Insect infestation of agricultural stored products is a significant challenge to food security across the globe. One common pest is Tribolium castaneum (red flour beetle). In a new approach to addressing the threat of these beetles, Direct Analysis in Real Time-High-Resolution Mass Spectrometry was used to examine infested and uninfested flour samples. These samples were then distinguished through statistical analysis techniques, including EDR-MCR, in order to highlight the important m/z values contributing to the differences in the flour profiles. A subset of these values responsible for the identification of infested flour (nominal m/z 135, 136, 137, 163, 211, 279, 280, 283, 295, 297, and 338) were further investigated, and compounds responsible for these masses included 2-(2-ethoxyethoxy)ethanol, 2-ethyl-1,4-benzoquinone, palmitic acid, linolenic acid and oleic acid. These results have the potential to lead to a rapid technique by which flour and other grains can be tested for insect infestation.