Objective: To establish a quality control method of Flos Scabiosaeby HPLC fingerprint and simultaneous determination of chlorogenic acid, galuteolin, isochlorogenic acid A and isochlorogenic acid C by the quantitative analysis of multi-components by single marker (QAMS) for evaluation of medicinal materials quality. Methods: Kromasil 100-5-C₁₈ column (250 mm×4.6 mm, 5 μm) was employed with a mobile phase of a cetonitrile-0.2% phosphoric acid solution with gradient elution. Detection wavelength was set at 350 nm (fingerprint) and 326 nm(QAMS), the column temperature was 30℃, the flow rate was 1.0 mL·min^-1, and the injection volume was 20 μL. The common mode of fingerprint was established using the determination of seventeen batches of Scabiosa Flos. Using isochlorogenic acid A as an internal reference, the relative correction factors among chlorogenic acid, galuteolin, isochlorogenic acid C and the internal reference were established, and the contents were calculated and compared to those obtained with external standard method. Results: HPLC fingerprint was established for the medicinal materials of Scabiosa Flos, 10 common peaks were labeled, and 7 chromatographic peaks were identified, which were chlorogenic acid, galuteolin, isochlorogenic acid A, apigenin-7-O-glucoside, isochlorogenic acid C, luteolin and apigenin, respectively. The established relative correction factors (RCF) had good reproducibility. No significant difference was found between the quantitative results of external standard method and QAMS. Conclusion: The application of HPLC fingerprint and QAMS method model can provide an effective method to evaluate the quality of Scabiosa Flos.