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Abstract

SIMULTANEOUS DETERMINATION OF ALOGLIPTINE, PIOGLITAZONE AND ITS ACTIVE METABOLITE IN HUMAN PLASMA BY LC-MS/MS METHOD AND ITS PHARMACOKINETIC APPLICATION

Vinutha K., K.P.R. Chowdary* and S.V.U.M. Prasad

ABSTRACT

A simple, rapid and sensitive liquid chromatography-tandem mass spectrometric (LC–MS/ MS) assay method has been developed and fully validated for the simultaneous quantification of Alogliptine, Pioglitazone and Hydroxy Pioglitazone. The analytes were extracted from human plasma via protein precipitation using acetonitrile. The reconstituted samples were chromatographed on a Alltima HP C18 column by using a 60:40 (v/v) mixture of acetonitrile and 10 mM ammonium acetate (pH 3.0) as the mobile phase at a flow rate of 1.1 mL/min. The calibration curves obtained were linear over the concentration range of 3.05-250.29 ng/mL for Alogliptine, 15–2500.50 ng/mL for Pioglitazone and 7–1500 ng/mL for Hydroxy Pioglitazone. The API-4000 LC–MS/MS in multiple reaction monitoring (MRM) mode was used for detection. The results of the intra- and inter-day precision and accuracy studies were well within the acceptable limits. All the analytes were found to be stable in a battery of stability studies. The method is precise and sensitive enough for its intended purpose. The developed assay method was successfully applied to a pharmacokinetic study in human volunteers.

Keywords: Alogliptine, Pioglitazone, Hydroxy Pioglitazone, Human plasma, LC-MS/MS, pharmacokinetic study.


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