The influence of small doses of ethanol on the urinary testosterone to epitestosterone ratio in men and women

Testosterone treatment stimulates the production of red blood cells and alters iron homeostasis. Thus, we investigated whether the ‘haematological module’ of the athlete biological passport (ABP) used by the World Anti‐Doping Agency can be used to indicate misuse of testosterone. Nineteen eugonadal men received intramuscular injections of either 250 mg Sustanon®, a blend of four testosterone esters, or placebo on days 0 and 21 in a randomized, placebo‐controlleddouble‐blind design. Urine samples and blood samples were collected twice pre‐treatment, at least 5 days apart, and on days 1, 3, 5, 10 and 14 post‐injections to assess steroidal and haematological biomarkers of the ABP. The steroidal profile was flagged suspicious in all Sustanon®‐treated subjects, whereas the haematological profile was flagged suspicious in six out of nine subjects. When both sensitivity and specificity were considered, reticulocyte percentage (RET%) appeared as the best marker of the haematological module for implying testosterone ester misuse. Atypical blood passport samples were used to select time points for further isotope‐ratio mass spectrometry (IRMS) analysis of testosterone and its metabolites in simultaneously collected urine. In addition to the testosterone (T) to epitestosterone (E) ratio, the RET% and OFF‐Score could help identify suspicious samples for more targeted IRMS testing. The results demonstrate that unexpected fluctuations in RET% can indicate testosterone doping if samples are collected 3–10 days after injection. From an anti‐doping perspective, the haematological and steroidal modules of the ABP should complement each other when planning targeted follow‐up testing and substantiating likely misuse of testosterone.

Section: Methodsmentioning

confidence: 53%

Changes in blood parameters after intramuscular testosterone ester injections – Implications for anti‐doping

Solheim

Mørkeberg²,

Dehnes

et al. 2020

“…Nine of the samples with EtG were from participants with undetectable T concentrations. As EtG may be associated with increased T concentrations, 15,16 the symbols showing the average T concentrations or T/E ratio from the participants with EtG concentrations in Figures 2 and 3 are white to illustrate that these values may be confounded by ethanol consumption. Removing these values from the average T concentration calculation did not change the significance level of differences in neither T concentration nor T/E ratio.…”

Section: Resultsmentioning

confidence: 99%

“…All the samples that may have been confounded by ethanol are marked with white dots, or squares around the dots in the figures. Notably, previous studies have shown that EtG concentrations <5 μg/ml also can exert effect on the urinary steroid profile and that 5αAdiol is sometimes also affected 15 …”

Section: Discussionmentioning

confidence: 99%

Urinary steroid profile in relation to the menstrual cycle

Schulze

Suominen

Bergström

et al. 2020

The interpretation of the steroidal module of the Athlete Biological Passport (ABP) in female athletes is complex due to the large variation of the endogenous urinary steroids. The menstrual cycle seems to be one of the largest confounders of the steroid profile. The duration of the different phases in the menstrual cycle differs between women and is difficult to predict only by counting days after menstruation. Here, we have determined the follicle, ovulation, and luteal phases, by assessing the menstrual hormones in serum samples collected from 17 healthy women with regular menses. Urine samples were collected three times per week during two consecutive cycles to measure the urinary steroid concentrations used in the ABP. The metabolite that was mostly affected by the menstrual phases was epitestosterone (E), where the median concentration was 133% higher in the ovulation phase compared to the follicle phase (p < 0.0001). The women with a large coefficient of variation (CV) in their first cycle also had a large CV in their second cycle and vice versa. The inter‐individual difference was extensive with a range of 11%–230% difference between the lowest and the highest T/E ratio during a cycle. In conclusion, E and ratios with E as denominator are problematic biomarkers for doping in female athletes. The timing of the sample collection in the menstrual cycle will have a large influence on the steroid profile. The results of this study highlight the need to find additional biomarkers for T doping in females.

“…Some of the measured T/E ratios in this administration are clearly influenced by alcohol. The current WADA technical document TD2018EAAS requires reporting of the conjugated metabolite of ethanol (EtG) above 5 μg/mL but recent publications have demonstrated that lower concentrations of EtG may also influence the T/E ratio 21 . Four subjects had a number of samples that contained EtG above 5 μg/mL.…”

Section: Discussionmentioning

confidence: 99%

“…ratio. 21 Four subjects had a number of samples that contained EtG above 5 μg/mL. The T/E ratios from these collections produced some of the highest values in the study and have been highlighted in Table 3.…”

Section: D2 a E A E A E A E A E C A E A E A A E A D4 A E A mentioning

confidence: 92%

Evaluation of longitudinal steroid profiling with the ADAMS adaptive model for detection of transdermal, intramuscular, and subcutaneous testosterone administration

Nair

Husk

Miller

et al. 2020

The steroidal module of the Athlete Biological Passport (ABP) has been used since 2014 for the longitudinal monitoring of urinary testosterone and its metabolites in order to identify samples suspicious for the use of synthetic forms of endogenous anabolic androgenic steroids (EAAS). Samples identified by the module may then be confirmed by isotope ratio mass spectrometry (IRMS) to establish clearly the exogenous origin of testosterone and/or metabolites in the sample. To examine the detection capability of the steroidal ABP model, testosterone administration studies were performed with various doses and three routes of administrationtransdermal, intramuscular, and subcutaneous with 15 subjects for each route of administration. Urine samples were collected before, during, and after administration and steroid profiles were analyzed using the steroidal ABP module in ADAMS. A subset of samples from each mode of administration was also analyzed by IRMS. The steroidal ABP module was more sensitive to testosterone use than population-based thresholds and with high dose administrations there was very good agreement between the IRMS results and samples flagged by the module. However, with low dose administration the ABP module was unable to identify samples where testosterone use was still detectable by IRMS analysis. The testosterone/epitestosterone (T/E) ratio was the most diagnostic parameter for longitudinal monitoring with the exception of low testosterone excretors for whom the 5α-androstane-3α, 17β-diol/epitestosterone (5αAdiol/E) ratio may provide more sensitivity.