The Grey Zone in Doping

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The Grey Zone in Doping THE GREY ZONE IN DOPING Van Thuyne Wim Thesis submitted in fulfilment of the requirements for the degree of Doctor (Ph.D.) in Medical Sciences Promotor: Prof. Dr. F.T. Delbeke Woord Vooraf Na deze pagina volgen nog vele pagina’s. Wanneer u deze allemaal doorleest zal u begrijpen dat ik deze onmogelijk heb kunnen schrijven zonder de hulp en inspiratie van anderen. Op mijn weg tot het bekomen van dit resultaat ben ik immers vele personen tegengekomen die elk op hun eigen manier bijgedragen hebben tot dit werk. De interesse van velen die steeds vroegen hoe het werk vorderde is steeds een stimulans geweest om door te blijven zetten. In eerste instantie wil ik dan ook mijn oprechte excuses aanbieden aan de personen die ik ongetwijfeld vergeet te vermelden in dit dankwoord. Allereerst wil ik mijn promotor, Prof. Dr. F.T. Delbeke hartelijk danken voor de vele mogelijkheden die hij mij geboden heeft de afgelopen jaren. Zonder uw begeleiding was ik nooit in staat geweest tot dit resultaat te komen. Ik hoop dat we onze goede samenwerking van de afgelopen jaren in de toekomst op dezelfde manier kunnen verderzetten. Peter, bureaugenoot gedurende de eerste jaren, bedankt om me op weg te helpen in de vele facetten die een doctoraat omvat. Hartelijk dank ook voor het geduldig beantwoorden van mijn vele vragen. Koen, de laatste twee jaar samen in ons “aquarium” zijn voorbijgevlogen. Het schrijven van dit werk werd iets gemakkelijker doordat je zelf ook aan je eigen thesis werkte. Ik wens je heel veel succes bij het afleggen van je eigen doctoraat in de nabije toekomst. Ann, “miss quality”, bedankt om steeds te blijven hameren op kwaliteitsvol onderzoek. Kris, Dominique, Trui, Pieter, Wim, Steven, ik wil jullie niet alleen bedanken voor de technische steun, maar ook voor de interesse in mijn onderzoek en de aangename sfeer die jullie dag na dag creëerden in het labo. Noël Desmet, hoewel je DoCoLab reeds enige tijd achter je hebt gelaten wil ik je toch bedanken voor het mij bijbrengen van de technische aspecten van GC/MS. Verder wil ik alle leden van de examencommissie, Prof. Dr. Verstraete, Prof. Dr. Pannier, Prof. Dr. Leroux-Roels, Prof. Dr.Bouckaert, Prof. Dr. De Brabander, Dr. Dr.h.c. Rogge, Dr. de Boer en Dr. Geyer bedanken voor hun opbouwende kritiek bij het tot stand komen van dit werk. Mijn dank gaat ook naar de Vlaamse Gemeenschap die mij een doctoraatsbeurs toekende gedurende een periode van vier jaar. Verder wil ik Medisch Verantwoord Sporten van de Vlaamse Gemeenschap, Doping Controle Nederland (DoCoNed), de International Wielerunie (UCI), de Koninklijke Belgische Wielrijdersbond (KBWB), het Belgisch Olympisch en Interfederaal Comité (BOIC), het Nederlands Centrum voor Dopingvraagstukken (NeCeDo) en alle andere nationale en internationale federaties bedanken voor het ter beschikking stellen van informatie en voor hun steun. Ook alle voedingssupplement producerende firma’s die door de jaren dat tikkeltje extra zekerheid en glans wilden geven aan hun producten wil ik van harte bedanken voor het vertrouwen die ze in DoCoLab stelden. Ik hoop dat we in de toekomst verder kunnen werken aan het streven naar de afwezigheid van (moedwillige) contaminatie in voedingssupplementen. Finaal wens ik natuurlijk enkele vrienden en mijn familie te bedanken. Eerst en vooral de vrienden van “WTC For Ever” voor het bezorgen van talloze uren aan ontspanning op en vooral ook naast de fiets. Weet dat ik het altijd eerlijk heb gespeeld en nooit zelf aan de verleiding van doping heb toegegeven. Mijn ouders, broer en zus (met partners en kroost), waar ik altijd bij terecht kon en steeds met veel belangstelling mijn doen en laten hebben gevolgd. Ik besef nog maar eens hoe belangrijk jullie zijn voor mij. En natuurlijk mijn lieve Klaar. Bedankt om sinds de dag waarop we elkaar ontmoetten elke dag weer als een zonnetje door mijn leven te wandelen. Bedankt ook om me de afgelopen maanden de mogelijkheid te geven volledig voor dit werk te gaan. Ik hoop dat ik in de nabije toekomst net hetzelfde kan betekenen voor jou. List of abbreviations List of abbreviations 6-OXO 4-androsten-3,6,17-trione 7-keto-DHEA 7-keto-dehydroepiandrosterone ANOVA Analysis of variances APCI Atmospheric pressure chemical ionisation ATP Association de tennis professionnelle CE Capillary electrophoresis CE Collision energy CF Communauté Française CI Chemical ionisation cmax Maximal concentration COX Cyclo-oxygenase DHEA Dehydroepiandrosterone DHT Dihydrotestosterone DI Diagnostic ion DoCoNed Dopingcontrole Nederland DSHEA Dietary Supplement Health and Education Act ECA-stack Ephedrine-caffeine-aspirin stack EI Electron ionisation List of abbreviations FDA Food and Drug Administration GC Gas chromatography GC/MS Gas chromatography – mass spectrometry GC/MS/MS