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 

作者机构：Department of Cardiologythe First Affiliated Hospital of Nanjing Medical UniversityNanjingJiangsu 210009China Department of Cardiologythe Affiliated Sir Run Run Hospital of Nanjing Medical UniversityNanjing Jiangsu 210009China Cardiovascular Centerthe Second Affiliated Hospital of Nanjing Medical UniversityNanjingJiangsu 210003China 

出 版 物：《The Journal of Biomedical Research》 (生物医学研究杂志（英文版）)

年 卷 期：2022年第36卷第2期

页      面：109-119页

核心收录：

学科分类：1006[医学-中西医结合] 07[理学] 1001[医学-基础医学(可授医学、理学学位)] 070302[理学-分析化学] 0703[理学-化学] 10[医学] 100602[医学-中西医结合临床] 

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. 

主　　题：clopidogrel drug metabolites liquid chromatography tandem mass spectrometry coronary artery disease 

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.