Welcome to DOPING.nl, the Anti-Doping Knowledge Center

This site has been established to host information about doping in the broadest sense of the word, and about doping prevention.

Initiator

The Anti-Doping Authority Netherlands (the Dutch Doping Authority for short) established this site and maintains it. The Doping Authority was founded in 1989 and it is one of the oldest NADOs in the world. Doping.nl was developed with financial support from the Dutch Ministry for Health, Welfare and Sport.

Goals

This website was established because of the importance that the Doping Authority and the Ministry attach to the dissemination of information relevant to doping prevention. Disclosing and supplying relevant information is one of the cornerstones in the fight against doping in sport. However, in practice, a significant amount of information is still not available, or only available to a limited group of users. We therefore decided to bring together all the relevant information in a single site: Doping.nl.

Activities

The Doping Authority aims to supply as much information through this website as possible on an ongoing basis. The information will be varied but will focus primarily on: WADA documents like the World Anti-Doping Code, the International Standards like the Prohibited List, Doping Regulations, scientific articles and abstracts, decisions by disciplinary bodies (mainly CAS decisions).As well as making documents available, the Doping Authority aims to supply searchable documents when possible, and to add relevant keywords to ensure easy access.
In the future, Doping.nl will also become a digital archive containing older information that is no longer available elsewhere.

Target readers

This site has been designed for use by anti-doping professionals such as National Anti-Doping Organisations and International Federations but also for students, journalists and other people interested in the subject.

More information explaining how to use this website can be found under "help".

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Analytical possibilities for the detection of stanozolol and its metabolites

24 Sep 2002

Analytical possibilities for the detection of stanozolol and its metabolites / S. Poelmans, K. De Wasch, H.F. De Brabander, M. Van De Wiele, D. Courtheyn, L.A. van Ginkel, S.S. Sterk, Ph. Delahaut, M. Dubois, R. Schilt, M. Nielen, J. Vercammen, S. Impens, R. Stephany, T. Hamoir, G. Pottie, C. Van Poucke, C. Van Peteghem. - (Analytica Chimica Acta 473 (2002) 1-2 (25 November); p. 39-47)

  • DOI: 10.1016/S0003-2670(02)00672-4


Abstract

In sports doping, as well in man as in horseracing, stanozolol (Stan) was abused and became the subject of metabolism research. Also in veterinary practice, stanozolol became an important misused anabolic steroid.

Like most other anabolic steroids, stanozolol has poor gas chromatographic behavior. It is difficult to detect in urine, because of low urinary excretion and renal clearance. This is due to the rapid metabolization, leading to low concentration levels of the parent compound found in urine. Therefore, most research studies have focused on the detection of its urinary metabolites.

For the identification of the metabolites, different methods of extraction and detection are described in the literature. These are reviewed in this article. Most authors use a hydrolysis to free the phase II metabolites. Extraction procedures vary from solid-phase extraction (SPE), liquid–liquid (L–L) extraction to immunoaffinity chromatography (IAC). For the final detection, the use of gas chromatography (GC)–mass spectrometry (MS) can be compared with liquid chromatography (LC)–MSn. Different metabolites are identified depending on the administration of stanozolol in the animal experiment (oral or intramuscular). Analyses for these analytes in other matrices are also briefly discussed.

Analysis of anabolic steroids using GC/MS with selected ion monitoring

1 Mar 1990

Analysis of anabolic steroids using GC/MS with selected ion monitoring / Bong Chul Chung, Hea-Young P. Choo, Tae Wook Kim, Khee Dong Eom, Oh Seung Kwon, Jawon Suh, Jongsoon Yang, Jongsei Park. - (Journal of Analytical Toxicology 14 (1990) 2 (March-April); p. 91-95)

  • PMID: 2325383
  • DOI: 10.1093/jat/14.2.91


Abstract

This study describes the use of gas chromatography/mass spectrometry with selected ion monitoring to screen 18 anabolic steroids banned by the International Olympic Committee. These anabolic steroids are analyzed in two fractions depending on their excretion pattern: nonconjugated (free) or conjugated fraction. The wet procedure of extracting steroids from urine consists of an initial isolation of lipophilic compounds on a column packed with Amberlite XAD-2 resin, followed by enzymatic hydrolysis with beta-glucuronidase from Escherichia coli. After extraction, the hydrolyzed steroids are derivatized to the corresponding trimethylsilyl ethers. The derivatized steroids are analyzed by gas chromatography/mass spectrometry with selected ion monitoring of their characteristic ions. It takes 12 and 26 min to run GC/MS and edit the raw data for nonconjugated and conjugated fractions respectively.

