Doping Control Analysis of Bovine Hemoglobin-Based Oxygen Therapeutics in Human Plasma by LC-Electrospray Ionization-MS/MS
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- 作者：Mario Thevis ; Rachel R. Ogorzalek Loo ; Joseph A. Loo ; and Wilhelm Schä ; nzer
- 刊名：Analytical Chemistry
- 出版年：2003
- 出版时间：July 15, 2003
- 年：2003
- 卷：75
- 期：14
- 页码：3287 - 3293
- 全文大小：102K
- 年卷期：v.75,no.14(July 15, 2003)
- ISSN：1520-6882
文摘
Since January 2000, hemoglobin-based oxygen carriers,such as Hemopure, belong to the list of prohibitedsubstances of the International Olympic Committee. Hemopure is based on bovine hemoglobin, which is intra-and intermolecularly cross-linked by glutaraldehyde unitscausing an average molecular weight of ~250 000. Bovineand human hemoglobins differ by 15% in amino acidsequence; hence, tryptic digestion of these proteinsgenerates species-common and -unique peptides. Thosespecific fragments originate from the 
- and 
-subunitsof hemoglobin, such as bovine Hb peptides 69-90 (2367.2Da) or 40-58 (2089.9 Da). By means of LC-MS/MS,peptides of human and bovine hemoglobin can be separated and identified, enabling the determination of compounds based on Hb of bovine origin and thus theadministration of oxygen carriers such as Hemopure.Blank plasma samples were spiked with Hemopure orhuman or bovine hemoglobin, filtered, enzymaticallydigested, and analyzed on an Agilent 1100 Series HPLCinterfaced to an Applied Biosystems API 2000 triplequadrupole mass spectrometer. In plasma aliquots of 50
L containing 50 g of Hemopure (1 mg/mL), peptidesof bovine hemoglobin were confirmed, and blank plasmasamples as well as 68 specimens of high-performanceathletes were tested with the developed procedure.
- and -subunitsof hemoglobin, such as bovine Hb peptides 69-90 (2367.2Da) or 40-58 (2089.9 Da). By means of LC-MS/MS,peptides of human and bovine hemoglobin can be separated and identified, enabling the determination of compounds based on Hb of bovine origin and thus theadministration of oxygen carriers such as Hemopure.Blank plasma samples were spiked with Hemopure orhuman or bovine hemoglobin, filtered, enzymaticallydigested, and analyzed on an Agilent 1100 Series HPLCinterfaced to an Applied Biosystems API 2000 triplequadrupole mass spectrometer. In plasma aliquots of 50L containing 50 g of Hemopure (1 mg/mL), peptidesof bovine hemoglobin were confirmed, and blank plasmasamples as well as 68 specimens of high-performanceathletes were tested with the developed procedure.