Department of Experimental and Clinical Toxicology, Institute of Experimental and Clinical Pharmacology and Toxicology, Saarland University, D-66421 Homburg (Saar), Germany.
J Mass Spectrom. 2012 Feb;47(2):253-62. doi: 10.1002/jms.2960.
3-Bromomethcathinone (3-BMC) and 3-Fluoromethcathinone (3-FMC) are two new designer drugs, which were seized in Israel during 2009 and had also appeared on the illicit drug market in Germany. These two compounds were sold via the Internet as so-called "bath salts" or "plant feeders." The aim of the present study was to identify for the first time the 3-BMC and 3-FMC Phase I and II metabolites in rat urine and human liver microsomes using GC-MS and LC-high-resolution MS (HR-MS) and to test for their detectability by established urine screening approaches using GC-MS or LC-MS. Furthermore, the human cytochrome-P450 (CYP) isoenzymes responsible for the main metabolic steps were studied to highlight possible risks of consumption due to drug-drug interaction or genetic variations. For the first aim, rat urine samples were extracted after and without enzymatic cleavage of conjugates. The metabolites were separated and identified by GC-MS and by LC-HR-MS. The main metabolic steps were N-demethylation, reduction of the keto group to the corresponding alcohol, hydroxylation of the aromatic system and combinations of these steps. The elemental composition of the metabolites identified by GC-MS could be confirmed by LC-HR-MS. Furthermore, corresponding Phase II metabolites were identified using the LC-HR-MS approach. For both compounds, detection in rat urine was possible within the authors' systematic toxicological analysis using both GC-MS and LC-MS(n) after a suspected recreational users dose. Following CYP enzyme kinetic studies, CYP2B6 was the most relevant enzyme for both the N-demethylation of 3-BMC and 3-FMC after in vitro-in vivo extrapolation.
3-溴甲卡西酮（3-BMC）和 3-氟甲卡西酮（3-FMC）是两种新型的设计药物，于 2009 年在以色列被缉获，并且也出现在德国的非法毒品市场上。这两种化合物通过互联网作为所谓的“浴盐”或“植物饲料”出售。本研究的目的是首次在大鼠尿液和人肝微粒体中使用 GC-MS 和 LC-高分辨率 MS（HR-MS）鉴定 3-BMC 和 3-FMC 的 I 相和 II 相代谢物，并使用 GC-MS 或 LC-MS 建立的尿液筛选方法测试其可检测性。此外，还研究了负责主要代谢步骤的人细胞色素 P450（CYP）同工酶，以突出由于药物相互作用或遗传变异而导致消费的潜在风险。为了实现第一个目标，在没有酶裂解结合物的情况下，在大鼠尿液样品中提取代谢物。通过 GC-MS 和 LC-HR-MS 分离和鉴定代谢物。主要的代谢步骤是 N-去甲基化、酮基还原为相应的醇、芳构化系统的羟化以及这些步骤的组合。通过 GC-MS 鉴定的代谢物的元素组成可以通过 LC-HR-MS 来确认。此外，使用 LC-HR-MS 方法鉴定了相应的 II 相代谢物。对于这两种化合物，在作者的系统毒理学分析中，使用 GC-MS 和 LC-MS(n) 均可以在疑似娱乐性使用者剂量后在大鼠尿液中检测到。在 CYP 酶动力学研究之后，CYP2B6 是 3-BMC 和 3-FMC 的 N-去甲基化最相关的酶，经过体外-体内外推。
