Gorynski Krzysztof, Bojko Barbara, Kluger Michael, Jerath Angela, Wąsowicz Marcin, Pawliszyn Janusz
Department of Chemistry, University of Waterloo, Waterloo, ON, Canada N2L 3G1; Department of Medicinal Chemistry, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toruń, Jurasza 2, 85-089 Bydgoszcz, Poland.
Department of Anesthesia and Pain Management, Toronto General Hospital, Toronto, ON, Canada M5G 2C4.
J Pharm Biomed Anal. 2014 Apr;92:183-92. doi: 10.1016/j.jpba.2014.01.026. Epub 2014 Jan 27.
A high-throughput method using solid-phase microextraction coupled to liquid chromatography-tandem mass spectrometry (SPME-LC-MS/MS) for determination of tranexamic acid and rocuronium bromide in human plasma was developed and validated. Standard analytical approaches employ acidification of the sample due to the instability of rocuronium bromide in collected plasma samples. However, acidification affects the binding equilibrium of the drug and consequently no information on the free/bound concentration can be obtained. Contrary to these protocols, the proposed method requires minimum sample handling and no ion pairing and/or derivatization procedure. A weak cation exchange coating was chosen as the best extracting phase for selected drugs, guaranteed a good recovery, minimum carry-over, reusability and reproducibility. SPME procedure met all Food and Drug Administration acceptance criteria for bioanalytical assays at three concentration levels, for both selected drugs. Post-extraction addition experiments showed that matrix effect was less than ±3%. Here, a weak cation exchange thin-film solid-phase microextraction (WCX TF-SPME) approach is presented, offering effective cleanup procedure and full quantitation of the drugs in plasma, undoubtedly one the most challenging matrices with regards to its complexity. In addition, the 96-well plate format of WCX TF-SPME system provides considerable advantages, such as high throughput analysis for up to 96 samples in 35min (22s/sample), requirement of small amounts of plasma samples (0.8mL), and a simple sample preparation protocol, all of which shows a promise for possible on-site application in hospitals to monitor concentrations of the drugs in close to real time.
建立并验证了一种采用固相微萃取结合液相色谱-串联质谱法（SPME-LC-MS/MS）测定人血浆中氨甲环酸和罗库溴铵的高通量方法。由于罗库溴铵在采集的血浆样本中不稳定，标准分析方法采用对样品进行酸化处理。然而，酸化会影响药物的结合平衡，因此无法获得有关游离/结合浓度的信息。与这些方案不同，所提出的方法需要最少的样品处理，且无需离子对和/或衍生化程序。选择弱阳离子交换涂层作为所选药物的最佳萃取相，确保了良好的回收率、最小的残留、可重复使用性和重现性。对于两种所选药物，SPME程序在三个浓度水平上均符合美国食品药品监督管理局生物分析检测的所有验收标准。萃取后添加实验表明基质效应小于±3%。本文提出了一种弱阳离子交换薄膜固相微萃取（WCX TF-SPME）方法，该方法提供了有效的净化程序，并能对血浆中的药物进行完全定量，血浆无疑是最具挑战性的复杂基质之一。此外，WCX TF-SPME系统的96孔板形式具有诸多显著优势，如在35分钟内可对多达96个样品进行高通量分析（每个样品22秒）、所需血浆样品量少（0.8mL）以及样品制备方案简单，所有这些都显示出有望在医院现场应用以近乎实时地监测药物浓度。
