Abstract: OBJECTIVE To establish an HPLC method for the determination of atractylenolide Ⅲ in bran-processed atractylodes and bran-processed Atractylodes Formula granule, and to establish the specific chromatogram of bran-processed Atractylodes Formula granule. METHODS The chromatographic separation was performed on a Welch Xtimate C₁₈ column (250 mm×4.6 mm, 5 μm). The mobile phase was a mixture of acetonitrile-water in gradient elution at a flow rate of 1.0 mL·min^-1. The detection wavelength was 224 nm and the column temperature was set at 30℃. RESULTS The linear range of atractylenolide Ⅲ in bran-processed atractylodes and bran-processed Atractylodes Formula granule was 0.031 3-0.782 4 μg, with average recovery of 102.0%(RSD=1.88%) and 101.0%(1.02%). The HPLC specific chromatogram of bran-processed Atractylodes Formula granule showed 8 characteristic peaks. CONCLUSION The method is reliable and accurate, and can provide a reference for the technical study and quality control of bran-processed Atractylodes Formula granule.