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A case-controlled study has compared urinary, blood plasma, and serum fluoride (F) levels of people living in endemic areas of the Thar Desert, Sindh, Pakistan, consuming groundwater with F concentrations as high as 4.00-10.00 mg/L with those consuming groundwater with low F levels of 0.30 mg/L. A total of 121 individuals from Samme Jo Tar Village of the Chachro Sub-District, Tharparker, and 121 controls from Gadap Town, Karachi, Pakistan, were selected for this study. Results indicated highly significant differences were present between the mean urinary F (12.90 versus 2.30 mg/L, p<0.001), plasma F (0.61 versus 0.25 mg/L, p<0.001) and serum F levels (0.75 versus 0.29 mg/L, p<0.001) of cases and controls. There was a strong positive correlation between plasma and serum F levels (r = 0.884), while moderate correlations were found between urine plus plasma and between urine plus serum (r = 0.576 and 0.621) F levels of cases, respectively.