A high performance liquid chromatography (HPLC) method was developed for fingerprint of Dipsacus asper. Analysis were carried out on a Zorbax C-18 column by gradient elution using 0.1% phosphoric acid and acetonitrile as the mobile phases. The column was maintained at 25 degrees C, the flow rate was 1 mL x min(-1), and the detection wavelength was set at 205 nm. Asperosaponin VI was selected as reference compound, Seventeen common peaks were selected, and the fingerprint with good precision, stability and repeatability was successfully used to evaluate quality of 24 batches of crude extracts of D. asper. Chemical characteristics of D. asper was analyzed by DAD detection and HPLC-MS techniques with an ESI source. The quasi-molecular ions of compounds in both negative and positive modes were observed for molecule mass information of 33 compounds, and the potential structures of 10 characteristic components were identified by study on the mass spectra of compounds and comparing with reference data and some of standards. The results indicate the HPLC fingerprint of D. asper will show more characters through identification of component structures using an HPLC-ESI-MS method, and will control the quality of D. asper more effectively and reasonable.