Metabolite markers for three synthetic tryptamines N-ethyl-N-propyltryptamine, 4-hydroxy-N-ethyl-N-propyltryptamine, and 5-methoxy-N-ethyl-N-propyltryptamine.
4-hydroxy-N-ethyl-N-propyltryptamine (4-OH-EPT)
5-methoxy-N-ethyl-N-propyltryptamine (5-MeO-EPT)
N-ethyl-N-propyltryptamine (EPT)
metabolite
microsomes
new psychoactive substances (NPS)
synthetic tryptamines
ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS)
Journal
Drug testing and analysis
ISSN: 1942-7611
Titre abrégé: Drug Test Anal
Pays: England
ID NLM: 101483449
Informations de publication
Date de publication:
09 Mar 2024
09 Mar 2024
Historique:
revised:
08
02
2024
received:
21
09
2023
accepted:
11
02
2024
medline:
9
3
2024
pubmed:
9
3
2024
entrez:
9
3
2024
Statut:
aheadofprint
Résumé
N-Ethyl-N-propyltryptamine (EPT), 4-hydroxy-N-ethyl-N-propyltryptamine (4-OH-EPT), and 5-methoxy-N-ethyl-N-propyltryptamine (5-MeO-EPT) are new psychoactive substances classified as tryptamines, sold online. Many tryptamines metabolize rapidly, and identifying the appropriate metabolites to reveal intake is essential. While the metabolism of 4-OH-EPT and 5-MeO-EPT are not previously described, EPT is known to form metabolites by indole ring hydroxylation among others. Based on general knowledge of metabolic patterns, 5-MeO-EPT is also expected to form ring hydroxylated EPT (5-OH-EPT). In the present study, the aim was to characterize the major metabolites of EPT, 4-OH-EPT, and 5-MeO-EPT, to provide markers for substance identification in forensic casework. The tryptamines were incubated with pooled human liver microsomes at 37°C for up to 4 h. The generated metabolites were separated and detected by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry analysis. The major in vitro EPT metabolites were formed by hydroxylation, N-dealkylation, and carbonylation. In comparison, 4-OH-EPT metabolism was dominated by double bond formation, N-dealkylation, hydroxylation, and carbonylation in vitro and hydroxylation or carbonylation combined with double bond loss, carbonylation, N-dealkylation, and hydroxylation in vivo. 5-MeO-EPT was metabolized by O-demethylation, hydroxylation, and N-dealkylation in vitro. The usefulness of the characterized metabolites in forensic casework was demonstrated by identification of unique metabolites for 4-OH-EPT in a human postmortem blood sample with suspected EPT or 4-OH-EPT intoxication.
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
© 2024 The Authors. Drug Testing and Analysis published by John Wiley & Sons Ltd.
Références
EMCDDA. European Drug Report 2022: Trends and Developments. EMCDDA; 2023. https://www.emcdda.europa.eu/system/files/publications/14644/TDAT22001ENN.pdf
Greene SL. Chapter 15-tryptamines. In: Dargan PI, Wood DM, eds. Novel Psychoactive Substances. Academic Press; 2013:363-381. doi:10.1016/B978-0-12-415816-0.00015-8
Shulgin AT, Shulgin A. Tryptamines I Have Known And Loved: The Continuation. Transform Press; 1997.
Varì MR, Pichini S, Giorgetti R, Busardò FP. New psychoactive substances-synthetic stimulants. WIREs Forensic Sci. 2019;1(2):e1197. doi:10.1002/wfs2.1197
Malaca S, Lo Faro AF, Tamborra A, Pichini S, Busardò FP, Huestis MA. Toxicology and analysis of psychoactive tryptamines. Int J Mol Sci. 2020;21(23):9279. doi:10.3390/ijms21239279
Titeler M, Lyon RA, Glennon RA. Radioligand binding evidence implicates the brain 5-HT2 receptor as a site of action for LSD and phenylisopropylamine hallucinogens. Psychopharmacology (Berl). 1988;94(2):213-216. doi:10.1007/BF00176847
Rickli A, Moning OD, Hoener MC, Liechti ME. Receptor interaction profiles of novel psychoactive tryptamines compared with classic hallucinogens. Eur Neuropsychopharmacol. 2016;26(8):1327-1337. doi:10.1016/j.euroneuro.2016.05.001
