Objective: To establish a HPLC method for the determination of liquiritin、 liquiritigenin and iso liquiritigenin in Licorice Mixture.Methods: A Cromasil C 18 column was used. the mobile phase was methanol 0.5% acetic acid for gradient elution. Liquiritin、 liquiritigenin and iso liquiritigenin were determined by dual wavelength, λ 1=276 nm,λ 2=372 nm.Results: The linear range of liquiritin was 50.0~1250.0ng, r= 0.9999; The average recovery was 98.30%, RSD =1.01%( n= 6). The linear range of liquiritigenin was 5.0~125.0 ng, r= 0.9998; The average recovery was 98.01%, RSD =0.68%( n= 6). The linear range of iso liquiritigenin was 3.0~6.0 ng, r= 0.9999. The average recovery was 98.37%, RSD= 0.89%( n= 6). Conclusion: This method is convenient, rapid and accurate. It can be used for quality control in production of Licorice Mixture.