Benthic diatom communities are used widely as indicators of river health due to their rapid response to changes in water quality. The ability for diatom-based indices to detect eutrophication has been well documented; however, an index designed specifically to detect herbicide impacts is yet to be established. This is required as herbicide contamination of rivers is common in agricultural regions and poses a potential threat to aquatic ecosystems. This study developed a new biomonitoring index (SPEAR herbicides) using benthic diatom communities to detect the toxic impacts of herbicides in rivers, and tested it across 14 rivers in the Great Barrier Reef catchment area, Australia. The new index uses diatom species traits to classify diatoms as either sensitive or tolerant to herbicides and calculates the fraction of sensitive taxa within a sample. The SPEAR herbicides index showed a decline in herbicide sensitive diatoms with increasing herbicide toxicity of the sites. The impacts of herbicide toxicity on the diatom community were only apparent after the wet season when aqueous herbicide concentrations typically peak and diatoms were able to recover during the dry season when herbicide concentrations were lower. SPEAR herbicides values had a negative relationship with the percentage of grazing and cropping in catchments but had a positive relationship with the percentage of conservation in catchments. SPEAR herbicides also had a negative relationship with co-occurring potential stressors such as nutrients and total suspended solids.