Carbohydrate-based oil-in-water emulsions for delivery of short-chain fatty acids : Doctor of Philosophy in Food Technology at Riddet Institute, Massey University, Palmerston North, New Zealand

Short-chain fatty acids (SCFAs) are important functional metabolites. There is clinical evidence to show that they are useful in the prevention of the metabolic syndrome, bowel disorders and certain types of cancer. Therefore, supplementation of SCFAs to the daily diet brings benefits to human health. However, SCFAs are small and water-soluble molecules that are quickly absorbed in the upper gastrointestinal tract. This project aimed to develop carbohydrate-based systems to deliver tripropionin (TP, glycerol tripropionate) and tributyrin (TB, glycerol tributyrate) as sources of propionic and butyric acids into the colon. Two types of emulsion systems were employed, i.e. surfactant-stabilised oil-in-water (O/W) emulsions (single and double-layer systems) and particle-stabilised O/W emulsions (Pickering emulsions). The systems were characterised in terms of structural stability, surface charge, rheological properties, lipolysis degree and release of SCFAs under a static in vitro gastrointestinal digestion and an in vivo study with ileal-cannulated pigs. In the screening experiments, several potential carbohydrate materials were explored, i.e. three modified starches (GUM, N46 and N-LOK), four pectins (PEC) and hydrophobically modified inulin (M-IN), to produce single-layer O/W emulsions. A double-layer O/W emulsion was also produced by combining whey protein isolate (WPI) and chitosan (CS) as the first and second layers, respectively. The capacity of emulsion systems for colon-targeted delivery of SCFAs was then tested using a static in vitro gastrointestinal digestion. The results show that PEC displayed the poorest emulsifying capacity amongst all investigated carbohydrates, leading to an emulsion droplets size (d32) of around 7.3 µm. However, PEC-based formulation was the best system for protection against gastric and intestinal conditions. On the other hand, other single-layer systems and the double-layer system proved to be unstable in the intestinal phase with a significant SCFA release. Deeper investigation on the emulsifying capacity showed that PEC stabilised the O/W emulsion mainly through steric effects. In addition, PEC had the ability to form thick layer around the O/W interface, which was evidenced by confocal laser scanning microscopy and the quantification of adsorbed PEC on the interface. In addition to the above systems, a Pickering O/W emulsion stabilised by hydrophobically modified cellulose nanocrystals (CNCs) was also investigated. The hydrophobic modification of CNCs was carried out, resulting in an increase in static water contact angle from 56o (untreated CNCs) to 80.2o (MCNCs). As a result, the emulsifying capacity of MCNCs was significantly improved. The emulsions prepared from MCNCs ≥ 0.20 wt% were stable against droplet coalescence for up to 4-week storage. In addition, the Pickering emulsions were prone to droplet flocculation at ionic strength ≥ 20 mM NaCl (pH 7.0) or pH < 4.0 (without addition of NaCl), which was due to the charge screening associated with the cellulose molecules at the surface. Similar droplet flocculation was also observed under in vitro gastric conditions, where the emulsions were exposed to low pH and high ionic strength. This gastric-induced structural changes improved physical strength of the emulsions and that enhanced resistance to bile-salt displacement and consequently delayed lipid digestion in the intestinal conditions. In addition, high desorption energy of the MCNC particles at O/W interface of the Pickering emulsion contributed to low lipolysis degree (30–35%). High proportions of SCFAs remaining after the intestinal digestion observed in both PEC and MCNC-based emulsions show a strong promise their use in the colon-targeted delivery of SCFAs. However, CNCs are currently not considered as food-grade materials; therefore, PEC was chosen for the in vivo study using female ileal-cannulated pigs. The in vivo study demonstrated significant higher intestinal lipolysis (~ 51–53%) and lower SCFA release (~ 15%) as compared to the in vitro digestion (~ 40 and 35% respectively). The main reason for the difference between the two models was the absorption of the SCFAs in the pig’s small intestine. However, high proportions of unhydrolysed triglycerides (~ 47–49%) and presence of oil droplets in the ileal-digesta demonstrated successful delivery of SCFAs. Based on the findings in this research, we propose the use of PEC-based emulsion for human trials by incorporating the system into a daily diet or dessert liquid/gel products, such as drinking milk or yogurt. We also believe that the application of MCNC-based Pickering emulsions for colon-target delivery of could be of interest if the regulatory status could be confirmed. The study identifies promising directions for researchers who are interested in improving gut health through delivery of SCFAs to the colon