A fast screening method for the detection of the abuse of hemoglobin-based oxygen carriers (HBOCs) in doping control

Artificial oxygen carriers (AOCs) can be abused by the athletes to improve their aerobic capacity. The AOCs produce a performance enhancing effect, especially in endurance sports. This article presents a method for the rapid screening of hemoglobin-based oxygen carriers (HBOCs) in blood samples. Common screening tests to reveal HBOC misuse by athletes are based on colorimetric detection since HBOC use causes discoloration of the plasma. In this communication we are presenting a different approach for HBOC detection using an hematological analyzer capable of measuring hemoglobin by two methods: a standard cyanmethemoglobin colorimetric method to calculate the amount of total hemoglobin (HGBtot) and a flow cytometric optical method to calculate the amount of hemoglobin within the red blood cells (HGBcell). Thanks to this dual contemporary hemoglobin measurement, the HGBdelta value (corresponding to free HGB) is automatically calculated by subtraction of HGBcell from HGBtot and can be used as a fast screening index of HBOC abuse. We tested the effectiveness of this approach using 68 normal blood samples with different basal HGB values fortified with three different HBOCs at varying concentrations. We evaluated the performance of the method by calculating the correlation between HGBcell and HGBtot values in normal samples. Finally we used a simple statistical approach to calculate a reliable HGBdelta cut-off value (0.35 g/dL) as a limit of decision to discriminate between a clear negative sample and a suspect sample to submit to a confirmation analysis.