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.


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.


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.


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".

Circulating microRNA-122 as Potential Biomarker for Detection of Testosterone Abuse

12 May 2016

Circulating microRNA-122 as Potential Biomarker for Detection of Testosterone Abuse / Olivier Salamin, Laetitia Jaggi, Norbert Baume, Neil Robinson, Martial Saugy, Nicolas Leuenberger. - (PLoS One 11 (2016) 5 (May); p. 1-13)

  • PMID: 27171140
  • PMCID: PMC4865044
  • DOI: 10.1371/journal.pone.0155248


MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression and thus influence many cellular and physiological processes. miRNAs are also present in cell-free body fluids such as plasma or serum, and these circulating miRNAs are very stable, sensitive, and specific biomarkers of pathophysiological states. In this study, we investigated whether circulating miRNAs could serve as biomarkers of exogenous testosterone administration. Misuse of testosterone as a performance-enhancing drug is thought to be widespread in sports. Detection of testosterone through the urinary steroid profile of the Athlete Biological Passport faces several obstacles, indicating that new biomarkers are required. To this end, we analyzed plasma miRNA levels by high-throughput quantitative real-time PCR. Plasma samples were obtained before and at several time points after transdermal and oral testosterone administration. Screening identified three potential candidate miRNAs that were altered by both routes of testosterone administration. Longitudinal monitoring of these candidates revealed that variation in two of them (miR-150 and miR-342), relative to the corresponding levels in control samples, was testosterone-independent. However, levels of the liver-specific miR-122 increased 3.5-fold 1 day after drug intake. Given that testosterone is metabolized by the liver, this observation suggests that miR-122 in cell-free fluids may be used as a sensitive biomarker of testosterone misuse via multiple dosing routes and could therefore be integrated into a blood-based multiparametric follow-up.

Erythropoietin as a performance-enhancing drug: Its mechanistic basis, detection, and potential adverse effects

22 Jan 2017

Erythropoietin as a performance-enhancing drug: Its mechanistic basis, detection, and potential adverse effects / Olivier Salamin, Tiia Kuuranne, Martial Saugy, Nicolas Leuenberger. - (Molecular and Cellular Endocrinology 464 (2018) 15 March; p. 75-87)

  • PMID: 28119134
  • DOI: 10.1016/j.mce.2017.01.033


Erythropoietin (EPO) is the main hormone regulating red blood cell (RBC) production. The large-scale production of a recombinant human erythropoietin (rHuEPO) by biotechnological methods has made possible its widespread therapeutic use as well as its misuse in sports. Since the marketing of the first epoetin in 1989, the development has progressed to the third-generation analogs. However, the production of rHuEPO is costly, and the frequent administration of an injectable formula is not optimal for compliance of therapeutic patients. Hence, pharmaceutical industries are currently developing alternative approaches to stimulate erythropoiesis, which might offer new candidates for doping purposes. The hypoxia inducible factors (HIF) pathway is of particular interest. The introduction of new erythropoiesis-stimulating agents (ESAs) for clinical use requires subsequent development of anti-doping methods for detecting the abuse of these substances. The detection of ESAs is based on two different approaches, namely, the direct detection of exogenous substances and the indirect detection, for which the effects of the substances on specific biomarkers are monitored. Omics technologies, such as ironomics or transcriptomics, are useful for the development of new promising biomarkers for the detection of ESAs. Finally, the illicit use of ESAs associates with multiple health risks that can be irreversible, and an essential facet of anti-doping work is to educate athletes of these risks. The aim of this review is to provide an overview of the evolution of ESAs, the research and implementation of the available detection methods, and the side effects associated with the misuse of ESAs.

