To develop a high-performance liquid chromatography (HPLC) method for simultaneous determination of hypocrellin A, hypocrellin B, and hypocrellin C.The separation was carried out on a Kromasil C18 (4.6 mm x 250 mm, 5 micrm) colum eluted with in mobile phases of water containing 0.5% glacial acetic acid and acetonitrile. The column temperature was 35 degrees C, and the flow rate was 1.0 mL x min(-1). The detection wavelength was set at 265 nm.The three compounds were well separated. Calibration curves of hypocrellin A, hypocrellin B, and hypocrellin C showed good linear relationship RSD > 2.0%. The average recoveries of the hypocrellin A, hypocrellin B, and hypocrellin C were 101.8%, 102.3%, 100.0%, respectively.The developed method is simple, accurate, and repeatable, and can be readily used as valid tool for the quality control of Hypocrella bambusae.