Abstract

The incorporation of dietary fibers into food formulations is a promising approach to developing new functional ingredients. In this study, we investigated the formation of Maillard conjugates using whey protein concentrate (WPC) combined with fructooligosaccharides (FOS) and xylooligosaccharides (XOS). These conjugates, innovatively utilized as emulsifiers for annatto seed oil, were subjected to in vitro gastrointestinal digestion to assess their chemical stability and effectiveness as encapsulating systems. Our study confirmed the occurrence of the Maillard reaction through different methods, including grafting degree (GD) assessment and sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) to evaluate molecular weight variations in the proteins. The key innovation of our research lies in demonstrating the conjugation process through carbohydrate mass balance analysis using High-Performance Anion Exchange Chromatography coupled with Pulsed Amperometric Detection (HPAEC-PAD). The GD for the WPC-XOS and WPC-FOS was 46 ± 3 % and 56 ± 3 %, respectively. Our findings suggest that sugar quantification could serve as a novel approach to confirming the formation of Maillard conjugates. The emulsions stabilized by these conjugates exhibited larger oil droplet sizes compared to those stabilized by WPC alone yet maintained high encapsulation efficiency (≥ 90 %). Notably, the WPC-XOS emulsion demonstrated kinetic stability after 30 days of storage at 4°C, while the WPC-FOS emulsion showed signs of creaming after just 16 days. In vitro gastrointestinal digestion revealed that while the conjugates were susceptible to enzymatic degradation, the FOS and XOS sugars displayed gastrointestinal resistance of up to 82 % and 83 %, respectively. This study highlights the potential of WPC-XOS conjugates not only as effective emulsifiers but also as functional ingredients that enhance the nutritional and antioxidant properties of food products, presenting a novel approach to functional food development.

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