Prior work in our lab has indicated that reduced Hydroxocobalamin (B12r) can be added to human blood and is able to convert carbon monoxide (CO) into carbon dioxide. This has great potential as a direct antidote to mitigate the toxic effects of CO poisoning which is a public health risk. In the first part of our work, we use highly specific wavelengths of light and Raman spectroscopy to study changes in Carboxyhemoglobin (COHb) between blood treated with oxygen and blood treated with oxygen and B12r in a flowing circuit of blood. Using Raman spectroscopy, we found that the addition of B12r hastens the conversion of the COHb Raman signals to Oxyhemoglobin (HbO2) Raman signals. In addition, the B12r absorbance of light energy within the Raman spectrum is an exploitable relationship that can be used to measure B12r presence in the blood. In part two of our study we focused on the neurobehavioral testing of rats injured by CO exposure, however, we were not able to find statistical differences in the behavioral tests between exposed and unexposed rats.

Based upon experimental evidence from the 1970’s we proposed that a reduced form of hydroxocobalamin should be capable of producing carbon dioxide (CO2) from carbon monoxide (CO) in blood, and that this conversion should be detectable. Using resonance raman spectroscopy we demonstrated that a mixture of hydroxocobalamin and ascorbic acid could create the reduced form of hydroxocobalamin. We used a closed-loop circulation system with a hollow-fiber membrane oxygenator to produce carboxyhemoglobin. Using sensitive gas monitoring equipment to the gas-out port of the oxygenator we analyzed the CO and CO2 concentrations coming from the oxygenator. The mixture of hydroxocobalamin and ascorbic acid caused a 5-fold increase in the CO2 concentration of the gas-out flow, in comparison to baseline and negative controls. These findings offer initial support for the potential use of a mixture of hydroxocobalamin and ascorbic acid as an injectable antidote for carbon monoxide poisoning.