A reversed-phase high-performance liquid chromatographic method with diode array detection was established to simultaneously determine the seven bioactive lignans in Herpetospermum caudigerum, namely ent-isolariciresinol (1), dehydrodiconiferyl alcohol (2), herpetrione (3), herpetin (4), herpetetrone (5), herpetotriol (6) amd herpetal (7). The HPLC assay was performed on a Restek Pinnacle DB C₁₈ column (250 × 4.6 mm, 5 µm) with gradient elution of acetonitrile and 0.1% phosphoric acid within 65 min. The detection wavelength was 240 nm. The flow-rate was 1.0 mL/min. All calibration curves showed good linearity (r² > 0.9998) within test ranges. The method was reproducible with intra- and inter-day variation of less than 1.98%. The method provided good accuracy with recoveries in the range 95.19-102.64% with RSDs less than 1.52%. The method was successfully applied to the quantification of seven constituents in 15 H. caudigerum samples collected from different cities. The results indicated that the developed assay could be considered as a suitable quality control method for H. caudigerum. Copyright © 2008 John Wiley & Sons, Ltd.