The work aims to investigate the effect of infestation of Tribolium castaneum and Tribolium confusum on the type, content and composition of volatile substances in wheat flour. Gas Chromatography-Ion Mobility Spectrometry (GC-IMS) was used for the qualitative and quantitative determination of the normal wheat flour and the wheat flour infested with Tribolium castaneum and Tribolium confusum. A total of 50 peaks were detected in the three kinds of wheat flour and 41 volatile substances were identified. Wheat flour infested with the T.castaneum contained high concentrations of hexanal, 2-ethylhexanol, nonanal, 2,6-dimethylpyridine, (E)-2-heptenal, 1-octen-3-one, dimethyl trisulfide compounds, heptanal, and furfural. In contrast, wheat flour infested with T.confusum contained high concentrations of 1-penten-3-ol, hexanol, 2-heptanone, pentanol, and 3-methyl butanol aldehydes. Compared to the control group, the contents of aldehydes and alcohols significantly increased in the wheat flour infested by two kinds of test pests. By analyzing the volatile substances in wheat flour and combining gas chromatography-ion mobility spectrometry, fingerprinting and principal component analysis, the differences in volatile components between normal wheat flour and wheat flour infested with T.castaneum and T.confusum are revealed. The results of this study provide a reference basis for predicting the occurrence of early pests during wheat flour storage.