Non-invasive measurements of hemoglobin + myoglobin, their oxygenation and NIR light pathlength in heart in vivo by diffuse reflectance spectroscopy

The existing non-invasive optical methods of the hemoglobin (Hb) and myoglobin (Mb) estimation in cardiac tissues imply knowledge of the light pathlength (L) when various modifications of Lambert-Beer law for either spectrophotometry or light diffuse reflectance is applied. For Hb and/or Mb quantification in tissue, a few invasive (biochemical) approaches were applied. For L (differential pathlength factor; DPF) determination in tissue, special optical methods were used. No approaches have been proposed to simultaneously and non-invasively determine Hb/Mb and L in cardiac or other muscle tissues. In the present study, the first derivative of the NIR diffuse reflectance spectrum is shown to be effective in simultaneous determination of Hb+Mb concentration (in mM) and L (in mm) in cardiac tissue in vivo. The results showed that measured in a few minutes in a normal pig heart in vivo the total Hb+Mb concentration was 0.9-1.2 mM of heme, tissue oxygen saturation parameter (OSP) was approximately 65%, and DPF at 700-965 nm was of 2.7-2.8. At the experimental ischemia, total [Hb+Mb] decreased by 25%, OSP reduced to zero, while DPF did not change. These results correlated with the previously published. The method may be applied during open-heart surgery, heart studies ex vivo or to any muscle tissue to continuously and non-invasively monitor the [Hb+Mb] content and oxygenation as well as L, which may reflect the changes in tissue structure.