Development and Validation of HPLC Method for the Estimation of Metformin HCL and Anagliptin in its Synthetic Mixture

Abstract

The goal of the current study was to develop and validate a reversed-phase high-performance liquid chromatography method using a C18 (250 × 4.6mm, 5μm) column for the measurement of metformin hydrochloride and anagliptin in bulk medication samples. The method is simple, quick, accurate, precise, selective and reproducible. A mobile phase of buffer and acetonitrile (ACN) (pH 3.0) 80:20 v/v was used to elute 20 μL injection volume at a flow rate of 1.0ml/min. UV detection at 232 nm for metformin hydrochloride and 247 nm for anagliptin was applied. There was good separation between the two components as seen by the retention time of anagliptin and metformin which were 5.51 and 2.38 minutes respectively. In compliance with International Conference on Harmonization (ICH) criteria, the procedure was verified for linearity, accuracy, precision, robustness, limit of detection (LOD), and limit of quantification (LOQ) as well as system suitability. Metformin and anagliptin had linear ranges of 40-160 μg/mL and 8-32 μg/mL, respectively, with recoveries in the range 101-110%. The LOD and LOQ for metformin hydrochloride were 0.96 and 2.91, and for anagliptin were 0.71 and 2.14, respectively. The proposed method may be used for routine quality control analyses of tablets and bulk.