Department of Analytical Chemistry, Silesian University of Technology, 7M. Strzody Street, 44-100 Gliwice, Poland.
J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Nov 1;938:86-95. doi: 10.1016/j.jchromb.2013.09.001. Epub 2013 Sep 8.
A sensitive and selective ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) method was developed for the fast, simultaneous quantification of three novel cardiac drugs (aliskiren, prasugrel and rivaroxaban) in human urine. Sample preparation was performed with microextraction with packed sorbent (MEPS), which is a miniaturization of solid phase extraction. The optimal conditions for MEPS extraction were obtained using C8 sorbent, small sample volumes and a short time period (about 3min for the entire sample preparation step). Chromatographic separation of the selected compounds was achieved in less than 1.5min on a Zorbax Rapid Resolution High Definition SB-C18 column using isocratic elution with 0.1% formic acid and acetonitrile (70:30, v/v) at a flow rate of 0.8mLmin(-1). The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring via an electrospray ionization source with positive ionization mode. The method was fully validated according to the latest recommendations of international guidelines. The lower limit of quantification was 5.0pgmL(-1) for aliskiren and rivaroxaban and 0.5pgmL(-1) for prasugrel. The intra- and inter-day precision was within 7.12% and the accuracy ranged from -7.54% to 4.17%. The mean extraction recoveries of the MEPSC8 methodology were found to be 98.3% for aliskiren, 100.3% for rivaroxaban and 99.9% for prasugrel. This MEPSC8-UHPLC-MS/MS method offers a fast, simple and precise way to determine selected novel cardiac drugs in human urine that could be applied to therapeutic drug monitoring and pharmacokinetic studies.
建立了一种灵敏、选择性的超高效液相色谱-串联质谱（UHPLC-MS/MS）方法，用于快速、同时定量测定人尿中的三种新型心脏药物（阿利克仑、普拉格雷和利伐沙班）。样品制备采用微萃取填充固相萃取（MEPS）进行，这是固相萃取的微型化。通过 C8 吸附剂、小体积样品和短时间（整个样品制备步骤约 3 分钟）获得 MEPS 萃取的最佳条件。在 Zorbax Rapid Resolution High Definition SB-C18 柱上，采用等度洗脱，以 0.1%甲酸和乙腈（70:30，v/v）为流动相，流速为 0.8mL/min，在不到 1.5min 内实现了所选化合物的色谱分离。通过电喷雾电离源和正电离模式进行多反应监测，在三重四极杆串联质谱仪上进行检测。该方法根据国际指南的最新建议进行了全面验证。阿利克仑和利伐沙班的定量下限为 5.0pg/mL，普拉格雷为 0.5pg/mL。日内和日间精密度均在 7.12%以内，准确度范围为-7.54%至 4.17%。MEPSC8 方法的平均提取回收率分别为阿利克仑 98.3%、利伐沙班 100.3%和普拉格雷 99.9%。该 MEPSC8-UHPLC-MS/MS 方法提供了一种快速、简单、精确的方法，可用于测定人尿中的选定新型心脏药物，可应用于治疗药物监测和药代动力学研究。
