Novel liquid chromatographic method for simultaneous estimation of pioglitazone and glimepiride in rat plasma by solid phase extraction: Application to preclinical pharmacokinetic studies

Prashant B. Musmade, Kranti B. Talole, Praful B. Deshpande, Arumugam Karthik, Shriram M. Pathak, Sureshwar Pandey, Nayanabhirama Udupa

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The need for a reliable bioanalytical method is of primary importance during preclinical studies. The aim of the present study was simultaneous determination of pioglitazone (CAS 111025-46-8) (PIO) and glimepiride (CAS 93479-97-1) (GLM) in plasma of rats. A high-performance liquid chromatographic method has been developed and validated using C18 column and UV detector. A mobile phase composed of acetonitrile and ammonium acetate buffer pH 4.5 in the ratio of 55:45%. The plasma samples clean-up was carried out using solid phase cartridges. The method was in the linear range of 50-8000 ng/mL for PIO and 50-2000 ng/mL for GLM. The coefficient of regression was found to be ≥0.99. Precision and accuracy were within the acceptable limits, as indicated by relative standard deviation varying from 1.5 to 6.1% for PIO and 3.1 to 7.0% for GLM whereas the accuracy ranged from 97.0 to 106.4% for PIO and 96.5 to 106.4% for GLM. The mean extraction recovery was found to be 90.2 ± 4.5, 76.8 ± 2.8 and 85.2 ± 5.2% for PIO, GLM and internal standard, respectively. Moreover, PIO and GLM were stable in plasma, up to 30 days of storage at -70 °C and after being subjected to bench top, autosampler, and three freeze-thaw cycles. The developed method was applied for preclinical pharmacokinetic studies.

Original languageEnglish
Pages (from-to)23-31
Number of pages9
JournalArzneimittel-Forschung/Drug Research
Issue number1
Publication statusPublished - 2011


All Science Journal Classification (ASJC) codes

  • Drug Discovery

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