An ultra-fast liquid chromatography－triple quadrupole－linear ion trap mass spectrometry （UFLC－QTRAP－MS/MS） method was developed for the simultaneous determination of 33 active constituents consisting of flavonoids，organic acids，nucleosides，and amino acids in Taxilli Herba．The stems branches and leaves of Taxilli Herba from different hosts were also compared and analyzed．The optimum conditions for the preparation of the test solution were selected by the Box－Behnken response surface method（BBD－RSM）．The target constituents were separated on an XBridge,®, C,18,（4.6 mm × 100 mm，3.5 μm） analytical column by gradient elution using 0.1% formic acid aqueous solution and methanol as mobile phases，then analyzed by mass spectrometry with an electrospray ion source（ESI） in multi reaction monitoring（MRM） mode，and finally quantified by the external standard method．The correlation analysis and independent sample ,t,-test were performed for the contents of the 33 target constituents．The results showed that the optimal extraction conditions were as follows：a methanol volume fraction of 70%，a liquid to material ratio （mL∶g） of 31∶1 and an extraction time of 32 min，under which the extraction yield for quercetrin reached 3.77 mg/g．The standard curves for 33 target constituents showed good linearity in the corresponding range with their correlation coefficients（,r,） not less than 0.999 0．The limits of detection（LODs） and limits of quantitation（LOQs） were in the range of 0.13－66.85 ng/mL and 0.42－222.83 ng/mL，respectively．The precision，repeatability and stability of this method were excellent．The average recoveries at low，medium and high spiked levels in real samples ranged from 98.0% to 101%，with the relative standard deviations（RSDs） all less than 4.0%．The correlation analysis revealed that the contents of active constituents were associated to a certain extent．A ,t,-test revealed that the contents of various active constituents in the stems branches and leaves of Taxilli Herba differed significantly，and the contents of each constituent，such as quercetin-3-O-glucuronide，hyperin，rutin，isoquercitrin，quercetrin and auicularin，were significantly higher in the leaves than in the stems and branches．The developed method is accurate and stable，which could provide a new method reference for the comprehensive evaluation and overall control on the quality of Taxilli Herba，and a certain reference basis for its rational exploitation．