OBJECTIVE： To establish a method for the simultaneous determination of shikonin， acetylshikonin and β，β-dimethylacrylshikonin in Arnebia euchroma. METHODS： RP-HPLC method was adopted. The determination was performed on Kromasil 100-5 C18 column with mobile phase consisted of acetonitrile-0.1％ formic acid solution（80 ∶ 20，V/V） at the flow rate of 1.0 mL/min. The detection wavelength was set at 516 nm， column temperature was 25 ℃， and sample size was 10 μL. RESULTS： The linear ranges of shikonin， acetylshikonin and β，β-dimethylacrylshikonin were 0.404-10.100 μg/mL（r＝0.999 8）， 5.350-107.000 μg/mL（r＝0.999 6）， 2.035-40.700 μg/mL（r＝0.999 8）， respectively. The limit of quantitation was 0.40，2.91，1.34 μg/mL， and the limit of detection was 0.12，0.87，0.40 μg/mL. RSDs of precision， stability and reproducibility tests were all lower than 2.0％ （n＝6）. The recovery rate were 99.12％-104.18％（RSD＝1.85％，n＝6），96.51％-100.21％（RSD＝1.43％，n＝6）， 98.11％-102.51％ （RSD＝1.42％，n＝6）， respectively. CONCLUSIONS： The method is simple， precise， stable and reproducible. It can be used for simultaneous determination of shikonin， acetylshikonin and β，β-dimethylacrylshikonin in A. euchroma.