The invention discloses an injectable biological adhesive, a preparation method and application thereof, and belongs to the technical field of high polymer materials and biological materials. According to the preparation method, saccharide macromolecules, namely glucan and chitosan oligosaccharide, with good biocompatibility are selected as matrixes and mixed and dispersed in an aqueous solution with dopamine and an oxidative cross-linking agent, and rapid gelatinization is performed to obtain the multi-cross-linked biological adhesive. The biological adhesive material obtained by the invention has the advantages of injectability, convenience in practical application and operation, capability of being applied to skin tissues, capability of being injected to the required amount according tothe required adhesion size and the like, high mechanical strength, no cytotoxicity and excellent tissue adhesion performance, can be applied to adhesion of various materials such as animal epidermis,in-vivo tissues and organs, glass, metals and oxides thereof, sealants, plastics and the like in clinical operations, and has wide application prospects in the fields of tissue adhesion and repair, hemostasis, packaging, sealing and the like in clinical medicine.