Department of Pharmaceutical Analysis and Nuclear Pharmacy, Faculty of Pharmacy, Comenius University, Odbojárov 10, SK-832 32 Bratislava, Slovak Republic; Toxicological and Antidoping Center, Faculty of Pharmacy, Comenius University, Odbojárov 10, SK-832 32 Bratislava, Slovak Republic.
Villa Labeco spol. s r. o., Chrapčiakova 1, 052 01 Spišská Nová Ves, Slovak Republic.
J Chromatogr A. 2014 Sep 5;1358:285-92. doi: 10.1016/j.chroma.2014.06.083. Epub 2014 Jul 1.
The advanced two dimensional isotachophoresis (ITP)-capillary zone electrophoresis (CZE) hyphenated with tandem mass spectrometry (MS/MS, here triple quadrupole, QqQ) was developed in this work to demonstrate analytical potentialities of this approach in the analysis of drugs in multicomponent ionic matrices. Pheniramine (PHM), phenylephrine (PHE), paracetamol (PCM) and their potential metabolic products were taken for the analysis by the ITP-CZE-ESI-QqQ technique working in hydrodynamically closed CE separation system and then a comparison with the conventional (hydrodynamically open) CZE-ESI-QqQ technique was made. The ITP-CZE-ESI-QqQ method was favorable in terms of obtainable selectivity (due to highly effective heart-cut analysis), concentration limits of detection (LOD at pgmL(-1) levels due to enhanced sample load capacity and ITP preconcentration), sample handling (on-line sample pretreatment, i.e. clean-up, preconcentration, preseparation), and, by that, possibilities for future automation and miniaturization. On the other hand, this experimental arrangement, in contrast to the CZE-ESI-QqQ arrangement supported by an electroosmotic flow, is principally limited to the analysis of uniformly (i.e. positively or negatively) charged analytes in one run without any possibilities to analyze neutral compounds (here, PCM and neutral or acidic metabolites of the drugs had to be excluded from the analysis). Hence, these general characteristics should be considered when choosing a proper analytical CE-MS approach for a given biomedical application. Here, the analytical potential of the ITP-CZE-ESI-QqQ method was demonstrated showing the real time profiles of excreted targeted drugs and metabolite (PHM, PHE, M-PHM) in human urine after the administration of one dose of Theraflu(®) to the volunteers.
本工作中发展了一种先进的二维等速电泳（ITP）-毛细管区带电泳（CZE）与串联质谱（MS/MS，此处为三重四极杆，QqQ）联用技术，以展示该方法在多组分离子基质中药物分析中的应用潜力。本研究采用 ITP-CZE-ESI-QqQ 技术对非那根（PHM）、苯肾上腺素（PHE）、扑热息痛（PCM）及其潜在代谢产物进行分析，并与传统（水力开放）CZE-ESI-QqQ 技术进行了比较。与传统 CZE-ESI-QqQ 技术相比，ITP-CZE-ESI-QqQ 方法在可获得的选择性（由于高效的中心切割分析）、检测限浓度（由于增强的样品负载能力和 ITP 预浓缩，在 pgmL(-1) 水平达到检测限）、样品处理（在线样品预处理，即净化、预浓缩、预分离）方面具有优势，并且具有未来自动化和微型化的可能性。另一方面，与基于电渗流支持的 CZE-ESI-QqQ 排列相比，这种实验排列原则上仅限于在一次运行中分析均匀（即正或负）带电的分析物，而没有分析中性化合物的可能性（此处，PCM 和药物的中性或酸性代谢物必须从分析中排除）。因此，在为给定的生物医学应用选择适当的分析 CE-MS 方法时，应考虑这些一般特征。此处，通过对志愿者给予一剂Theraflu(®)后尿液中排泄的靶向药物和代谢物（PHM、PHE、M-PHM）的实时分析，展示了 ITP-CZE-ESI-QqQ 方法的分析潜力。
