Biopharmaceutics and Pharmacokinetics Division, Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Sciences, Iran.
J Anal Toxicol. 2013 Jul-Aug;37(6):357-61. doi: 10.1093/jat/bkt039. Epub 2013 May 31.
3,4-Methylenedioxymethamphetamine (MDMA) is one of the most commonly abused illicit drugs in the world. We developed a rapid and simple high-performance liquid chromatography with a fluorescence (FL) detector method to determine MDMA and its metabolites, such as 3,4-methylenedioxyamphetamine (MDA), 4-hydroxy-3-methoxyamphetamine (HMA) and its main unstable metabolite 3,4-dihydroxymethamphetamine (HHMA) besides the internal standards, in a perfusion medium. The separation of analytes was performed at 25°C on a Chromolith® C18 (100 × 4.6 mm) column from Merck (Darmstadt, Germany) without any derivatization. The FL detector wavelength was fixed at 285 nm for excitation and at 320 nm for emission. Acetonitrile:phosphate buffer (0.02 M) at pH = 3 (5:95 v/v) was used as the mobile phase. The elution order was HHMA, HMA, MDA and MDMA with a retention time of 1.7, 2.6, 6.1 and 7.4 min, respectively. The method was validated according to the FDA bioanalytical method validation guideline. The limits of quantifications (LOQs) obtained for MDMA, MDA, HMA and HHMA were 1, 1, 1.5 and 5 ng/mL, respectively. The repeatability of relative standard deviation was <11% (except for LOQs). This method was applied successfully to determine MDMA and its metabolites in rat liver perfusion samples. To our knowledge, this is the first method introduced for the determination of HHMA as a free form with an FL detector.
3,4-亚甲二氧基甲基苯丙胺（MDMA）是世界上最常滥用的非法药物之一。我们开发了一种快速而简单的高效液相色谱法，结合荧光（FL）检测器，用于测定 MDMA 及其代谢物，如 3,4-亚甲二氧基苯丙胺（MDA）、4-羟基-3-甲氧基苯丙胺（HMA）及其主要不稳定代谢物 3,4-二羟基甲基苯丙胺（HHMA），以及内标物。在 Merck（德国达姆施塔特）的 Chromolith® C18（100 × 4.6 毫米）柱上，在 25°C 下无需衍生化即可进行分析物的分离。FL 检测器的波长固定在激发时为 285nm，发射时为 320nm。乙腈：磷酸盐缓冲液（0.02 M，pH = 3）（5:95v/v）用作流动相。洗脱顺序为 HHMA、HMA、MDA 和 MDMA，保留时间分别为 1.7、2.6、6.1 和 7.4 分钟。该方法根据 FDA 生物分析方法验证指南进行了验证。MDMA、MDA、HMA 和 HHMA 的定量限（LOQ）分别为 1、1、1.5 和 5ng/mL。相对标准偏差的重复性<11%（LOQ 除外）。该方法成功应用于大鼠肝灌注样品中 MDMA 及其代谢物的测定。据我们所知，这是首次使用 FL 检测器以游离形式测定 HHMA 的方法。
