Confirmation and quantification of hemoglobin-based oxygen carriers in equine and human plasma by hyphenated liquid chromatography tandem mass spectrometry.

This study presents a method for detecting and measuring the amount of oxygen-carrying agents in the blood of horses and humans. These agents include Oxyglobin, Hemopure, and Hemolink, which could potentially be misused by athletes for performance enhancement.

Overview of the Research

• Researchers have developed a technique to detect, confirm, and quantify the presence of certain hemoglobin-based oxygen carriers, namely Oxyglobin (OXY), Hemopure (HMP), and Hemolink (HML) in the blood of horses and humans.
• This study has been conducted in the context of potential misuse of these products by athletes to improve performance through enhanced oxygen delivery to tissues.

Detection and Quantitation Procedure

• The scientists extracted OXY, HMP, and HML from the plasma of equines or humans using a method known as solid-phase extraction.
• They then digested the extracts with an enzyme called trypsin and subjected the resulting digests to a technique called Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS).
• Specific tryptic peptides derived from the bovine and human hemoglobin chains were targeted for detection and quantification.

Results and Findings from the Study

• The researchers successfully detected and quantified OXY and HMP in the plasma samples using specific peptides from the alpha chain of the bovine hemoglobin. In contrast, HML was quantified using peptides derived from the human hemoglobin alpha chain.
• The limit of detection and confirmation, and the range of quantification, for each of the targeted substances in both equine and human plasma was determined.
• The OXY substance was found to remain stable for over 30 days at temperatures of -20 and -70 degrees C.
• When a very low dose of OXY was administered to horses, it was detected and quantified in equine plasma up to 24 hours after administration, and its presence was confirmed up to 12 hours after administration.

Implications of the Study

• The development of this reliable detection and quantification method can help prevent the misuse of these oxygen-carrying agents by athletes, supporting efforts to maintain integrity and fairness in competitive sports.
• Additionally, the procedure could have applications in medical fields, ensuring proper dosage and monitoring of these substances when they are used for treatments such as anemia.