Development and validation of the liquid chromatographic method for simultaneous estimation of metformin, pioglitazone, and glimepiride in pharmaceutical dosage forms

Abstract

Introduction: A simple, precise, and accurate HPLC method for simultaneous estimation of metformin hydrochloride (MET), pioglitazone hydrochloride (PIO), and glimepiride (GLIMP) was developed and validated. Materials and Methods: Chromatographic separation of the drugs was performed by using a Phenomenex-ODS-3 (C-18) column (250 × 4.60 mm, 5 μm) with a mobile phase consisting of methanol:acetonitrile:15 mM potassium dihydrogen phosphate (pH 4) in the proportion of 40:35:25 (v/v) at a flow rate of 1 ml/min. Detection was carried out using a UV-SPD-10AVP detector at 240 nm. Results: The retention time for MET, PIO, and GLIMP were 2.85 ± 0.03 min, 4.52 ± 0.03 min, and 7.08 ± 0.02min, respectively. Parameters such as linearity (0.2–50 μg/ml for MET, 0.2-30 μg/ml for PIO, and GLIMP, respectively), precision (intra-day % RSD was 1.01-3.24 and inter-day % RSD was 1.54-4.09 for MET; intra-day % RSD was 1.03-2.09 and inter-day % RSD was 2.26-3.10 for PIO; and intra-day% RSD was 1.00-3.15 and inter-day % RSD was 1.58-3.07 for GLIMP), accuracy (99.66 ± 0.14 for MET, 98.46 ± 0.40 for PIO, and 98.62 ± 0.39 for GLIMP), specificity and robustness were calculated in accordance with ICH guidelines. Conclusions: The method was proved to be simple, rapid, precise, accurate, and cost effective.