Evaluation of epiandrosterone as a long‐term marker of testosterone use

Self Cite

The steroidal module of the Athlete Biological Passport (ABP) has been used since 2014 for the longitudinal monitoring of urinary testosterone and its metabolites to identify samples suspicious for the use of synthetic forms of Endogenous Anabolic Androgenic Steroids (EAAS). Multiple recent studies have suggested that monitoring of blood parameters may provide enhanced detectability of exogenous testosterone administration. Transdermal and intramuscular testosterone administration studies were carried out in 15 subjects, and the effect on blood steroidal levels, hematological parameters, and gonadotropins was evaluated. Serum testosterone and dihydrotestosterone levels increased while gonadotropin levels were suppressed after administration. A modest increase in reticulocytes was also observed. The blood parameters that were responsive to the administrations were combined into several linear discriminant models targeting both administration (on) and washout (off) phases. The models were effective in detecting the large dose intramuscular administration but were less successful in the detection of the lower dose transdermal application. The blood profiling models may provide complementary value but do not appear to be substantially more advantageous than longitudinal urinary profiling.

Section: Resultsmentioning

confidence: 99%

“…Urine samples from subjects 1, 2, 6, 8, and 10 were also analyzed by GC/C/IRMS, and all subjects had more collections that were adverse by IRMS than identified by either profiling model. 31…”

Section: Comparison With Urinary Steroidal Abp Valuesmentioning

confidence: 99%

Evaluation of blood parameters by linear discriminant models for the detection of testosterone administration

Nair

Sharpe

Husk³

et al. 2021

Self Cite

“…This design allowed for a highly complex multivariate analysis of T treatment effects on serum and urine steroid profiles explaining 25.7% and 31%, respectively, of total variation. The models identified two main com- The present study investigated the effects of T administration on the carbon isotope ratio of T and 5α-diol, both shown previously to be the most informative for administration of transdermal T gel, 52,71 with the effects on 5α-diol due to the prominence of 5α-reductase expression in skin acting on T during its transdermal passage. Previous studies have investigated the potential of epiandrosterone sulfate as a long-term biomarker for androgen doping in men 8,52,72 including use of a single-oral dose of 50-mg DHEA in nine men 72 or transdermal T gel in two men taking 50-mg T applied topically for 7 days 8 and five men taking 40-mg T applied topically for 14 days.…”

Section: Discussionmentioning

confidence: 99%

Detection of testosterone microdosing in healthy females

Savkovic

Desai

et al. 2022

Self Cite

The ready detectability of synthetic androgens by mass spectrometry (MS)-based antidoping tests has reoriented androgen doping to using testosterone (T), which must be distinguished from its endogenous counterpart making detection of exogenous T harder. We investigated urine and serum steroid and hematological profiling individually and combined to determine the optimal detection model for T administration in women. Twelve healthy females provided six paired blood and urine samples over 2 weeks prior to treatment consisting of 12.5-mg T in a topical transdermal gel applied daily for 7 days. Paired blood and urine samples were then obtained at the end of treatment and Days 1, 2, 4, 7, and 14 days later. Compliance with treatment and sampling was high, and no adverse effects were reported. T treatment significantly increased serum and urine T, serum dihydrotestosterone (DHT), urine 5α-androstane-3α,17β-diol (5α-diol) epitestosterone (E), and urine T/E ratio with a brief window of detection (2-4 days) as well as total and immature (medium and high fluorescence) reticulocytes that remained elevated over the full 14 posttreatment days. Carbon isotope ratio MS and the OFF score and Abnormal Blood Profile score (ABPS) were not discriminatory. The optimal multivariate model to identify T exposure combined serum T, urine T/E ratio with three hematological variables (% high fluorescence reticulocytes, mean corpuscular hemoglobin, and volume) with the five variables providing 93% correct classification (4% false positive, 10% false negatives). Hence, combining select serum and urine steroid MS variables with reticulocyte measures can achieve a high but imperfect detection of T administration to healthy females.

“…[10][11][12][13][14] In recent years, the diagnostic relevance of intact phase II metabolites was increasing, for example, shown for exogenous AASs like stanozolol and clostebol as well as for epiandrosterone-sulfate as a marker for misuse of testosterone and related endogenous steroids. [15][16][17][18][19][20][21][22][23] The investigation of the HepG2 cells as an adequate in vitro model to study and generate metabolites for doping analysis was the aim of the present study. The exogenous AAS metandienone, for which the biotransformation has been well studied over the last decades, was chosen as model substance.…”

Section: Introductionmentioning

confidence: 99%

“…Though Westerink and Schoonen as well as Gerets et al showed phase II enzyme expression in HepG2 cells, previous studies focusing steroid biotransformation investigated testosterone phase I metabolism solely 10–14 . In recent years, the diagnostic relevance of intact phase II metabolites was increasing, for example, shown for exogenous AASs like stanozolol and clostebol as well as for epiandrosterone‐sulfate as a marker for misuse of testosterone and related endogenous steroids 15–23 …”

Section: Introductionmentioning

confidence: 99%

HepG2 as promising cell‐based model for biosynthesis of long‐term metabolites: Exemplified for metandienone

Zschiesche¹,

Chundela²,

Thieme³

et al. 2021

In order to detect the abuse of substances in sports, the knowledge of their metabolism is of undisputable importance. As in vivo administration of compounds faces ethical problems and might even not be applicable for nonapproved compounds, cell‐based models might be a versatile tool for biotransformation studies. We coincubated HepG2 cells with metandienone and D3‐epitestosterone for 14 days. Phase I and II metabolites were analyzed by high‐performance liquid chromatography (HPLC)–tandem mass spectrometry and confirmed by gas chromatography–mass spectrometry (GC–MS). The metandienone metabolites formed by HepG2 cells were comparable with those renally excreted by humans. HepG2 cells also generated the two long‐term metabolites 17β‐hydroxymethyl‐17α‐methyl‐18‐nor‐androst‐1,4,13‐trien‐3‐one and 17α‐hydroxymethyl‐17β‐methyl‐18‐nor‐androst‐1,4,13‐trien‐3‐one used in doping analyses, though in an inverse ratio compared with that observed in human urine. In conclusion, we showed that HepG2 cells are suitable as model for the investigation of biotransformation of androgens, especially for the anabolic androgenic steroid metandienone. They further proved to cover phase I and II metabolic pathways, which combined with a prolonged incubation time with metandienone resulted in the generation of its respective long‐term metabolites known from in vivo metabolism. Moreover, we showed the usability of D3‐epitestosterone as internal standard for the incubation. The method used herein appears to be suitable and advantageous compared with other models for the investigation of doping‐relevant compounds, probably enabling the discovery of candidate metabolites for doping analyses.