Institute of Forensic Medicine, Forensic Toxicology, Medical Center, University of Freiburg, Faculty of Medicine, Albertstraße 9, 79104 Freiburg, Germany.
Hermann Staudinger Graduate School, University of Freiburg, Hebelstr. 27, 79104 Freiburg, Germany.
J Anal Toxicol. 2022 May 20;46(5):540-548. doi: 10.1093/jat/bkab038.
Rising numbers of psychoactive tryptamine derivatives have become available on the drug market over the last decade, making these naturally occurring or synthetically manufactured compounds highly relevant for forensic analyses. One of these compounds is 5-methoxy-N,N-dimethyltryptamine (5-MeO-DMT), a constituent of the dried poison of Incilius alvarius (Colorado River toad), which has a history of ritual and/or recreational use. Still, comprehensive and validated qualitative as well as quantitative analytical data on the psychoactive components of this poison are scarce. In this study, samples of the poison of Incilius alvarius were collected from live toads in the Sonoran Desert, Arizona (USA), and analyzed with a set of complementary methods. Acetone/water (70/30, v/v) proved to be the solvent of choice for the extraction of tryptamine derivatives. Trace compounds were enriched, and overload with 5-MeO-DMT was prevented by chromatographic separation of 5-MeO-DMT prior to qualitative analyses. The method for quantification was validated. Attenuated total reflection-Fourier transform infrared spectroscopy was suitable to identify 5-MeO-DMT as the main tryptamine in samples of the poison. The combined evaluation of analytical data gained from gas chromatography-mass spectrometry (GC-MS), high-performance liquid chromatography-quadrupole time-of-flight high-resolution MS (HPLC-qToF-HRMS) and HPLC-MS-MS confirmed the presence of 5-MeO-DMT, 5-MeO-N-methyltryptamine, 5-MeO-tryptamine, 5-MeO-tryptophol, 2-(5-methoxy-1H-indol-3-yl)-acetic-acid (5-MIAA), 5-HO-N-methyltryptamine, bufotenin, DMT and tryptophan. For the first time, 5-MeO-tryptamine and two positional isomers of hydroxylated MeO-DMT were detected in the poison of Incilius alvarius. The tryptamine present in the highest concentrations was 5-MeO-DMT (mean ± SD: 410,000 ± 30,000 μg/g). Mean concentrations of 5-MeO-tryptamine (490 ± 260 μg/g), 5-HO-N-methyltryptamine (270 ± 120 μg/g), bufotenin (2,800 ± 1,900 μg/g) and DMT (250 ± 80 μg/g) showed a relatively high variability between individual samples. The comprehensive analytical reference data of Incilius alvarius poison presented here might prove useful for forensic chemists.
在过去十年中，市场上出现了越来越多的具有致幻作用的色胺衍生物，使得这些天然存在或合成制造的化合物在法医分析中非常重要。其中一种化合物是 5-甲氧基-N,N-二甲基色胺（5-MeO-DMT），它是干燥的 Incilius alvarius（科罗拉多河蟾蜍）毒液的成分之一，具有仪式和/或娱乐用途的历史。尽管如此，关于这种毒液的精神活性成分的全面和经过验证的定性和定量分析数据仍然很少。在这项研究中，从亚利桑那州索诺兰沙漠（美国）的活蟾蜍中采集了 Incilius alvarius 毒液样本，并使用一系列互补的方法进行了分析。丙酮/水（70/30，v/v）被证明是提取色胺衍生物的首选溶剂。通过在定性分析之前对 5-MeO-DMT 进行色谱分离，可以富集痕量化合物并防止 5-MeO-DMT 过载。该定量方法已得到验证。衰减全反射-傅里叶变换红外光谱（ATR-FTIR）适合于鉴定样品中的 5-MeO-DMT 作为主要色胺。通过气相色谱-质谱联用（GC-MS）、高效液相色谱-四极杆飞行时间高分辨率质谱（HPLC-qToF-HRMS）和 HPLC-MS-MS 获得的分析数据的综合评估证实了 5-MeO-DMT、5-MeO-N-甲基色胺、5-MeO-色胺、5-MeO-色氨酸、2-（5-甲氧基-1H-吲哚-3-基）-乙酸（5-MIAA）、5-HO-N-甲基色胺、蟾毒色胺、DMT 和色氨酸的存在。这是首次在 Incilius alvarius 毒液中检测到 5-MeO-色胺和两种羟基化 MeO-DMT 的位置异构体。存在浓度最高的色胺是 5-MeO-DMT（平均值±SD：410,000±30,000μg/g）。5-MeO-色胺（490±260μg/g）、5-HO-N-甲基色胺（270±120μg/g）、蟾毒色胺（2,800±1,900μg/g）和 DMT（250±80μg/g）的平均浓度在个体样本之间显示出相对较高的变异性。本文提供的综合分析参考数据可能对法化学家有用。
