Abstract: To establish HPLC dual-wavelength fingerprint of Artemisia rupestris L. , and provide a scientific basis for the improvement of its quality specifications. The separation was performed on an Agilent Zorbax SB-C₁₈(4. 6 mm×250 mm, 5 μm)column maintained at 32 ℃, with methanol-0. 2% formic acid-water gradient elution at flow rate of 1. 0 mL/min and the UV detection wavelength at both 245 and 325 nm. The sample injection volume was 20 μL. The fingerprint of Artemisia rupestris L. was established. 7 and 8 common peaks were found, respectively, of which 5 common peaks were identified, and the similarity among 9 batches of Artemisia rupestris L. and the fingerprints of control was over 0. 9. The HPLC dual-wavelength fingerprint of Artemisia rupestris L. was established for the first time, providing a new scientific basis for its identification and quality control.