The invention discloses a preparing method for pig thrombin freeze-dried powder. The preparing method comprises the following steps that anticoagulant pig plasma and gel are mixed and stirred for adsorption and pass through a column for elution, and a prothrombin solution is obtained; normal saline is added to rabbit brain powder, then the prothrombin solution and CaCl2 are added for zymogen activation, a crude thrombin enzyme solution is obtained, subjected to ultrafiltration for desalination and concentrated, viral inactivation is carried out, an obtained crude enzyme solution passes through DEAE-Sepharose Fast Flow chromatographic column, elution is carried out, target peaks are collected, and pig thrombin is obtained; the pig thrombin is added with mannitol or dextran 40, filtering, sterilizing, subpackaging, freeze drying and vacuum tamponing are carried out, an aluminum plastic combined cover is rolled, and a pig thrombin freeze-dried product is obtained; the freeze-dried product is subjected to dry heat treatment at 100 DEG C for secondary virus inactivation, and the pig thrombin freeze-dried powder is obtained after packaging. The specific activity of pig thrombin in the product is not lower than 130 U/mg, the whole process is simple in step and easy to implement, the product safety is improved, and the preparing method is suitable for industrial production.