Development and Validation of RP-HPLC Method for Simultaneous Estimation of Teneligliptin, Pioglitazone Hydrochloride and Metformin Hydrochloride in Bulk and Their Pharmaceutical Dosage Form

Abstract

The pharmaceutical combined dosage forms of teneligliptin, pioglitazone hydrochloride and metformin hydrochloride are used for the treatment of type II diabetes mellitus. A reverse-phase high-performance liquid chromatography method has been developed for simultaneous estimation of teneligliptin, pioglitazone hydrochloride and metformin hydrochloride available in combined tablet dosage form. The method was developed using Kromasil 100 C18 column (150 mm x 4.5 mm x 5 μm), and mobile phase composition of acetonitrile:10 mM phosphate buffer (pH 3.5): methanol in the ratio of (30:50:20 v/v/v). The flow rate was adjusted at 0.8 ml/min for isocratic elution and detection was performed at 238 nm with UV detector. The retention time for metformin hydrochloride, teneligliptin and pioglitazone hydrochloride was found to be 1.74 min, 2.77 min and 5.98 min, respectively. The method was validated as per ICH guidelines. The linearity range was found to be 5-25 μg/ml for teneligliptin and pioglitazone hydrochloride and 150-750 μg/ml for metformin hydrochloride. The percent recovery was found to 101.9% for teneligliptin, 102.6% for pioglitazone hydrochloride and 99.7% for metformin hydrochloride, which indicate that method is accurate. The %RSD was found to be less than 2%, which indicates the developed method is precise. The proposed methods were successfully applied for the quantification of teneligliptin, pioglitazone hydrochloride and metformin hydrochloride in pharmaceutical formulations without any interference from excipients.