Abstract: OBJECTIVE To establish HPLC fingerprints of Mongolian medicine of the flos of Scabiosa comosa Fisch. METHODS Hypersil-ODS column(4.6 mm×300 mm, 5 μm) was adopted, gradient elution was applied with mobile phase A (0.1% phosphoric acid solution) and mobile phase B(acetonitrile). The flow rate was 1.0 mL·min^-1, the detective wavelength was 254 nm, the column temperature was 30 ℃ and injection volume was 10 μL. RESULTS HPLC fingerprints of different source of the flos of Scabiosa comosa Fisch were established. Ten peaks were observed on the HPLC fingerprints and the semblance was calculated. The results of methodological study met the technical requirements for fingerprint. CONCLUSION This method is accurate, reproducible and provides a scientific basis for quality evaluation of the flos of Scabiosa comosa Fisch.