A comparison study of the structures and properties of starches regenerated from 1-ethyl-3-methylimidazolium acetate ([Emim][OAc]) using different anti-solvents (water, ethanol, or both water and ethanol) was conducted. The starch regenerated with water presented a V-type crystalline structure whereas the one regenerated in ethanol displayed amorphous structure. Moreover, when an ethanol-water-ethanol method was used for regeneration, the product showed a weak V-type crystalline structure. SAXS and FTIR were also used to investigate the molecular order of native and regenerated starches. With water used for regeneration, the aggregation and rearrangement of starch molecules occurred more easily. The increased enzyme resistance and thermal stability of regenerated starch with water could be ascribed to the rearrangement of molecular chains forming an aggregated structure with some degree of order. The reconstitution of starch molecules during regeneration with different anti-solvents changed the multiscale structures and properties of the starch. (c) 2018 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2018, 56, 1231-1238