Department of Physical Pharmacy and Pharmacokinetics, Poznan University of Medical Sciences, 6 Swiecickiego Street, 60-781 Poznań, Poland.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Apr 15;878(13-14):1013-8. doi: 10.1016/j.jchromb.2010.02.033. Epub 2010 Mar 6.
Fast and reproducible Capillary Zone Electrophoresis (CZE) method for the quantification of (+)-S clopidogrel carboxylic acid metabolite in human fluids was elaborated for the first time. Optimal buffer and CZE conditions were established to obtain the complete separation of clopidogrel, its metabolite and piroxicam (internal standard), during one analytical run. Finally, resolution of the analytes was obtained in an uncoated silica capillary filled with a phosphate buffer of pH 2.5. The analytes were isolated from plasma and urine samples using solid phase extraction (SPE). Validation of the CZE method was carried out. The calibration curve of clopidogrel was linear in the range of 0.5-10.0mg/L in plasma and urine, whereas for (+)-S carboxylic acid metabolite linearity was confirmed in the range of 0.25-20.0mg/L in plasma and 0.25-10.0mg/L in urine. Intra- and inter-day precision and accuracy were repeatable. LOD and LOQ were also estimated. SPE recovery of the analytes from plasma and urine was comparable and greater than 80%. The validated method was successfully applied in pharmacokinetic investigations of (+)-S carboxylic acid metabolite of clopidogrel following the oral administration of clopidogrel to patients prior to percutaneous coronary intervention.
首次详细阐述了用于定量检测人液中（+）-S 氯吡格雷羧酸代谢物的快速且可重现的毛细管区带电泳（CZE）方法。优化了缓冲液和 CZE 条件，以便在一次分析运行中完全分离氯吡格雷、其代谢物和吡罗昔康（内标）。最后，在填充 pH 2.5 磷酸盐缓冲液的未涂层硅胶毛细管中获得了分析物的分辨率。使用固相萃取（SPE）从血浆和尿液样品中分离分析物。对 CZE 方法进行了验证。氯吡格雷在血浆和尿液中的校准曲线在 0.5-10.0mg/L 范围内呈线性，而（+）-S 羧酸代谢物在血浆中的线性范围为 0.25-20.0mg/L，在尿液中的线性范围为 0.25-10.0mg/L。日内和日间精密度和准确度均具有可重复性。还估算了 LOD 和 LOQ。从血浆和尿液中 SPE 回收分析物的效率相当且大于 80%。该验证方法成功应用于经皮冠状动脉介入术前给予氯吡格雷后氯吡格雷（+）-S 羧酸代谢物的药代动力学研究。
