An ultrahigh performance liquid chromatographic(UPLC) method was developed for the determination of paeonol(Pae),thymol(Thy),honokiol(Hon),magnolol(Mag) and glycyrrhetinic acid(Gly) in toothpaste.The sample was ultrasonically extracted with 90% methanol.After centrifugation and filtration,the supernatant was separated on a Waters ACQUITY UPLCHSS C18 column(2.1 mm×100 mm,1.8 μm) by gradient elution at a flow rate of 0.3 mL/min,using acetonitrile and 0.1% formic acid(pH 2.8) as mobile phases.The ultraviolet detection wavelength was set at 275 nm and 250 nm,respectively.Results indicated that the five botanic active constituents had good linear relationships in the range of 2-100 mg/L with their correlation coefficients(r) more than 0.999.The limits of detection and quantitation were in the ranges of 0.2-1.0 mg/kg and 0.8-3.5 mg/kg,respectively.The average recoveries at four spiked levels ranged from 90.5% to 99.4% with their relative standard deviations(RSDs) of 0.7%-5.1%.With the characteristics of rapidness,good repeatability,accuracy and sensitivity,the method was applied in the analysis on botanic active constituents in real toothpaste samples.