Spray water quality is an important consideration for optimizing herbicide efficacy. Hard water cations in the carrier water can reduce herbicide performance. Greenhouse studies were conducted to evaluate the influence of hard water cations and the use of ammonium sulfate (AMS) on the efficacy of 2,4-D choline and premixed 2,4-D choline plus glyphosate for giant ragweed, horseweed, and Palmer amaranth control. Carrier water hardness was established at 0, 200, 400, 600, 800, or 1,000 mg L−1 using CaCl2 and MgSO4, and each hardness level consisted of without or with AMS at 10.2 g L−1. One-third of the proposed use rates of 2,4-D choline at 280 g ae ha−1 and 2,4-D choline plus glyphosate at 266 plus 283 g ae ha−1, respectively, were applied in the study. An increase in carrier water hardness showed a linear trend for reducing 2,4-D choline and 2,4-D choline plus glyphosate efficacy on all weed species evaluated in both studies. The increase in water hardness level reduced giant ragweed control with 2,4-D choline and the premix formulation of 2,4-D choline plus glyphosate to a greater extent without AMS than it did with AMS in the spray solution. Increases in water hardness from 0 to 1,000 mg L−1 reduced weed control 20% or greater with 2,4-D choline. Likewise, the efficacy of the premixed 2,4-D choline plus glyphosate was reduced 21% or greater with increased water hardness from 0 to 1,000 mg L−1. The addition of AMS improved giant ragweed, horseweed, and Palmer amaranth control ≥ 17% and ≥ 10% for 2,4-D choline and 2,4-D choline plus glyphosate application, respectively. The biomass of all weed species was reduced by ≥ 8% and ≥ 5% with 2,4-D choline and 2,4-D choline plus glyphosate application, respectively, when AMS was added to hard water.