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

© 2021 The Authors. Drug Testing and Analysis published by John Wiley & Sons Ltd..

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.

Medienart:

E-Artikel

Erscheinungsjahr:

2021

Erschienen:

2021

Enthalten in:

Zur Gesamtaufnahme - volume:13

Enthalten in:

Drug testing and analysis - 13(2021), 6 vom: 12. Juni, Seite 1169-1177

Sprache:

Englisch

Beteiligte Personen:

Savill, Ryan [VerfasserIn]
Baues, Helge [VerfasserIn]
Voigt, Emmely [VerfasserIn]
Zierau, Oliver [VerfasserIn]
Thieme, Detlef [VerfasserIn]
Keiler, Annekathrin Martina [VerfasserIn]

Links:

Volltext

Themen:

3XMK78S47O
Cell culture
Comparative Study
Doping
Journal Article
LC-MS/MS
Metabolism
Testosterone

Anmerkungen:

Date Completed 06.12.2021

Date Revised 14.12.2021

published: Print-Electronic

Citation Status MEDLINE

doi:

10.1002/dta.3009

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM320886484