Near-infrared spectra (1300-2500 nm) collected from lysed blood solutions were shown to correlate with the pH of the solutions measured potentiometrically. Cross-validated partial least-squares (PLS) models were developed from these spectral data, which provided standard error of prediction (SEP) values below 0.05 pH units for a pH range of 1.0 (6.8-7.8). Experiments were designed to eliminate possible correlation between pH and other components in the blood in order to ensure that variations in the spectral data correlated to pH were due to hydrogen ion changes only. Further work was performed to discern the primary source of pH information in the lysed blood spectra by using spectra collected from plasma and histidine solutions. The blood, plasma, and histidine data sets were compared with the use of loading vectors from principal component analysis (PCA). These loading vectors show that variations in the spectra of the titrated amino acid histidine mimic those seen in lysed blood, but not those seen in plasma. These results suggest that histidine residues of hemoglobin are providing the spectral variation necessary for pH modeling in the lysed blood solutions. It is further shown that the observed pH-sensitive histidine bands do not arise from the exchangeable proton on the imidazole ring of histidine; rather they arise from the variation in the C-H bonds of the C2 and/or the C4 carbons of the imidazole ring as they are influenced by the titration of the nitrogen-bound proton of the imidazole ring.
M. Kathleen Alam, James E. Franke, Thomas M. Niemczyk, John D. Maynard, Mark R. Rohrscheib, M. Ries Robinson, and R. Philip Eaton, "Characterization of pH Variation in Lysed Blood by NearInfrared Spectroscopy," Appl. Spectrosc. 52, 393-399 (1998)
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