Field and greenhouse trials have shown that ultra-low-volume applications (ULV) of glyphosate (N-(phosphonomethyl)glycine] exhibit a greater phytotoxic effect on barley (Hordeum vulgare L. ‘Bonanza′) at a given rate than do high-diluent-volume applications with a conventional hydraulic sprayer. Three greenhouse studies and one field trial were conducted to evaluate the influence of individual drop makeup on the effectiveness of ULV applications. Barley plants were treated at the four-leaf stage with one, three, or nine drops (1 μl each) of glyphosate and a non-ionic surfactant with the concentration of the drops adjusted so that the total amount of active ingredient applied to each plant was constant. Application of a single, concentrated drop was significantly more effective in reducing total shoot growth than were applications of more dilute drops in greater number. When the dilute drop was supplemented with additional surfactant, phytotoxicity was restored, provided there was a sufficiently high concentration of glyphosate in the drop. With the substitution of a commercial formulation of glyphosate, the response of barley to variations in drop number and drop concentrations followed a similar trend. The response of field-grown plants did not differ from plants grown under controlled conditions.