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Capillary electrophoretic analysis of whole blood samples for hemoglobin‐based oxygen carriers without the use of immunoprecipitation
Author(s) -
Harrison Christopher R.,
Vydha Srilatha
Publication year - 2012
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201100506
Subject(s) - hemoglobin , whole blood , capillary electrophoresis , chromatography , chemistry , blood substitute , oxygen , medicine , biochemistry , surgery , organic chemistry
Hemoglobin‐based oxygen carriers ( HBOC s) are blood substitutes, synthesized by polymerizing hemoglobin, which are being developed and investigated as alternatives to blood for medical purposes. However, due to their ability to increase the oxygen carrying capacity when taken by healthy individuals, HBOC s have been used as a doping agent among endurance athletes and are included in the World Anti‐Doping Agency's Prohibited List. To maintain the fairness of competitions and continue the battle against doping, it is essential to be able to detect HBOC s if present in an athlete's blood. To achieve this goal, it is necessary to differentiate HBOC s from the native hemoglobin and to do so in a cost and time effective manner. We have developed a rapid capillary zone electrophoresis ( CZE ), UV absorbance, method capable of detecting HBOC s, in whole blood samples, at levels below those considered necessary to provide a performance enhancement. Our approach to the analysis for HBOC s utilizes the whole blood sample, not just the plasma, and does not require the use of immunoprecipitants to ensure accurate analysis. By lysing the red blood cells and using centrifugal filtration, followed by our CZE separation, we are able to effectively distinguish between native hemoglobin and HBOC s. Through this method, we have been able to reliably detect concentrations of HBOC s at the equivalent of 5.5 g/L, the equivalent to a 3.5% increase in blood hemoglobin concentration for an athlete.