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.
Wellenberg K Simon, Tanja M Gampfer, Wagmann Lea, Schippers Philip, Herrmann Jennifer, Müller Rolf, Westphal Folker, Markus R Meyer
British Journal of Clinical Pharmacology June 16, 2025 DOI: 10.1002/bcp.70140 (opens in new tab)
Study at a glance
AI-extracted from the abstract| Characteristics | Experimental study with zebrafish embryo model and human microdosing Pilot study Peer reviewed |
|---|---|
| Population | Zebrafish embryos and human participants |
| Interventions | Amphetamine cocaine LSD MDMA methamphetamine THC MDMB-CHMICA MDPPP |
| Duration | 24 h |
| Topics | Microdosing |
| Keywords | Lc‐hrms/ms Drugs of abuse New psychoactive substances Xenobiotic metabolism Animal model Drug analysis Toxicology screening Zebrafish model zebrafish Zebrafish embryos Research model Biological model Drug metabolism metabolism Drug pharmacokinetics Metabolite identification Biomarker discovery Substance identification Drug testing Lc-hrms/ms |
| Citations | 1 |
| Key findings | Zebrafish embryo metabolism followed by human microdosing can identify main human urine metabolites for drugs of abuse, though challenges with low doses and bioavailability remain. |
Abstract
The aim of this study was to develop a novel workflow to identify human urine biomarkers for drugs of abuse and new psychoactive substances. Metabolites of amphetamine, cocaine, LSD, MDMA, methamphetamine, THC, MDMB-CHMICA, and MDPPP were first identified in a zebrafish embryo (ZE) metabolism study followed by comparison to most abundant human metabolites in literature. Finally, metabolites were confirmed by human microdosing (HMD). ZEs 4 days post fertilization were exposed via immersion in Danieau's medium containing the compound or by injection into the caudal vein. After euthanization and freeze-drying, the ZEs were extracted using methanol. HMD was performed by oral administration of individual compound solutions according to HMD guidelines. Urine was collected spontaneously over 24 h and extracted via solid-phase extraction with and without conjugate cleavage. Samples were then analysed using reversed-phase liquid chromatography coupled to high resolution tandem mass spectrometry. Overall, both ZE and HMD allowed identification of main human urine metabolites as described in literature. Exceptions for HMD were LSD due to the low applied dose and the cannabinoids, probably due to low oral bioavailability. Furthermore, ZE generally produced more metabolites compared to HMD, including conjugates. The proposed workflow of ZE exposure followed by HMD can provide quick and reliable data for prediction of analytical biomarkers for urinary drug screening. Following the initial identification of metabolites in ZE, human metabolites can afterwards be confirmed by HMD study. However, there are still challenges regarding HMD such as different routes of administration and detectability of applied low doses.
Comparable studies
Other preclinical and animal studies on microdosing, most cited first.
| Study | Year | Design | Participants |
|---|---|---|---|
| Psilocin and ketamine microdosing: effects of subchronic intermittent microdoses in the elevated plus-maze in male Wistar rats Wistar rats | 2018 | Experimental study | n = 40 |
| Repeated low doses of psilocybin increase resilience to stress, lower compulsive actions, and strengthen cortical connections to the paraventricular thalamic nucleus in rats. Rats | 2023 | Experimental study | |
| Repeated microdoses of LSD do not alter anxiety or boldness in zebrafish. Zebrafish (Danio rerio) | 2024 | Experimental study | |
| Application of microarray patches for the transdermal administration of psychedelic drugs in micro-doses. Excised neonatal porcine skin and rats | 2025 | Experimental study | |
| Toward Predicting Acute Myeloid Leukemia Patient Response to 7 + 3 Induction Chemotherapy via Diagnostic Microdosing Paired sensitive and resistant cancer cell lines, AML cell lines, and 10 viably... | 2018 | Preclinical cell line study with a pilot retrospective analysis of primary patient samples | n = 10 |
Citations in the library
Cites 6
- Human Pharmacology of MDMA Therapeutic Drug Monitoring March 19, 2004
- The effects of microdose LSD on time perception: a randomised, double-blind, placebo-controlled trial Psychopharmacology November 26, 2018
- Pharmacokinetics and Concentration-Effect Relationship of Oral LSD in Humans The International Journal of Neuropsychopharmacology June 24, 2015
- Absorption, Distribution, Metabolism and Excretion Pharmacogenomics of Drugs of Abuse Pharmacogenomics February 1, 2011
- Metabolism of lysergic acid diethylamide (LSD): an update Drug Metabolism Reviews July 3, 2019
and 1 more in the library