This study aimed to prepare three types of chitosan-gelatin films crosslinked using Schiff base reactions and EDC/NHS-citric acid (EMC: 1-ethyl-3-(3- 3-dimethylaminopropyl) carbodiimide/N-hydroxyisocinnamic acid) and glutaraldehyde, and jenipharin to produce three types of crosslinked chitosan-gelatin films (CSCA, CSGTA, CSGP), and compared and analyzed their mechanical and chemical properties. The CSGTA film, which had the optimal composition, showed significantly improved mechanical durability with tensile strength increasing from 35.5 to 82.7 MPa, elastic modulus from 34.5 to 79.2 MPa, and swelling degree from 1013 to 130%. Next, thrombin, a blood coagulation protein, was coated at a concentration of 640 µg/cm², and the immobilization efficiency was confirmed through EDAX analysis. The whole-blood clotting time was drastically reduced from 280 s to 16.5 s, and the APTT results confirmed the activation of the intrinsic pathway. In an MTT assay using L-929 cells, the cell survival rate of all cross-linked films was over 70%, meeting the ISO 10993-5 standard. Finally, the patch, which integrates the cross-linked film with a hydrocolloid adhesive layer, enables immediate adhesion and sealing without additional tape, demonstrating hemostatic performance equivalent to or better than SURGIFLO® and excellent elasticity and adhesion. Accelerated aging tests according to ASTM F1980 standards predicted storage stability for over 12 months at 25°C. These results suggest that the thrombin-coated cross-linked chitosan-gelatin hydrocolloid patch can be utilized as a next-generation hemostatic solution for non-compression bleeding control in surgical, emergency, and battlefield environments.

Schiff Base Crosslinking, EDC/NHS-Citric Acid, Thrombin Coating, Hydrocolloid Patch, Hemostatic Performance, Biocompatibility