Improving Detection of Endogenous Anabolic Steroids misuse by measuring endogenous Sulfate metabolites 2 (IDEASS 2)

Code: 19D03RV 

The detection of EAS abuse is currently performed using the steroid profile, however improvements are needed to prolong detection times (DTs). Results of a previous WADA (17D09RV) demonstrated that ratios between some sulfate metabolites significantly prolonged the DTs of oral T administration in Caucasian volunteers with respoec to T/E ratio. Some of the ratios were elevated up to the lsat sample collected several days later. Steroid sulfates have not been evaluated comprehensively in Asian population after oral T administration. And dermal administration has not been evaluation in Caucasian or Asian populations.

The objective of the present project is to continue the evaluation of sulfate metabolites to prolong the detectability of T misuse. The research will be focused in the following specific objectives:
- To verify the usefulness of sulfate markers in urine in Asian population after oral T administration.
- To verify the usefulness of sulfate markers in urine in Asian and Caucasian populations after dermal T administration.
- To identify androstanediol sulfate 1, which was one of the most useful markers identified in the previous study.
- To develop an initial testing procedure to quantify the relevant steroid sulfates in urine on a routine basis.

Main findings

Testosterone (T) and its metabolites are also excreted in urine as sulfate conjugates. The aim of this project was to evaluate sulfate metabolites as a markers of T administration, and the study was focused in two specific objectives: behaviour of sulfate markers after dermal T administration, and after oral T administration in Asian volunteers.

Regarding dermal administration, T gel was administered for three consecutive days (100 mg/day) to six Caucasian and six Asian volunteers. Androsterone sulfate/testosterone sulfate (A-S/T-S), epiandrosterone sulfate/epitestosterone sulfate (epiA-S/E-S), epiA-S/T-S and etiocholanolone sulfate/epitestosterone sulfate (Etio-S/E-S) provided the most consistent detectability for all volunteers and populations, with detection times ranging from 12 h to 2 days after the last administered dose. Additional ratios improved detectability to up to 5 days after the last administered dose, but only in particular cases. In general, sensitivity was similar or better than the conventional T/E ratio of the steroid profile.

Regarding oral T, a clinical study was performed with administration of a single oral dose (80 mg) to 13 Asian volunteers (11 del/del UGT2B17 genotype, 2 del/ins genotype). Some sulfate ratios, specifically androstanediol sulfate 1 (DiolS1)/ dihydroandrosterone sulfate (DHAS) and epiAS/DHAS, provided detection up to 5-6 days after administration in most of the volunteers. In all volunteers, the detection times using sulfate markers were significantly longer than those provided using T/E ratio (up to 24-36 h) and IRMS markers (up to 24-48h). Steroid sulfate ratios provide significantly better detection times for oral T administration than all other markers described up to now.

In summary, endogenous steroid sulfates improve the detection of T administration and data available indicate that steroid sulfates might complement and improve the current steroid profile in the detection of T administration.