A new high performance liquid chromatographic method was applied for the determination of hyperin in Acanthopanax senticosus and A sessiliflorus. The stationary phase used was $eta$-Bondapak $C_{18}$ reverse-phase column and a mobile phase program was a gradient of acetonitrile and distilled water at a flow rate of 1.0 ml/min. Hyperin was detected at 210 nm, and the analysis was successfully carried out within 20 min. Hyperin was detected in the one year-grown and two years-grown stem of A senticosus (0.47 and 0.13 mg/g, respectively) and A sessiliflorus (0.14 and 0.03 mg/g, respectively). Hyperin was detected in the main and branch root of A sessiliflorus (0.30 and 0.09 mg/g, respectively). But there is no detection of hyperin in the main and branch root of A senticosus.