The content of hyperin in Acanthopanax species was determined by high performance liquid chromatography (HPLC). Hyperin was quantified by a reverse-phase column with elution program [initially gradient solvent (acetonitrile : water = 85 : 15 to 80 : 20 for 20 min), then isocratic solvent (acetonitrile : water = 80 : 20 for 20 min), and finally gradient solvent (acetonitrile : water = 80 : 20 to 65 : 35 for 20 min)]. UV detection was conducted at 210 nm. The content of hyperin in the fruits of Acanthopanax was measured in the species A. chiisanensis (2.04 mg/g), A. sessiliflorus (1.13 mg/g), A. divaricatus (0.98 mg/g), A. koreanum (0.75 mg/g) and A. senticosus (0.05 mg/g). The content of hyperin in A. chiisanensis was higher than that of other Acanthopanax species.