Studies on the metabolism and the detectability of 4-methyl-amphetamine and its isomers 2-methyl-amphetamine and 3-methyl-amphetamine in rat urine using GC-MS and LC-(high-resolution)-MSn
Studies on the metabolism and the detectability of 4-methyl-amphetamine and its isomers 2-methyl-amphetamine and 3-methyl-amphetamine in rat urine using GC-MS and LC-(high-resolution)-MSn
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DOI：
10.1007/s00216-013-7595-5
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发表时间：
2014-03-01
影响因子：
4.3
通讯作者：
Maurer, Hans H.
中科院分区：
文献类型：
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作者：
Welter, Jessica;Meyer, Markus R.;Maurer, Hans H.
4-甲基苯丙胺(1-(4-甲基苯基)丙烷-2-胺；4-MA)及其异构体2-甲基苯丙胺(2-MA)和3-甲基苯丙胺(3-MA)属于苯丙胺类兴奋剂和新的精神活性物质。几项研究表明，在释放去甲肾上腺素和多巴胺方面的能力相似，但在释放5-羟色胺方面的能力高于苯丙胺。2013年3月，欧盟理事会根据欧洲毒品和毒瘾监测中心的风险评估报告决定在整个欧盟范围内实施管制，该报告记录了4-MA作为苯丙胺在非法市场上出售，并在几起致命案件中被发现。因此，在临床和法医毒理学中，4-MA及其异构体应纳入药物检测范围。这项工作的目的是研究尿样中各异构体的新陈代谢和可检测性。在代谢研究中，大鼠的尿样通过固相萃取分离，没有和在酶裂解偶联物之后都是如此。用气相色谱-质谱仪(GC-MS)和/或高效液相-高分辨-线性离子捕集质谱仪(LC-HR-MSN)对乙酰化后的I相代谢产物和LC-HR-MSN进行分离鉴定。从已鉴定的I相和II相代谢物推断出以下主要代谢途径：芳香族羟化、苯甲基的羟基化然后氧化成相应的羧酸、侧链的羟基化以及羟基和羧基的葡萄糖醛酸化和/或硫酸盐化。CYP2D6参与了芳香族羟基化反应。最后，使用作者的GC-MS和LC-MSN标准尿液筛查方法，可以确认大鼠尿中摄入了常用剂量的MAS。区分异构体以确认特定异构体的摄取是可能的，只需在大鼠尿中进行额外的检查。
4-Methyl-amphetamine (1-(4-methylphenyl)propane-2-amine; 4-MA) and its isomers 2-methyl-amphetamine (2-MA) and 3-methyl-amphetamine (3-MA) belong to the group of amphetamine-type stimulants and of new psychoactive substances. Several studies showed similar potencies in releasing noradrenalin and dopamine, but higher potencies in releasing serotonin than amphetamine. In March 2013, the EU Council decided on an EU-wide control based on the European Monitoring Centre for Drugs and Drug Addiction risk assessment report documenting that 4-MA was sold as amphetamine on the illicit market and detected in several fatal cases. Therefore, 4-MA and its isomers should be covered by drug testing in clinical and forensic toxicology. The aims of the presented work were to study the metabolism and detectability of each isomer in urine samples. For metabolism studies, rat urine samples were isolated by solid-phase extraction without and after enzymatic cleavage of conjugates. The phase I metabolites were separated and identified after acetylation by gas chromatography-mass spectrometry (GC-MS) and/or liquid chromatography-high resolution-linear ion trap mass spectrometry (LC-HR-MSn) and the phase II metabolites by LC-HR-MSn. From the identified phase I and II metabolites, the following main metabolic pathways were deduced: aromatic hydroxylation, hydroxylation of the phenylmethyl group followed by oxidation to the corresponding carboxylic acid, hydroxylation of the side chain, and glucuronidation and/or sulfation of the hydroxy and carboxy groups. CYP2D6 was involved in the aromatic hydroxylation. Finally, the intake of a commonly used dose of the MAs could be confirmed in rat urine using the authors' GC-MS and the LC-MSn standard urine screening approaches. Differentiation of the isomers to confirm the intake of a specific isomer was possible with an additional workup in rat urine.