A high performance liquid chromatography (HPLC) method was developed for the separation of secondary metabolites and quantitative analysis of hinokiol from cell suspension cultures of Cephalotaxus fortunei. The samples were prepared by extraction using methanol followed by partitioning between ammonium hydroxide and chloroform. The HPLC separation was achieved on an Apollo C 18 column (250 mm × 4.6 mm, 5 μm) with gradient elution using methanol and water at 1mL/min and 30 °c. The detection was carried out at 290 nm. A good linear correlation between the hinokiol peak area and mass concentration was observed over the mass concentration range of 0.012 5-0.2 g/L. The proposed method was applied to the determination of hinokiol in the actual samples with recoveries of 87. 2% - 94. 7% and with the relative standard deviations of 0. 9% - 4. 2%. This method is reliable and reproducible and is suitable for the analysis of hinokiol in plant cell cultures.