Background: Dachaihu Granule, commonly used for treating cholecystitis, is derived from a famous traditional Chinese formula named Dachaihu Decoction. No analytical method has been reported for simultaneous determination of 10 bioactive compounds for quality control in Dachaihu Granule so far. Objective: To develop a high-performance liquid chromatographic (HPLC) method with diode array detector (DAD) for simultaneous determination of 10 bioactive compounds (paeoniflorin, aloe-emodin, rhein, emodin, chrysophanol, physcion, naringin, hesperidin, neohesperidin, and baicalin) in traditional Chinese medicine Dachaihu Granule. Materials and Methods: The samples were separated on a Kromasil C 18 (250 × 4.6 mm,i.d. with 5.0 μm particle size)column with multi-wavelength detection method by a gradient elution using acetonitrile (A) and 0.2% acetic acid (B) as the mobile phase. The column temperature was maintained at 30°C and the detection wavelength was set at 230 nm for paeoniflorin, 254 nm for aloe-emodin, rhein, emodin, chrysophanol, and physcion, 280 nm for naringin, hesperidin, neohesperidin, and baicalin. Results: The developed method provided satisfactory precision and the accuracy of this method was in the range from 94.0% to 103.1%, all of the 10 compounds showed good linearity (r > 0.999) in a detected concentration range. Conclusion: The validated method was successfully applied to the simultaneously of these active components in Dachaihu Granule from different production batches.