Blood oxyhemoglobin saturation measurements by blue-green spectral shift

Previous work describing a resilient method for measuring oxyhemoglobin saturation using the blue-green spectral shift was performed using cell free hemoglobin solutions. Hemoglobin solution and whole blood sample spectra measured under similar conditions in a spectrophotometer are used here to begin evaluating the impact of cellular scattering on this method. The blue-green spectral shift with changing oxyhemoglobin saturation was preserved in these blood samples and the blue-green spectral shift was relatively unaffected by physiological changes in blood pH (6.6, 7.1, and 7.4), path length through blood (100 and 200 microm), and blood hematocrit (19 to 48%). The packaging of hemoglobin in red blood cells leads to a decreased apparent path length through hemoglobin, and an overall decrease in scattering loss with increasing wavelength from 450 to 850 nm. The negative slope of the scattering loss in the 476 to 516-nm range leads to a +3.0 nm shift in the oxyhemoglobin saturation calibration line when the blue-green spectral minimum in these blood samples was compared to cell free hemoglobin. Further research is needed to fully evaluate the blue green spectral shift method in cellular systems including in vivo testing.