New insights for identification of doping with recombinant human erythropoietin micro-doses after high hydration.
Identifieur interne : 001A48 ( Main/Exploration ); précédent : 001A47; suivant : 001A49New insights for identification of doping with recombinant human erythropoietin micro-doses after high hydration.
Auteurs : L. Martin [France] ; M. Ashenden [Australie] ; J. Bejder [Danemark] ; M. Hoffmann [Danemark] ; N. Nordsborg [Danemark] ; K. Karstoft [Danemark] ; J. Morkeberg [Danemark] ; K. Sharpe [Australie] ; F. Lasne [France] ; A. Marchand [France]Source :
- Drug testing and analysis [ 1942-7611 ] ; 2016.
Descripteurs français
- KwdFr :
- Détection d'abus de substances (), Focalisation isoélectrique (), Humains, Protéines recombinantes (métabolisme), Protéines recombinantes (sang), Protéines recombinantes (urine), Électrophorèse sur gel de polyacrylamide (), Érythropoïétine (métabolisme), Érythropoïétine (sang), Érythropoïétine (urine).
- MESH :
English descriptors
- KwdEn :
- MESH :
- chemical , blood : Erythropoietin, Recombinant Proteins.
- chemical , metabolism : Erythropoietin, Recombinant Proteins.
- methods : Electrophoresis, Polyacrylamide Gel, Isoelectric Focusing, Substance Abuse Detection.
- chemical , urine : Erythropoietin, Recombinant Proteins.
- Humans.
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.
DOI: 10.1002/dta.2004
PubMed: 27390252
Affiliations:
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Martin</name><affiliation wicri:level="1"><nlm:affiliation>Analysis Department-Agence Française de Lutte contre le Dopage (AFLD), Châtenay-Malabry, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Analysis Department-Agence Française de Lutte contre le Dopage (AFLD), Châtenay-Malabry</wicri:regionArea><wicri:noRegion>Châtenay-Malabry</wicri:noRegion><wicri:noRegion>Châtenay-Malabry</wicri:noRegion></affiliation></author><author><name sortKey="Ashenden, M" sort="Ashenden, M" uniqKey="Ashenden M" first="M" last="Ashenden">M. Ashenden</name><affiliation wicri:level="1"><nlm:affiliation>SIAB Research Consortium, Gold Coast, QLD, Australia.</nlm:affiliation><country xml:lang="fr">Australie</country><wicri:regionArea>SIAB Research Consortium, Gold Coast, QLD</wicri:regionArea><wicri:noRegion>QLD</wicri:noRegion></affiliation></author><author><name sortKey="Bejder, J" sort="Bejder, J" uniqKey="Bejder J" first="J" last="Bejder">J. Bejder</name><affiliation wicri:level="1"><nlm:affiliation>Department of Nutrition, Exercise and Sports, University of Copenhagen, Denmark.</nlm:affiliation><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Nutrition, Exercise and Sports, University of Copenhagen</wicri:regionArea><wicri:noRegion>University of Copenhagen</wicri:noRegion></affiliation></author><author><name sortKey="Hoffmann, M" sort="Hoffmann, M" uniqKey="Hoffmann M" first="M" last="Hoffmann">M. Hoffmann</name><affiliation wicri:level="1"><nlm:affiliation>Department of Nutrition, Exercise and Sports, University of Copenhagen, Denmark.</nlm:affiliation><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Nutrition, Exercise and Sports, University of Copenhagen</wicri:regionArea><wicri:noRegion>University of Copenhagen</wicri:noRegion></affiliation></author><author><name sortKey="Nordsborg, N" sort="Nordsborg, N" uniqKey="Nordsborg N" first="N" last="Nordsborg">N. Nordsborg</name><affiliation wicri:level="1"><nlm:affiliation>Department of Nutrition, Exercise and Sports, University of Copenhagen, Denmark.</nlm:affiliation><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Nutrition, Exercise and Sports, University of Copenhagen</wicri:regionArea><wicri:noRegion>University of Copenhagen</wicri:noRegion></affiliation></author><author><name sortKey="Karstoft, K" sort="Karstoft, K" uniqKey="Karstoft K" first="K" last="Karstoft">K. Karstoft</name><affiliation wicri:level="1"><nlm:affiliation>The Centre of Inflammation and Metabolism and the Centre for Physical Activity Research, Rigshospitalet, University of Copenhagen, Denmark.</nlm:affiliation><country xml:lang="fr">Danemark</country><wicri:regionArea>The Centre of Inflammation and Metabolism and the Centre for Physical Activity Research, Rigshospitalet, University of Copenhagen</wicri:regionArea><wicri:noRegion>University of Copenhagen</wicri:noRegion></affiliation></author><author><name sortKey="Morkeberg, J" sort="Morkeberg, J" uniqKey="Morkeberg J" first="J" last="Morkeberg">J. Morkeberg</name><affiliation wicri:level="1"><nlm:affiliation>Metabolic Mass Spectrometry Facility, Rigshospitalet, Denmark.</nlm:affiliation><country xml:lang="fr">Danemark</country><wicri:regionArea>Metabolic Mass Spectrometry Facility, Rigshospitalet</wicri:regionArea><wicri:noRegion>Rigshospitalet</wicri:noRegion></affiliation></author><author><name sortKey="Sharpe, K" sort="Sharpe, K" uniqKey="Sharpe K" first="K" last="Sharpe">K. Sharpe</name><affiliation wicri:level="4"><nlm:affiliation>Statistical Consulting Centre, Department of Mathematics and Statistics, University of Melbourne, VIC, Australia.