Schappler Julie, Guillarme Davy, Prat Josiane, Veuthey Jean-Luc, Rudaz Serge
Laboratory of Pharmaceutical Analytical Chemistry, School of Pharmaceutical Sciences - EPGL, University of Geneva, Geneva, Switzerland.
Electrophoresis. 2008 May;29(10):2193-202. doi: 10.1002/elps.200700464.
Due to its selectivity and sensitivity, CE coupled to MS (CE-MS) has evolved as a useful analytical tool for determining drugs and metabolites in biological samples. A generic CE-ESI/MS method was developed for the enantioselective determination of basic compounds in plasma. The use of protein precipitation (PP) prior to a hydrodynamic injection (HD) was well adapted to high-concentration samples (>1 ppm) and allowed high throughput. In contrast, the combination of liquid-liquid extraction (LLE) and electrokinetic injection (EK) was time-consuming but did allow detection at the ppb level. Both approaches were fully validated according to ICH guidelines and SFSTP protocols for two pharmaceutical compounds (ecstasy and methadone (MTD)). Deuterated internal standards (IS) in the analytical procedures were used and good quantitative performance was obtained in terms of trueness and precision (repeatability and intermediate precision) since accuracy profiles were within the acceptance limits (30% for biological assay). Methods were linear over the concentration range of 0.50-175 ng/mL and 0.25-5 microg/mL for LLE-EK and PP-HD procedures, respectively. The LLE-EK methodology was finally successfully applied to quantitation of ecstasy and MTD in real cases obtained from toxicology.
由于其选择性和灵敏度，毛细管电泳与质谱联用（CE-MS）已发展成为一种用于测定生物样品中药物和代谢物的有用分析工具。开发了一种通用的CE-电喷雾电离/质谱（CE-ESI/MS）方法用于对血浆中的碱性化合物进行对映体选择性测定。在流体动力学进样（HD）之前使用蛋白沉淀（PP）非常适合高浓度样品（>1 ppm），并可实现高通量。相比之下，液液萃取（LLE）和电动进样（EK）的组合虽然耗时，但确实能够检测到ppb水平的物质。根据国际协调会议（ICH）指南和法国毒物学和药物化学学会（SFSTP）方案，对两种药物化合物（摇头丸和美沙酮（MTD））的这两种方法均进行了全面验证。在分析过程中使用了氘代内标（IS），并且在准确性和精密度（重复性和中间精密度）方面获得了良好的定量性能，因为准确度曲线在可接受范围内（生物测定为30%）。对于LLE-EK和PP-HD方法，方法分别在0.50 - 175 ng/mL和0.25 - 5 μg/mL的浓度范围内呈线性。LLE-EK方法最终成功应用于从毒理学获得的实际案例中摇头丸和美沙酮的定量分析。
