High Performance Liquid Chromatographic Determination of Inactive Carboxylic Acid Metabolite of Clopidogrel in Human Serum: Application to a Bioequivalence Study

Authors

  • Amir Farshchi School of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, Iran Department of Pharmocoeconomy and Pharmaceutical Management, Shcool of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran
  • Gholamreza Bahrami School of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, Iran
  • Golbarg Ghiasi School of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, Iran Department of Pharmocoeconomy and Pharmaceutical Management, Shcool of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran
Abstract:

        A sensitive and rapid method is described for determination of clopidogrel carboxylic acid (CCA), the inactive metabolite of the antiplatelet agent clopidogrel in human serum. The analytical procedure involves liquid-liquid extraction of the analyte and an internal standard (phenytoin) with ethyl acetate. A mobile phase consisting of 0.05 M phosphate buffer containing triethylamine (0.5 ml/l; pH 5.7) and acetonitrile (56:44; v/v) was used and chromatographic separation was achieved using a C18 analytical column at detector wavelength of 220 nm. The calibration curves were linear over a concentration range of 0.05-10 μg/ml of CCA in human serum. The total run time of analysis was 5.5 min. and the lower limits of detection (LOD) and quantification (LOQ) were 0.02 and 0.05 μg/ml, respectively. The method validation was carried out in terms of specificity, sensitivity, linearity, precision, accuracy and stability. The validated method was applied in a randomized cross-over bioequivalence study of two different clopidogrel preparations in 24 healthy volunteers.

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Journal title

volume 5  issue 4

pages  231- 238

publication date 2009-10-01

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