Immobilized β-lactoglobulin on a HPLC-column: a rapid way to determine protein—flavour interactions

To study the interaction of β-lactoglobulin (BLG) with flavour compounds a fast screening methodology was developed. BLG (variant AB, pure A and pure B) was immobilized onto a silica support, filled into a column and combined with a HPLC-system. A total of 24 different flavour compounds were injected and their retention times determined at different pHs and protein concentrations. The binding constant for each compound was calculated from the retention times and the protein concentration of the column. This simple system allows the rapid screening of many flavour compounds under a variety of external conditions like pH, salt content and flavour concentration. This procedure also permits the study of competitive effects with several flavour compounds in the solution.