New Validated Stability Indicating RP-HPLC Method for Simultaneous Estimation of Metformin and Alogliptin in Human Plasma

Abstract

A validated new stability indicating RP-HPLC method for the quantitative determination of metformin and alogliptin in human plasma was developed as per US-FDA guidelines. The drug was spiked in the plasma and extracted with mobile phase by precipitation method. The extracted analyte was injected into X-Terra C18 (4.6 × 150 mm, 3.5 μm, Make: ACE) or equivalent, maintained at 25°C temperature and effluent was monitored at 235 nm. The mobile phase was consisted of sodium dihydrogen ortho phosphate [pH 4.0]:acetonitrile [HPLC Grade] (70:30 v/v). The flow rate was maintained at 1.0 mL/min. The calibration curve for metformin and alogliptin was linear from 300.0 to 700.0 μg/mL (r2=0.997) and 7.5 to 17.5 μg/mL (r2=0.998) respectively. The inter-day and intra-day precision was found to be within limits. The Lower limit of quantification (LLOQ) for metformin and alogliptin were 5.936 and 1.983 μg/mL respectively. The average % recovery for metformin and alogliptin were 100.17 and 99.40-99.55% respectively and reproducibility was found to be satisfactory. This RP-HPLC method is suitable for determining the concentration of metformin and alogliptin in human plasma and it can applied for routine analysis for determination of the metformin and alogliptin from dosage form during pharmacokinetic study.