Rasmussen Louise Bang, Olsen Kristine Høje, Johansen Sys Stybe
Department of Pharmaceutics and Analytical Chemistry, The Danish University of Pharmaceutical Sciences, Denmark.
J Chromatogr B Analyt Technol Biomed Life Sci. 2006 Oct 2;842(2):136-41. doi: 10.1016/j.jchromb.2006.05.011. Epub 2006 Jun 21.
The enantioselective composition of the amphetamines is of interest, as the enantiomers show differences in their pharmacological effects and several methods for chiral separation of amphetamines have been described. Only a few methods have used whole blood as matrix and none of these separates both classic amphetamines (amphetamine and methamphetamine) and designer amphetamines (MDA, MDMA and MDEA). The aim of this study was, therefore, to develop a method for enantioselective analysis of AM, MA, MDA, MDMA, and MDEA in whole blood. The amphetamines were extracted from 0.5 g of whole blood by liquid-liquid extraction. After derivatization with R-MTPCl, the resulting diastereomers were separated by GC on a HP-5MS column and detected by SIM-MS. R-MTPCl was used as derivatization reagent because of the stability of this reagent and good separation of these analytes. Through the method, development time and temperature of the derivatization were optimized, and by admixture of 0.02% triethylamine it became possible to detect the amphetamines in adequately low concentrations as more analytes were derivatized. The method was validated and it was linear from 0.004 to 3 microg/g per enantiomer. The accuracy was within 91-115%, while the repeatability and reproducibility were < or =15% R.S.D. A method suitable for enantioselective separation and analysis of the amphetamines has been achieved, and the method was applied to analysis of whole blood samples originating from traffic and criminal cases and post mortem cases.
苯丙胺类药物的对映体组成备受关注，因为其对映体在药理作用上存在差异，且已有多种苯丙胺类药物的手性分离方法被报道。仅有少数方法使用全血作为基质，且这些方法均无法同时分离经典苯丙胺类药物（苯丙胺和甲基苯丙胺）和设计型苯丙胺类药物（3,4－亚甲基二氧基苯丙胺、3,4－亚甲基二氧基甲基苯丙胺和3,4－亚甲基二氧基乙基苯丙胺）。因此，本研究的目的是开发一种用于全血中苯丙胺、甲基苯丙胺、3,4－亚甲基二氧基苯丙胺、3,4－亚甲基二氧基甲基苯丙胺和3,4－亚甲基二氧基乙基苯丙胺对映体选择性分析的方法。通过液－液萃取从0.5 g全血中提取苯丙胺类药物。用R－MTPCl进行衍生化后，所得非对映异构体在HP－5MS柱上通过气相色谱分离，并用选择离子监测质谱法检测。使用R－MTPCl作为衍生化试剂是因为该试剂稳定性好且能对这些分析物实现良好分离。通过该方法，对衍生化的时间和温度进行了优化，并且通过加入0.02%的三乙胺，由于更多分析物被衍生化，使得能够检测到浓度足够低的苯丙胺类药物。该方法经过验证，每个对映体在0.004至3 μg/g范围内呈线性。准确度在91%至115%之间，而重复性和重现性的相对标准偏差≤15%。已实现一种适用于苯丙胺类药物对映体选择性分离和分析的方法，并将该方法应用于交通、刑事案件及尸检案件的全血样本分析。
