Abstract
We have investigated the influence of oral miconazole administration on the urinary concentrations of endogenous anabolic androgenic steroids of doping relevance, specifically considering all these compounds routinely monitored in doping control analysis, in the framework of the steroidal module of the “athlete biological passport”, and other steroids, including dehydroepiandrosterone, 5α-dihydrotestosterone, and the hydroxylated metabolites recently proposed as additional markers of the intake of testosterone-related steroids (16α-hydroxy-androsterone, 16α-hydroxy-etiocholanolone, 6β-hydroxy-androsterone, 6β-hydroxy-etiocholanolone, 7α-hydroxy-dehydroepiandrosterone, and 7β-hydroxy-dehydroepiandrosterone). Urinary concentrations of the final metabolic products of the glucocorticoid biosynthetic pathways (11β-hydroxy-androsterone and 11β-hydroxy-etiocholanolone, the formerly used as an endogenous reference compound for the gas chromatography–combustion-isotope ratio mass spectrometry confirmation analysis) were also monitored. Two healthy Caucasian volunteers exhibiting physiologically high testosterone/epitestosterone ratios and elevated concentrations of the main target steroids were selected for the study. Miconazole was administered orally (500 mg/day) for 1 week. Multiple urine samples were collected for 1 week before and during the treatment, and analyzed according to a validated analytical procedure based on gas chromatography–electron ionization-mass spectrometry in selected ion monitoring mode. Our results indicated that oral administration of miconazole decreased the urinary concentrations of androsterone, and to a lesser extent, of etiocholanolone (both detected as the sum of free and glucuronated steroids), and consequently the androsterone/testosterone and androsterone/etiocholanolone ratios. Furthermore, the urinary concentrations of 16α-hydroxy-etiocholanolone, 16α-hydroxy-androsterone, 7β-hydroxy-dehydroepiandrosterone, 6β-hydroxy-etiocholanolone, 7α-hydroxy-dehydroepiandrosterone, 6β-hydroxy-androsterone, 11β-hydroxy-androsterone, and 11β-hydroxy-etiocholanolone were significantly suppressed. This evidence suggests the potential intake of miconazole whenever the urinary steroid profile is characterized by abnormally low concentrations of the above-mentioned steroids.
Similar content being viewed by others
References
The World Anti-Doping Code (2016) The 2016 prohibited list international standard. World Anti-Doping Agency, Montreal. http://www.wada-ama.org. Accessed 6 Jan 2016
Rapporto di attività di controllo antidoping, Ministero della Salute (2015) Direzione generale della prevenzione sanitaria, anno 2015 Gennaio-Giugno. http://www.salute.gov.it/imgs/C_17_pubblicazioni_2430_allegato.pdf. Accessed 2 Dec 2015
Wim VT, Delbeke FT (2008) Declared use of medication in sports. Clin J Sport Med 18:143–147
Corrigan B, Rymantas K (2003) Medication use in athletes selected for doping control at the Sydney Olympics (2000). Clin J Sport Med 13:33–40
Huang SH, Johnson K, Pipe AL (2006) The use of dietary supplements and medications by Canadian athletes at the Atlanta and Sydney Olympic Games. Clin J Sport Med 16:27–33
Tscholl P, Junge A, Dvorak J (2008) The use of medication and nutritional supplements during FIFA World Cups 2002 and 2006. Br J Sports Med 42:725–730
Geyer H, Schänzer W, Donike M (1992) Probenecid as masking agent in dope control—inhibition of the urinary excretion of steroid glucuronides. In: Donike M, Geyer H, Gotzmann A, Mareck-Engelke U, Rauth S (eds) 10th Cologne workshop on dope analysis, 7th to 12th June 1992. Sport und Buch Strauß, Köln, pp 141–145
Mareck U, Geyer H, Opfermann G, Thevis M, Schänzer W (2008) Factors influencing the steroid profile in doping control analysis. J Mass Spectrom 43:877–891
Ventura R, Segura J (2010) Masking and manipulation. In: Thieme D, Hemmersbach P (eds) Doping in sports, handbook of experimental pharmacology 195. Springer, Berlin, pp 327–354
Botrè F, de la Torre X, Donati F, Mazzarino M (2014) Narrowing the gap between the number of athletes who dope and the number of athletes who are caught: scientific advances that increase the efficacy of antidoping tests. Br J Sports Med 48:833–836
Botrè F (2015) Masking and unmasking strategies in sport doping. In: Georgakopoulos K, Alsayrafi M (eds) Advances and challenges in antidoping analysis. Future Sciences, London, pp 167–182
Mazzarino M, de la Torre X, Fiacco I, Palermo A, Botrè F (2014) Drug-drug interaction and doping, part 1: an in vitro study on the effect of non-prohibited drugs on the phase I metabolic profile of toremifene. Drug Test Anal 6:482–491
