Objective:To isolate and purify trypsin hydrolysates of Eupolyphaga sinensis by solvent extraction method and sephadex gel column chromatography

and initially identified anticoagulate active components and chemical composition. Method: Rypsin hydrolysates of E. sinensis were prepared by trypsin hydrolysis method

with Active partial thromboplastin time (APTT) as index

anticoagulant active site IV was obtained by solvent extraction method

and purified trypsin hydrolysates were obtained by sephadex G-10 gel column chromatography;Coomassie brilliant blue G-250 was used to quantitative analysis of purified trypsin hydrolysates

and range of molecular weight distribution of separating portion was detected by MALDI-TOF-MS. Result: The composition IV which separated from trypsin hydrolysates of E. sinensis had strong anti-coagulate activity

and the activity was in a positive correlation with the concentration of hydrolysates;After purified by Sephadex G-25

the mass fraction of protein in IV site was 5.5%

which was determined by Coomassie brilliant blue G-250;Purified material was detected by MALDI-TOF-MS after further desalted with Sephadex G-10 gel column

its relative molecular weight range of three groups of polypeptide compounds was identified 850 to 910 Da. Conclusion: It was feasible for separation and purification trypsin hydrolysates of E. sinensis by gel column chromatography

anticoagulant active component was small molecular peptide of less than 1 000 Da.