Objective To establish a method based on ultrahigh performance liquid chromatography-tandem mass spectrometry for rapid determination of eight sweeteners in glycyrrhizin.Methods The sample is ultrasonically extracted with 20 mL pure water for 30 min, passed through 0.22 μm microporous filter membrane, separated through the Waters HSS T3 column, subjected to gradient elution with 0.2% (V/V) ammonia solution-methanol, and detected under the negative mode of electrospray ionization.Results The eight sweeteners showed a good linear relationship between peak area and concentration within the range of 0.1~20.0 μg/mL, with correlation coefficients greater than 0.99. The limits of quantification are within the range of 0.2~1.0 μg/g. The average recoveries of sample spiked are within the range of 85.8%~103.0%, and the relative standard deviations are in the range of 2.4%~4.6%.Conclusion This method is simple, rapid, and suitable for determination of the content of 8 sweeteners in glycyrrhizin.