Abstract: To a method for the simultaneous quantification of five ingredients in Malu Huangxian Miture is established. Ultra-high performance liquid chromatography-photodiode array detector(UPLC-DAD) was performed on a column of Waters BEH(Φ1.7μm,100 mm×2.10 mm) at the custom wavelengths including 232 nm for 0~3 min, 274 nm for 3~7 min, and at 249 nm for 7~10 min. Methanol-1% acetic acid was used as the mobile phase using gradient elution at a flow rate of 0.61 mL/min. The contents of paeoniflorin, baicalin, baicalein, wogonin and ammonium glycyrrhetate were simultaneously detected by a single injection. Good linear correlation were observed(r=0.9991~0.9999) and the average recovery rates were 99.70%~100.70%. The proved method is simple, accurate, reliable and repeatable that is suitable for the quality control of Malu Huangxian Mixture.