The pharmaceuticalisation of 'healthy' ageing: Testosterone enhancement for longevity

12 Feb 2021

The pharmaceuticalisation of 'healthy' ageing : Testosterone enhancement for longevity / Matthew Dunn, Kyle J.D. Mulrooney, Cynthia Forlini, Katinka van de Ven, Mair Underwood. - (International Journal of Drug Policy (2021) 103159 (12 February))

  • PMID: 33583680
  • DOI: 10.1016/j.drugpo.2021.103159


Abstract

The United Nations estimates that the world's population will reach 8.5 billion by 2030, and the populations of most countries are expected to grow older. This is case for many developed countries, including Australia, the United Kingdom, Canada, the United States of America, and member states of the European Union. Older cohorts will comprise a larger proportion of overall populations, driven in part by our increases in life expectancy. An ageing population poses challenges for governments; notably, older people tend to have multiple, chronic health conditions which can place a burden of health budgets. At the same time, we are witnessing a shift in how we respond to the health needs of our populations, with global drug policy acknowledging that some substances are contributing to increased morbidity and mortality (e.g. opioids) while others may have beneficial therapeutic effects (e.g. psylocibin, cannabis). There is general agreement that as men age their levels of testosterone decrease, and there is some evidence to suggest that there have been population-level declines in testosterone which are not associated with age. Anecdotally, testosterone is accessed by men seeking to self-medicate in the belief that they are experiencing low testosterone levels. There has also been a rise in anti-ageing clinics in the United States, providing access to testosterone replacement therapy (TRT). The non-medical use of testosterone can result in a number of adverse health events, including complications from the use of black market or underground products. Placing testosterone under a new prescribing regime may address some of these concerns, but is society ready for this change, and if so, what would this regime look like? This paper will explore the issue of how society responds to enhancement for longevity, or how we increasingly use pharmaceuticals to address and prevent illness, with a specific focus on testosterone and testosterone deficiency.

Detection of anabolic steroids in head hair

20 Aug 1998

Detection of anabolic steroids in head hair / Xin-Sheng Deng, Akira Kurosu, Derrick J. Pounder. - (Journal of forensic sciences 44 (1999) 2 (March); p. 343-346)

  • PMID: 10097359
  • DOI: 10.1520/JFS14460J


Abstract

We developed a gas chromatography/mass spectrometry method for detection and quantitation of anabolic steroids in head hair. Following alkaline digestion and solid-phase extraction, the MO-TMS derivatives gave a specific fragmentation pattern with EI ionization. For stanozolol, the TMS-HFBA derivative showed several diagnostic ions. For androstanolone, mestanolone (methylandrostanolone), and oxymetholone two chromatographic peaks for cis and trans isomers of derivatives were seen. Recoveries were 35 to 45% for androstanolone, oxymetholone, chlorotestosterone-acetate, dehydromethyltestosterone, dehydrotestosterone, fluoxymesterone, mestanolone, methyltestosterone, and nandrolone; 52% for mesterolone, trenbolone; 65% for bolasterone; 24% for methenolone and 17% for stanozolol. Limits of detection were 0.002 to 0.05 ng/mg and of quantitation were 0.02 to 0.1 ng/mg. Seven white male steroid abusers provided head hair samples (10 to 63 mg) and urine. In the hair samples, methyltestosterone was detected in two (confirmed in urine); nandrolone in two (also confirmed in urine); dehydromethyltestosterone in four (but not found in urine); and clenbuterol in one (but not in urine). Oxymethalone was found in urine in one, but not in the hair. One abuser had high levels of testosterone: 0.15 ng/mg hair, and 1190 ng/mL urine. We conclude that head hair analysis has considerable potential for the detection and monitoring of steroid abuse.

