Lanz M, Brenneisen R, Thormann W
Department of Clinical Pharmacology, University of Bern, Switzerland.
Electrophoresis. 1997 Jun;18(6):1035-43. doi: 10.1002/elps.1150180628.
Using capillary zone electrophoresis with a phosphate buffer at pH 2.5 containing 30 mM (2-hydroxypropyl)-beta-cyclodextrin as chiral selector, the simultaneous separation of the enantiomers of 3,4-methylenedioxymethamphetamine (MDMA or Ecstasy) and its two metabolites 4-hydroxy-3-methoxymethamphetamine (HMMA) and 3,4-methylenedioxyamphetamine (MDA) in human urine is reported. The assay described is based upon enzymatic hydrolysis of conjugated HMMA (major urinary metabolite) and solid-phase extraction followed by injection of a few nL of the extract onto a 50 microm internal diameter (ID) fused-silica capillary of 60 cm length. Solutes are detected via on-column absorbance at 195 nm. For 375 ng/mL drug levels, intraday and interday imprecision is < 4%. With 5 mL urine samples, the detection limit is in the 20-50 ng/mL range. Via analysis of the urines of two patients, the metabolism of MDMA is demonstrated to be enantioselective, with significantly higher urinary amounts of R-(-)-MDMA being excreted compared to S-(+)-MDMA. Within 72h after drug administration one patient was determined to excrete 42.28 and 10.16% of the racemic MDMA dose (1.5 mg/kg body weight) as R-(-) and S-(+)-MDMA enantiomers, respectively. Corresponding values for the second subject were found to be 28.63 and 9.34%. The metabolism of the enantiomers of the two metabolites showed interindividual differences. The first and second detected HMMA enantiomers represented 3.79 and 5.42% (first subject) and 8.51 and 4.36% (second), respectively, of the administered MDMA dose. For the MDA enantiomers, corresponding values were 2.44, 1.76, 0.75, and 0.79%, respectively.
采用毛细管区带电泳法，以pH 2.5的磷酸盐缓冲液为背景电解质，其中含有30 mM（2-羟丙基）-β-环糊精作为手性选择剂，实现了在人尿液中对3,4-亚甲基二氧基甲基苯丙胺（摇头丸或迷魂药）及其两种代谢物4-羟基-3-甲氧基甲基苯丙胺（HMMA）和3,4-亚甲基二氧基苯丙胺（MDA）对映体的同时分离。所描述的分析方法基于对结合型HMMA（主要尿液代谢物）的酶促水解和固相萃取，然后将几纳升提取物注入内径为50微米、长度为60厘米的熔融石英毛细管中。通过在195 nm处的柱上吸光度检测溶质。对于375 ng/mL的药物水平，日内和日间不精密度<4%。对于5 mL尿液样本，检测限在20 - 50 ng/mL范围内。通过对两名患者尿液的分析，证明摇头丸的代谢具有对映体选择性，与S-(+)-摇头丸相比，R-(-)-摇头丸的尿排泄量显著更高。给药后72小时内，一名患者分别以R-(-)和S-(+)-摇头丸对映体形式排泄了外消旋摇头丸剂量（1.5 mg/kg体重）的42.28%和10.16%。第二名受试者的相应值分别为28.63%和9.34%。两种代谢物对映体的代谢存在个体差异。检测到的第一种和第二种HMMA对映体分别占给药摇头丸剂量的3.79%和5.42%（第一名受试者）以及8.51%和4.36%（第二名受试者）。对于MDA对映体，相应值分别为2.44%、1.76%、0.75%和0.79%。
