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

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The Journal of Biomedical Research 2022年第2期36卷 109-119页

作者： Lei Xu Ran Li Juan Li Zhou Dong Jiaxin Zong Chuchu Tan Zekang Ye Lu Shi Xiaoxuan Gong Chunjian Li Department of Cardiology the First Affiliated Hospital of Nanjing Medical UniversityNanjingJiangsu 210009China Department of Cardiology the Affiliated Sir Run Run Hospital of Nanjing Medical UniversityNanjing Jiangsu 210009China Cardiovascular Center the Second Affiliated Hospital of Nanjing Medical UniversityNanjingJiangsu 210003China

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.

关键词： clopidogrel drug metabolites liquid chromatography tandem mass spectrometry coronary artery disease

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