Mazzarino M, de la Torre X, Fiacco I, Botrè F (2014) Drug-drug interaction and doping, part 2: an in vitro study on the effect of non-prohibited drugs on the phase I metabolic profile of stanozolol. Drug Test Anal 6:969–977
World Anti-Doping Agency (2014) Decision limits for the confirmatory quantification of threshold substances (WADA Technical Document TD2014DL). http://www.wada-ama.org. Accessed 6 Jan 2016
Mareck U, Schultze G, Geyer H, Schänzer W (2002) The appearance of urinary 19-norandrosterone during pregnancy. Eur J Sport Sci 2:1–7
Palermo A, Alessi B, Botrè F, Torre X, Fiacco I, Mazzarino M (2015) In vitro evaluation of the effects of anti-fungals, benzodiazepines and non-steroidal anti-inflammatory drugs on the glucuronidation of 19-norandrosterone: implications on doping control analysis. Drug Test Anal. doi:10.1002/dta.1897
Sottas PE, Saudan C, Schweizer C, Baume N, Mangin P, Saugy M (2008) From population- to subject-based limits of T/E ratio to detect testosterone abuse in elite sports. Forensic Sci Int 174:166–172
Rane A, Ekström L (2012) Androgens and doping tests: genetic variation and pit-falls. Br J Clin Pharmacol 74:3–15
Jakobsson J, Ekström L, Inotsume N, Garle M, Lorentzon M, Ohlsson C, Rane A (2006) Large differences in testosterone excretion in Korean and Swedish men are strongly associated with a UDP-glucuronosyl transferase 2B17 polymorphism. J Clin Endocrinol Metab 91:687–693
Donike M, Rauth S, Mareck-Engelke U, Geyer H, Nitschke R (1994) Evaluation of longitudinal studies, the determination of subject based reference ranges of the testosterone/epitestosterone ratio. In: Donike M, Geyer H, Gotzmann A, Mareck-Engelke U, Rauth S (eds) 11th Cologne workshop on dope analysis. Sport und Buch Strauß, Köln, pp 33–39
Donike M, Mareck-Engelke U, Rauth S (1995) Statistical evaluation of longitudinal studies, part 2: the usefulness of subject based reference ranges. In: Donike M, Geyer H, Gotzmann A, Mareck-Engelke U, Rauth S (eds) 12th Cologne workshop on dope analysis. Sport und Buch Strauß, Köln, pp 157–165
Sottas PE, Robinson N, Rabin O, Saugy M (2011) The athlete biological passport. Clin Chem 57:969–976
Saugy M, Lundby C, Robinson N (2014) Monitoring of biological markers indicative of doping: the athlete biological passport. Br J Sports Med 48:827–832
Sottas PE, Baume N, Saudan C, Schweizer C, Kamber M, Saugy M (2007) Bayesian detection of abnormal values in longitudinal biomarkers with an application to T/E ratio. Biostatistics 8:285–296
Van Renterghem P, Van Eenoo P, Van Thuyne W, Geyer H, Schänzer W, Delbeke FT (2008) Validation of an extended method for the detection of the misuse of endogenous steroids in sports, including new hydroxylated metabolites. J Chromatogr B 876:225–235
Van Renterghem P, Van Eenoo P, Sottas PE, Saugy M, Delbeke F (2010) Subject-based steroid profiling and the determination of novel biomarkers for DHT and DHEA misuse in sports. Drug Test Anal 2:582–588
Van Renterghem P, Van Eenoo P, Delbeke FT (2010) Population based evaluation of a multi-parametric steroid profiling on administered endogenous steroids in single low dose. Steroids 75:1047–1057
Van Renterghem P, Van Eenoo P, Sottas PE, Saugy M, Delbeke F (2011) A pilot study on subject-based comprehensive steroid profiling: novel biomarkers to detect testosterone misuse in sports. Clin Endocrinol 75:134–140
Parr MK, Schänzer W (2010) Detection of the misuse of steroids in doping control. J Steroid Biochem 121:528–537
Van Renterghem P, Van Eenoo P, Geyer H, Schänzer W, Delbeke FT (2010) Reference ranges for urinary concentrations and ratios of endogenous steroids, which can be used as markers for steroid misuse, in a Caucasian population of athletes. Steroids 75:154–163
Kuuranne T, Saugy M, Baume N (2014) Confounding factors and genetic polymorphism in the evaluation of individual steroid profiling. Br J Sports Med 48:848–855
World Anti-Doping Agency (2015) Endogenous anabolic androgenic steroids measurement and reporting, WADA Technical Document TD2016EAAS. http://www.wada-ama.org. Accessed 6 Jan 2016
Thevis M, Geyer H, Mareck U, Flenker U, Schänzer W (2007) Doping control analysis of the 5-reductase inhibitor finasteride: determination of its influence on urinary steroid profiles and detection of its major urinary metabolite. Ther Drug Monit 29:236–247
Feldman D (1986) Ketoconazole and other imidazole derivatives as inhibitors of steroidogenesis. Endoc Rev 7:409–420
