To develop a method for simultaneous determination of astilbin，scutellarin，scutellarein，luteolin，apigenin，and isorhamnetin in Migan Mixture by high-performance liquid chromatography(HPLC）.Methods:Separation was performed on an Agilent ZORBAX SB-C18 column(4.6 mm×250 mm，5 μm) at 23 ℃.The mobile phases were methanol(A)-0.4% phosphoric acid(B) with gradient elution.The flow rate was 1.0 mL·min-1 and the detection wavelength was 320 nm.Results:Astilbin，scutellarin，scutellarein，luteolin，apigenin，and isorhamnetin showed good linearity between the injection volume and the peak area at 0.144~1.44 μg(r=0.999 9)，0.196~1.96 μg(r=0.999 7)，0.076~0.76 μg(r=0.999 3)，0.034~0.34 μg(r=0.999 3)，0.032~0.32 μg(r=0.999 8)，and 0.026~0.26 μg(r=0.999 5)，respectively.The average recoveries were 95.6%，96.3%，94.5%，92.3%，97.8%，and 95.2%，with RSDs of 1.7%，1.5%，2.2%，1.2%，2.0%，and 1.6%，respectively.Conclusion:This method is simple，stable，and repeatable，which can be used to determine the content of six flavonoids in Migan Mixture by HPLC.