A pilot study on subject-based comprehensive steroid profiling: novel biomarkers to detect testosterone misuse in sports

Context Until now, the testosterone/epitestosterone (T/E) ratio is the main marker for the detection of testosterone (T) misuse in athletes. As this marker can be influenced by a number of confounding factors, additional steroid profile parameters indicating T misuse can provide substantiating evidence of doping with endogenous steroids. The evaluation of a steroid profile is currently based upon population statistics. As large inter-individual variations exist, a paradigm shift towards subject-based references is ongoing in doping analysis. Objective Proposition of new biomarkers for the detection of testosterone in sports using extensive steroid profiling and an adaptive model based upon Bayesian inference. Subjects Six healthy male volunteers were administered with testosterone undecanoate. Population statistics were performed upon steroid profiles from 2014 male Caucasian athletes participating in official sport competition. Design An extended search for new biomarkers in a comprehensive steroid profile combined with Bayesian inference techniques as used in the athlete biological passport resulted in a selection of additional biomarkers that may improve detection of testosterone misuse in sports. Results Apart from T/E, 4 other steroid ratios (6 alpha-OH-androstenedione/16 alpha-OH-dehydroepiandrostenedione, 4-OH-androstenedione/16 alpha-OH-androstenedione, 7 alpha-OH-testosterone/7 beta-OH-dehydroepiandrostenedione and dihydrotestosterone/5 beta-androstane-3 alpha, 17 beta-diol) were identified as sensitive urinary biomarkers for T misuse. These new biomarkers were rated according to relative response, parameter stability, detection time and discriminative power. Conclusion Newly selected biomarkers were found suitable for individual referencing within the concept of the Athlete's Biological Passport. The parameters showed improved detection time and discriminative power compared to the T/E ratio. Such biomarkers can support the evidence of doping with small oral doses of testosterone.