Abstract: OBJECTIVE To explore hemostatic, coagulant and anti-inflammatory effect and the extraction mechanism of the separation-purification of the flavonoids extracts from persimmon(FEP) in aqueous two-phase systems(ATPS) of PEG 600 and (NH₄)₂SO₄. METHODS The yield rate of flavonoids from persimmon was taken as an index, to optimize the conditions such as the concentration of PEG 600, the concentration of (NH₄)₂SO₄, temperature, amounts of MgCl₂ and pH were studied by single factor experiments and response surface methodology(RSM) with a three-factorthe concentration of PEG 600, the concentration of (NH4)2SO4, pH and three-level experiment design were developed by Box-Behnken central composite design method. Furthermore, hemostatic, coagulant and anti-inflammatory effect of FEP were studied by breaking tail method, capillary tubes method and the ear swelling method induced by dimethylbenzene in KM mice and results were compared to Panax notoginseng powder and aspirin. RESULTS The result showed that the optimum condition was as follows: 12% PEG 600, 28% (NH₄)₂SO₄, pH 8 and the yield rate of flavonoids reached 95.57%. The FEP could shorten bleeding time(BT), coagulation time and restrain ear swelling induced by dimethylbenzene. CONCLUSION The purification method for flavonoid by ATPS is easy, efficient, and moderate which can keep the bioactivity. The FEP is a potential product added in oral daily necessities and oral drugs as active ingredient.