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Lonicera japonica Thunb.(Caprifoliaceae plants) is one of the most common traditional Chinese medicines and used for treating various diseases,including arthritis,diabetes mellitus,fever and infections.The leaves of L.Japonica is not fully research,it is necessary to develope an efficient method to separate and purify active compounds.The scavenging ability on ROS free radical such as O2￣? and ?OH of the n-butanol extract from leaves of L.Japonica in vitro were determined by flow injection chemiluminescence.When the concentration was 2.0 mg/mL,the clearance rate of O2￣? was 97.03%.And the clearance rate of ?OH was 95.57% at the concentration of 4.0 mg/mL.n-butanol extracts have good ability of natural antioxidant and possess a certain value of applications.Then,HSCCC together with preparative HPLC were used for the separation of flavonoid glycosides and caffeoylquinic acid derivatives.1.0 g of the n-butanol extract was firstly isolated by HSCCC using a two-phase solvent system composed of methyl tert-butyl ether-n-butanol-acetonitrile-water (0.5% acetic acid) (2: 2: 1: 5,v/v),yielding five fractions F1,F2,F3,F4,F5 (collected from the column after the separation).F1 is the mixture of chlorogenic acid,lonicerin and rutin.This mixture was separated by preparatve HPLC successfully.F2 and F3 were rhoifolin and luteoloside.F4 (mixed with 3,4-O-dicaffeoylquinic acid and hyperoside) and F5 (mixed with 3,5-O-dicaffeoylquinic acid and 4,5-O-dicaffeoylquinic acid) were separated by preparatve HPLC successfully.Combination these two methods,10.9 mg of chlorogenic acid (1),30.7 mg of lonicerin (2),16.7 mg of rutin (3),21.5 mg of rhoifolin (4),32.0 mg of luteoloside (5),20.3 mg of 3,4-O-dicaffeoylquinic acid (6),18.2 mg of hyperoside (7),24.7 mg of 3,5-O-dicaffeoylquinic acid (8) and 26.1 mg of 4,5-O-dicaffeoylquinic acid (9) were yielded,with the purities of 99.5%,98.7%,99.3%,94.3%,96.1%,97.1%,97.4%,96.9% and 97.8%,respectively,as determined by HPLC.The structures of these compounds were identified by ESI-MS,1H NMR and 13C NMR.Being a kind of support-free all-liquid partition chromatography,HSCCC can eliminate irreversible adsorption of sample on the solid support.It is an excellent enrich method for separation similar polity compounds.