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

Lei Xu; Ran Li; Juan Li; Zhou Dong; Jiaxin Zong; Chuchu Tan; Zekang Ye; Lu Shi; Xiaoxuan Gong; Chunjian Li

doi:10.7555/JBR.36.20210125

Volume 36 Issue 2

Mar. 2022

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The Journal of Biomedical Research > 2022 > 36(2): 109-119. > DOI: 10.7555/JBR.36.20210125

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

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

Lei Xu^{1, 2},
Ran Li¹,
Juan Li^{1, 3},
Zhou Dong¹,
Jiaxin Zong¹,
Chuchu Tan¹,
Zekang Ye¹,
Lu Shi¹,
Xiaoxuan Gong^1, ,,
Chunjian Li^1, ,

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

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.
- clopidogrel,
- drug metabolites,
- liquid chromatography tandem mass spectrometry,
- coronary artery disease

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References (26)

References

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Corresponding author: Chunjian Li, lijay@njmu.edu.cn

1.
Department of Cardiology, the First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210009, China

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