Rajfer J, Sikka SC, Rivera F, Handelsman J (1986) Mechanism of inhibition of human testicular steroidogenesis by oral ketoconazole. J Clin Endocr Metab 63:1193–1198
Rendic S (1996) Human cytochrome P450 (CYP) enzymes in doping control: metabolism, interactions, adverse effects. In: Donike M, Geyer H, Gotzmann A, Mareck-Engelke U, Rauth S (eds) Recent advances in doping analysis. Proceedings of the 13th Cologne workshop on dope analysis. Sport und Buch Strauß, Köln, pp 13–53
Touchette MA, Chandrasekar PH, Milad MA, Edwards DJ (1992) Contrasting effects of fluconazole and ketoconazole on phenytoin and testosterone disposition in man. Br J Clin Pharmacol 34:75–78
Kicman AT, Oftebro H, Walker C, Norman N, Cowan DA (1993) Potential use of ketoconazole in a dynamic endocrine test to differentiate between biological outliers and testosterone use by athletes. Clin Chem 39:1798–1803
Oftebro H, Jensen J, Mowinckel P, Norli HR (1994) Establishing a ketoconazole suppression test for verifying testosterone administration in the doping control of athletes. J Clin Endocr Metab 78:973–977
Ayub M, Levell MJ (1987) Inhibition of testicular 17α-hydroxylase and 17, 20-lyase but not 3β-hydroxysteroid dehydrogenase-isomerase or 17β-hydroxysteroid oxidoreductase by ketoconazole and other imidazole drugs. J Steroid Biochem 28:521–531
Kicman AT (2010) Biochemical and physiological aspects of endogenous androgens. In: Thieme D, Hemmersbach P (eds) Doping in sports, handbook of experimental pharmacology 195. Springer, Berlin, pp 25–64
Niwa T, Shiraga T, Takagi A (2005) Effect of antifungal drugs on cytochrome P450 (CYP) 2C9, CYP2C19, and CYP3A4 activities in human liver microsomes. Biol Pharm Bull 28:1805–1808
Niwa T, Inoue-Yamamoto S, Shiraga T, Takagi A (2005) Effect of antifungal drugs on cytochrome P450 (CYP) 1A2, CYP2D6, and CYP2E1 activities in human liver microsomes. Biol Pharm Bull 28:1813–1816
World Anti-Doping Agency (2015) Detection of synthetic forms of endogenous anabolic androgenic steroids by GC-C-IRMS, (WADA Technical Document TD2016 IRMS). http://www.wada-ama.org. Accessed 6 Jan 2016
Mazzarino M, Abate MG, Alocci R, Rossi F, Stinchelli R, Molaioni F, de la Torre X, Botrè F (2011) Urine stability and steroid profile: towards a screening index of urine sample degradation for anti-doping purpose. Anal Chim Acta 683:221–226
Mazzarino M, Braganò MC, de la Torre X, Molaioni F, Botrè F (2011) Relevance of the selective oestrogen receptor modulators tamoxifen, toremifene and clomiphene in doping field: endogenous steroids urinary profile after multiple oral doses. Steroids 76:1400–1406
Leinonen A, Kuuranne T, Moisander T, Rautava K (2007) Artificial urine as sample matrix for calibrators and quality controls in determination of testosterone to epitestosterone ratio. In: Schanzer W, Geyer H, Gotzmann A, Mareck U (eds) Recent advances in doping analysis, vol 15. Sport und Buch Strauß, Köln, pp 401–404
Lamberts SW, Bons EG, Bruining HA, de Jong FH (1987) Differential effects of the imidazole derivatives etomidate, ketoconazole and miconazole and of metyrapone on the secretion of cortisol and its precursors by human adrenocortical cells. J Pharmacol Exp Ther 240:259–264
Loose DS, Kan PB, Hirst MA, Marcus RA, Feldman D (1983) Ketoconazole blocks adrenal steroidogenesis by inhibiting cytochrome P450-dependent enzymes. J Clin Invest 71:1495–1499
Mazzarino M, de la Torre X, Fiacco I, Khevenhüller F, Botrè F (2013) Effects of ketoconazole on the excretion kinetics of methandienone. An in vivo study. In: Donike M, Geyer H, Gotzmann A, Mareck-Engelke U (eds) Recent advances in doping analysis. 21th Cologne workshop on dope analysis. Sport und Buch Strauß, Köln, pp 34–40
Acknowledgments
This project has been supported in part by a research grant of the World Anti-Doping Agency (Project Code: 13D14MM).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflicts of interest.
Ethical approval
All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. The administration studies were approved by the local ethical committee (Approval Code: Prot. 1055/2014 CE Lazio 1). Informed consent was obtained from all individual participants included in the study.
Rights and permissions
About this article
Cite this article
Palermo, A., Botrè, F., de la Torre, X. et al. Drug-drug interactions and masking effects in sport doping: influence of miconazole administration on the urinary concentrations of endogenous anabolic steroids. Forensic Toxicol 34, 386–397 (2016). https://doi.org/10.1007/s11419-016-0325-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11419-016-0325-x