Objective: To establish a method for the determination of acetylshikonin in the Hairy Roots of Arnebia euchroma (Royle) Johnst. Method: The RP-HPLC system consisting of YMC-Pack ODS-A C18 column(4.6 mm×150 mm

5 μm) and a mixture of acetonitrile solution and water solution and formic acid solution(700 ∶300 ∶0.5)as the mobile phase was adopted; the detection wavelength was 275 nm; the flow rate was set at 1.0 mL·min-1 and the column temperature was at 35 ℃. Result: The linear range of acetylshikonin was 51.55-1 031.0 mg·L-1(r=0.9 999). The average recovery (n=9) was 99.81%. Conclusion: The method is quick

simple and reproducible for the determination of acetylshikonin in Hairy roots of A. euchroma.