The moral disengagement in doping scale

13 Feb 2016

The moral disengagement in doping scale / Maria Kavussanu, Antonis Hatzigeorgiadis, Anne-Marie Elbe, Christopher Ring. - (Psychology of Sport and Exercise 24 (2016) May; p. 188-198)

  • DOI: 10.1016/j.psychsport.2016.02.003


Abstract

Statement of problem

The use of banned substances to enhance performance occurs in sport. Therefore, developing valid and reliable instruments that can predict likelihood to use banned substances is important.

Method

We conducted three studies. In Study 1, football players (N = 506) and athletes from a variety of team sports (N = 398) completed the Moral Disengagement in Doping Scale (MDDS). In Study 2, team sport athletes (N = 232) completed the MDDS and questionnaires measuring moral disengagement in sport, doping attitudes, moral identity, antisocial sport behavior, situational doping temptation, and task and ego goal orientations. A week later, a subsample (n = 102) completed the MDDS and indicated their likelihood to use a banned substance in a hypothetical situation. In Study 3, athletes (N = 201) from a variety of individual sports completed the MDDS and indicated their likelihood to use a banned substance in a hypothetical situation.

Results

The results of Study 1 showed that a one-factor model fitted the data well, and the scale had measurement invariance across males and females. In Study 2, we provided evidence for convergent, concurrent, discriminant, and predictive validity, as well as test-rest reliability, of the scale. In Study 3, doping moral disengagement was positively related with reported likelihood and temptation to use a banned substance. The scale exhibited very good internal consistency across the three studies.

Conclusions

In conclusion, the MDDS can be used to measure moral disengagement in doping in team and individual sports.

Annual banned-substance review: analytical approaches in human sports drug testing - [2019-2020]

12 Nov 2020

Annual banned-substance review: analytical approaches in human sports drug testing / Mario Thevis, Tiia Kuuranne, Hans Geyer. - (Drug Testing and Analysis 13 (2021) 1 (January); p. 8-35)

  • PMID: 31724288
  • DOI: 10.1002/dta.2969


Contents:

  • Inroduction
  • Anabolic Agent
    • Anabolic-androgenic steroids
    • Initial testing procedures: Comprehensive screening, metabolism studies
    • Steroid profiling in urine and serum
    • Confirmatory testing procedures – IRMS
    • Other anabolic agents
  • Peptide Hormones, Growth Factors, Related Substances, and Mimetics
    • Erythropoietin-receptor agonists and hypoxia-inducible factor (HIF) activating agents
    • Growth hormone, its fragments and releasing factors, chorionic gonadotrophin and luteinizing hormone (LH)
  • β2‐Agonists
  • Hormone and Metabolic Modulators
  • Diuretics and other Masking Agents, Stimulants
  • Corticoids and cannabinoids
  • Manipulation of blood and blood components
  • Gene Doping
  • Conclusion


Abstract

Analytical chemistry‐based research in sports drug testing has been a dynamic endeavor for several decades, with technology‐driven innovations continuously contributing to significant improvements in various regards including analytical sensitivity, comprehensiveness of target analytes, differentiation of natural/endogenous substances from structurally identical but synthetically derived compounds, assessment of alternative matrices for doping control purposes, and so forth. The resulting breadth of tools being investigated and developed by anti‐doping researchers has allowed to substantially improve anti‐doping programs and data interpretation in general. Additionally, these outcomes have been an extremely valuable pledge for routine doping controls during the unprecedented global health crisis that severely affected established sports drug testing strategies. In this edition of the annual banned‐substance review, literature on recent developments in anti‐doping published between October 2019 and September 2020 is summarized and discussed, particularly focusing on human doping controls and potential applications of new testing strategies to substances and methods of doping specified the World Anti‐Doping Agency's 2020 Prohibited List.