Gas chromatography – tandem mass spectrometry GPC Gel permeation chromatography HPLC High performance liquid chromatography I.D. Internal diameter IAAF International Association of Athletics Federations ILAC International Laboratory Accreditation Cooperation IOC International Olympic Committee IS Internal standard ISO International Organisation for Standardisation KBWB Koninklijke Belgische Wielrijdersbond (Royal Belgian Cyclist Federation) LC/MS Liquid chromatography – mass spectrometry LC/MS/MS Liquid chromatography – tandem mass spectrometry LOD Limit of detection LOQ Limit of quantification m/z Mass/charge MAOI Monoamine oxydase inhibitor List of abbreviations MBTFA N-methyl-bistrifluotoacetamide MDA 3,4-methylenedioxyamphetamine MDEA 3,4-methylenedioxyethylamphetamine MDMA 3,4-methylenedioxymethamphetamine MeOH methanol MRPL Minimum required performance limit MS mass spectrometry MS/MS Tandem mass spectrometry MSD Mass selective detector MSn Multiple mass spectrometry MSTFA N-methyl-N-trimethylsilyltrifluoroacetamide NADO National doping organisation NARL National Analytical Reference Laboratory NeCeDo Nederlands centrum voor dopingvraagstukken NPD Nitrogen-Phosphorus detector NPN Natuur- en gezondheidsproducten Nederland NSAID Non-steroidal anti-inflammatory drug NZVT The Netherlands Security System Nutritional Supplements ODS Octadecylsilica List of abbreviations OFN Oxygen free nitrogen PI Precursor ion RRT Relative retention time RSD Relative standard deviation RSDmax Maximal relative standard deviation RT Retention time SD Standard deviation SIM Selected ion monitoring SSRI Selective seretonine re-uptake inhibitor T/E Testosterone-epitestosterone ratio TCA Tricyclic antidepressant TIC Total ion chromatogram TLC Thin Layer Chromatography TMS Trimethylsilyl TSP Thermo Separation Products TUE Therapeutic use exemption U.S. United States UCI Union Cycliste International UEFA Union of European Football Associations UK United Kingdom List of abbreviations UV Ultra-violet VlGem Vlaamse Gemeenschap WADA World Anti-Doping Agency Outline of the study Outline of the study Anti-doping programs seek to preserve what is intrinsically valuable about sport. This intrinsic value is often referred to as “the spirit of sport”, which is the celebration of the human spirit, body and mind and is characterised by values such as ethics, fair play, honesty, health, fun, team work, courage and many others. Doping, which is not only the detection of prohibited substances but also its possession and use, failing or tampering a doping test or refusing a control, is considered as fundamentally against the spirit of sport. As a result of this definition, lists of prohibited substances and methods have been developed to keep the spirit of sport as pure as possible. Since the introduction of the first doping test in the early 1960’s and the development of anti-doping rules by the UCI in the same period, doping rules have been extensively revised by the IOC and currently by WADA. These rules are very extensive and include numerous classes defined as doping agents, such as anabolic steroids, stimulants, diuretics and EPO, and also prohibited methods as blood doping. Doping control generally operates a strict liability principle. In other words, the presence of a banned substance, or its metabolite, in a urine or body fluid constitutes and doping offence. In a refinement of this principle, several substances now have a requirement for laboratories to report their presence only above a specified urinary concentration. Some of these substances are regulated by a urinary threshold level because they are, or may be, endogenous. Examples of those substances are testosterone and 19-norandrosterone, the major metabolite of nandrolone. Other substances can be present as a constituent in the daily diet, such as caffeine and ephedrine, and are only restricted when large urinary concentrations indicate the deliberate intake of this substance to enhance performance. In an attempt to run faster, jump higher or throw further athletes are always seeking for means to accomplish this goal. Since the mid 1990’s the use of diverse types of nutritional supplements has become very popular in an attempt to optimise performance. The most important factor behind the explosive growth of the supplement market was the passage of the Dietary Supplement Health and Education Act (DSHEA) which makes it possible to market numerous products as nutritional supplements as long as no statements are made that these Outline of the study products prevent, treat or cure a specific disease. Other factors behind the growth of this market is the easy access through the internet and the ever-improving results and record performances of top-level athletes who are promoting the use of supplements. However,
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