In this study we determined the fluoride content in drinking water and hair of 12-year-old schoolchildren from different Serbian municipalities, i.e. Valjevo, Veliko Gradiste, Kacarevo and Vranjska Banja. The analyses were performed using composite fluoride ion-selective electrode. Average fluoride levels were 0.10, 0.15, 0.79 and 11 ppm in well water, 0.07, 0.10, 0.17 and 0.15 ppm in tap water, 19.3, 21.5, 25.4, and 32.5 ppm in hair samples, in Valjevo, Veliko Gradiste, Kacarevo and Vranjska Banja, respectively. Correlation analysis indicated statistically significant positive relationship between fluoride in wells water and fluoride in hair, for all municipalities: correlation coefficients were 0.54 (p < 0.05), 0.89, 0.97 and 0.99 (p < 0.001), in Vranjska Banja, Valjevo, Veliko Gradiste, and Kacarevo, respectively. Positive correlation was obtained also between fluoride in tap water and hair samples in all regions under the study, with statistical significance only in Valjevo municipality, p < 0.05. Dental examination of schoolchildren confirmed dental fluorosis only in the region of Vranjska Banja. Moreover, in endemic fluorotic region of Vranjska Banja, positive and statistically significant correlations were confirmed between fluoride in well water and dental fluorosis level (r = 0.61; p < 0.01) and additionally between fluoride in hair and dental fluorosis level (0.62; p < 0.01). The primary findings from this study have shown that fluoride content in hair is highly correlated with fluoride content in drinking water and dental fluorosis level, indicating that hair may be regarded as biomaterial of high informative potential in evaluating prolonged exposure to fluorides and to individuate children at risk of fluorosis regardless of the phase of teeth eruption.