Development and validation of an HPLC-MS/MS method to determine clopidogrel in human plasma.

Journal: Acta pharmaceutica Sinica. B
Published Date: Dec 15, 2015

Abstract

A quantitative method for clopidogrel using online-SPE tandem LC-MS/MS was developed and fully validated according to the well-established FDA guidelines. The method achieves adequate sensitivity for pharmacokinetic studies, with lower limit of quantifications (LLOQs) as low as 10 pg/mL. Chromatographic separations were performed on reversed phase columns Kromasil Eternity-2.5-C18-UHPLC for both methods. Positive electrospray ionization in multiple reaction monitoring (MRM) mode was employed for signal detection and a deuterated analogue (clopidogrel-d 4) was used as internal standard (IS). Adjustments in sample preparation, including introduction of an online-SPE system proved to be the most effective method to solve the analyte back-conversion in clinical samples. Pooled clinical samples (two levels) were prepared and successfully used as real-sample quality control (QC) in the validation of back-conversion testing under different conditions. The result showed that the real samples were stable in room temperature for 24 h. Linearity, precision, extraction recovery, matrix effect on spiked QC samples and stability tests on both spiked QCs and real sample QCs stored in different conditions met the acceptance criteria. This online-SPE method was successfully applied to a bioequivalence study of 75 mg single dose clopidogrel tablets in 48 healthy male subjects.

Authors

  • Gangyi Liu
    Central Laboratory, Shanghai Clinical Center, Chinese Academy of Sciences/Central Laboratory, Shanghai Xuhui Central Hospital, Shanghai 200031, China.
  • Chunxia Dong
    Central Laboratory, Shanghai Clinical Center, Chinese Academy of Sciences/Central Laboratory, Shanghai Xuhui Central Hospital, Shanghai 200031, China.
  • Weiwei Shen
    Central Laboratory, Shanghai Clinical Center, Chinese Academy of Sciences/Central Laboratory, Shanghai Xuhui Central Hospital, Shanghai 200031, China.
  • Xiaopei Lu
    Central Laboratory, Shanghai Clinical Center, Chinese Academy of Sciences/Central Laboratory, Shanghai Xuhui Central Hospital, Shanghai 200031, China.
  • Mengqi Zhang
    Central Laboratory, Shanghai Clinical Center, Chinese Academy of Sciences/Central Laboratory, Shanghai Xuhui Central Hospital, Shanghai 200031, China.
  • Yuzhou Gui
    State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Science, Shanghai 200233, China.
  • Qinyi Zhou
    Central Laboratory, Shanghai Clinical Center, Chinese Academy of Sciences/Central Laboratory, Shanghai Xuhui Central Hospital, Shanghai 200031, China.
  • Chen Yu
    Central Laboratory, Shanghai Clinical Center, Chinese Academy of Sciences/Central Laboratory, Shanghai Xuhui Central Hospital, Shanghai 200031, China.

Keywords

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