Τίτλος:
Stabilization of human urine doping control samples: IV. Human chorionic gonadotropin
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
The presence of proteolytic enzymes in urine samples, coming from exogenous or endogenous sources, enhances the cleavage of human chorionic gonadotropin (hCG). Moreover, elevated temperatures occurring occasionally during the delayed transportation of sport urine samples, favor the nicking of the hCG molecule. The aim of the current study, funded by the World Anti-Doping Agency (WADA), was the application of a stabilization mixture in athletes' urine samples to chemically inactivate proteolytic enzymes coming from exogenous or endogenous sources so as to prevent the degradation of hCG. The stabilization mixture applied, already tested for the stabilization of endogenous steroids and recombinant erythropoietin (rEPO), was a combination of antibiotics, antimycotic substances, and protease inhibitors. Incubation experiments were conducted in the presence or absence of the stabilization mixture in urine aliquots spiked with six proteases (first series of experiments) and one microorganism associated with urinary tract infections (UTI) (second series of experiments). Intact hCG levels were evaluated by using the EIAgen Total hCG kit. In the first series of experiments, hCG levels were reduced in the untreated aliquots following incubation at 37 °C. The addition of the chemical stabilization mixture prevented degradation of hCG induced by four of the proteases applied. In the second series of experiments, no significant difference was found in urine inoculated with E. coli, between aliquots treated with chemical mixture and the untreated aliquots. The addition of the proposed chemical stabilization mixture improves the quality of athletes' urine samples against possible deterioration due to high temperatures or attempts of proteolytic manipulation. © 2010 Springer-Verlag.
Συγγραφείς:
Tsivou, M.
Dimopoulou, H.A.
Georgakopoulos, D.G.
Koupparis, M.A.
Atta-Politou, J.
Georgakopoulos, C.G.
Περιοδικό:
Analytical and Bioanalytical Chemistry
Λέξεις-κλειδιά:
Chemical mixtures; Chemical stabilization; Doping control; Doping control analysis; E. coli; Elevated temperature; Endogenous steroids; High temperature; Human chorionic gonadotropins; Human urine; Protease inhibitor; Proteases; Proteolytic enzyme; Recombinant erythropoietin (rEpo); Urinary tract infections; Urine; Urine sample; World Anti-Doping Agency, Body fluids; Catalysts; Chemical analysis; Degradation; Doping (additives); Enzymes; Escherichia coli; Experiments; Microorganisms; Mixtures, Stabilization, chorionic gonadotropin, conference paper; doping; enzyme immunoassay; human; standard; urine, Chorionic Gonadotropin; Doping in Sports; Humans; Immunoenzyme Techniques; Reference Standards, Escherichia coli
DOI:
10.1007/s00216-010-4033-9