Feasibility of a liquid-phase microextraction sample clean-up and liquid chromatographic/mass spectrometric screening method for selected anabolic steroid glucuronides in biological samples

2 Jan 2003

Feasibility of a liquid-phase microextraction sample clean-up and liquid chromatographic/mass spectrometric screening method for selected anabolic steroid glucuronides in biological samples / Tiia Kuuranne, Tapio Kotiaho, Stig Pedersen-Bjergaard, Knut Einar Rasmussen, Antti Leinonen, Steven Westwood, Risto Kostiainen. - (Journal of Mass Spectrometry 38 (2003) 1 (January); p. 16-26)

  • PMID: 12526002
  • DOI: 10.1002/jms.393


Abstract

Anabolic androgenic steroids (AAS) are metabolized extensively in the human body, resulting mainly in the formation of glucuronide conjugates. Current detection methods for AAS are based on gas chromatographic/mass spectrometric (GC/MS) analysis of the hydrolyzed steroid aglycones. These analyses require laborious sample preparation steps and are therefore time consuming. Our interest was to develop a rapid and straightforward method for intact steroid glucuronides in biological samples, using liquid-phase microextraction (LPME) sample clean-up and concentration method combined with liquid chromatographic/tandem mass spectrometric (LC/MS/MS) analysis. The applicability of LPME was optimized for 13 steroid glucuronides, and compared with conventional liquid-liquid extraction (LLE) and solid-phase extraction (SPE) procedures. An LC/MS/MS method was developed for the quantitative detection of AAS glucuronides, using a deuterium-labeled steroid glucuronide as the internal standard. LPME, owing to its high specificity, was shown to be better suited than conventional LLE and SPE for the clean-up of urinary AAS glucuronides. The LPME/LC/MS/MS method was fast and reliable, offering acceptable reproducibility and linearity with detection limits in the range 2-20 ng ml(-1) for most of the selected AAS glucuronides. The method was successfully applied to in vitro metabolic studies, and also tested with an authentic forensic urine sample. For a urine matrix the method still has some unsolved problems with specificity, which should be overcome before the method can be reliably used for doping analysis, but still offering additional and complementary data for current GC/MS analyses.

Speeding up the screening of steroids in urine: development of a user-friendly library

11 Sep 2013

Speeding up the screening of steroids in urine : development of a user-friendly library / M. Galesio, H. López-Fdez, M. Reboiro-Jato, Silvana Gómez-Meire, D. Glez-Peña, F. Fdez-Riverola, Carlos Lodeiro, M.E. Diniz, J.L. Capelo. - (Steroids 78 (2013) 12-13 (11 December); p. 1226-1232)

  • PMID: 24036418
  • DOI: 10.1016/j.steroids.2013.08.014


Abstract

This work presents a novel database search engine - MLibrary - designed to assist the user in the detection and identification of androgenic anabolic steroids (AAS) and its metabolites by matrix assisted laser desorption/ionization (MALDI) and mass spectrometry-based strategies. The detection of the AAS in the samples was accomplished by searching (i) the mass spectrometric (MS) spectra against the library developed to identify possible positives and (ii) by comparison of the tandem mass spectrometric (MS/MS) spectra produced after fragmentation of the possible positives with a complete set of spectra that have previously been assigned to the software. The urinary screening for anabolic agents plays a major role in anti-doping laboratories as they represent the most abused drug class in sports. With the help of the MLibrary software application, the use of MALDI techniques for doping control is simplified and the time for evaluation and interpretation of the results is reduced. To do so, the search engine takes as input several MALDI-TOF-MS and MALDI-TOF-MS/MS spectra. It aids the researcher in an automatic mode by identifying possible positives in a single MS analysis and then confirming their presence in tandem MS analysis by comparing the experimental tandem mass spectrometric data with the database. Furthermore, the search engine can, potentially, be further expanded to other compounds in addition to AASs. The applicability of the MLibrary tool is shown through the analysis of spiked urine samples.

