Malaca Sara, Huestis Marilyn A, Lattanzio Leonardo, Marsella Luigi T, Tagliabracci Adriano, Carlier Jeremy, Busardò Francesco P
Unit of Forensic Toxicology, Section of Legal Medicine, Department of Excellence of Biomedical Sciences and Public Health, Marche Polytechnic University, Via Tronto 10/a, 60126 Ancona, Italy.
Institute of Emerging Health Professions, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Metabolites. 2022 Jul 29;12(8):705. doi: 10.3390/metabo12080705.
Tryptamine intoxications and fatalities are increasing, although these novel psychoactive substances (NPS) are not controlled in most countries. There are few data on the metabolic pathways and enzymes involved in tryptamine biotransformation. 4-acetoxy-,-diisopropyltryptamine (4-AcO-DiPT) is a synthetic tryptamine related to 4-hydroxy-,-diisopropyltryptamine (4-OH-DiPT), 4-acetyloxy-,-dipropyltryptamine (4-AcO-DPT), and 4-acetoxy-,-dimethyltryptamine (4-AcO-DMT). The aim of this study was to determine the best 4-AcO-DiPT metabolites to identify 4-AcO-DiPT consumption through human hepatocyte metabolism and high-resolution mass spectrometry. 4-AcO-DiPT metabolites were predicted in silico with GLORYx freeware to assist in metabolite identification. 4-AcO-DiPT was incubated with 10-donor-pooled human hepatocytes and sample analysis was performed with reversed-phase liquid chromatography coupled with high-resolution tandem mass spectrometry (LC-HRMS/MS) in positive- and negative-ion modes. Software-assisted LC-HRMS/MS raw data mining was performed. A total of 47 phase I and II metabolites were predicted, and six metabolites were identified after 3 h incubation following ester hydrolysis, -glucuronidation, -sulfation, -oxidation, and -dealkylation. All second-generation metabolites were derived from the only first-generation metabolite detected after ester hydrolysis (4-OH-DiPT). The metabolite with the second-most-intense signal was 4-OH-iPT-sulfate followed by 4-OH-DiPT-glucuronide, indicating that glucuronidation and sulfation are common in this tryptamine's metabolic pathway. 4-OH-DiPT, 4-OH-iPT, and 4-OH-DiPT--oxide are suggested as optimal biomarkers to identify 4-AcO-DiPT consumption.
尽管大多数国家并未对这些新型精神活性物质（NPS）进行管控，但色胺中毒及死亡事件却日益增多。关于色胺生物转化所涉及的代谢途径及酶的数据很少。4-乙酰氧基-α,α-二异丙基色胺（4-AcO-DiPT）是一种与4-羟基-α,α-二异丙基色胺（4-OH-DiPT）、4-乙酰氧基-α,α-二丙基色胺（4-AcO-DPT）以及4-乙酰氧基-α,α-二甲基色胺（4-AcO-DMT）相关的合成色胺。本研究的目的是通过人肝细胞代谢及高分辨率质谱法确定用于识别4-AcO-DiPT摄入情况的最佳4-AcO-DiPT代谢产物。利用GLORYx免费软件对4-AcO-DiPT代谢产物进行了计算机模拟预测，以辅助代谢产物鉴定。将4-AcO-DiPT与人肝细胞10供体混合孵育，并采用反相液相色谱与高分辨率串联质谱联用（LC-HRMS/MS）在正离子和负离子模式下进行样品分析。进行了软件辅助的LC-HRMS/MS原始数据挖掘。共预测出47种I相和II相代谢产物，孵育3小时后，经酯水解、β-葡萄糖醛酸化、硫酸化、氧化和脱烷基化作用，鉴定出6种代谢产物。所有第二代代谢产物均源自酯水解后检测到的唯一第一代代谢产物（4-OH-DiPT）。信号强度第二高的代谢产物是4-OH-iPT-硫酸盐，其次是4-OH-DiPT-葡萄糖醛酸苷，这表明葡萄糖醛酸化和硫酸化在该色胺的代谢途径中较为常见。4-OH-DiPT、4-OH-iPT和4-OH-DiPT-α-氧化物被建议作为识别4-AcO-DiPT摄入情况的最佳生物标志物。
