HPLC-MS/MS method for the simultaneous determination of clopidogrel, its carboxylic acid metabolite and derivatized isomers of thiol metabolite in clinical samples
HPLC-MS/MS method for the simultaneous determination of clopidogrel, its carboxylic acid metabolite and derivatized isomers of thiol metabolite in clinical samples
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DOI：
10.1016/j.jchromb.2012.11.005
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发表时间：
2012-12-12
影响因子：
3
通讯作者：
Glowka, Franciszek
中科院分区：
文献类型：
--
作者：
Karazniewicz-Lada, Marta;Danielak, Dorota;Glowka, Franciszek
建立了同时测定氯吡格雷（CLP）及其羧酸衍生物（CLPM）的高效液相色谱-串联质谱（HPLC-MS/MS）法。衍生硫醇代谢物异构体MP-H3和活性MP-H4。采用乙腈简单蛋白沉淀法从血浆样品中提取CLP、CLPM、MP-H3和MP-H4异构体以及内标吡罗昔康。在HPLC Zorbax Plus C18柱上，通过以水和乙腈（均含有0.1%（v/v）甲酸）进行梯度洗脱来分离分析物。在三重四极杆MS上进行分析物的检测，通过电喷雾离子化进行多反应监测。发现在250 μ L血浆中制备的分析物的校准曲线在以下范围内呈线性：CLP为0.25-5.00 ng/mL，MP-H3和MP-H4异构体为0.25-50.00 ng/mL，CLPM为50- 10，000 ng/mL。CLP、MP-H3、MP-H4的定量下限为0.25 ng/mL，CLPM的定量下限为50.00 ng/mL。试验内和试验间精密度，表示为相对标准偏差，
A fast and reproducible HPLC-MS/MS method was developed for the simultaneous determination of clopidogrel (CLP), its carboxylic acid derivative (CLPM). derivatized thiol metabolite isomers MP-H3 and the active MP-H4 in incurred human plasma. CLP, CLPM, MP-H3 and MP-H4 isomers together with the internal standard piroxicam were extracted from plasma samples using a simple protein precipitation with acetonitrile. The analytes were separated on HPLC Zorbax Plus C18 column via gradient elution with water and acetonitrile, both containing 0.1% (v/v) formic acid. Detection of the analytes were performed on a triple-quadrupole MS with multiple-reaction-monitoring via electrospray ionization. Calibration curves of the analytes prepared in 250 mu L plasma were found to be linear in ranges: 0.25-5.00 ng/mL for CLP, 0.25-50.00 ng/mL for MP-H3 and MP-H4 isomers and 50-10,000 ng/mL for CLPM. The lower limit of quantitation was 0.25 ng/mL for CLP, MP-H3, MP-H4 and 50.00 ng/mL for CLPM. Intra- and inter-assay precision, expressed as relative standard deviation, was