The aim of this thesis was to develop a reliable analysis method for determination of protein-flavour binding and to explore aspects of binding of flavour compounds to milk proteins. Solid-phase microextraction (SPME) technique was developed and was shown to be relatively sensitive and efficient for monitoring flavour-protein interactions.
SPME conditions (the size and the type of the fibre, the extraction times and the size of container used for SPME) were optimized. 100 and 30um polydimethylsiloxane (PDMS) fibres were chosen; extraction times of 5 min and 30 seconds were applied for equilibrium and dynamic extraction, respectively and 4 ml HPLC vials were selected for SPME extraction. The SPME technique developed in this study showed good repeatability, with the maximum deviation being ~4.0%. The technique could be used to determine flavour-protein binding in a system in which more than 10% of the total flavour binds to the protein.
The SPME technique developed in this study is a reliable technique to study milk protein and flavour binding. The studies on the effects of pH on binding parameters and the binding position mapping work improve our understanding of the mechanism of flavour binding to shey proteins.