Temperature-Dependent Near-Infrared Spectra of Bovine Serum Albumin in Aqueous Solutions: Spectral Analysis by Principal Component Analysis and Evolving Factor Analysis

Abstract

Fourier transform near-infrared (FT-NIR) spectra have been measured for bovine serum albumin (BSA) in an aqueous solution (pH 6.8) with a concentration of 5.0 wt % over a temperature range of 45–85 °C. Not only conventional spectral analysis methods, such as second-derivative spectra and difference spectra, but also chemometrics, such as principal component analysis (PCA) and evolving factor analysis (EFA), have been employed to analyze the temperature-dependent NIR spectra in the 7500–5500 and 4900–4200 cm⁻¹ regions of the BSA aqueous solution. Intensity changes of bands in the 7200–6600 cm⁻¹ and 4650–4500 cm⁻¹ regions in the difference spectra indicate variations of the hydration and secondary structure of BSA in the aqueous solution, respectively. The plot of a band intensity at 7080 cm⁻¹ in the different spectra shows a clear turning point at 63 °C, revealing that a significant change in the hydration occurs at about 63 °C. The forward and backward eigenvalues (EVs) from EFA suggest that marked changes in the hydration and secondary structure of BSA take place in the temperature ranges of 61–65 °C and 59–63 °C, respectively. In addition, the temperature of 71 °C marked in the EFA plots may correspond to the onset temperature of increase in the intermolecular β-sheet structure.