Novel Erythropoietic Agents: A Threat to Sportsmanship

27 Apr 2007

Novel Erythropoietic Agents : A Threat to Sportsmanship / Wolfgang Ernst Bernard Jelkmann. - (Medicina Sportiva 11 (2007) 2; p. 32-42)


The mass of hemoglobin (Hb) is an important determinant of aerobic power. Red blood cell production is stimulated by
the glycoprotein erythropoietin (EPO). Recombinant human EPO (rHuEPO) engineered in Chinese hamster ovary (CHO)
cell cultures (Epoetin alfa and Epoetin beta) and its hyperglycosylated analogue Darbepoetin alfa are known to be misused by athletes. The drugs can be detected in urine by isoelectric focusing, because the pattern of their glycans differs from that of endogenous EPO. However, doping control is becoming much more difficult, as various novel erythropoiesis stimulating agents (ESAs) are - or are to come - on the market. Gene-activated EPO (Epoetin delta) from human fibrosarcoma cells (HT-1080) has been approved in the European Union. rHuEPO (Epoetin omega) produced in transformed baby hamster kidney (BHK) cells is also available. ESAs to come include Biosimilars of the established Epoetins, long-acting pegylated Epoetin beta (CERA), EPO fusion proteins, synthetic erythropoiesis stimulating protein (SEP) and peptidic (Hematide) as well as non-peptidic EPO mimetics. Furthermore, small orally active drugs that are capable of stimulating endogenous EPO production are in preclinical or clinical trials. These include stabilizers of the hypoxia-inducible transcription factors (HIF) that bind to the EPO gene enhancer and GATA inhibitors which prevent GATA from suppressing the EPO gene promoter. It is crucial to inform the athletes and their supporting staff on the potential health risks.

Untargeted profiling of urinary steroid metabolites after testosterone ingestion: opening new perspectives for antidoping testing

20 Nov 2014

Untargeted profiling of urinary steroid metabolites after testosterone ingestion: opening new perspectives for antidoping testing / Julien Boccard, Flavia Badoud, Nicolas Jan, Raul Nicoli, Carine Schweizer, François Pralong, Jean-Luc Veuthey, Norbert Baume, Serge Rudaz, Martial Saugy. - (Bioanalysis 6 (2014) 19; p. 2523-2536)

  • PMID: 25411696
  • DOI: 10.4155/bio.14.200


Aim: Antidoping procedures are expected to greatly benefit from untargeted metabolomic approaches through the discovery of new biomarkers of prohibited substances abuse.

Results: Endogenous steroid metabolites were monitored in urine samples from a controlled elimination study of testosterone undecanoate after ingestion. A platform coupling ultra-high pressure LC with high-resolution quadrupole TOF MS was used and high between-subject metabolic variability was successfully handled using a multiblock data analysis strategy. Links between specific subsets of metabolites and influential genetic polymorphisms of the UGT2B17 enzyme were highlighted.

Conclusion: This exploratory metabolomic strategy constitutes a first step toward a better understanding of the underlying patterns driving the high interindividual variability of steroid metabolism. Promising biomarkers were selected for further targeted study.

Blood manipulation: current challenges from an anti-doping perspective

6 Dec 2013

Blood manipulation : current challenges from an anti-doping perspective / Jakob Mørkeberg. - (Sports Medicine in Hematology (2013) 1 (6 December); p. 627-631)

  • PMID: 24319242
  • DOI: 10.1182/asheducation-2013.1.627


The delivery of oxygen is the limiting factor during whole-body endurance exercise in well-trained individuals, so manipulating the amount of hemoglobin in the blood results in changes in endurance exercise capacity. Athletes began using novel erythropoiesis-stimulating agents well before they were approved for medical use. Older manipulation practices, such as autologous blood transfusions or the administration of first-generation recombinant human erythropoietins, are still widely abused due to challenges in their detection. More recent performance enhancement maneuvers include efforts to mask doping and to induce increased endogenous erythropoietin expression. Confessions by athletes have revealed an ongoing yet extremely sophisticated modus operandi when manipulating the blood. In this review, weaknesses in detection methods and sample collection procedures are scrutinized and strategies developed to circumvent the test system discussed.

