Despite many efforts into the study of fluids interaction in low salinity water flooding, they are not probing the basics of transport phenomena between the involved phases. This work is aimed to bring new understanding of fluid–fluid interaction during low salinity water flooding through a series of organized experiments in which a crude oil sample with known properties was kept in contact with different brine solutions of various ionic strengths. Measuring brine pH, conductivity and crude oil viscosity and density for a period of 45 days illustrates the strong effect of the contact time and ionic strength on the dissociation of polar components and physical properties of the crude oil and brine. Besides, the interfacial tension (IFT) measurements show that the interfacial interactions are affected by several competitive interfacial processes. By decreasing the ionic strength of the brine, the solubility of naphthenic acids in the aqueous solution increases, and hence, the conductivity and the pH of the aqueous phase decrease. To verify this important finding, UV–Vis spectroscopy and 1H NMR analysis were also performed on aged brine samples. Notably, there is an ionic strength of brine in which the lowest IFT is observed, while the other physical properties are remained relatively unchanged.