Journal of Chromatography B: Biomedical Sciences and Applications
High doses of alcohol increase urinary testosterone-to-epitestosterone ratio in females
Abstract
The effect of alcohol (1.2 and 2.0 g/kg) on the urinary testosterone-to-epitestosterone (T/E) ratio was studied by two experiments each conducted with four healthy females and males. The intake of 2.0 g/kg of ethanol within 5 h in the evening significantly increased plasma testosterone concentration and ratio of T/E in urine collected next morning in females. The results suggest that alcohol increases the T/E ratio more in females than in males. The effect of high doses of alcohol on urinary T/E ratio must be kept in mind when doping tests are performed during training periods.
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Recent Advances in Doping Analysis (2)
Cited by (31)
Influence of ethanol consumption and food intake on serum concentrations of endogenous steroids
2024, SteroidsSteroid biosynthesis and biotransformation are based on a cascade of enzymatic processes being highly sensitive to various external influences. Amongst those, ethanol was shown to affect testosterone metabolism. For doping analyses, athlete steroid profiles comprise seven urinary steroid metabolites, of which relevant ratios are significantly increased following ethanol consumption. This effect is presumably based on the lack of hepatic NAD+-coenzyme as a consequence of ethanol oxidation. Only recently, testosterone (T) and androstenedione (A4) blood profiles have been introduced as additional approach for doping control. However, a potential influence of ethanol intake on testosterone biosynthesis and thus on blood steroid profiles has not been investigated so far. Therefore, steroid concentrations from 10 males and 10 females receiving an ethanol infusion up to a breath alcohol concentration of 0.5 mg/L which was hold as a plateau for two hours were conducted. Blood samples were drawn every 15 min for steroid quantification.
An ethanol-dependent T/A4 increase up to 385 % resulting from A4 suppression was observed in 14 volunteers. In addition, we observed sporadic A4 increases coinciding with cortisol and ACTH pulses pointing to a meal-induced adrenal stimulation. While testosterone levels in males showed diurnal variation solely, testosterone levels in some females were found to be susceptible to ethanol- and ACTH-dependent perturbations, which is thought to be due to its predominant adrenal synthesis in females.
In conclusion, the results of the present study emphasize the importance of blood sampling at a sufficient time interval from food and ethanol intake. This is of interest if T and A4 are used for diagnostics in doping control.
Biological factors influencing urinary anabolic steroid concentrations in doping control analysis
2009, Science and SportsRecenser les facteurs biologiques qui peuvent modifier les concentrations urinaires de certains stéroïdes anabolisants lors des contrôles antidopage.
D’après les statistiques de l’Agence mondiale antidopage, la prise de stéroïdes anabolisants constitue la principale infraction dans le monde sportif. L’utilisation de substances naturellement produites par l’organisme (testostérone, nandrolone) rend de plus en plus difficile l’interprétation des analyses urinaires. De plus, de nombreux facteurs biologiques (âge, alimentation, cycle menstruel, prise de contraceptifs oraux…) peuvent modifier leurs concentrations chez les sportifs.
La recherche de marqueurs indirects du dopage à travers la mise en place d’un passeport biologique offre une alternative intéressante à la méthode de dépistage utilisée actuellement. L’objectif n’est plus de détecter la présence de substances exogènes dans les matrices biologiques, mais plutôt d’examiner les paramètres biologiques qui peuvent être modifiés par la prise du produit dopant.
To review the biological factors that may influence the urinary concentrations of anabolic steroids in doping controls.
According to the World Anti-Doping Agency statistics, the use of anabolic steroids represents the main offence carried out by the athletes. The use of substances naturally produced by the body (such as testosterone, nandrolone) makes the interpretation of urine analyses very difficult. Furthermore, many biological factors (age, diet, menstrual cycle, oral contraceptives…) may modify these concentrations.
The search of indirect markers of doping through the setting of the biological passport offers a powerful alternative to the current anti-doping method. Detecting the presence of the exogenous substances in biological matrices would not be the main purpose in that kind of strategy, but rather the examination of the spectra of biological variances that can be affected by doping substances.
The idea of the presence of androgens in females may sound peculiar as androgens generally refer to male hormones. Although produced in small amounts in women, androgens have direct and significant effects on many aspects of female physiology. Moreover, androgens are precursors to estrogens, which are the predominant female sex hormones.
The measurement of androgens in blood is important in the diagnosis of both gonadal and adrenal functional disturbances, as well as monitoring subsequent treatments. The accuracy of such measurements is crucial in sports medicine and doping control. Therefore, the concentration of androgens in female subjects is frequently measured.
Analysing such compounds with accuracy is especially difficult, costly and time consuming. Therefore, laboratories widely use direct radioimmunoassay kits, which are often insensitive and inaccurate.
It is especially complicated to determine the level of androgens in women, as the concentration is much lower compared to the concentration found in males. Additionally, the amount of androgens in fluids tends to decrease with aging.