Glennon RA, Titeler M, McKenney JD. Evidence for 5-HT2 involvement in the mechanism of action of hallucinogenic agents. Life Sci. 1984;35(25):2505-2511. doi:10.1016/0024-3205(84)90436-3
Halberstadt AL, Nichols DE. Chapter 4.7-serotonin and serotonin receptors in hallucinogen action. In: Müller CP, Jacobs BL, eds. Handbook of Behavioral Neuroscience. Vol. 21. Elsevier; 2010:621-636. doi:10.1016/S1569-7339(10)70103-X
Tittarelli R, Mannocchi G, Pantano F, Romolo FS. Recreational use, analysis and toxicity of tryptamines. Curr Neuropharmacol. 2015;13(1):26-46. doi:10.2174/1570159X13666141210222409
Jovel A, Felthous A, Bhattacharyya A. Delirium due to intoxication from the novel synthetic tryptamine 5-MeO-DALT. J Forensic Sci. 2014;59(3):844-846. doi:10.1111/1556-4029.12367
Grafinger KE, Hädener M, König S, Weinmann W. Study of the in vitro and in vivo metabolism of the tryptamine 5-MeO-MiPT using human liver microsomes and real case samples. Drug Test Anal. 2018;10(3):562-574. doi:10.1002/dta.2245
Boland DM, Andollo W, Hime GW, Hearn WL. Fatality due to acute alpha-methyltryptamine intoxication. J Anal Toxicol. 2005;29(5):394-397. doi:10.1093/jat/29.5.394
Martin R, Schürenkamp J, Pfeiffer H, Köhler H. A validated method for quantitation of psilocin in plasma by LC-MS/MS and study of stability. Int J Leg Med. 2012;126(6):845-849. doi:10.1007/s00414-011-0652-8
Yan X, Xiang P, Zhao Y, Yu Z, Yan H. Determination of 5-MeO-DIPT in human urine using gas chromatography coupled with high-resolution Orbitrap mass spectrometry. J Anal Toxicol. 2020;44(5):461-469. doi:10.1093/jat/bkaa005
Anastos N, Barnett NW, Pfeffer FM, Lewis SW. Investigation into the temporal stability of aqueous standard solutions of psilocin and psilocybin using high performance liquid chromatography. Sci Justice. 2006;46(2):91-96. doi:10.1016/S1355-0306(06)71579-9
Martin R, Schürenkamp J, Pfeiffer H, Lehr M, Köhler H. Synthesis, hydrolysis and stability of psilocin glucuronide. Forensic Sci Int. 2014;237:1-6. doi:10.1016/j.forsciint.2014.01.006
Schüller M, McQuade TA-P, Bergh MS-S, Pedersen-Bjergaard S, Øiestad EL. Determination of tryptamine analogs in whole blood by 96-well electromembrane extraction and UHPLC-MS/MS. Talanta Open. 2023;7:100171. doi:10.1016/j.talo.2022.100171
Lindenblatt H, Krämer E, Holzmann-Erens P, Gouzoulis-Mayfrank E, Kovar K. Quantitation of psilocin in human plasma by high-performance liquid chromatography and electrochemical detection: comparison of liquid-liquid extraction with automated on-line solid-phase extraction. J Chromatogr B Biomed Appl. 1998;709(2):255-263. doi:10.1016/S0378-4347(98)00067-X
Michely JA, Brandt SD, Meyer MR, Maurer HH. Biotransformation and detectability of the new psychoactive substances N,N-diallyltryptamine (DALT) derivatives 5-fluoro-DALT, 7-methyl-DALT, and 5,6-methylenedioxy-DALT in urine using GC-MS, LC-MSn, and LC-HR-MS/MS. Anal Bioanal Chem. 2017;409(6):1681-1695. doi:10.1007/s00216-016-0117-5
Caspar AT, Gaab JB, Michely JA, Brandt SD, Meyer MR, Maurer HH. Metabolism of the tryptamine-derived new psychoactive substances 5-MeO-2-Me-DALT, 5-MeO-2-Me-ALCHT, and 5-MeO-2-Me-DIPT and their detectability in urine studied by GC-MS, LC-MSn, and LC-HR-MS/MS. Drug Test Anal. 2018;10(1):184-195. doi:10.1002/dta.2197
Manier SK, Felske C, Zapp J, Eckstein N, Meyer MR. Studies on the in vitro and in vivo metabolic fate of the new psychoactive substance N-ethyl-N-propyltryptamine (EPT) for analytical purposes. J Anal Toxicol. 2020;45(2):195-202. doi:10.1093/jat/bkaa060
Tanaka R, Kawamura M, Hakamatsuka T, Kikura-Hanajiri R. Identification of six tryptamine derivatives as designer drugs in illegal products. Forensic Toxicol. 2021;39(1):248-258. doi:10.1007/s11419-020-00556-5
EMCDDA. European Database on New Drugs. EMCDDA. Accessed December 19, 2023. https://ednd2.emcdda.europa.eu/ednd/login
