Chitosan (CHI) is a versatile biological material that is well known for its hemostatic properties. This preliminary study evaluated several self-assembling hydrophobically modified chitosan (HM-CHI) sponges to determine their efficacy on hemostasis. Fourier transform infrared (FT-IR) spectroscopy was used to determine the successful graft of dodecyl groups onto the nitrogen atoms of CHI molecules. A platelet aggregation assay revealed that HM-CHI accelerated the platelet aggregation rate. Fluorescence spectroscopy showed that the HM-CHI changed the structure of fibrinogen in blood. Activated partial thromboplastin time, prothrombin time, fibrinogen time, and thromboelastographic assays were used to explore the effect of HM-CHI on the autologous blood coagulation pathway. Finally, a hemostatic sponge was made with HM-CHI and freeze-dried zeolite composite film and was applied to the rat femoral artery hemostasis model. A hemostasis time of 86 AE 5 sec was achieved, which was significantly better than the one composed with pure CHI. The experimental results of the HM-CHI hemostatic materials are inspiring and will encourage the research and development of such materials. HM-CHI may be a strong candidate as a safe and effective hemostatic material.