Abstract: In this study, the effect of three self-assembly sequences on the structural properties, three-dimensional (3D) printing characteristics, storage stability and freeze-thaw stability of high internal phase Pickering emulsions (HIPPEs) stabilized by ternary aggregates of lactoferrin, epigallocatechin-3-gallate (EGCG) and low methoxylated pectin was evaluated. The results showed that self-assembly sequence directly affected the ability of ternary aggregates to stabilize HIPPEs. The particle sizes of the ternary aggregates were between 1.7 and 2.9 μm, and they showed different network structures and surface wettabilities (between 55.62° and 68.15°). Optical microrheology and texture analysis showed that the decreasing order of the viscoelasticity, hardness, adhesiveness and chewiness of HIPPEs was lactoferrin-EGCG-low methoxylated pectin > lactoferrin-low methoxylated pectin-EGCG > low methoxylated pectin-EGCG-lactoferrin. The HIPPE stabilized by lactoferrin-EGCG-low methoxylated pectin was solid-like and more suitable for 3D printing. The microstructure showed that these HIPPEs were oil-in-water Pickering emulsions. It is particularly noteworthy that the HIPPE stabilized by lactoferrin-EGCG-low methoxylated pectin showed excellent long-term storage stability and freeze-thaw stability, which is of great significance for its potential application in cold chain logistics. In conclusion, when using ternary complexes to stabilize HIPPEs, self-assembly sequence is an important factor that cannot be ignored.

Key words: ternary aggregates; high internal phase Pickering emulsions; self-assembly sequence; storage stability