</nlm:affiliation><country xml:lang="fr">Australie</country><wicri:regionArea>Statistical Consulting Centre, Department of Mathematics and Statistics, University of Melbourne, VIC</wicri:regionArea><orgName type="university">Université de Melbourne</orgName><placeName><settlement type="city">Melbourne</settlement><region type="état">Victoria (État)</region></placeName></affiliation></author><author><name sortKey="Lasne, F" sort="Lasne, F" uniqKey="Lasne F" first="F" last="Lasne">F. Lasne</name><affiliation wicri:level="1"><nlm:affiliation>Analysis Department-Agence Française de Lutte contre le Dopage (AFLD), Châtenay-Malabry, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Analysis Department-Agence Française de Lutte contre le Dopage (AFLD), Châtenay-Malabry</wicri:regionArea><wicri:noRegion>Châtenay-Malabry</wicri:noRegion><wicri:noRegion>Châtenay-Malabry</wicri:noRegion></affiliation></author><author><name sortKey="Marchand, A" sort="Marchand, A" uniqKey="Marchand A" first="A" last="Marchand">A. Marchand</name><affiliation wicri:level="1"><nlm:affiliation>Analysis Department-Agence Française de Lutte contre le Dopage (AFLD), Châtenay-Malabry, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Analysis Department-Agence Française de Lutte contre le Dopage (AFLD), Châtenay-Malabry</wicri:regionArea><wicri:noRegion>Châtenay-Malabry</wicri:noRegion><wicri:noRegion>Châtenay-Malabry</wicri:noRegion></affiliation></author></analytic><series><title level="j">Drug testing and analysis</title><idno type="eISSN">1942-7611</idno><imprint><date when="2016" type="published">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Electrophoresis, Polyacrylamide Gel (methods)</term><term>Erythropoietin (blood)</term><term>Erythropoietin (metabolism)</term><term>Erythropoietin (urine)</term><term>Humans</term><term>Isoelectric Focusing (methods)</term><term>Recombinant Proteins (blood)</term><term>Recombinant Proteins (metabolism)</term><term>Recombinant Proteins (urine)</term><term>Substance Abuse Detection (methods)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Détection d'abus de substances ()</term><term>Focalisation isoélectrique ()</term><term>Humains</term><term>Protéines recombinantes (métabolisme)</term><term>Protéines recombinantes (sang)</term><term>Protéines recombinantes (urine)</term><term>Électrophorèse sur gel de polyacrylamide ()</term><term>Érythropoïétine (métabolisme)</term><term>Érythropoïétine (sang)</term><term>Érythropoïétine (urine)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>Erythropoietin</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Erythropoietin</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Electrophoresis, Polyacrylamide Gel</term><term>Isoelectric Focusing</term><term>Substance Abuse Detection</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Protéines recombinantes</term><term>Érythropoïétine</term></keywords><keywords scheme="MESH" qualifier="sang" xml:lang="fr"><term>Protéines recombinantes</term><term>Érythropoïétine</term></keywords><keywords scheme="MESH" type="chemical" qualifier="urine" xml:lang="en"><term>Erythropoietin</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Humans</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Détection d'abus de substances</term><term>Focalisation isoélectrique</term><term>Humains</term><term>Protéines recombinantes</term><term>Électrophorèse sur gel de polyacrylamide</term><term>Érythropoïétine</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">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.</div></front></TEI><affiliations><list><country><li>Australie</li><li>Danemark</li><li>France</li></country><region><li>Victoria (État)</li></region><settlement><li>Melbourne</li></settlement><orgName><li>Université de Melbourne</li></orgName></list><tree><country name="France"><noRegion><name sortKey="Martin, L" sort="Martin, L" uniqKey="Martin L" first="L" last="Martin">L. Martin</name></noRegion><name sortKey="Lasne, F" sort="Lasne, F" uniqKey="Lasne F" first="F" last="Lasne">F. Lasne</name><name sortKey="Marchand, A" sort="Marchand, A" uniqKey="Marchand A" first="A" last="Marchand">A. Marchand</name></country><country name="Australie"><noRegion><name sortKey="Ashenden, M" sort="Ashenden, M" uniqKey="Ashenden M" first="M" last="Ashenden">M. Ashenden</name></noRegion><name sortKey="Sharpe, K" sort="Sharpe, K" uniqKey="Sharpe K" first="K" last="Sharpe">K. Sharpe</name></country><country name="Danemark"><noRegion><name sortKey="Bejder, J" sort="Bejder, J" uniqKey="Bejder J" first="J" last="Bejder">J. Bejder</name></noRegion><name sortKey="Hoffmann, M" sort="Hoffmann, M" uniqKey="Hoffmann M" first="M" last="Hoffmann">M. Hoffmann</name><name sortKey="Karstoft, K" sort="Karstoft, K" uniqKey="Karstoft K" first="K" last="Karstoft">K. Karstoft</name><name sortKey="Morkeberg, J" sort="Morkeberg, J" uniqKey="Morkeberg J" first="J" last="Morkeberg">J. Morkeberg</name><name sortKey="Nordsborg, N" sort="Nordsborg, N" uniqKey="Nordsborg N" first="N" last="Nordsborg">N. 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