Hepcidin as a potential biomarker for blood doping

18 Oct 2016

Hepcidin as a potential biomarker for blood doping / Nicolas Leuenberger, Emanuele Bulla, Olivier Salamin, Raul Nicoli, Neil Robinson, Norbert Baume, Martial Saugy. - (Drug Testing and Analysis 9 (2017) 7 (July); p. 1093-1097)

  • PMID: 27758046
  • DOI: 10.1002/dta.2122


The concentration of hepcidin, a key regulator of iron metabolism, is suppressed during periods of increased erythropoietic activity. The present study obtained blood samples from 109 elite athletes and examined the correlations between hepcidin and markers of erythropoiesis and iron metabolism (i.e., haemoglobin, erythropoietin (EPO), ferritin, erythroferrone (ERFE), and iron concentration). Furthermore, an administration study was undertaken to examine the effect of recombinant human EPO (rhEPO) delta (Dynepo™) on hepcidin concentrations in healthy male volunteers. The effects on hepcidin were then compared with those on reticulocyte percentage (Ret%) and ferritin concentration. There was a significant positive correlation between hepcidin and ferritin, iron, and haemoglobin levels in athletes, whereas hepcidin showed an inverse correlation with ERFE. Administration of rhEPO delta reduced hepcidin levels, suggesting that monitoring hepcidin may increase the sensitivity of the Athlete Biological Passport (ABP) for detecting rhEPO abuse.

Erythropoiesis-Stimulating Agents

18 Oct 2010

Erythropoiesis-Stimulating Agents / Steve Elliot. - Chapter 4; p. 55-74

Published in:

Hematopoietic Growth Factors in Oncology. - Boston, MA, Springer. - (Cancer Treatment and Research book series (CTAR); volume 157)

  • DOI: 10.1007/978-1-4419-7073-2_4
  • Print ISBN 978-1-4419-7072-5
  • Online ISBN 978-1-4419-7073-2


Erythropoiesis is the process whereby erythroid progenitor cells differentiate and divide, resulting in increased numbers of red blood cells (RBCs). RBCs contain hemoglobin, the main oxygen carrying component in blood. The large number of RBCs found in blood is required to support the prodigious consumption of oxygen by tissues as they undergo oxygen-dependent processes. Erythropoietin is a hormone that when it binds and activates Epo receptors resident on the surface of cells results in stimulation of erythropoiesis. Successful cloning of the EPO gene allowed for the first time production of recombinant human erythropoietin and other erythropoiesis stimulating agents (ESAs), which are used to treat anemia in patients. In this chapter, the control of Epo levels and erythropoiesis, the various forms of ESAs used commercially, and their physical and biological properties are discussed.

Red blood cell populations and membrane levels of peroxiredoxin 2 as candidate biomarkers to reveal blood doping

23 May 2012

Red blood cell populations and membrane levels of peroxiredoxin 2 as candidate biomarkers to reveal blood doping / Cristina Marrocco, Valeria Pallotta, Angelo D'alessandro, Gilda Alves, Lello Zolla. - (Blood Transfusion 10 (2012) Supplement 2 (May); p. s71-s77)

  • PMID: 22890272
  • PMCID: PMC3418622
  • DOI: 10.2450/2012.011S


Background: Blood doping represents one main trend in doping strategies. Blood doping refers to the practice of boosting the number of red blood cells (RBCs) in the bloodstream in order to enhance athletic performance, by means of blood transfusions, administration of erythropoiesis-stimulating substances, blood substitutes, natural or artificial altitude facilities, and innovative gene therapies. While detection of recombinant EPO and homologous transfusion is already feasible through electrophoretic, mass spectrometry or flow cytometry-based approaches, no method is currently available to tackle doping strategies relying on autologous transfusions.

