Identification of darbepoetin alfa in human plasma by liquid chromatography coupled to mass spectrometry for doping control.
Int J Sports Med
; 30(2): 80-6, 2009 Feb.
Article
em En
| MEDLINE
| ID: mdl-19177313
ABSTRACT
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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Detecção do Abuso de Substâncias
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Cromatografia Líquida de Alta Pressão
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Eritropoetina
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Dopagem Esportivo
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Hematínicos
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
Idioma:
En
Revista:
Int J Sports Med
Ano de publicação:
2009
Tipo de documento:
Article
País de afiliação:
Estados Unidos