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  • ISSN 1674-8301
  • CN 32-1810/R
Volume 36 Issue 2
Mar.  2022
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Lei Xu, Ran Li, Juan Li, Zhou Dong, Jiaxin Zong, Chuchu Tan, Zekang Ye, Lu Shi, Xiaoxuan Gong, Chunjian Li. Simultaneous determination of clopidogrel, 2-oxo-clopidogrel, and the thiol metabolite of clopidogrel in human plasma by LC-MS/MS[J]. The Journal of Biomedical Research, 2022, 36(2): 109-119. DOI: 10.7555/JBR.36.20210125
Citation: Lei Xu, Ran Li, Juan Li, Zhou Dong, Jiaxin Zong, Chuchu Tan, Zekang Ye, Lu Shi, Xiaoxuan Gong, Chunjian Li. Simultaneous determination of clopidogrel, 2-oxo-clopidogrel, and the thiol metabolite of clopidogrel in human plasma by LC-MS/MS[J]. The Journal of Biomedical Research, 2022, 36(2): 109-119. DOI: 10.7555/JBR.36.20210125
shu

Simultaneous determination of clopidogrel, 2-oxo-clopidogrel, and the thiol metabolite of clopidogrel in human plasma by LC-MS/MS

  • 1.

    Department of Cardiology, the First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210009, China

  • 2.

    Department of Cardiology, the Affiliated Sir Run Run Hospital of Nanjing Medical University, Nanjing Jiangsu 210009, China

  • 3.

    Cardiovascular Center, the Second Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210003, China

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  • Corresponding author:

    Chunjian Li and Xiaoxuan Gong, Department of Cardiology, the First Affiliated Hospital of NanjingMedical University, 300 Guangzhou Road, Nanjing, Jiangsu 210009, China. Tels: +86-25-68303824 and +86-18851727059, E-mails: lijay@njmu.edu.cn and xiaoxuangong@sina.com

  • Received Date: July 25, 2021
  • Revised Date: December 11, 2021
  • Accepted Date: December 30, 2021
  • Available Online: February 27, 2022
    Graphical Abstract
    • Abstract
  • Clopidogrel is a pro-drug which needs two-step metabolism to produce the active thiol metabolite. This study aimed to explore an efficient method to simultaneously determine the plasma clopidogrel, 2-oxo-clopidogrel (2-Oxo-CLP), and the clopidogrel active metabolite (CAM). A high-throughput liquid chromatography tandem mass spectrometry (LC-MS/MS) was therefore developed. The analytes were extracted from plasma by using methyl tert-butyl ether (MTBE). Chromatographic separation was performed on a C18 column under an isocratic elution, accompanied with acetonitrile and deionized water containing 0.1% formic acid. After optimizing the condition of LC-MS/MS, a stable linearity was observed in the standard curves over the concentration ranges of 0.05 to 50.0 ng/mL for clopidogrel, 0.5 to 50.0 ng/mL for 2-Oxo-CLP, and 0.5 to 100 ng/mL for clopidogrel active metabolite derivative (CAMD). The retention time was 4.78 minutes, 3.79 minutes, 3.59 minutes, and 4.82 minutes for clopidogrel, 2-Oxo-CLP, CAMD, and internal standard, respectively. Both the relative standard deviation and the relative error were within the requirement of operating criteria. No significant degradation of clopidogrel, 2-Oxo-CLP, and CAMD occurred under different storage conditions. This method was successfully validated in 3 patients with coronary artery disease. The results showed that the current LC-MS/MS method was efficient for simultaneously detecting clopidogrel, 2-Oxo-CLP, and CAM with fine linearity, accuracy, precision, and stability.
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    • References (26)
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