A reverse-phase system of HPLC using a linear gradient of acetonitrile and deionized water was developed for the quantification lignans, eleutherosides B and E, in Acanthopanax senticosus. The HPLC system consisted of linear gradient of acetonitrile and deionized water, and UV/VIS detection was set at 210 nm. Both eleutherosides B and E contents in different parts of A. senticosus were determined. As a result, the contents of eleutherosides B and E were measured in the leaves (trace amounts and 0.029 mg/g, respectively), stems (0.107 and 1.015 mg/g, respectively), roots (0.026 and 0.390 mg/g, respectively), and fruits (0.022 and 0.043 mg/g, respectively). Moreover, eleutherosides B and E in the water extract were found 0.011 and 0.171 mg/g, respectively.