Development and validation of HPLC method for the determination of pioglitazone in human plasma

Abstract

Pioglitazone is widely used for the management of type–II diabetes mellitus. The objective of the present study was to develop a simple and cost-effective HPLC method for the quantification of pioglitazone in human plasma. The mobile phase comprises of Acetonitrile, 0.1 M ammonium acetate and glacial acetic acid (25:25:1 v/v/v) at a flow rate of 1.2 mL/min., using Macherey-Nagel Column C18, (dimensions: 5 μm; 250 × 4.6mm) with a guard column. The UV detector was set at 269nm. The method was validated according to FDA guidelines. The present method showed good linearity (R2=0.9998) from 0.1 to 2.0µg/ml standards, with a limit of detection 0.1 µg/ml. Intra-day accuracy and precision in terms of %CV (range: 93.33% to 100.4% and 3.8% to 9.2%) and interday accuracy and precision (range: 94.1% to 102.7% and 4.8% to 9.6%) were in agreement with FDA guidelines. Freeze thaw stability showed that the plasma samples could be stored for one month at -20oC without any appreciable degradation. The present method was successfully applied to the blood samples obtained from one volunteer after oral administration of 30 mg pioglitazone tablet. Some preliminary pharmacokinetic parameters were calculated. It is concluded that the present method could be conveniently used for the routine analysis of pioglitazone blood samples obtained in pharmacokinetics studies.