Silver nanoparticles (AgNPs) are frequently found in everyday products and, as a consequence, their release into the environment cannot be avoided. Once in aquatic systems, AgNPs interact with natural constituents and undergo different transformation processes. Therefore, it is important to characterize and quantify AgNPs in environmental waters in order to understand their behavior, their transformation, and their associated toxicological risks. However, the coexistence of ionic silver (Ag⁺) with AgNPs in aquatic systems is one of the greatest challenges for the determination of nanosilver. Ion-exchange resins can be used to separate Ag⁺ from AgNPs, taking advantage of the different charges of the species. In this work, Dowex 50W-X8 was used to separate Ag⁺ and AgNPs in order to easily determine AgNP concentrations using inductively coupled plasma optical emission spectroscopy. The separation methodology was successfully applied to river water samples with different ratios of Ag⁺ and AgNPs. However, the methodology is not useful for wastewater samples. The described methodology also demonstrated an improvement in the determination of the particle size of AgNPs present in river waters by single particle inductively coupled plasma mass spectrometry when a significant amount of Ag⁺is also present.