Efficacy of needle-free administration of recombinant human growth hormone in adults with growth hormone deficiency

2 Feb 2006

Efficacy of needle-free administration of recombinant human growth hormone in adults with growth hormone deficiency Alberto M. Pereira, Agatha A. van der Klaauw, Hans P.F. Koppeschaar, Jan W.A. Smit, Sjoerd W. van Thiel, Jaap van Doorn, Nienke R. Biermasz, Ferdinand Roelfsema, Johannes A. Romijn. - (British Journal of Clinical Pharmacology 61 (2006) 4 (April); p. 451-455)

  • PMID: 16542206
  • PMCID: PMC1885043
  • DOI: 10.1111/j.1365-2125.2006.02585.x


Abstract

Aim: Needle-free administration of recombinant human growth hormone (rhGH) is effective in the treatment of growth hormone deficiency (GHD) in children, but has not been studied in adult patients. Therefore, we evaluated the efficacy of needle-free administration of rhGH in adults with GHD.

Methods: Insulin-like growth factor-I (IGF-I) concentrations were compared in newly diagnosed patients with GHD (n = 21) and in patients previously treated by subcutaneous injection of rhGH (switchers, n = 34), at baseline, 12 months and 24 months.

Results: In the new patients, IGF-I standard deviation scores (SDS) increased from - 1.82 +/- 0.46 to + 0.75 +/- 0.33 at 12 months and to + 0.65 +/- 0.41 at 24 months (P < or = 0.001 vs. baseline). In switchers, IGF-I SDS remained unchanged with values of + 0.98 +/- 0.32 at baseline, + 0.87 +/- 0.23 at 12 months and + 0.73 +/- 0.29 at 24 months (P = 0.696 vs. baseline). In new patients, the rhGH dose was 0.46 +/- 0.03 mg day(-1) at 12 months and 0.47 +/- 0.03 mg day(-1) at 24 months. In switchers, the rhGH dose was 0.53 +/- 0.04 mg day(-1) at baseline (s.c. injection), 0.52 +/- 0.03 mg day(-1) at 12 months and 0.48 +/- 0.03 mg day(-1) at 24 months (NS between the different time points). There was no difference in the dose of rhGH at 12 and 24 months between the two groups. Side-effects were generally minor and consisted of local tissue reactions.

Conclusion: Administration of rhGH by needle-free, transdermal injection is effective in maintaining IGF-I concentrations in the normal range for age in adults with GHD, and is as effective as traditional subcutaneous injection of rhGH.

Alternative long-term markers for the detection of methyltestosterone misuse

2 Nov 2012

Alternative long-term markers for the detection of methyltestosterone misuse / C Gómez, O.J. Pozo, J. Marcos, J. Segura, R. Ventur. - (Steroids 78 (2013) 1 (January); p. 44-52)

  • PMID: 23127819
  • DOI: 10.1016/j.steroids.2012.10.008


Abstract

Methyltestosterone (MT) is one of the most frequently detected anabolic androgenic steroids in doping control analysis. MT misuse is commonly detected by the identification of its two main metabolites excreted as glucuronide conjugates, 17α-methyl-5α-androstan-3α,17β-diol and 17α-methyl-5β-androstan-3α,17β-diol. The detection of these metabolites is normally performed by gas chromatography-mass spectrometry, after previous hydrolysis with β-glucuronidase enzymes, extraction and derivatization steps. The aim of the present work was to study the sulphate fraction of MT and to evaluate their potential to improve the detection of the misuse of the drug in sports. MT was administered to healthy volunteers and urine samples were collected up to 30days after administration. After an extraction with ethyl acetate, urine extracts were analysed by liquid chromatography tandem mass spectrometry using electrospray ionisation in negative mode by monitoring the transition m/z 385 to m/z 97. Three diol sulphate metabolites (S1, S2 and S3) were detected. Potential structures for these metabolites were proposed after solvolysis and mass spectrometric experiments: S1, 17α-methyl-5β-androstan-3α,17β-diol 3α-sulphate; S2, 17β-methyl-5α-androstan-3α,17α-diol 3α-sulphate; and S3, 17β-methyl-5β-androstan-3α,17α-diol 3α-sulphate. Synthesis of reference compounds will be required in order to confirm the structures. The retrospectivity of these sulphate metabolites in the detection of MT misuse was compared with the obtained with previously described metabolites. Metabolite S2 was detected up to 21days after MT administration, improving between 2 and 3 times the retrospectivity of the detection compared to the last long-term metabolite of MT previously described, 17α-hydroxy-17β-methylandrostan-4,6-dien-3-one.

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