The effects of pH, cation valence, and ionic strength (IS) on the stability and aggregation behavior of zinc oxide nanoparticles (ZnO NPs) were investigated in this study. Results showed that ZnO NPs were most prone to aggregation at the isoelectric point (pH = 8.7), with an aggregation rate (ΔD/Δt) of 30.1. ZnO NPs showed a greater propensity for dissolution at lower pH (pH < 7), and Zn2+ was more rapidly released into the aqueous phase in acidic solutions than neutral or alkaline conditions. The C/C0 of ZnO NPs was about 21.56% and remained stable in acidic solution of pH 4.0. Additionally, slow sedimentation with a C/C0 ratio of 95.0% was observed due to an increase in repulsive interactions between nanoparticles under pH = 10. The effect of cations on the ΔD/Δt of ZnO NPs decreased in strength as follows: Ca2+ > Mg2+ > K+ > Na+. High-valence metal cations (Ca2+, Mg2+) were more competitively adsorbed onto the surface of ZnO NPs with a hydrogen atom due to Coulomb's law, increasing the zeta potential and stabilizing the suspension of ZnO NPs at IS < 10 mM. Furthermore, compression of the electric double layer (EDL) became stronger than electrostatic adsorption with increasing IS, reaching a maximum ΔD/Δt of 23.3 (Ca2+, pH = 7, IS = 1 M). The C/C0 ratio of ZnO NPs decreased from 100% to 56.5% (Na+), 52.2% (K+), 45.2% (Mg2+), and 40.1% (Ca2+) at pH = 7 and an IS of 0.5 M. In addition to the cation valence, the hydration forces and ionic radii of the metal cations might be other factors that affected the interactions of metal cations with ZnO NPs. Finally, the total interaction energy between ZnO NPs was calculated using the Derjaguin-Landau-Verwey-Overbeek (DLVO) theoretical formula, and the calculated results were in agreement with the experimental outcomes under various aquatic environmental conditions.