Background: A high-performance liquid chromatography fingerprint of different variants of Phyllostachys pubescens was developed for the source discrimination and quality control of Phyllostachys pubescens medicines produced in Zhejiang Province, China. Materials and Methods: Samples of Phyllostachys pubescens collected from nine cultivars in Zhejiang Province were used to establish the fingerprint. Taking the chromatographic peak (isoorientin) as the reference peak, the chromatographic column was Sunfire C 18 ODS (250 mm×4.6 mm, 5 ΅m) at a column temperature of 25°C. The mobile phase was acetonitrile 0.8% acetic acid water solution (in a gradient elution mode) at a flow rate of 1 ml/min. The detection wavelength was 330 nm. Results: 20 common peaks were established in the chromatographic fingerprints of Phyllostachys pubescens. And then by comparing the UV and standard substance with those of the authentic standards and literature, eight main peaks in the fingerprints were identified. Data of nine cultivars were evaluated statistically using similarity analysis, hierarchical cluster analysis (HCA) in order to establish a similarity standard of fingerprint for quality control of Phyllostachys pubescens, then to classify the samples. The similarity indexes were all above 0.95 between reference chromatogram and each sample's chromatogram. Conclusion: in this study, the methods established are reliable, and could be used to evaluate the quality and to identify the species of Phyllostachys pubescens in the future.