Development of criteria for the detection of adrenosterone administration by gas chromatography‐mass spectrometry and gas chromatography‐combustion‐isotope ratio mass spectrometry for doping control

Abstract

Adrenosterone (androst-4-ene-3,11,17-trione, 11-oxoandrostenedione) is an endogenous steroid hormone that has been promoted as a dietary supplement capable of reducing body fat and increasing muscle mass. It is proposed that adrenosterone may function as an inhibitor of the 11beta-hydroxysteroid dehydrogenase type 1 enzyme (11beta-HSD1), which is primarily responsible for reactivation of cortisol from cortisone. The urinary metabolism of adrenosterone was investigated, after a single oral administration in two male subjects, by gas chromatography-mass spectrometry (GC-MS) and gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). Substantially increased excretion of 11beta-hydroxyandrosterone, 11beta-hydroxyetiocholanolone, 11-oxoandrosterone and 11-oxoetiocholanolone was observed. Minor metabolites such as 3alpha,17beta-dihydroxy-5beta-androstan-11-one, 3alpha-hydroxyandrost-4-ene-11,17-dione and 3alpha,11beta-dihydroxyandrost-4-en-17-one were also identified. The exogenous origin of the most abundant adrenosterone metabolites was confirmed by GC-C-IRMS according to World Anti-Doping Agency criteria. Through analysis of a reference population data set obtained from urine samples provided by elite athlete volunteers (n = 85), GC-MS doping control screening criteria are proposed: 11beta-hydroxyandrosterone concentration greater than 10 000 ng/mL (specific gravity adjusted to 1.020) or 11beta-hydroxyandrosterone/11beta-hydroxyetiocholanolone ratio greater than 20.Urine samples fulfilling these screening criteria may be subjected to GC-C-IRMS analysis for confirmation of adrenosterone administration.

Multiple mechanisms of ligand interaction with the human organic cation transporter, OCT2

“…5; Table 2) were, arguably, consistent with their presence within structural subclusters identified by Kido et al (24) as being effectively free of known transported substrates of OCT2. Adrenosterone, an endogenous steroid hormone that has been promoted as a dietary supplement capable of reducing body fat and increasing muscle mass (8), is a representative member of cluster III, which consists primarily of neutral sterols. It is relevant to note, therefore, that previous studies of corticosterone's inhibition of OCT2-mediated transport described behavior inconsistent with a competitive mechanism, including evidence of a noncompetitive mode of interaction (5,52).…”

Recent developments in the use of isotope ratio mass spectrometry in sports drug testing

“…Two laboratories reported data using collected samples, which were investigated for CIR and which gave a significant difference in δ values between athletes from different countries but which showed that the Δ values were not influenced [19,76]. Investigations on the misuse of 4-androstene-3,11,17-trione, a nutritional supplement [77], and a comprehensive method to detect cortisone misuse, again using oxidation of target analytes, were published [78]. Another research focus was on EpiT and its metabolites 5aEpiD and 5bEpiD, which provided data for another RefPop (n=74), again showing differences between the genders in their CIR [79], and the first extensive results on the 2 H/ 1 H ratio of endogenous urinary steroids were reported [80].…”

A review of designer anabolic steroids in equine sports

“…In humans, 11‐adrenosterone is an endogenous steroid produced predominately in the adrenal cortex that exerts a mild anabolic effect (Fig. ) . It has been reported as a component of dietary supplements such as 11‐OXO (ErgoPharm) and is marketed as a ‘selective cortisol modulator’ rather than as an anabolic agent.…”