title: New insights for identification of doping with recombinant human erythropoietin micro-doses after high hydration.
abstract: To minimize the chances of being caught after doping with recombinant human erythropoietins (rhEPO), athletes have turned to new practices using micro-doses and excess fluid ingestion to accelerate elimination and decrease the probability of detection. Our objective was to test the sensitivity of detection by validated methods (IEF: isoelectric focusing; SDS-PAGE: sodium dodecyl sulfate polyacrylamide gel electrophoresis) when such practices are used. First, after a three-week rhEPO boost period and 10 days of wash out, detection of a single 900 IU micro-dose of Eprex® was evaluated in healthy male subjects. After an injection in the evening, urine and plasma samples were collected the following morning. Half of the subjects then drank a bolus of water and new samples were collected 80 min later. Interestingly, rhEPO was detected in 100% of the samples even after water ingestion. A second similar protocol was then performed with a single injection of a micro-dose of rhEPO (500 IU or 900 IU), without a prior rhEPO boost. In addition, urine and plasma samples were also collected 15 and 20 h post rhEPO administration. Once again drinking water did not affect the rate of detection. Urine appeared a better matrix to detect micro-doses after 10 h, enabling between 92% and 100% of identification at that time. The rate of identification decreased rapidly thereafter, in particular for the 500 IU micro-dose. However IEF analysis still resulted in 71% identification of rhEPO in urine after 20 h. These results could help to define a better strategy for controlling and identifying athletes using rhEPO micro-doses. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: Analysis Department-Agence Française de Lutte contre le Dopage (AFLD), Châtenay-Malabry, France.
authors.forename: L
authors.initials: L
authors.lastname: Martin
authors.affiliation: SIAB Research Consortium, Gold Coast, QLD, Australia.
authors.forename: M
authors.initials: M
authors.lastname: Ashenden
authors.affiliation: Department of Nutrition, Exercise and Sports, University of Copenhagen, Denmark.
authors.forename: J
authors.initials: J
authors.lastname: Bejder
authors.affiliation: Department of Nutrition, Exercise and Sports, University of Copenhagen, Denmark.|Institute of Sports Science and Clinical Biomechanics, University of Southern Denmark, Denmark.
authors.forename: M
authors.initials: M
authors.lastname: Hoffmann
authors.affiliation: Department of Nutrition, Exercise and Sports, University of Copenhagen, Denmark.
authors.forename: N
authors.initials: N
authors.lastname: Nordsborg
authors.affiliation: The Centre of Inflammation and Metabolism and the Centre for Physical Activity Research, Rigshospitalet, University of Copenhagen, Denmark.
authors.forename: K
authors.initials: K
authors.lastname: Karstoft
authors.affiliation: Metabolic Mass Spectrometry Facility, Rigshospitalet, Denmark.
authors.forename: J
authors.initials: J
authors.lastname: Morkeberg
authors.affiliation: Statistical Consulting Centre, Department of Mathematics and Statistics, University of Melbourne, VIC, Australia.
authors.forename: K
authors.initials: K
authors.lastname: Sharpe
authors.affiliation: Analysis Department-Agence Française de Lutte contre le Dopage (AFLD), Châtenay-Malabry, France.
authors.forename: F
authors.initials: F
authors.lastname: Lasne
authors.affiliation: Analysis Department-Agence Française de Lutte contre le Dopage (AFLD), Châtenay-Malabry, France.
authors.forename: A
authors.initials: A
authors.lastname: Marchand
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004921:Erythropoietin / Q000097:blood* / Q000378:metabolism / Q000652:urine*; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods*; D011994:Recombinant Proteins / Q000097:blood* / Q000378:metabolism / Q000652:urine*; D015813:Substance Abuse Detection / Q000379:methods*
chemical_list: D011994:Recombinant Proteins; D004921:Erythropoietin
keywords: EPO; IEF; SDS-PAGE; doping; micro-doses