Humans commonly apply chemicals to manage agroecosystems. If those chemicals influence the behaviour or survival of non-target arthropods, the food web could be altered in unintended ways. Glyphosate-based herbicides are among the most ubiquitous pesticides used around the world, yet little is known about if and how they might affect the success of terrestrial predatory arthropods in agroecosystems. In this study, we quantified the effects of a commercial formulation of a glyphosate-based herbicide on the activity of three predatory arthropod species that inhabit agricultural fields in the eastern United States. We also measured the survival of the most common species. We tested the reactions of the wolf spider, Pardosa milvina, to either direct application (topical) or contact with a treated substrate (residual). We quantified the reactions of a larger wolf spider, Hogna helluo, and a ground beetle, Scarites quadriceps, to a compound (topical plus residual) exposure. Pardosa milvina reduced locomotion time and distance under topical herbicide exposure, but increased speed and non-locomotory activity time on exposed substrate. Both H. helluo and S. quadriceps increased non-locomotory activity time under compound herbicide exposure. Over a period of 60 days post-exposure, residually exposed P. milvina exhibited lower survivorship compared to topically exposed and control groups. Thus, exposure of terrestrial arthropods to glyphosate-based herbicides affects their behaviour and long-term survival. These results suggest that herbicides can affect arthropod community dynamics separate from their impact on the plant community and may influence biological control in agroecosystems.