Quantification of endogenous steroid sulfates and glucuronides in human urine after intramuscular administration of testosterone esters

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open access

Highlights

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A LC-MS method was developed for the urinary analysis of conjugated steroids in their intact form.

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Changes in urinary levels of phase II metabolites are prominent after administration of testosterone esters.

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A new potential marker for the intake of exogenous testosterone was identified.

Abstract

For an effective detection of doping with pseudo-endogenous anabolic steroids, the urinary steroid profile is of high value. In this work, the aim was to investigate steroid metabolism disruption after exogenous intramuscular administration of different testosterone esters. The investigation focused on both sulfo – and glucoro conjugated androgens. A single intramuscular injection of either 1000 mg testosterone undecanoate (Nebido®) or a mixture of 30 mg testosterone propionate, 60 mg testosterone phenylpropionate, 60 mg testosterone isocaproate, and 100 mg testosterone decanoate (Sustanone®), was given to six healthy volunteers. Urine was collected throughout a testing period of 60 days. A LC-MS method was developed and validated for the analysis of eight conjugated steroids in their intact form. The results show that urinary changes in both sulfo – and glucuro conjugated steroid levels are prominent after the injection of testosterone esters. A promising potential marker for the intake of exogenous testosterone is the combined ratio of epitestosterone sulfate/epitestosterone glucuronide to testosterone sulfate/testosterone glucuronide ((ES/EG)/(TS/TG)) as a complementary biomarker for testosterone abuse. This represents a new piece of evidence to detect testosterone doping, representing a new approach and being independent from the metabolic connections of the markers in the steroid passport.

Keywords

Doping analysis

Urinary steroid profile

Phase II metabolites

Steroid sulfates

Steroid glucuronides

© 2020 The Authors. Published by Elsevier Inc.