SARCOSYL‐PAGE: a new method for the detection of MIRCERA‐ and EPO‐doping in blood

Reichel, Christian; Abzieher, Friedrich; Geisendorfer, Thomas

doi:10.1002/dta.97

Cited by 78 publications

(107 citation statements)

References 32 publications

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“…[21] For HBOCs, visual inspection of plasma and quantification of haemoglobin were used as screening methods. Samples with altered plasma colour (whole blood spiked with >0.6% Hemopure) had their haemoglobin concentration measured using a Sysmex XT-2000i instrument.…”

Section: Biological Assaysmentioning

confidence: 99%

“…The XXXI Summer Olympic Games (August [3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21]2016) and the XV Paralympic Games (September 7-18, 2016) were held in Rio de Janeiro, marking the first time that these events were organised in South America. Doping control analyses have been conducted in Brazil since 1989, were incorporated into the International Olympic Committee (IOC) accreditation in 2002, and were absorbed by the World Anti-Doping Agency (WADA) accreditation system in 2005.…”

Section: Introduction and Facility Descriptionmentioning

confidence: 99%

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Doping control analysis at the Rio 2016 Olympic and Paralympic Games

Pereira

Sardela

Padilha

et al. 2017

Drug Testing and Analysis

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This paper summarises the results obtained from the doping control analyses performed during the Summer XXXI Olympic Games (August 3-21, 2016) and the XV Paralympic Games (September 7-18, 2016). The analyses of all doping control samples were performed at the Brazilian Doping Control Laboratory (LBCD), a World Anti-Doping Agency (WADA)-accredited laboratory located in Rio de Janeiro, Brazil. A new facility at Rio de Janeiro Federal University (UFRJ) was built and fully operated by over 700 professionals, including Brazilian and international scientists, administrative staff, and volunteers. For the Olympic Games, 4913 samples were analysed. In 29 specimens, the presence of a prohibited substance was confirmed, resulting in adverse analytical findings (AAFs). For the Paralympic Games, 1687 samples were analysed, 12 of which were reported as AAFs. For both events, 82.8% of the samples were urine, and 17.2% were blood samples. In total, more than 31 000 analytical procedures were conducted. New WADA technical documents were fully implemented; consequently, state-of-the-art analytical toxicology instrumentation and strategies were applied during the Games, including different types of mass spectrometry (MS) analysers, peptide, and protein detection strategies, endogenous steroid profile measurements, and blood analysis. This enormous investment yielded one of the largest Olympic legacies in Brazil and South America.

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Section: Biological Assaysmentioning

confidence: 99%

Section: Introduction and Facility Descriptionmentioning

confidence: 99%

Doping control analysis at the Rio 2016 Olympic and Paralympic Games

Pereira

Sardela

Padilha

et al. 2017

Drug Testing and Analysis

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“…The methods often require an additional purification and/or concentration step before urine and plasma samples analysis [14,15]. Endogenous and recombinant HuEPO glycosylation can be distinguished by differences in charge [16,17], isoelectric point [18][19][20], molecular mass [21][22][23] or interaction with lectins, e.g. wheat germ agglutinin (WGA) [24,25] for the glycoproteins.…”

Section: Introductionmentioning

confidence: 99%

Detection of recombinant human EPO administered to horses using MAIIA lateral flow isoform test

Lönnberg

Bondesson

Cormant³

et al. 2012

Anal Bioanal Chem

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Doping of horses with recombinant human erythropoietin (rHuEPO) to illegally enhance their endurance capacity in horseracing has been reported during the last years. This leads to increased blood viscosity which can result in sudden death and is of concern for the horse welfare. Additionally, the horse can start production of rHuEPO antibodies, which cross-reacts with endogenous equine EPO and can lead to severe anaemia and even death. In this study, a novel micro-chromatographic method, EPO WGA MAIIA, has been tested for the capability in plasma and urine samples to detect administration of erythropoiesis-stimulating agents, like the rHuEPO glycoprotein varieties Eprex and Aranesp, to horses. After administration of 40 IU Eprex kg(-1) day(-1) to seven horses during 6 days, the presence of Eprex in horse plasma was detected up to 2-5 days after last injection. In urine samples collected from two horses, Eprex was detected up to 3 days. A single injection of Aranesp (0.39 μg/kg) was detected up to 9 days in plasma and up to 8 days, the last day of testing, in the urine sample. The LC-FAIMS-MS/MS system, with 1 day reporting time, confirmed the presence of Eprex up to 1 day after last injection for six out of seven horses and the presence of Aranesp up to 5 days after last injection in plasma samples. The MAIIA system showed to be a promising tool with high sensitivity and extremely short reporting time (1 h).

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“…Exchanging SDS with SARCOSYL in the sample and running buffers is reported to solve the problem without compromising for the other rhEPOs. 96 As mentioned, information derived from blood analyses can assist in the detection and/or deterrence of blood doping. Several hematologic indices (number of red blood cells, blood hemoglobin concentration, Hct, ret count, number of hypochromic macrocytes) and biochemical parameters (serum EPO and sTfr concentrations) change significantly after rhEPO intake.…”

Section: Alternative Approaches To the Direct Recombinant Human Erythmentioning

confidence: 99%

Applications and Biomonitoring Issues of Recombinant Erythropoietins for Doping Control

Tsitsimpikou¹,

Kouretas²,

Tsarouhas³

et al. 2011

Therapeutic Drug Monitoring

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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.

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SARCOSYL‐PAGE: a new method for the detection of MIRCERA‐ and EPO‐doping in blood

Cited by 78 publications

References 32 publications

Doping control analysis at the Rio 2016 Olympic and Paralympic Games

Doping control analysis at the Rio 2016 Olympic and Paralympic Games

Detection of recombinant human EPO administered to horses using MAIIA lateral flow isoform test

Applications and Biomonitoring Issues of Recombinant Erythropoietins for Doping Control

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