Jiubiying,the dried barks of Ilex rotunda Thunb.(Aquifoliaceae) has been used as herbal tea and traditional Chinese drug for the functions of heat-clearing,detoxifying,dehumidification,and odynolysis.Pedunculoside and syringin are the two main bioactive components.In order to obtain more components for the new drug development,we try to isolate and purify some chemical constituents from Jiubiying by high-speed counter-current chromatography (HSCCC).The two-phase solvent system was composed of ethyl acetate-n-butanol-water at an optimized volume ratio of 1:6:7 (v/v/v).19.6 Mg dunculoside with a purity of 92.4%,10.6 mg syringin with a purity of 94.1%,and 3 other compounds were obtained from 300 mg ethanol extract by one run of TBE-300A HSCCC machine (Shanghai Tauto Biotech,Shanghai,China).Their structures were identified on the basis of MS,IR,UV,and extensive NMR studies.