A Simple and Rapid method to Determine Pioglitazone from Human Plasma by Liquid Chromatography Tandem Mass Spectrometry

Abstract

A simple, rapid, specific and sensitive liquid chromatography tandem mass spectrometric method has been developed and validated for the estimation of Pioglitazone from 100µL of human plasma. Pioglitazone is extracted from human plasma by Liquid Liquid Extraction. Nevirapine was used as an internal standard. Detection was performed using API 2000 PE SCIEX with Electron Spray Ionisation (ESI) source in positive polarity. The detection transition for Pioglitazone is 357.100→ 134.100 and for Nevirapine is 267.100 → 226.100 (Fig A and B). Chromatographic separation of analyte and internal standard were carried out using a reverse phase KROMASIL, C18, 5µM (4.5x150mm) at a flow rate of 0.500mL/min. The mobile Phase is composed of Acetonitrile: 10mM Ammonium Acetate (90:10%v/v). The assay of Pioglitazone is linear over the range of 18.058ng/ml to 1099.99ng/ml (Fig C) with a precision <8.78% and the limit of quantification in plasma for Pioglitazone was 18.058ng/mL. Mean extraction recovery obtained was 96.94%. Samples are stable at room temperature for 12 hrs, processed samples were stable at least for 12 hrs and also stable at four freeze–thaw cycles. The method has been used to perform pharmacokinetic and bioequivalence studies in humans.