Identification of Darbepoetin Alfa in Human Plasma by Liquid Chromatography Coupled to Mass Spectrometry for Doping Control
| Autor: | Fuyu Guan, C. E. Uboh, Birksz E, Chen J, Lawrence R. Soma |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Doping in Sports
chemistry.chemical_classification PNGase F Detection limit Spectrometry Mass Electrospray Ionization Analyte Chromatography Darbepoetin alfa Physical Therapy Sports Therapy and Rehabilitation Peptide Trypsin Tandem mass spectrometry Mass spectrometry Substance Abuse Detection chemistry Hematinics medicine Humans Orthopedics and Sports Medicine Erythropoietin Chromatography High Pressure Liquid medicine.drug |
| Zdroj: | International Journal of Sports Medicine. 30:80-86 |
| ISSN: | 1439-3964 0172-4622 |
| DOI: | 10.1055/s-0028-1103283 |
| Popis: | Darbepoetin alfa (DPO) or Novel Erythropoiesis Erythropoiesis Stimulating Protein (NESP), an analog of recombinant human erythropoietin (rhEPO), is abused as a blood doping agent along with the latter in human sports. This paper describes a new method for unequivocal identification of DPO in human plasma. The analyte was extracted from plasma by immunoaffinity separation with anti-rhEPO antibodies, digested by trypsin followed by PNGase F, and analyzed by liquid chromatography coupled to tandem mass spectrometry. The deglycosylated tryptic peptide, T (9), was employed in DPO identification using liquid chromatographic retention time and major product ions of the T (9) peptide. The limit of detection of this method for DPO was 0.1 ng/mL in plasma, and that of identification was 0.2 ng/mL. This method is definitive and devoid of false positive results, providing "mass fingerprints" for identification of DPO in human plasma samples. Although this method is not applicable to identification of rhEPO in human plasma because it cannot differentiate rhEPO from endogenous EPO, it is the first successful attempt towards establishing a reliable and selective method for definitive identification of exogenously administered EPOs in doping control analyses. |
| Databáze: | OpenAIRE |
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