An efficient method for the isolation and preparation of two anthocyanins，cyaniding 3-O-（β-D-giucopyranoside）（compound 1） and cyaniding 3-O-（6″-O-malonyl-β-D-glucopyranoside）（compound 2），from Echinacea purpurea flower by high speed counter-current chromatography（HSCCC） was established.The fresh flower was extracted with 60% ethanol（containing 0.1% HCl） at low temperature and dark conditions.2.1 g of crude sample for HSCCC was prepared after extracted with ethyl acetate and passed through an adsorbing column（2 cm× 30 cm，containing 100 mL D101 macroporous resin）.Then water-n-butanol-methyl tert-butyl ether-acetonitrile-trifluoroacetic acid（6∶3∶2∶1∶0.001） was used as the two-phase solvent system，in which the upper phase was used as the stationary phase，while the lower phase was used as the mobile phase，the flow rate was set at 2.0 mL/min，and the revolution speed was 850 r/min.9.8 mg compound 1 with purity of 95.1% and 14.3 mg compound 2 with purity of 98.2% were obtained from 160 mg crude sample by one-step，as determined by high-performance liquid chromatography（HPLC）.The chemical structures were identified by mass spectrometry（MS） and nuclear magnetic resonance imaging（NMR）.Additionally，the antioxidant activities of the two compounds were also evaluated by the method of 1,1-diphenyl-2-picrylhydrazyl（DPPH?） radical scavenging assay with Vc as the control group.The results of antioxidative activity test showed that the two compounds have strong capacity of scavenging DPPH（EC50＜10 mg/L）.Both of their EC50 were lower than that of Vc，and the scavenging ability of compound 1 was stronger than that of compound 2.