A gradient high performance liquid chromatography(HPLC)coupled with UV-vis photodiode array detection(DAD)was developed for the separation of rutin,hyperoside,and quercetin from the flavoind extracts from hawthorn powder.The flavoinds were extracted using the solvent of 70% ethanol(by volume).Chromatographic separation was performed in DevelosilC30 column(250mm×4.6mm ID,5μm)with an eluent consisting of acetonitrile and 0.4% Phosphate at a flow rate of 1.0mL/min,the eluate was monitored at 360nm.The result showed that separation and identification of rutin,hyperoside,quercetin,and other components were completed in 30min.The linear ranges of rutin,hyperoside,and quercetin were 0.0050~0.1600mg/mL(R2=0.9999),0.0016~0.0500mg/mL(R2=0.9996)and 0.0005~0.0150mg/mL(R2=0.9997),respectively.This method was precise and accurate(RSD 0.48%~0.78%),the average recoveries ranged from 92.36% to 98.64%,and can be used for the precise quantification of the three flavoinds and other flavoinds of dry plant material such as hawthorn power.