Cell culture as a toolbox to generate phase I metabolites for antidoping screening

Publikation: Beitrag in FachzeitschriftForschungsartikelBeigetragenBegutachtung

Beitragende

Abstract

The knowledge of the biotransformation of compounds prohibited by the World Anti Doping Agency is of high concern as doping analyses are mostly based on the detection of metabolites instead of the parent compounds abused by athletes. While the self-administration of doping-relevant compounds is from an ethical point of view a rather problematic method to investigate metabolism, the usage of cell culture systems allows for studies on biotransformation in vitro. Five cell culture models with different tissue origin (liver, ovary, skin, kidney, and testis) were comparatively incubated with testosterone and epitestosterone as well as with the synthetic testosterone derivatives 17α-methyltestosterone and 4-chlorotestosterone to investigate the impact of synthetic modifications on phase I metabolic pathways. Cell culture supernatants were analyzed by high-performance liquid chromatography-tandem mass spectrometry. All cell lines possessed the default steroid phase I biotransformation reactions. The highest conversion rate was observed in ovarian (BG-1) and liver cells (HepG2). For BG-1 and skin cells (HaCaT), the 5α-reductase products 5α-dihydrotestosterone (for both) and 5α-androstane-3α/β,17β-diol (for BG-1 solely) were found to be prevailing after testosterone incubation. In kidney (COS-1) and HepG2 cells, the 17β-hydroxysteroid dehydrogenase activity was predominant as supported by the observation that the 17α-OH (epitestosterone) and the methyl group (17α-methyltestosterone) impeded the conversion rate in these cell lines. In conclusion, future work should extend the characterization of the BG-1 and HepG2 cells on phase II metabolic pathways to examine whether they are suitable models for the generation of metabolite reference collections comparable to those obtained by human excretion studies.

Details

OriginalspracheEnglisch
Seiten (von - bis)1169-1177
Seitenumfang9
FachzeitschriftDrug testing and analysis
Jahrgang13
Ausgabenummer6
PublikationsstatusVeröffentlicht - Juni 2021
Peer-Review-StatusJa

Externe IDs

Scopus 85100951501
ORCID /0000-0002-2157-4711/work/142251650

Schlagworte

Schlagwörter

  • Animals, COS Cells, Cell Culture Techniques, Cell Line, Chlorocebus aethiops, Chromatography, High Pressure Liquid/methods, Doping in Sports/prevention & control, HaCaT Cells, Hep G2 Cells, Humans, Tandem Mass Spectrometry/methods, Testosterone/analogs & derivatives