An analytical approach to reveal the addition of heat-denatured whey proteins in lab-scale cheese making

A simple analytical procedure for the detection of self-aggregated heat-denatured whey proteins (HDWP) in model cheeses was developed. The principle of the approach lies in the solubilization of the cheese matrix by a sodium citrate solution (0.2 M, pH 7.0) resulting in the dissociation of the casein micelles and the insolubilization of HDWP aggregates, which are collected in the pellet after a centrifugation step. The reliability of the procedure was tested in lab-scale cheeses from peroxidase-positive pasteurized milk with different protein-based ingredients (microparticulated whey protein concentrate, milk protein concentrate, whey protein isolate and Ricotta cheese) at concentrations ranging from 0.2 to 1.2% protein (w/v on cheese milk). A linear relationship between the amount of the HDWP added to cheese milk and that recovered from model cheeses was observed. Heat-damage indicators, furosine and lysinoalanine, showed levels in the experimental cheese samples not related with added HDWP, but represented a source of information on the ingredients other than liquid milk. Overall, in the model cheeses, the proposed method was an easy-to-apply and reliable tool for the evaluation of the presence of HDWP-based products. Further investigation is required for the application to real cheeses and for the evaluation of possible interferences from proteolysis during ripening.