Uchiyama N, Miyazawa N, Kawamura M, Kikura-Hanajiri R, Goda Y. Analysis of newly distributed designer drugs detected in the products purchased in fiscal year 2008. Yakugaku Zasshi. 2010;130(2):263-270. doi:10.1248/yakushi.130.263
Klein AK, Chatha M, Laskowski LJ, et al. Investigation of the structure-activity relationships of psilocybin analogues. ACS Pharmacol Transl Sci. 2021;4(2):533-542. doi:10.1021/acsptsci.0c00176
Halberstadt AL, Chatha M, Klein AK, et al. Pharmacological and biotransformation studies of 1-acyl-substituted derivatives of d-lysergic acid diethylamide (LSD). Neuropharmacology. 2020;172:107856. doi:10.1016/j.neuropharm.2019.107856
Glatfelter GC, Naeem M, Pham DNK, et al. Receptor binding profiles for tryptamine psychedelics and effects of 4-propionoxy-N,N-dimethyltryptamine in mice. ACS Pharmacol Transl Sci. 2023;6(4):567-577. doi:10.1021/acsptsci.2c00222
Kamata T, Katagi M, Kamata HT, et al. Metabolism of the psychotomimetic tryptamine derivative 5-methoxy-N,N-diisopropyltryptamine in humans: identification and quantification of its urinary metabolites. Drug Metab Dispos. 2006;34(2):281-287. doi:10.1124/dmd.105.005835
Kamata T, Katagi M, Tsuchihashi H. Metabolism and toxicological analyses of hallucinogenic tryptamine analogues being abused in Japan. Forensic Toxicol. 2010;28(1):1-8. doi:10.1007/s11419-009-0087-9
Fabregat-Safont D, Barneo-Muñoz M, Martinez-Garcia F, Sancho JV, Hernández F, Ibáñez M. Proposal of 5-methoxy-N-methyl-N-isopropyltryptamine consumption biomarkers through identification of in vivo metabolites from mice. J Chromatogr A. 2017;1508:95-105. doi:10.1016/j.chroma.2017.06.010
Michely JA, Helfer AG, Brandt SD, Meyer MR, Maurer HH. Metabolism of the new psychoactive substances N,N-diallyltryptamine (DALT) and 5-methoxy-DALT and their detectability in urine by GC-MS, LC-MSn, and LC-HR-MS-MS. Anal Bioanal Chem. 2015;407(25):7831-7842. doi:10.1007/s00216-015-8955-0
Shen HW, Jiang XL, Winter JC, Yu AM. Psychedelic 5-methoxy-N,N-dimethyltryptamine: metabolism, pharmacokinetics, drug interactions, and pharmacological actions. Curr Drug Metab. 2010;11(8):659-666. doi:10.2174/138920010794233495
Bergh MS, Bogen IL, Wohlfarth A, Wilson SR, Øiestad ÅML. Distinguishing between cyclopropylfentanyl and crotonylfentanyl by methods commonly available in the forensic laboratory. Ther Drug Monit. 2019;41(4):519-527. doi:10.1097/FTD.0000000000000617
Øiestad EL, Johansen U, Øiestad ÅML, Christophersen AS. Drug screening of whole blood by ultra-performance liquid chromatography-tandem mass spectrometry. J Anal Toxicol. 2011;35(5):280-293. doi:10.1093/anatox/35.5.280
Bruni PS, Grafinger KE, Nussbaumer S, König S, Schürch S, Weinmann W. Study of the in vitro and in vivo metabolism of 4-HO-MET. Forensic Sci Int. 2018;290:103-110. doi:10.1016/j.forsciint.2018.06.037
Wagmann L, Manier SK, Meyer MR. Can the intake of a synthetic tryptamine be detected only by blood plasma analysis? A clinical toxicology case involving 4-HO-MET. J Anal Toxicol. 2022;46(5):567-572. doi:10.1093/jat/bkab062
Zhai W, Li L, Zhao J, et al. Tentative identification of in vitro metabolites of O-acetylpsilocin (psilacetin, 4-AcO-DMT) by UHPLC-Q-Orbitrap MS. Drug Test Anal. 2022;14(7):1300-1309. doi:10.1002/dta.3255
Malaca S, Huestis MA, Lattanzio L, et al. Human hepatocyte 4-acetoxy-N,N-diisopropyltryptamine metabolite profiling by reversed-phase liquid chromatography coupled with high-resolution tandem mass spectrometry. Metabolites. 2022;12(8):705. doi:10.3390/metabo12080705
Carlier J, Malaca S, Huestis MA, Tagliabracci A, Tini A, Busardò FP. Biomarkers of 4-hydroxy-N,N-methylpropyltryptamine (4-OH-MPT) intake identified from human hepatocyte incubations. Expert Opin Drug Metab Toxicol. 2022;18(12):831-840. doi:10.1080/17425255.2022.2166826