Apostolou Constantinos, Kousoulos Constantinos, Dotsikas Yannis, Loukas Yannis L
Laboratory of Pharmaceutical Analysis and Bioequivalence Services (GLP Compliant), Department of Pharmaceutical Chemistry, School of Pharmacy, University of Athens, Panepistimioupoli Zografou GR-157 71, Athens, Greece.
Biomed Chromatogr. 2008 Dec;22(12):1393-402. doi: 10.1002/bmc.1071.
In the present study, hydrophilic interaction liquid chromatography (HILIC) and reversed-phase liquid chromatography (RPLC) combined with tandem mass spectrometric detection (MS/MS) were evaluated and compared for the determination of donepezil, cetirizine and loratadine in human plasma, in terms of sensitivity and sample preparation procedure. A retention study for the above compounds of various polarities was performed, using both C(18) and silica columns, with several aqueous-organic mobile phase ratios, in order to investigate their retention mechanism profile under HILIC and RPLC. Both chromatographic conditions were compared for chromatographic analysis of plasma samples processed with a liquid-liquid extraction (LLE) method for donepezil determination, resulting in significantly higher sensitivity under HILIC. Furthermore, HILIC and RPLC were compared for direct injection, and novel methods including LLE, solid-phase extraction and protein precipitation protocols were developed. Direct injection technique significantly reduced sample preparation time, increasing at the same time method sensitivity. The current study contributes to broadening the range of analyzable compounds by HILIC-MS/MS to molecules of medium polarity.
在本研究中，就灵敏度和样品制备程序而言，对亲水作用液相色谱（HILIC）和反相液相色谱（RPLC）结合串联质谱检测（MS/MS）用于测定人血浆中多奈哌齐、西替利嗪和氯雷他定进行了评估和比较。使用C(18)柱和硅胶柱，采用几种水-有机流动相比例，对上述不同极性的化合物进行保留研究，以研究它们在HILIC和RPLC下的保留机制特征。比较了两种色谱条件用于采用液-液萃取（LLE）法处理的血浆样品中多奈哌齐测定的色谱分析，结果表明HILIC下灵敏度显著更高。此外，比较了HILIC和RPLC的直接进样情况，并开发了包括LLE、固相萃取和蛋白沉淀方案在内的新方法。直接进样技术显著减少了样品制备时间，同时提高了方法灵敏度。本研究有助于将HILIC-MS/MS可分析化合物的范围拓宽至中等极性分子。
