Revealing the interactions of water with cryoprotectant and protein by near-infrared spectroscopy.

Taking formamide (FA) as a model compound of protein, the water structure in the ternary mixtures of dimethyl sulfoxide (DMSO)-water-FA was studied by near-infrared (NIR) spectroscopy. The interaction of DMSO and water, and the effect of FA on the interaction, were analyzed with the help of chemometric methods. Continuous wavelet transform (CWT) was used to enhance the resolution of the spectra. A peak at 6437 cm depicting the interaction of DMSO and water through hydrogen bonding (SO…HO) was observed in the transformed spectra. When FA exists in the mixture, the intensity of the peak decreases with the increase of formamide content, showing that FA may replace the water to form the hydrogen bond of SO and HN. In addition, temperature-dependent NIR spectroscopy was used to analyze the effect of the three components on the spectral variation with temperature. Analyzing the spectral data by alternating trilinear decomposition (ATLD) and multiple linear regression, two varying spectral features were obtained that are related to water and DMSO, but no spectral feature was found that significantly varies with the content of FA. The result implies that DMSO is still the key component to prevent the water from icing, although FA may reduce slightly the anti-freezing effect.