Summary. An ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry UPLC-QTOF-MS method has been developed for qualitative and quantitative analysis major compounds in Saussurea eopygmaea Hand-Mazz, which has long been used as a traditional Tibetan medicine. This method was validated to be sensitive, precise, and accurate with the limits of detection of compounds 2, 3, 4, 6, and 7 with 0.67-1.90 μg mL −1 , the overall intra-day and inter-day variations less than 8.45%, and the overall recovery over 93.8%, respectively. The correlation coefficients (R 2 ) of the calibration curves were higher than 0.998. In addition, by comparison MS and MS/MS spectra with those of authentic compounds and literatures, a total of 14 main peaks were identified within 6 min. These results demonstrate that this approach has the potential for quality control of S. eopygmaea and other Tibetan herbal medicines.