An improved HPLC-DAD–ESI-MSn method has been developed to simultaneously quantify eight major compounds in Saussurea tridactyla Sch.-Bip. ex Hook. f. which has long been used as a traditional Tibetan medicine. This method was validated to be sensitive, precise and accurate with the LODs of 0.11–5.01 μg/ml, the overall intra-day and inter-day variations less than 2.70%, and the overall recovery over 98.0%, respectively. The correlation coefficients (r2) of the calibration curves were higher than 0.991. This newly established method was successfully applied to reveal the difference in the chemical profiles and contents of these analyses in S. tridactyla from different localities. In addition, by comparison UV and MS spectra with those of authentic compounds and literatures, a total of fourteen peaks were identified. It can be concluded that this method was effective to ensure the safety and efficacy consistency of S. tridactyla, and can be applied to other traditional Tibetan medicinal plants from different resources in Tibet.