UPLC-MS/MS法同时测定人血浆中氯吡格雷及其代谢物的浓度  

Simultaneous Determination of Clopidogrel and Its Metabolites in Human Plasma by UPLC-MS/MS

作　　者：孙增先[1] 王海东[1] 倪善红[1] 程聪[1] 刘乃丰[2] 

机构地区：[1]连云港市第一人民医院I临床药学中心,江苏连云港222002 [2]东南大学附属中大医院,南京210009

出　　处：《中国药房》2015年第35期4942-4945,共4页China Pharmacy

基　　金：连云港市社会发展计划项目(No.SH1216)

摘　　要：目的：建立同时测定人血浆中氯吡格雷（CLO）及其活性代谢产物（CATM）、非活性代谢产物（CCAM）浓度的方法,并用于药动学研究。方法：取烷化剂2-溴-3′-甲氧基苯乙酮（MPB）保护的血浆样品,经乙腈沉淀蛋白后,以卡马西平为内标,采用超高效液相色谱-串联质谱（UPLC-MS/MS）法测定。色谱柱为Waters ACQUITY UPLC HSS T3,流动相为水（含0.1%甲酸）-乙腈（含0.1%甲酸）,梯度洗脱,流速为0.50 ml/min。采用电喷雾电离源（ESI）,多反应监测（MRM）方式进行正离子监测,用于定量分析的离子对分别为m/z 322.1→211.8（CLO）、m/z 504.1→155.0（CATM烷化衍生物,CATMD）、m/z 308.3→198.0（CCAM）、m/z 273.2→194.3（内标）。结果：CLO、CATMD、CCAM血药浓度分别在0.03-20.00、0.30-200.00、10.00-10 000.00 ng/ml范围内线性关系良好;日内、日间RSD〈15%,相对误差（RE）为-3.5%-5.7。5名健康受试者单剂量口服CLO 300 mg后,CLO、CATM、CCAM的cmax分别为（7.89±5.46）、（15.58±8.08）、（8 023.33±1 047.39）ng/ml,tmax分别为（1.25±0.43）、（1.25±0.43）、（1.67±0.29）h,t1/2分别为（2.31±0.61）、（0.64±0.08）、（6.53±2.55）h,AUC0-t分别为（17.19±14.59）、（21.39±9.58）、（30 648.85±8 026.63）ng·h/ml。结论：该方法操作简便、灵敏度高、分析时间短,适用于CLO及其代谢物血药浓度测定及药动学研究。OBJECTIVE：To establish the method for the simultaneous determination of clopidogrel（CLO） and its active metabolites（CATM）and inactive metabolites（CCAM）,and to conduct pharmacokinetic study. METHODS：The plasma sample had been derivatized by 2-bromine-3′-methoxy acetophenone（MPB）,and was precipitated by acetonitrile. Using carbamazepine as internal standard,UPLC-MS/MS was adopted. The separation was performed on Waters ACQUITY UPLC HSS T3 column with mobile phase consisted of water（containing 0.1% formic acid）-acetonitrile（containing 0.1% formic acid）using a gradient elution program at the flow rate of 0.50 ml/min. The ESI was equipped and quantitative analysis was operated in positive ion and MRM mode. The mass transition ion-pairs were followed as m/z 322.1→211.8（CLO）,m/z 504.1→155.0（the alkylation derivatives of CATM,CATMD）,m/z 308.3→198.0（CCAM）,m/z 273.2→194.3（internal standard）. RESULTS：The linear calibration curves for CLO,CATMD and CCAM were obtained in the concentration range of 0.03-20.00 ng/ml,0.30-200.00ng/ml and 10.00-10 000.00 ng/ml in plasma,respectively;intra-day and inter-day RSD for them were all less than 15%,and relative error（RE）ranged from-3.5% to 5.7%. Main pharmacokinetic parameters of CLO,CATMD and CCAM in 5 healthy volunteers after oral administration of CLO 300 mg were as follows：cmaxwere（7.89±5.46）,（15.58±8.08）,（8 023.33±1 047.39）ng/ml;tmaxwere（1.25 ± 0.43）,（1.25 ± 0.43）,（1.67 ± 0.29）h;t1/2were（2.31 ± 0.61）,（0.64 ± 0.08）,（6.53 ± 2.55）h;AUC0-twere（17.19±14.59）,（21.39±9.58）,（30 648.85±8 026.63）ng·h/ml. CONCLUSIONS：The established method is sensitive,rapid and convenient,which is suitable for pharmacokinetic study and plasma concentration determination of CLO and its metabolites.

关 键 词：氯吡格雷 活性代谢产物 非活性代谢产物 超高效液相色谱-串联质谱法 血药浓度 药动学 

分 类 号：R969.1[医药卫生—药理学]