be more than 2000.All validation parameters were within the acceptable range. The developed method was successfully applied for the estimation of Metformin hydrochloride and Glipizide in bulk and pharmaceutical dosage forms. Keywords: Metformin hydrochloride, Glipizide, RP-HPLC, Validation, Simultaneous estimation, ICH guidelines.