Abstract: Pulsatilla chinensis is one of the most commonly used Chinese herbal medicines in treatment of gastrointestinal diseases of poultry and livestock which has the efficacy of clearing heat and antibacterial activity. Polysaccharide is one of the crucial effective components in PulsatiUa chinensis. In this study, the polysaccharide content in Pulsatilla chinensis was determined by phenol-sulfuric acid method; the influencing factors associated with the extraction rate of polysaccharide in PulsatiUa chinensis were assessed by single factor experiment. The results revealed that the comminution mesh number for Pulsatilla chinensis was 60, the soaking duration was 90 min, the feed solution proportion was 1：50, the extracting temperature was 95℃, the extracting duration was 90 min, and the extracting times was 2. Under this condition, the highest extracting rate of polysaccharide was obtained from Pulsatilla chinensis. The results of verification experiment showed that the highest extracting rate of polysaccharide reached （21.7±0.55 ） %. This study lays a foundation for the scaled production of polysaccharide from Puls atilla chine ns is .