Event DAS-40278-9 maize expresses the aryloxyalkanoate dioxygenase-1 enzyme, which was originally identified in the soil bacterium Sphingobium herbicidovorans. This enzyme degrades 2,4-dichlorophenoxyacetic acid (2,4-D) and aryloxyphenoxypropionate herbicides (e.g., haloxyfop, cyhalofop, quizalofop, etc.); therefore, plants that contain this enzyme are tolerant to these herbicides. We employed the substantial equivalence approach to investigate the compositional safety of event DAS-40278-9 maize. A total of 82 different compositional analyses were conducted to evaluate the equivalence of event DAS-40278-9 and conventional maize. Analyte levels within the transgenic entries were either within literature ranges for non-transgenic maize or statistically indistinguishable from the non-transgenic near-isogenic hybrid, thus indicating substantial equivalence between event DAS-40278-9 and its conventional counterpart. These results agree with dozens of published studies for other transgenic events where input traits were found to have a negligible effect on crop composition compared with traditional breeding methods.