Abstract: AimTo investigate the chemical constituents of the secondary metabolites of the roots of Daphne genkwa.MethodsThe roots of D.genkwa were extracted with 95% ethanol at 60-70 ℃ for 7 days to obtain the crude extract.The crude extract was purified by silica gel and Sephadex LH-20 column chromatography as well as the HPLC techniques.The structures of the isolates were elucidated by combined spectroscopic methods including 1D and 2D NMR, MS, UV, IR and CD.ResultsThree new biflavonoids were isolated from the ethanol extract of the roots of D.genkwa and their structures were identified as daphnodorin H-3-methyl ether (1), daphnodorin H-3″-methyl ether (2) and daphnodorin G-3″-methyl ether (3).ConclusionCompounds 1, 2 and 3 are three new biflavonoids.