Abstract: OBJECTIVE To establish the UHPLC specific chromatogram and determine 2,3,5,4-tetrahydroxyl diphenylethylene-2-O-β-D-glucoside, rheum emodinand rheochrysidin of Polygoni Multiflori Caulis. To evaluate the quality of Polygoni Multiflori Caulis based on principal component analysis(PCA) and cluster analysis(CA). METHODS UHPLC analysis was performed on a shiseido C₁₈ column(2.1 mm×100 mm, 2.7 μm). The mobile phase was composed of 0.1% phosphoric acid and acetonitrile with gradient elution. And the flow rate was 0.3 mL·min^-1, the detection wavelength was 254 nm. The SPSS 20.0 software was used for PCA and CA analysis. RESULTS The method for specific chromatogram was established six characteristic peaks. The similarities of 40 batches of Polygoni Multiflori Caulis were 0.655-0.998. The content of 2,3,5,4-tetrahydroxyl diphenylethylene-2-O-β-D-glucoside, rheum emodin, rheochrysidin in different batches ranged from 0.01 to 1.96 mg·g^-1, from 0.02 to 3.15 mg·g^-1, and from 0.01 to 1.48 mg·g^-1. PCA and CA results showed that all of Polygoni Multiflori Caulis were classified into 3 categories, and reflected the quality characteristics of different batches. CONCLUSION The UHPLC specific chromatogram coupled with PCA and CA can objectively and effectively assess the quality of Polygoni Multiflori Caulis.