Identify the interactions between phytochemicals and proteins in the complicated food matrix.

Phytochemicals usually mix with food proteins in our regular diet. Unexpected interactions may lead to changes in bioaccessibility, bioactivity, and bioavailability of phytochemicals. However, our understanding of these interactions between phytochemical and food proteins is limited because of the experimental restrictions. Here, we used pulse-proteolysis to conduct the unfolding equilibrium and dose-dependent experiments on the food proteins for the first time. The interaction between epigallocatechin gallate (EGCG) and caseins was identified in the complicated food matrix, whole milk. Another food proteome, soymilk, was also optimized for identifying the binding targets of EGCG and caffeine. Among the identified interactions, the mixing of milk with coffee generates the most prominent masking effect of 46.61 ± 3.86% relative to the calculated antioxidant capacity. Our results demonstrated that pulse proteolysis is applicable for identifying the interactions between phytochemicals and proteins in the complicated food matrix.