Water demand for urban, industrial, and agricultural purposes is a major concern in developed and third world countries. A careful evaluation for an appropriate and sustainable use of water resources is a priority. Geochemical processes can lead to measurable variations of the aquatic environment, which can be studied through the analysis of the dissolved solutes. Even if this review is not meant to be exhaustive, it is intended to give a view on the importance of environmental isotopes in the context of groundwater quality assessments. This is done by briefly recalling some basic notions for each described system, followed by relevant applications and reports about some significant case studies. This review includes well-established isotopic systematics, such as those of O and H in water, C in dissolved inorganic carbon (DIC), S and O in sulfates, and N and O in nitrates and those of boron and Sr, which in the last lustrums have found large application in the field of water geochemistry. This chapter ends with some examples related to nontraditional isotopes, i.e., Fe, Cr, and Cu, in order to highlight the potential of the environmental isotopes to trace sources, fate, and behavior of different solutes and metals in surface water and groundwater.