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

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

机构地区：[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

出　　处：《The Journal of Biomedical Research》2022年第2期109-119,共11页生物医学研究杂志（英文版）

基　　金：This work was supported by National Natural Science Funding of China(Grant No.81170181);the Jiangsu Province's Key Provincial Talents Program(Grant No.ZDRCA2016013);the Second Level of 333 High Level Talent Training Project in Jiangsu Province(Grant No.BRA2019099);Special Fund for Key R&D Plans(Social Development)of Jiangsu Province(Grant No.BE2019754);a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutes.

摘　　要：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 

分 类 号：O657.63[理学—分析化学] R969.1[理学—化学]