Nano-sized titanium dioxide in the aquatic environment has a potential impact on the environment and human health. In this study, the impact of pH value, dissolved organic matter (DOM) and divalent cations (Ca2+) on the stability of titanium dioxide nanoparticles (nano-TiO2) in an aqueous environment was investigated in batch tests. The results showed that the particle size of nano-TiO2 was not sensitive to pH value but was inversely proportional to zeta potential. The nano-TiO2 becomes more stable with surface zeta potential, accompanied by small particle size and high dispersion. In the presence of DOM, the particle size was smaller and the stability of nano-TiO2 could be enhanced. This might be a synergistic effect of the ligand exchange and electrostatic force. Particle size increased with the addition of Ca2+ and the stability decreased.