OBJECTIVE： To establish the HPLC fingerprints of Lycianthes biflora. METHODS： HPLC was performed on the column of Dima ODS-C18 with mobile phase of methanol-water （gradient elution） at a flow rate of 1.0 ml/min， the detection wavelength was 370 nm， column temperature was 30 ℃，and injection volume was 20 μl. Using chromatographic peak 2， the HPLC chromatograms of 11 batches were determined， and common peak identification and similarity evaluation were conducted by using Fingerprint Similarity Evaluation System of Traditional Chinese Medicine （version A in 2004）. RESULTS： There were 10 common peaks in the HPLC chromatograms of 11 batches， the similarity was no lower than 0.965. According to the verification， the HPLC chromatograms of 11 batches and reference patterns had a good consistency. CONCLUSIONS： The established fingerprint is specific， stable， and can be used for one of the main basis of quality evaluation of L. biflora.