Metabolism and cytotoxicity studies of the two hallucinogens 1cP-LSD and 4-AcO-DET in human liver and zebrafish larvae models using LC-HRMS/MS and a high-content screening assay.
Tanja M Gampfer, Victoria Schütz, Philip Schippers, Sari Rasheed, Jonas Baumann, Lea Wagmann, Benedikt Pulver, Folker Westphal, Veit Flockerzi, Rolf Müller, Markus R Meyer
Journal of Pharmaceutical and Biomedical Analysis August 1, 2024 DOI: 10.1016/j.jpba.2024.116187 (opens in new tab)
Study at a glance
AI-extracted from the abstract| Characteristics | Preclinical study Peer reviewed |
|---|---|
| Population | Pooled human liver S9 fraction, recombinant human isozymes, zebrafish larvae, and human hepatoma cell line HepG2 |
| Interventions | 1cP-LSD 4-AcO-DET |
| Topics | LSD |
| Keywords | Cytotoxicity Hepg2 Lc-hrms/ms New psychoactive substances Zebrafish larvae Preclinical research Hallucinogens Drug metabolism Toxicology testing |
| Citations | 11 |
| Key points | 1cP-LSD and 4-AcO-DET were metabolized in vitro and in vivo, with no cytotoxicity observed in HepG2 cells, but 4-AcO-DET with a CYP inhibitor altered two parameters at very high concentrations. |
Abstract
The continuous emergence of new psychoactive substances (NPS) attracted a great deal of attention within recent years. Lately, the two hallucinogenic NPS 1cP-LSD and 4-AcO-DET have appeared on the global market. Knowledge about their metabolism to identify potential metabolic targets for analysis and their cytotoxic properties is lacking. The aim of this work was thus to study their in vitro and in vivo metabolism in pooled human liver S9 fraction (pHLS9) and in zebrafish larvae (ZL) by means of liquid chromatography-high-resolution tandem mass spectrometry. Monooxygenases involved in the initial metabolic steps were elucidated using recombinant human isozymes. Investigations on their cytotoxicity were performed on the human hepatoma cell line HepG2 using a multiparametric, fluorescence-based high-content screening assay. This included measurement of CYP-enzyme mediated effects by means of the unspecific CYP inhibitor 1-aminbenzotriazole (ABT). Several phase I metabolites of both compounds and two phase II metabolites of 4-AcO-DET were produced in vitro and in vivo. After microinjection of 1cP-LSD into the caudal vein of ZL, three out of seven metabolites formed in pHLS9 were also detected in ZL. Twelve 4-AcO-DET metabolites were identified in ZL after exposure via immersion bath and five of them were found in pHLS9 incubations. Notably, unique metabolites of 4-AcO-DET were only produced by ZL, whereas 1cP-LSD specific metabolites were found both in ZL and in pHLS9. No toxic effects were observed for 1cP-LSD and 4-AcO-DET in HepG2 cells, however, two parameters were altered in incubations containing 4-AcO-DET together with ABT compared with incubations without ABT but in concentrations far above expected in vivo concentration. Further investigations should be done with other hepatic cell lines expressing higher levels of CYP enzymes.
Comparable studies
Other preclinical and animal studies on LSD, most cited first.
Citations in the library
Cited by 4
- In vitro metabolic fate of 1-[3-(trimethylsilyl)propanoyl] lysergic acid diethylamide (1S-LSD), a silicon-containing LSD analog. Forensic Toxicology July 29, 2025
- Metabolic fate of drugs of abuse and new psychoactive substances: A pilot study on a novel workflow using a zebrafish embryo model combined with human microdosing. British Journal of Clinical Pharmacology June 16, 2025
- In Vitro Metabolism of 1‐Benzoyl‐Lysergic Acid Diethylamide (1Bz‐LSD) and Identification of a Deethylated Metabolite (1Bz‐LAE) Using a Synthesized Reference Standard Drug Testing and Analysis February 17, 2026
- Toxicokinetics and analytical toxicology of four N, N-dimethyltryptamine derivatives studied in human in vitro systems and in vivo by means of zebrafish embryos. Archives of Toxicology September 21, 2026
Cites 10
- Pharmacokinetics and Pharmacodynamics of Lysergic Acid Diethylamide in Healthy Subjects Clinical Pharmacokinetics February 14, 2017
- Is LSD toxic? Forensic Science International March 1, 2018
- Pharmacokinetics of N,N-dimethyltryptamine in Humans. European Journal of Drug Metabolism and Pharmacokinetics May 1, 2023
- Pharmacologic Activity of Substituted Tryptamines at 5-Hydroxytryptamine (5-HT)2A Receptor (5-HT2AR), 5-HT2CR, 5-HT1AR, and Serotonin Transporter. Journal of Pharmacology and Experimental Therapeutics January 20, 2023
- Return of the lysergamides. Part VI: Analytical and behavioural characterization of 1‐cyclopropanoyl‐d‐lysergic acid diethylamide (1CP‐LSD) Drug Testing and Analysis March 16, 2020
and 5 more in the library