Method Development and Validation of Metformine, Pioglitazone and Glibenclamide in Tablet Dosage Form by using RP-HPLC

Abstract

Reverse phase high performance liquid chromatographic method has developed for the simultaneous estimation of metformine, pioglitazone and glibenclamide in tablet dosage for using gliclazide as an internal standard. The separation was achieved at ambient temperature with low pressure gradient mode by using 5 μ size phenomenex luna CN (100R 250×4.60 (mm) column with mobile phase containing acetonitrile, water and buffer (0.5% potassium dihydrogen phosphate) pH 2.5 adjusted with orthophosphoic acid in the ratio of 60:20:20. The flow rate was 1 mL min-1 and eluent was monitored at 230 nm by using UV detector. The selected chromatographic conditions effectively separated Metformine, pioglitazone and Glibenclamide with the retention time of 2.2, 2.8 and 5.8 min respectively. The linearity range for metformine, pioglitazone and glibenclamide is found in the range of 50-300 μg mL-1, 1.5-9.0 μg mL-1 and 0.5- 3.0 μg mL-1 respectively. The developed method was found to be accurate, simple, specific and reproducible. It can also be used for routine quality control analysis of these antidiabetic drugs in combinational dosage forms.