Objective:To establish the fingerprint chromatogram of anthraquinones compounds in Radix et Rhizoma Rhei. Methods: HPLC with ?Bondapak-C 18 [3.9mm?300mm,10?m] column was used, the CH 3CN-0.5% H 3PO 4 (gradient elution) as a mobile phase and detection wavelength at 430nm. Results: 12 peaks were separated on the HPLC fingerprint of anthraquinones compounds in Rhubarb. Conclusion: The method is simple and accurate with a good reproducibility and can be used for a quality control method for Radix et Rhizoma Rhei.