Objective: To establish a new RP-HPLC method for simultaneous determination of the seven naphthoquinones in crude extract of Arnebia euchroma

namely lithospermum acid

shikonin

acetyl shikonin

deoxyshikonin

isobutyryl shikonin

β

β-dimethylacry shikonin and isovalerylshikonin. Method: Agilent ZORBAX SB-C18(4.6 mm×250 mm

5 μm) column was used with acetonitrile-pure water (with 0.03% formic acid) as mobile phase in gradients elution. The flow rate was 1 mL·min-1

and the UV detector was monitored at 254

516 nm. Result: All the calibration curves of the seven naphthoquinones showed good linearity in test ranges (r>0.999 1); and the overall recoveries were in the range of 97%-103% (n=3) with RSD less than 3%. The overall of RSD of precision test were less than 3%. A HPLC method for simultaneous determination of the seven naphthoquinones in crude extract of A. euchroma was established. Conclusion: The established HPLC method is rapid

accurate

credible and repeatable for the determination of the seven water-soluble (lithospermum acid) and liposoluble (the rest of six components) components simultaneously

and provide a more comprehensive scientific basis for the quality evaluation of A. euchroma.