RP-HPLC Method Development and Validation for Quantitative Analysis of Metformin and Gliclazide in Pharmaceutical Formulations

Abstract

A rapid and sensitive isocratic reversed phase high performance liquid chromatographic method has been developed for quantitative analysis of metformin hydrochloride and gliclazide in bulk as well as pharmaceutical dosage forms. The method was validated according to FDA, ICH and USP guidelines with respect to accuracy, precision, specificity and linearity. For metformin, the method was developed by using the mobile phase comprising of sodium dihydrogen phosphate solution (NaH2PO4) (pH 3.0) and acetonitrile (90:10, v/v) at a flow rate of 0.5 mL/min over C18 bonded silica column (3.9 x 300 mm, 5 µm) at ambient temperature. On the other hand, the same mobile phase was used in the ratio of 20:80 (v/v) at a flow rate of 0.6 mL/min for gliclazide. The recovery was found to be more than 97% for metformin and 102% for gliclazide that demonstrated the accuracy of the protocol. Intraday and interday precisions of the new method were less than the maximum allowable limit (%RSD 2.0) specified by the USP, ICH and FDA. The method revealed linear response with correlation coefficient value of 0.999 in both cases.