Sensitive and Rapid Determination of Gliclazide in Human Plasma by UPLC-MS/MS and its Application to a Bioequivalence Study
Darshan V. Chaudhary, Daxesh P. Patel, Jaivik V. Shah, Mallika Sanyal, Puran Singhal, Pranav S. Shrivastav
Corresponding Author : Pranav S. Shrivastav,
Department of Chemistry, School of Sciences, Gujarat University, Ahmedabad- 380009, India.
Email ID : firstname.lastname@example.org
Received : 2014-09-27 Accepted : 2014-10-28 Published : 2014-10-28
Abstract : An ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method has been developed for the determination of gliclazide in human plasma using gliclazide-d4 as the internal standard (IS). The plasma samples were prepared by protein precipitation with acetonitrile employing 50 ÂµL human plasma. Chromatography was performed on Acquity UPLC BEH C18 (50 mm Ã— 2.1 mm, 1.7 Âµm) analytical column under isocratic conditions using a mobile phase which consisted of 0.1% formic acid in water-acetonitrile (10:90, v/v). The MRM transitions for gliclazide (m/z 324.2 â†’ 127.3) and gliclazide-d4 (m/z 328.2 â†’ 127.4) were monitored on a triple quadrupole mass spectrometer, operating in the positive ionization mode. The method was validated over a dynamic concentration range of 1.0-2000 ng/mL for gliclazide. Matrix effect was assessed by post-column analyte infusion and the mean extraction recovery was 95.7 % across six quality control levels. Stability of gliclazide in plasma was evaluated under different conditions like bench top, auto sampler, dry and wet extract, freeze-thaw and long term stability. The method was applied to a bioequivalence study with 30 mg gliclazide tablet formulation in 28 healthy subjects under fasting. Further, the assay reproducibility was confirmed by reanalysis of 129 incurred samples.
Keywords : UPLC-MS/MS; protein precipitation; sensitive; bioequivalence study
Citation : Pranav S. Shrivastav, et al (2014), Sensitive and Rapid Determination of Gliclazide in Human Plasma by UPLC-MS/MS and its Application to a Bioequivalence Study. J. of Modern Drug Discovery and Drug Delivery Research. V2I1. DOI : 10.5281/zenodo.999187
Copyright : Pranav S. Shrivastav, et al (2014), Sensitive and Rapid Determination of Gliclazide in Human Plasma by UPLC-MS/MS and its Application to a Bioequivalence Study. J. of Modern Drug Discovery and Drug Delivery Research. V2I1.
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