Abstract: OBJECTIVE To establish HPLC fingerprints of ethanolic extraction part of Toosendan Fructus. METHODS Agilent zorbax SB-C₁₈ column (4.6 mm×150 mm, 5 μm) was adopted. A gradient elution was applied with mobile phase A (methanol and acetonitrile 1∶10) and mobile phase B (0.4% phosphoric acid solution). The flow rate was 1.0 mL·min-1, the detective wavelength was 360 nm, and the column temperature was 25 ℃. RESULTS HPLC fingerprints of ethanolic extraction part of Toosendan Fructus was established. Twenty peaks were observed on the HPLC fingerprints of Toosendan Fructus and 3 of them were identified. CONCLUSION The method developed in this study is accurate with high repeatability, and available for the quality control of Toosendan Fructus and its decoction pieces.