Background: In China, rhizome of Curcuma wenyujin (CW) is used to improve blood stasis syndrome-related diseases for many years. Nonsteamed, steamed, and boiled with vinegar rhizomes of CW can be used as three different traditional Chinese medicine s, named as Pian-Jiang-Huang (PJH), Sheng-E-Zhu (SEZ), and Cu-E-Zhu (CEZ), respectively. After processing, the therapeutic effects have changed. Objective: In order to illustrate the effective substance of the three kinds of rhizome of CW, an ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS) method coupled with statistical approach was developed. Materials and Methods: Fresh rhizomes of CW were processed into PJH, SEZ, and CEZ according to Chinese pharmacopoeia 2015. UPLC-Q/TOF-MS coupled with chemometric analysis was applied to compare the differences in chemical profiles of the three kinds of rhizome of CW. Results: In this study, 21 compositions in the rhizome of CW were identified. The one-way ANOVA of relative intensities of the 21 compositions was processed by SPSS software version 20.0. The column superposed graph of the relative intensities of 21 components shows that the total relative intensities of the 21 components in SEZ are the topmost and that of PJH is about the same as CEZ, but the proportion of each component is quite differ from each sample. Principal component analysis and orthogonal partial least squares discrimination analysis were processed by Simca-p14.1 software. Finally, five ions including curcumenone, curcumol, curzerenone, furanodiene, and germacrone were discovered to be the Q-Makers of the three kinds of rhizome of CW. Conclusion: This study provides a reliable research basis for the clinical application of the three kinds of rhizome of CW.