Fluorescence imaging of spatial location of lipids and proteins during digestion of protein-stabilized oil-in-water emulsions: A simulated gastrointestinal tract study

- Protein-fluorescent dye conjugate was synthesized.
- Changes in location of proteins during digestion of O/W emulsions were visualized.
- Proteins remained attached to the lipid droplet in the mouth and stomach phases.
- After exposure to small intestine phase, conjugates tended to aggregate together.

A whey protein isolate-rhodamine B conjugate (WPI-RB) was synthesized to visualize changes in the location of a protein emulsifier in oil-in-water emulsions during digestion. An oil-soluble dye (Nile Red) was used to visualize changes in the lipid phase during digestion. Protein-labeled and lipid-labeled emulsions were passed through a simulated gastrointestinal tract consisting of mouth, stomach, and intestinal phases, and changes in protein and lipid location and morphology were monitored using confocal laser scanning microscopy. The proteins remained attached to the lipid droplet surfaces in the mouth and stomach, but formed large aggregates in the small intestine. The lipid droplets were highly flocculated in the mouth, highly coalesced in the stomach, and fully digested in the small intestine. Our results show that conjugation of fluorescence dyes to protein emulsifiers is useful for direct visualization of their location, as well as for understanding the behavior of food emulsions within the gastrointestinal tract.