Abstract: Isolation and prepare high-purity flavonoid from lotus leaves by High-speed counter-current chromatography and research antioxidant activity. Results showed that:The two-phase solvent system composed of N-hexane:ethyl acetate:methanol:water=1:6:1:6 and N-butanol:methanol:water=5:1:4.The quality of the three target components(C₁,C₂ and C₃)was obtained as 12.4,5.9 and 2.1 mg and Check the purity by HPLC was 92.67%,90.43%,87.52%. The three target components were likely to be orientin,isoquercitrin and quercetin by standard control experiment.The three target components showed good antioxidant activity,the half-value clearance(IC₅₀)of hydroxyl radical was 0.94,0.85,0.76 mg/mL,the half-value of DPPH free radical scavenging rate(IC₅₀)were 0.39,0.28,0.06 mg/mL,the ABTS⁺ free radical Half of the clearance rate(IC₅₀)were 0.56,0.47,0.29 mg/mL.