Effective hemorrhage control is indispensable for life-threatening emergencies in defense fields and civilian trauma. During major injuries, hemostatic agents are applied externally to mimic and accelerate the natural hemostasis process. Commercially available topical hemostatic agents are associated with several limitations, e.g., burning sensation, necrosis, futile in severe injuries, and high costs of the products. In the present study, we developed silk fibroin fiber-based formulations and evaluated their use as a cost-effective potential hemostatic agent with shortened clotting time. Silk fiber-based powder was produced following the alkaline hydrolysis process, wherein Bombyx mori silk fibroin fibers were treated with sodium hydroxide (NaOH) solution that randomly chopped the silk microfibers. Physicochemical reaction parameters, e.g., reaction temperature, molarity of NaOH solution, and incubation time, were optimized to achieve the maximum yield of microfibers. The surface properties of alkaline hydrolyzed silk microfibers (AHSMf) were analyzed by field emission scanning electron microscopy and energy dispersive X-ray studies. The water uptake capacity of AHSMf and the change in pH and temperature (∼30 °C) during blood clotting were analyzed. Further, the hemostatic potential of AHSMf was evaluated by an in vitro whole blood clotting assay using both goat and human blood. The in vitro studies demonstrated a reduced blood clotting time (CT = 20-30 s), prothrombin time (PT = ∼27%), and activated partial thromboplastin time (APTT = ∼14%) in the presence of AHSMf when compared to silk hydrogel powder (devoid of NaOH). Thus, the developed AHSMf could be a promising material to serve as a potential hemostatic agent.