In this study, an efficient composite hemostatic material (DA-diatom-T) was prepared, using a polydopamine layer as a linker to immobilize thrombin on the surface of diatom biosilica. DA-diatom-T retained the porous structure of the diatom with high water absorption capacity, which can absorb 31 times its own weight of water. The thrombin activity of DA-diatom-T was as high as 5.81 U mg-1 that could be maintained at 67% after 30 days at room temperature. DA-diatom-T exhibited non-toxicity to mouse fibroblast cell lines, favorable hemocompatibility and fast procoagulant ability. DA-diatom-T could promote the initiation of the coagulation process and increase platelet activity and blood clot strength to form a physical barrier at the wound. In an in vivo study, DA-diatom-T could significantly reduce the clotting time and reduce the bleeding volume. The above results showed that DA-diatom-T had potential as a new hemostatic material.