Materials and methods: We exploited an in vitro model of autologous transfusion through a 1:10 dilution of concentrated RBCs after 30 days of storage upon appropriate dilution in freshly withdrawn RBCs from the same donor. Western blot towards membrane Prdx2 and Percoll density gradients were exploited to assess their suitability as biomarkers of transfusion.

Results: Membrane Prdx2 was visible in day 30 samples albeit not in day 0, while it was still visible in the 1:10 dilution of day 30 in day 0 RBCs. Cell gradients also highlighted changes in the profile of the RBC subpopulations upon dilution of stored RBCs in the fresh ones.

Discussion: From this preliminary in vitro investigation it emerges that Prdx2 and RBC populations might be further tested as candidate biomarkers of blood doping through autologous transfusion, though it is yet to be assessed whether the kinetics in vivo of Prdx2 exposure in the membrane of transfused RBCs will endow a sufficient time-window to allow reliable anti-doping testing.

Applications and biomonitoring issues of recombinant erythropoietins for doping control

1 Feb 2011

Applications and biomonitoring issues of recombinant erythropoietins for doping control / Christina Tsitsimpikou, Demetrios Kouretas, Konstantinos Tsarouhas, Kenneth Fitch, Demetrios A. Spandidos, Aristides Tsatsakis. - (Therapeutic Drug Monitoring 33 (2011) 1 (February); p. 3-13)

  • PMID: 21099742
  • DOI: 10.1097/FTD.0b013e31820032c4


The biochemical actions and side effects of recombinant erythropoietins (rhEPOs), their analogs and mimetics, their misuse as doping agents, and the principal analytical strategies developed to identify them in athletes' biologic fluids are reviewed. Patients who experience a range of pathologies have benefited from the administration of rhEPOs to correct severe anemia. Currently, monitoring the biologic effect of rhEPO in patients under treatment is by measuring the hemoglobin concentration. However, it may be valuable to directly monitor the actual levels of the administered drug and determine a dose-dependent correlation with any clinical adverse effect observed. This may permit the adoption of a patient-specific administration regime. Currently, the method of detecting EPO approved for doping control is an isoelectric-focusing, double-blotting, chemiluminescence assay based on charge differences between isoforms of rhEPOs and endogenous EPO in urine. The advantages and limitations of this method are presented. A new approach using sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a complementary tool to the established method is discussed. The application of matrix-assisted laser desorption/ionization mass spectrometry and liquid chromatography combined with tandem mass spectrometry for the direct detection of the rhEPO molecules may prove to be promising. Indirect evidence of rhEPO abuse by athletes is based on the analysis of blood parameters (hemoglobin hematocrit, reticulocytes, macrocytes, etc) and serum markers (concentration of EPO and serum transferrin receptors, etc). Enrichment of the screened parameters with gene or biochemical markers revealing altered erythropoiesis and adoption of longitudinal monitoring of athletes' hematologic and biochemical parameters could also be a complementary approach in the fight against doping.

Techniques of deceptive communication about doping

31 May 2021

Techniques of deceptive communication about doping / Marcel Reinold. - (European Journal for Sport and Society (2021) 31 May)

  • DOI: 10.1080/16138171.2021.1930944


The decision to dope almost inevitably implies the decision to engage in deceptive communication. We, therefore, analysed six autobiographies of deceiving cyclists and identified six communication techniques which deceivers routinely apply: (1) moralisation without personal criticism, (2) exaggerating the intensity of anti-doping policies, (3) victimisation, (4) playing down the extent of the doping problem, (5) omitting narrative details, and (6) pretending lack of doping-relevant knowledge. These techniques help deceivers to present themselves as compliant with the anti-doping system and appear credible in their commitment against drugs. Furthermore, it helps them to allay suspicion, prevent falsification, and generally manage the flow of destructive information in a way that avoids leakage and detection. Though there is no reliable cue to deceit, knowledge about deceivers’ communication techniques might increase awareness and help to ask critical questions.

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