Variability of the urinary and blood steroid profiles in healthy and physically active women with and without oral contraception

Abstract

The steroidal module of the athlete biological passport (ABP) targets the use of pseudo‐endogenous androgenous anabolic steroids in elite sport by monitoring urinary steroid profiles. Urine and blood samples were collected weekly during two consecutive oral contraceptive pill (OCP) cycles in 15 physically active women to investigate the low urinary steroid concentrations and putative confounding effect of OCP. In urine, testosterone (T) and epitestosterone (E) were below the limit of quantification of 1 ng/ml in 62% of the samples. Biomarkers' variability ranged between 31% and 41%, with a significantly lesser variability for ratios (except for T/E [41%]): 20% for androsterone/etiocholanolone (p < 0.001) and 25% for 5α‐androstane‐3α,17β‐diol/5ß‐androstane‐3α,17β‐diol (p < 0.001). In serum, markers' variability (testosterone: 24%, androstenedione: 23%, dihydrotestosterone: 19%, and T/A4: 16%) was significantly lower than in urine (p < 0.001). Urinary A/Etio increased by >18% after the first 2 weeks (p < 0.05) following withdrawal blood loss. In contrast, serum T (0.98 nmol/l during the first week) and T/A4 (0.34 the first week) decreased significantly by more than 25% and 17% (p < 0.05), respectively, in the following weeks. Our results outline steroidal variations during the OCP cycle, highlighting exogenous hormonal preparations as confounder for steroid concentrations in blood. Low steroid levels in urine samples have a clear negative impact on the subsequent interpretation of steroid profile of the ABP. With a greater analytical sensitivity and lesser variability for steroids in healthy active women, serum represents a complementary matrix to urine in the ABP steroidal module.

Annual banned‐substance review 16th edition—Analytical approaches in human sports drug testing 2022/2023

“…The ABP was recently complemented by the inclusion of two serum steroid markers, that is, T and androst‐4‐ene‐3,17‐dione (Adione; A4), to further strengthen the ABPs capability of detecting the administration of endogenous AAS, especially in females 130 . The intraindividual stability of urinary steroid profiles was found to be less robust in female than in male athletes, 34 and despite the use of concentrations and concentration ratios of a variety of steroidal analytes such as T, epitestosterone (EpiT), androsterone (A), etiocholanolone (E), 5α‐androstane‐3α,17β‐diol (5αAdiol), and 5β‐androstane‐3α,17β‐diol (5βAdiol), several confounding factors as for instance time of day, in‐ versus out‐of‐competition sample collection, and hormonal contraception have been shown to affect urinary steroid profile data, 35 and consensus exists in the added value of serum steroid analyses in the context of sports drug testing 36 . Accounting for confounding factors in ABP result interpretations is vital and likewise is the optimization of analytical tests.…”