The invention discloses a fibroin medical biological adhesive and a preparation method thereof, and belongs to the field of biomaterial preparation and biomedical application. The adhesive is preparedby uniformly mixing a fibroin solution and a polyphenolic polymer solution at room temperature. The fibroin biological adhesive has the advantages of high adhesion strength, high adhesion speed, safety, no toxicity, good biocompatibility and low price in a humid environment. The adhesive is simple to operate and easy to grasp, and suture removal and other secondary operations are not required after an operation to relieve the pain of a patient. The small or even scar-free wound surfaces have a cosmetic effect. The adhesive can be used on almost all wound surfaces, has the effects of bonding tissues, covering the wound surfaces, filling gaps and voids, stopping bleeding, inhibiting bacteria and the like, can be used as a hemostatic adhesive material for tissues such as skins, organs, bloodvessels, nerves, muscles, mucous membranes, bones, joints and eyes, and has extensive clinical application prospects.