The hydrochemical interaction between groundwater and lakewater influences the composition of water that percolates downward from the surficial aquifer system through the underlying intermediate confining unit and recharges the Upper Floridan aquifer along highlands in Florida. The ⁸⁷Sr/⁸⁶Sr ratio along with the stable isotopes, D, ¹⁸O, and ¹³C were used as tracers to study the interaction between groundwater, lakewater, and aquifer minerals near Lake Barco, a seepage lake in the mantled karst terrane of northern Florida. Upgradient from the lake, the ⁸⁷Sr/⁸⁶Sr ratio of groundwater decreases with depth (mean values of 0.71004, 0.70890, and 0.70852 for water from the surficial aquifer system, intermediate confining unit, and Upper Floridan aquifer, respectively), resulting from the interaction of dilute oxygenated recharge water with aquifer minerals that are less radiogenic with depth. The concentrations of Sr²⁺ generally increase with depth, and higher concentrations of Sr²⁺ in water from the Upper Floridan aquifer (20–35 μg/L), relative to water from the surficial aquifer system and the intermediate confining unit, result from the dissolution of Sr-bearing calcite and dolomite in the Eocene limestone. Dissolution of calcite [δ¹³C= −1.6permil(‰)] is also indicated by an enriched δ¹³C_DIC(-8.8 to -11.4 ‰) in water from the Upper Floridan aquifer, relative to the overlying hydrogeologic units (δ¹³C_DIC< -16‰).

Groundwater downgradient from Lake Barco was enriched in¹⁸O and D relative to groundwater upgradient from the lake, indicating mixing of lakewater leakage and groundwater. Downgradient from the lake, the ⁸⁷Sr/⁸⁶Sr ratio of groundwater and aquifer material become less radiogenic and the Sr²⁺ concentrations generally increase with depth. However, Sr²⁺ concentrations are substantially less than in upgradient groundwaters at similar depths. The lower Sr²⁺concentrations result from the influence of anoxic lakewater leakage on the mobility of Sr²⁺ from clays. Based on results from mass-balance modeling, it is probable that cation exchange plays the dominant role in controlling the ⁸⁷Sr/⁸⁶Sr ratio of groundwater, both upgradient and downgradient from Lake Barco. Even though groundwater from the three distinct hydrogeologic units displays considerable variability in Sr concentration and isotopic composition, the dominant processes associated with the mixing of lakewater leakage with groundwater, as well as the effects of mineral-water interaction, can be ascertained by integrating the use of stable and radiogenic isotopic measurements of groundwater, lakewater, and aquifer minerals.