Fluoride (F) contamination in groundwater can be problematic to human health. This study evaluated the concentration of fluoride in groundwater resources of Isfahan Province, the central plateau of Iran, and its related health issues to the inhabitant populations. For this purpose, 573 drinking groundwater samples were analyzed in 2016 by using the spectrophotometric method. Non-carcinogenic health risks due to F exposure through consumption of drinking water were assessed using the US EPA method. In addition, the associated zoning maps of the obtained results were presented using geographic information system (GIS). The results indicated that F content in drinking water ranged from 0.02 to 2.8 mg/L. The F contents were less than 0.50 mg/L in 63% of the drinking groundwater samples, 0.51-1.5 mg/L in 33.15%, and higher than 1.5 mg/L in 3.85% (Iran and World Health Organization guidelines) of the drinking groundwater samples. The F levels in the west and the south groundwater resources of the study areas were lower than 0.5 mg/L, which is within the recommended values for controlling dental caries (0.50-1.0 mg/L). Therefore, these places require more attention and more research is needed to increase F intake for health benefit. The HQ index for children, teens and male and female adults had health hazards (HQ > 1) in 51, 17, 28, and 18 of samples, respectively. Groundwater resources having a risk of more than one were located in the counties of Nayin, Natanz, and Ardestan. So, in these areas, there are potential risks of dental fluorosis. The most vulnerable groups were children. The F levels must be reduced in this region to decrease endemic fluorosis.