Groundwater quality is a consequence of cumulative effects of natural and anthropogenic processes occurring in unsaturated and saturated zone, which, in certain conditions, can lead to elevated concentrations of chemical substances in groundwater. In this paper, the concept of determining the ambient background value of a chemical substance in groundwater was applied, because the long-term effects of human activity influence the increase in concentrations of substances in the environment. The upper limits of ranges of ambient background values were estimated for targeted chemical substances in four groundwater bodies in the Pannonian region of Croatia, according to the demands of the EU Groundwater Directive. The selected groundwater bodies are typical, according to the aquifer typology, for the Pannonian region of Croatia. Probability plot (PP), the modified Lepeltier method, as well as the simple pre-selection method, were used in this paper, depending on a number of chemical data in analysed data sets and in relation to the proportion of