The invention relates to a dentin collagen fiber demineralization material and a preparation method and an application thereof. In particular, the invention relates to a conjugate of water-soluble chitosan and an amino carboxylic acid metal chelating agent, and a use thereof for dental demineralization, preferably for dentin demineralization. The invention also discloses the dentin collagen fiberdemineralization material and the preparation method and the application thereof. The material is prepared from 0.5-1 part by mass of water-soluble chitosan, 10-30 parts by mass of EDTA and 0.8-1.5 parts by mass of a crosslinking agent. The invention also relates to a dental demineralization composition including the conjugate and the material. The conjugate, material and composition can be used for dentin demineralization treatment before dental adhesive repair. When being used as a dentin collagen demineralization agent, the conjugate, material and composition have excellent selective extrafibrous demineralization performance, good biocompatibility, effective bacteriostasis, inhibition of endogenous matrix proteinase activity and extremely low cytotoxicity.