Analysis of hemoglobin-based oxygen carriers by CE-UV/Vis and CE-ESI-TOF/MS.
Blood doping involves the use of products that enhance the uptake, transport, or delivery of oxygen to the blood. One approach uses artificial oxygen carriers, known as hemoglobin-based oxygen carriers (HBOCs). This study describes an analytical strategy based on CE for detecting intact HBOCs in plasma samples collected for doping control. On-capillary detection was performed by UV/Vis at 415 nm, which offered detection selectivity for hemoproteins (such as hemoglobin and HBOCs). On-line ESI-MS detection with a TOF analyzer was further used to provide accurate masses on CE peaks and to confirm the presence of HBOCs. An immunodepletion sample preparation step was mandatory prior to analysis, in order to remove most abundant proteins that interfered with CE separation and altered the ESI process. This analytical method was successfully applied to plasma samples enriched with Oxyglobin, a commercially available HBOC used for veterinary purposes. Detection limits of 0.20 and 0.45 g/dL were achieved in plasma for CE-UV/Vis at 415 nm and CE-ESI-TOF/MS, respectively.
Staub
Aline
A
School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Geneva, Switzerland.|Swiss Centre for Applied Human Toxicology, University of Geneva, Geneva, Switzerland.
Rudaz
Serge
S
School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Geneva, Switzerland.|Swiss Centre for Applied Human Toxicology, University of Geneva, Geneva, Switzerland.
Saugy
Martial
M
Swiss Centre for Applied Human Toxicology, University of Geneva, Geneva, Switzerland.|Swiss Laboratory for Doping Analysis, Centre Universitaire Romand de Médecine Légale, Epalinges, Switzerland.
Veuthey
Jean-Luc
JL
School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Geneva, Switzerland.|Swiss Centre for Applied Human Toxicology, University of Geneva, Geneva, Switzerland.
Schappler
Julie
J
School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Geneva, Switzerland.|Swiss Centre for Applied Human Toxicology, University of Geneva, Geneva, Switzerland.
D001802:Blood Substitutes / Q000737:chemistry*; D004300:Doping in Sports; D019075:Electrophoresis, Capillary / Q000379:methods*; D006454:Hemoglobins / Q000737:chemistry*; D006801:Humans; D007163:Immunosorbent Techniques; D011506:Proteins / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013056:Spectrophotometry, Ultraviolet
D001802:Blood Substitutes; D006454:Hemoglobins; D011506:Proteins; C402601:hemoglobin glutamer-200