This research aims to investigate the active constituents of Rhizoma Smilacis Glabrae and its anti-uric acid effect on mice.The active constituents was extracted by ethanol reflux,separated and purified by multiple chromatography.The structure of the obtained product was identified by NMR.A mouse model with high blood uric acid was established and combined with xanthine oxidase in vitro inhibition experiment to screening the active constituents.Results showed that 70% ethanol eluate from the macroporous resin extracted from Rhizoma Smilacis Glabrae had the most obvious anti-uric acid effect on mice (P<0.01).After separation by MCI and ODS column,five compound monomers were further separated:astilbin (1),Taxifolin-3-O-α-L-rhamnopyranosyl-α-L-rhamnopyranoside (2),engeletin (3),isoengeletin (4),4-(pyrrolo［3,2-d］pyrimidin-4-yl)pheno (5).Compounds 2-5 had significant inhibitory effect on xanthine oxidase activity (P<0.01);Compound 5 showed the strongest inhibitory effect with an inhibitory rate of 80.47%.