Analyses of hormone concentrations are influenced by a myriad of factors. The factors influencing the outcome of these tests can be divided into in vivo preanalytical factors (e.g., aging, chronobiological rhythms, diet, menstrual cycle, physical exercise, etc.), in vitro preanalytical factors (e.g., specimen collection, equipment, transport, storage, etc.) and as mentioned before, analytical factors. To improve the value of these tests, the strongly influencing factors must be controlled. This can be accomplished using standardised assays and specimen collection procedures. In general, sufficient attention is not given to the preanalytical (biological) factors, especially in the measurement of androgens in females. Biological factors (non-pathological factors) that may influence the outcome of these tests in female subjects have received little attention and are the topic of the present review.
Follow-up of the urinary steroid profils in the sportmen doping control
2008, Revue Francophone des LaboratoiresDétecter et confirmer l’administration d’un stéroïde androgène anabolisant qui est présent normalement dans les fluides biologiques pose un défi analytique tout particulier. Par l’étude minutieuse des altérations, parfois subtiles, du profil des métabolites urinaires, des marqueurs d’utilisation ont été identifiés. Leur mesure précise par CG-SM lors des analyses de routine est essentielle afin de déterminer en premier lieu les fourchettes de valeurs normales des diverses populations ciblées, et également la stabilité de ces paramètres chez les individus, sous toutes les conditions physiologiques naturelles et propres aux sportifs d’élite. L’analyse par CG/C/SMRI permettra ultimement la confirmation de l’origine exogène des métabolites urinaires caractéristiques. Cependant, comme les laboratoires analysent des échantillons sans connaître l’identité de l’athlète, l’analyse CG/C/SMRI n’étant initiée qu’après l’obtention d’une donnée anormale par rapport aux valeurs de référence des populations, il demeure que l’utilisation de testostérone, notamment, peut très certainement demeurer indécelable. Un suivi individuel doit être rapidement adopté afin de repérer les variations anormales et ainsi soit procéder aux analyses de confirmation par SMRI ou cibler l’athlète pour de plus fréquents contrôles.
Detecting and confirming the administration of an anabolic androgenic steroid that could be present normally in the human body is a complex task. Markers of utilisation, parameters of the urinary steroid profile were identified. Their precise measurement by GC/MS in routine operations is essential to establish reference ranges in the athletic population, to determine their individual stability, and to detect anomalies in an athlete's steroid profile. GC/C/IRMS analyses are generally employed to confirm the exogenous origin of the urinary marker having exceeded the population reference range. Nonetheless, those methods are not foolproof and the utilisation of testosterone may well remain undetected. A strategy based upon individual athlete's steroid profiling should be adopted in order to pick up variations that would trigger further IRMS analysis and investigations.
Analysis of anabolic steroids by partial filling micellar electrokinetic capillary chromatography and electrospray mass spectrometry
2004, Journal of Chromatography AA partial filling micellar electrokinetic capillary chromatography (PF-MEKC) separation of six anabolic androgenic steroids (androstenedione, metandienone, fluoxymesterone, methyltestosterone, 17-epimetandienone and testosterone) is introduced. The method utilises a mixed micellar solution consisting of sodium dodecyl sulphate (SDS) and sodium taurocholate. The analytes are detected with a photodiode array detector at 247 nm wavelength. Methyltestosterone is used as internal standard. The detection limits were 39 μg/L for androstenedione, 40 μg/L for testosterone, 45 μg/L for fluoxymesterone, 45–90 μg/L for 17-epimetandienone, 59 μg/L for methyltestosterone and 90 μg/L for metandienone. Linear correlation between concentration (0.1–5.0 mg/L) and detector response was obtained with r2 of 0.994 for fluoxymesterone, 0.998 for 17-epimetandienone and 0.999 for androstenedione, metandienone and testosterone. In addition, ionisation of the investigated compounds in electrospray mass spectrometry (ESI-MS) was studied in positive ion mode. The most intense signal (100%) was the protonated molecular ion [M+H]+, except for 17-epimetandienone, which gave its strongest signal at m/z corresponding to [M−H2O+H]+. Finally, separation and identification of fluoxymesterone, androstenedione and testosterone by PF-MEKC–ESI-MS is described. This is the first use of PF-MEKC and PF-MEKC–ESI-MS assays for anabolic androgenic steroids.
Epitestosterone
2003, Journal of Steroid Biochemistry and Molecular BiologyEpitestosterone has been identified as a natural component of biological fluids of several mammals including man. For a long time it was believed that it is a metabolite without any hormonal activity and without any marked relationship to the hormonal state in health and disease. Neither the biosynthetic pathway nor the site of its formation in man have been unequivocally confirmed to date. It apparently parallels the formation of testosterone (T), but on the other hand its concentration is not influenced by exogenous administration of testosterone. This fact creates the basis of the present doping control of testosterone abuse. In 1989 an observation was presented in a dermatological study that epitestosterone exerts an effect counteracting the action of testosterone on flank organ of Syrian hamster. Further studies showed that a complex action consisting of competitive binding of epitestosterone to androgen receptor, of inhibition of testosterone biosynthesis and its reduction to dihydrotestosterone and of antigonadotropic activity could be demonstrated in rat, mice and human tissues. It can be presumed that epitestosterone as a natural hormone can contribute to the regulation of such androgen dependent events as, e.g. the control of prostate growth or body hair distribution.