Evaluating alcohol effects on Blood Steroid Profile by simultaneous measurement of EAAS doping and alcohol consumption markers via LC-MS

Code: 242D07FP

The longitudinal monitoring of Testosterone and Androstenedione blood concentrations was introduced in the Steroidal Module of Athlete Biological Passport in 2023 as a complementary tool to the urinary module. Although this approach has proven to increase the detection capability of endogenous anabolic androgenic steroids doping, limited information is currently available regarding confounding factors of blood steroid profiling. In this context, alcohol consumption is one of the exogenous confounding factors in urinary steroid analysis, but its influence on blood steroid profile has not yet been investigated in detail. The aim of this project is therefore to fill this gap by firstly focusing on the development of a novel LC-MS method for the simultaneous quantification of Testosterone, Androstenedione, and two markers of alcohol consumption, Ethyl glucuronide and Ethyl sulfate. Mass spectrometric parameters will be fine-tuned to achieve maximal sensitivity, and chromatographic conditions will be optimized to find the best compromise between resolution and analysis time. Once the developed method is validated according to WADA's requirements and transferred to a WADA-accredited laboratory, it will be employed in both laboratories to analyze real serum samples collected during a dedicated clinical study, allowing for inter-laboratory comparison. The clinical study will involve 20 healthy volunteers, who will be administered a moderate dose of alcohol. For each participant, both alcohol and placebo will be administered in two separate sessions. Serum and urine samples will be collected before the intervention and at 5 consecutive time points during the day of administration, with an additional sample collected the morning after the intervention. The results obtained from serum samples analyzed with the novel method and from urine samples analyzed using the routine method of WADA-accredited laboratory will then be investigated to characterize the effect of alcohol consumption on both parts of the Steroidal Module.