Cargando…
Human Hepatocyte 4-Acetoxy-N,N-Diisopropyltryptamine Metabolite Profiling by Reversed-Phase Liquid Chromatography Coupled with High-Resolution Tandem Mass Spectrometry
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-N,N-diisopropyltryptamine (4-AcO-DiPT) is a sy...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9413566/ https://www.ncbi.nlm.nih.gov/pubmed/36005577 http://dx.doi.org/10.3390/metabo12080705 |
_version_ | 1784775780953227264 |
---|---|
author | Malaca, Sara Huestis, Marilyn A. Lattanzio, Leonardo Marsella, Luigi T. Tagliabracci, Adriano Carlier, Jeremy Busardò, Francesco P. |
author_facet | Malaca, Sara Huestis, Marilyn A. Lattanzio, Leonardo Marsella, Luigi T. Tagliabracci, Adriano Carlier, Jeremy Busardò, Francesco P. |
author_sort | Malaca, Sara |
collection | PubMed |
description | 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-N,N-diisopropyltryptamine (4-AcO-DiPT) is a synthetic tryptamine related to 4-hydroxy-N,N-diisopropyltryptamine (4-OH-DiPT), 4-acetyloxy-N,N-dipropyltryptamine (4-AcO-DPT), and 4-acetoxy-N,N-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, O-glucuronidation, O-sulfation, N-oxidation, and N-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-N-oxide are suggested as optimal biomarkers to identify 4-AcO-DiPT consumption. |
format | Online Article Text |
id | pubmed-9413566 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-94135662022-08-27 Human Hepatocyte 4-Acetoxy-N,N-Diisopropyltryptamine Metabolite Profiling by Reversed-Phase Liquid Chromatography Coupled with High-Resolution Tandem Mass Spectrometry Malaca, Sara Huestis, Marilyn A. Lattanzio, Leonardo Marsella, Luigi T. Tagliabracci, Adriano Carlier, Jeremy Busardò, Francesco P. Metabolites Article 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-N,N-diisopropyltryptamine (4-AcO-DiPT) is a synthetic tryptamine related to 4-hydroxy-N,N-diisopropyltryptamine (4-OH-DiPT), 4-acetyloxy-N,N-dipropyltryptamine (4-AcO-DPT), and 4-acetoxy-N,N-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, O-glucuronidation, O-sulfation, N-oxidation, and N-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-N-oxide are suggested as optimal biomarkers to identify 4-AcO-DiPT consumption. MDPI 2022-07-29 /pmc/articles/PMC9413566/ /pubmed/36005577 http://dx.doi.org/10.3390/metabo12080705 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Malaca, Sara Huestis, Marilyn A. Lattanzio, Leonardo Marsella, Luigi T. Tagliabracci, Adriano Carlier, Jeremy Busardò, Francesco P. Human Hepatocyte 4-Acetoxy-N,N-Diisopropyltryptamine Metabolite Profiling by Reversed-Phase Liquid Chromatography Coupled with High-Resolution Tandem Mass Spectrometry |
title | Human Hepatocyte 4-Acetoxy-N,N-Diisopropyltryptamine Metabolite Profiling by Reversed-Phase Liquid Chromatography Coupled with High-Resolution Tandem Mass Spectrometry |
title_full | Human Hepatocyte 4-Acetoxy-N,N-Diisopropyltryptamine Metabolite Profiling by Reversed-Phase Liquid Chromatography Coupled with High-Resolution Tandem Mass Spectrometry |
title_fullStr | Human Hepatocyte 4-Acetoxy-N,N-Diisopropyltryptamine Metabolite Profiling by Reversed-Phase Liquid Chromatography Coupled with High-Resolution Tandem Mass Spectrometry |
title_full_unstemmed | Human Hepatocyte 4-Acetoxy-N,N-Diisopropyltryptamine Metabolite Profiling by Reversed-Phase Liquid Chromatography Coupled with High-Resolution Tandem Mass Spectrometry |
title_short | Human Hepatocyte 4-Acetoxy-N,N-Diisopropyltryptamine Metabolite Profiling by Reversed-Phase Liquid Chromatography Coupled with High-Resolution Tandem Mass Spectrometry |
title_sort | human hepatocyte 4-acetoxy-n,n-diisopropyltryptamine metabolite profiling by reversed-phase liquid chromatography coupled with high-resolution tandem mass spectrometry |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9413566/ https://www.ncbi.nlm.nih.gov/pubmed/36005577 http://dx.doi.org/10.3390/metabo12080705 |
work_keys_str_mv | AT malacasara humanhepatocyte4acetoxynndiisopropyltryptaminemetaboliteprofilingbyreversedphaseliquidchromatographycoupledwithhighresolutiontandemmassspectrometry AT huestismarilyna humanhepatocyte4acetoxynndiisopropyltryptaminemetaboliteprofilingbyreversedphaseliquidchromatographycoupledwithhighresolutiontandemmassspectrometry AT lattanzioleonardo humanhepatocyte4acetoxynndiisopropyltryptaminemetaboliteprofilingbyreversedphaseliquidchromatographycoupledwithhighresolutiontandemmassspectrometry AT marsellaluigit humanhepatocyte4acetoxynndiisopropyltryptaminemetaboliteprofilingbyreversedphaseliquidchromatographycoupledwithhighresolutiontandemmassspectrometry AT tagliabracciadriano humanhepatocyte4acetoxynndiisopropyltryptaminemetaboliteprofilingbyreversedphaseliquidchromatographycoupledwithhighresolutiontandemmassspectrometry AT carlierjeremy humanhepatocyte4acetoxynndiisopropyltryptaminemetaboliteprofilingbyreversedphaseliquidchromatographycoupledwithhighresolutiontandemmassspectrometry AT busardofrancescop humanhepatocyte4acetoxynndiisopropyltryptaminemetaboliteprofilingbyreversedphaseliquidchromatographycoupledwithhighresolutiontandemmassspectrometry |