Objective: To establish the HPLC fingerprint analysis methods for Madder and to provide the reference for quality evaluation of Madder. Method: Phenomenex ODS C18(4.60 mm×250 mm

4 μm)column was used and the methanol-0.2% H3PO4 water was chosen as the mobile phase in a gradient mode. The column temperature was at 25℃ and the detection wavelength was at 275 nm.The detection time was 80 min and the flow rate was 0.8 mL· min-1. Result: Taking mollugin as the reference peak

eleven common peaks were selected as the fingerprint peaks of Madder. Conclusion: This method is simple and accurate with a good reproducibility. It provides a reference standard for the quality control of Madder.