The color stability of anthocyanins was shown to improve with addition of whey proteins (WP). The goal of this study was to investigate the binding mechanisms of purple corn, grape and black carrot anthocyanin extracts to native and preheated WP (40-80 °C, 3.6 μM) at a pH of 3 using fluorescence quenching spectroscopy. The fluorescence spectra were collected with an excitation wavelength of 280 nm at 25 °C, 35 °C and 45 °C. The quenching data were analyzed by using the Stern-Volmer equation. The fluorescence intensity of WP decreased (up to 73%) and its λmax increased (by ~5 nm) with increasing anthocyanin concentration (0-100 μM). The quenching data showed that the interaction between anthocyanin extracts and WP was a static quenching process. Thermodynamic analysis showed their binding was mainly through hydrophobic interactions. Their binding affinity was higher for preheated WP than native WP and decreased gradually with increasing preheating temperature. Black carrot anthocyanin extract had the lowest binding affinity with WP, likely due to the larger molecular structure. These results help better understand the protection mechanism of native and preheated WP on anthocyanin color stability, expanding the application of anthocyanins as food colorants that better withstand processing and storage.
Keywords: anthocyanin color stability; anthocyanin-whey protein interaction; fluorescence quenching; preheating treatment; thermodynamic analysis.