Flavanoids are known to interact with proteins to form complexes whose properties depend on the structure of both the flavanoids and the protein. In this study, the fluorescent probe binding method (fluorimetry analysis) and isothermal titration calorimetry (ITC) analysis were used to characterize binding interactions of green tea (GT) flavanoids and milk proteins. The hydrophobicity of the surface sites of milk proteins was estimated using the reconstituted milk-green tea and casein-green tea systems. Reconstituted milk-green tea and casein-green tea samples were prepared with different solid-non-fat (SNF) and casein (Cn) concentrations, respectively. It was observed that the number of surface hydrophobic sites decreased in the presence of GT flavanoids for all SNF and Cn concentrations. The decrease in protein surface hydrophobicity was explained by the hydrophobic binding between milk proteins and GT flavanoids. The binding enthalpies obtained from ITC analysis implied that interaction was non-covalent between catechin and beta-casein. (C) 2009 Elsevier Ltd. All rights reserved.