[9] Selective herbicides were unavailable in the 1950s to protect soybean and cotton (2,4-DNP could have been used but had to be exactingly applied lest it destroy the crops), so Lilly Research Laboratories screened ~2000 compounds from 1958 to 1980 blindly looking for a result.

Trifluralin was initially thought a failure, yet the plots stayed free of weeds weeks later.

It is unclear why trifluralin's exotic 4-trifluoromethyl was tested so early (1960), before more common candidates such as fluoro, bromo, or iodo.

This resistance is especially hard to evolve for weeds to tubulin disrupting herbicides because both α-tubulin and β-tubulin must mutate, as imbalance between their expressions is potentially lethal.

Supposedly, the mechanism of prosulfocarb-resistance is inverse to trifluralin resistance, requiring lower metabolism of herbicide, rather than greater.

[18] Trifluralin has been banned in the European Union since 20 March 2008, primarily due to high toxicity to aquatic life.

[2] Application rates vary, such as 0.8-3.0 L of 480 g/L formulation per hectare, typically diluted with water,[5] and other compatible herbicides, e.g. isoproturon,[2] to be sprayed in one go.

[23] Selectivity is possible even on susceptible crops, by sowing below the herbicide band, and shallower germinating weeds will be controlled.

[5] Trifluralin breaks down into many products as it degrades, ultimately being incorporated into soil-bound residues or converted to carbon dioxide (mineralized).

This environmental degradation process has been reported for many structurally related herbicides (dinitroanilines) as well as a variety of explosives such as TNT and picric acid.

[2] Repeated annual application shows steady and continuous decline in soil and does not accumulate, even applied well in excess of recommended rates.

Research on humans remains unconvincing, but EPA animal toxicity data "supports the possible carcinogenicity" of trifluralin.

[30] Trifluralin on mammalian ovaries (tested in mice, at 150 mg/kg/day) showed no effect on oocyte quality, but may induce a stress response in ovarian somatic cells.

It, and other dinitroanilines, are tubulin-binding agents with selective antileishmanial properties, leishmania being the parasite causing the disease, which killed 60,000 people in 2001.

Research into expanding's trifluralin's medical use is stymied by its low water solubility and easy sublimation.

[37] Commercial formulations have included trifluralin mixtures with: linuron, napropamide, metribuzin, clomazone,tebutam, bromoxynil and ioxynil, isoproturon, terbutryn, trietazine, neburon and isoxaben.