The paper established an HPLC method for the determination of berberine hydrochloride in fat-soluble Qianliean suppositories. The HPLC was performed on a Kromasil C18 column (4.6 mm x 200 mm, 5 μm), using the mobile phase of acetonitrile-0.1% phosphoric acid solution (50 : 50) (0.1 g sodium dodecyl sulfate was dissolved in100 mL mobile phase), with the detection wavelength of 350 nm. Results showed that the calibration curve was linear in the range of 20-200 μg·ml-1. The average recovery was 98.9% and RSD was 0.5%. In conclusion, this method was sensitive, accurate, simple, and could be used for determination of berberine hydrochloride in fat-soluble Qianliean suppositories.