Abstract
Testosterone (T) and related androgens are performance enhancing drugs (PEDs) abused by some athletes to gain competitive advantage. To monitor unauthorized androgen abuse, doping control programs use mass spectrometry (MS) to detect androgens, synthetic anabolic-androgenic steroids (AASs) and their metabolites in an athlete’s urine. AASs of unknown composition will not be detected by these procedures. Since AASs achieve their anabolic effects by activating the Androgen Receptor (AR), cell-based bioassays that measure the effect of a urine sample on AR activity are under investigation as complementary, pan-androgen detection methods. We evaluated an AR BioAssay as a monitor for androgen activity in urine pre-treated with glucuronidase, which releases T from the inactive T-glucuronide that predominates in urine. AR BioAssay activity levels were expressed as ‘T-equivalent’ concentrations by comparison to a T dose response curve. The T-equivalent concentrations of androgens in the urine of hypogonadal participants supplemented with T (in whom all androgenic activity should arise from T) were quantitatively identical to the T measurements conducted by MS at the UCLA Olympic Analytical Laboratory (0.96 ± 0.22). All 17 AASs studied were active in the AR BioAssay; other steroids were inactive. 12 metabolites of 10 commonly abused AASs, which are used for MS monitoring of AAS doping because of their prolonged presence in urine, had reduced or no AR BioAssay activity. Thus, the AR BioAssay can accurately and inexpensively monitor T, but its ability to monitor urinary AASs will be limited to a period immediately following doping in which the active AASs remain intact.
Highlights
The abuse of unapproved performance enhancing drugs (PEDs) remains a barrier to fair athletic competition [1,2,3,4]
That Androgen Receptor (AR) BioAssay consists of a HeLa cell line that stably expresses the yellow fluorescent protein (FP) (YFP)-tagged AR together with an mCherry-NLS-mCherry construct
Anti-androgens, commonly used in the clinical setting and unlikely to be abused as PEDs, showed the predicted ability to counteract androgen activation of the AR BioAssay
Summary
The abuse of unapproved PEDs remains a barrier to fair athletic competition [1,2,3,4]. PED abuse is currently monitored by unannounced testing [5, 6]. The abuse of PEDs, androgens, is common amongst non-elite athletes and by those in the general population seeking to improve body image [7, 8]. Rates are higher in youths: in 2013, 2.9% of US grade 12 males self-reported androgen abuse over the prior twelve months [10]. The current rate of androgen abuse amongst US grade 12 males is down substantially from a peak of 8% in the early 2000s [10, 12], AAS abuse by adolescents and adults remains troubling [13] and unmonitored
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