The effect of time of day (TOD) on the activity of six common POST herbicides was investigated in field trials from 2007 to 2009 at two locations in southwestern Ontario. Percentage weed control was assessed following application of bentazon, chlorimuron-ethyl, fomesafen, glyphosate, imazethapyr, or quizalofop-p-ethyl applied at 3-h intervals from 6:00 A.M. to midnight, when weeds averaged 15 cm tall. The effect of time of day varied with weed species, but weed control was generally reduced when herbicides were applied at 6:00 A.M., 9:00 P.M., and midnight. Herbicide activity on velvetleaf was most frequently reduced, especially for chlorimuron-ethyl, glyphosate, and imazethapyr. Control of common ragweed with glyphosate and imazethapyr was also affected by the timing of application, and pigweed species only showed an effect with glyphosate. Variation in temperature, relative humidity, and dew presence/absence at different times of the day, as well as morphological/physiological characteristics such as weed size at time of application and diurnal leaf movement in response to light intensity, may account for the variation in weed control at different times of the day. Significant soybean yield loss was not observed in this study, but may occur if herbicide efficacy is severely reduced by application at inappropriate times of day. These results provide valuable information for growers, and suggest that POST herbicides are most effective when applied midday, rather than in the early morning or late evening.