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Background: The simultaneous determination of multiple components in a sample is an important factor in the quality control of traditional Chinese medicines and can give an indication of potential clinical applications. Introduction: A rapid and sensitive method has been introduced for the simultaneous quantitative analysis of eight bioactive flavonoid constituents from Scutellariae Radix using ultra-high performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry. Methods: The separation was performed on a Waters Acquity UPLC C18 column (2.1 mm×100 mm, 1.7 μm), under optimized mass spectrometry conditions, with a flow rate of 0.3 mL/min. The column temperature was maintained at 35°C and the injection volume was 3 μL. Results: The method showed a good linear relationship of each component; all R2 values were above 0.9990 in the experiment. The RSDs of the precision test, repeatability test, stability test and recovery test were all not more than 2.86 %. We found that the total percentage amounts of the eight flavonoids were 22.19%, 18.63% and 10.86% in Raw Scutellariae Radix (RSR), Wine Scutellaria Radix (WSR) and Scutellaria Radix Charcoal (SRC) respectively. Conclusion: The method was successfully applied to the simultaneous determination of the eight bioactive flavonoids of Raw Scutellariae Radix, Wine Scutellaria Radix and Scutellaria Radix Charcoal.