Longitudinal Profiling of Endogenous Steroids in Blood Using the Athlete Biological Passport Approach

Abstract

Context Detection of endogenous anabolic androgenic steroids (EAAS), like testosterone (T), as doping agents has been improved with the launch of the Steroidal Module of the Athlete Biological Passport (ABP) in urine samples. Objective To target doping practices with EAAS, particularly in individuals with low level of biomarkers excreted in urine, by including new target compounds measured in blood. Design T and T/Androstenedione (T/A4) distributions were obtained from 4 years of anti-doping data and applied as priors to analyze individual profiles from two T administration studies in female and male subjects. Setting Anti-doping laboratory. Elite athletes (n=823) and male and female clinical trials subjects (n=19 and 14, respectively). Intervention(s) Two open-label administration studies were carried out. One involved a control phase period followed by patch and then oral T administration in male volunteers and the other followed female volunteers during three menstrual cycles with 28 days of daily transdermal T application during the second month. Main Outcome Measure(s) Serum samples were analyzed for T and A4 and the performance of a longitudinal, ABP-based approach was evaluated for T and T/A4. Results An ABP-based approach set at a 99% specificity flagged all female subjects during the transdermal T application period and 44% of subjects three days after the treatment. Testosterone showed the best sensitivity (74%) in response to transdermal T application in males. Conclusions Inclusion of T and T/A4 as markers in the Steroidal Module can improve the performance of the ABP to identify T transdermal application, particularly in females.

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

“…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), 5αandrostane-3α,17β-diol (5αAdiol), and 5β-androstane-3α,17β-diol (5βAdiol), several confounding factors as for instance time of day, inversus 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 approaches but with much lower analytical effort. 43 It was stated that the methodology might have a limited range of optimal performance in the light of, for example, ion suppression effects, but in consideration of the commonly observed endogenous steroid concentrations, the arguably limited range might well be sufficient for the intended purpose.…”

“…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.…”

Influence of multiple human chorionic gonadotropin administrations on serum and urinary steroid Athlete Biological Passport profiles in males

“…8 After exogenous administration of T, the T/E ratio will rise 9 and samples with T/E ratios outside of an individual's limits can be flagged and further analyzed, such as assessing urinary steroid carbon isotope signature with isotope ratio mass spectrometry (IRMS) to determine the origin of the compound or for the presence of steroid esters in blood. While monitoring athletes' urinary T/E ratio for doping control purposes has been used for decades, the serum steroidal module of the ABP was introduced just recently to increase the sensitivity of the ABP in individuals with low levels of urinary T and E. 10,11 This module tracks serum T and androstenedione (A4), a metabolic precursor to T, and the T/A4 ratio. 8 Like the urinary module, samples outside of the individual's limits can be flagged for further, more detailed analysis.…”