A systematic study on the colloidal behavior of uncoated and polyvinylpyrrolidone (PVP) coated TiO2 engineered nanomaterials (ENMs) in simulated aqueous media is herein reported, in which conditions representative for natural waters (pH, presence of divalent electrolytes (i.e. Ca2+/Mg2+ and SO4 2−), of natural organic matter (NOM) and of suspended particulate matter (SPM)) were systematically varied. The colloidal stability of the different dispersions was investigated by means of Dynamic and Electrophoretic Light Scattering (DLS and ELS) and Centrifugal Separation Analysis (CSA), and a global stability index based on these three techniques was developed. The index allows to quantitatively classify the nano-based dispersions according to their colloidal stability affected by the different parameters studied. This multimethod approach clearly identifies inorganic SPM followed by divalent electrolytes as the main natural components destabilizing TiO2 ENMs upon entering in simulated natural waters, while it highlights a moderate stabilization induced by NOM, depending mainly on pH. Moreover, the PVP coating was found to attenuate the influence of these parameters on the colloidal stability. The obtained results show how the global stability index developed is influenced by the complexity of the system, suggesting the importance of combining the information gathered from all the techniques employed to better elucidate the fate and behavior of ENMs in natural surface waters.