An ultra-performance liquid chromatography coupled with quadrupole time of flight tandem mass spectrometry (UPLC/Q-TOF-MS) was first employed to establish the chromatographic fingerprint and thereby identify the major components of Yuanhu Zhitong tablet (YZT), a traditional Chinese medicine. Fingerprint was established by analyzing 15 different batches of YZT, in which we obtained 17 common peaks with 18 constituents. Fifteen components were identified in the fingerprint with reference to the standards and literature data, including protopine, -allocryptopine, coptisine, tetrahydropalmatine, tetrahydroberberine, corydaline, palmatine, berberine, byakangelicol, dehydrocorydaline, oxypeucedanin, imperatorin, phelloptorin, isoimperatorin, and osthole. Each of these was separated by using a Waters Acquity UPLC BEH C18 (2.1x50mm, 1.7 mu m) column with linear gradient elution of 0.2% formic acid water-acetonitrile. Meanwhile, the hierarchical cluster analysis was applied to evaluate the variation of the YZTs based on the characteristics of the common peaks in the UPLC/Q-TOF-MS profiles. This established method with high sensitivity and high speed may become a powerful tool for quality evaluation of YZT and related preparation.