To establish the UPLC fingerprint of Oldenlandia corymbosa from different regions and to distinguish it from Oldenlandia diffusa.UPLC procedure was performed on an ACQUITY UPLC BEH C18 (50 mm x 2. 1 mm, 1. 7 µm) column and eluted with a mobile phase consisted of methanol-l % acetic acid at a flow rate of 0. 2 mL/min. The column temperature was 30 °C . The detection wavelength was 254 nm. A matrix was constructed for similarity evaluation, cluster analysis and principle component analysis.The collected samples had a good similarity. A specificity fingerprint chromatogram was produced and 15 common peaks were designated. Samples were divided into four groups.It is a reliable and available method for specific identification of Oldenlandia corymbosa and for distinguishing Oldenlandia corynbosa and Oldenlandia diffusa.