Reversed-phase high-performance liquid chromatography on-line with second derivative ultraviolet spectroscopy as a tool for the identification of beta-casein

摘要

The aim of the study was to develop and improve methods for the identification of beta-casein (beta-CN) on the basis of the spectroscopic parameters A and B characterizing the position of selected extrema of UV spectra second derivatives, and the parameter C describing the absorbance ratio at different wavelengths. The effects of environmental factors, i.e. temperature and concentrations of beta-casein (beta-CN) solutions, were determined. The temperature increase was accompanied by an increase in the retention time of beta-CN. No differences were found between second derivatives of spectra obtained at 20 and 30 °C. The values of the parameters A and B did not differ significantly within the temperature range applied. The parameter B should not bedetermined for beta-CN solutions at concentrations lower than 1.0 mg/ml. The values of the parameter C were changing with temperature.