The relationship between mercury (Hg) concentrations in freshwater biota and trophic position, as defined by stable nitrogen isotope ratios (δ15N), was examined in 6 lakes in northwestern Ontario. The heavier isotope of nitrogen (15N) increases an average of 3 parts per thousand (‰) from prey to predator and is used as a measure of an organism’s trophic position. Dorsal muscle from lake trout, burbot, walleye, northern pike, white sucker, lake cisco, lake whitefish, and yellow perch was analyzed for Hg and δ15N using flameless atomic absorption and mass spectrometry respectively. Within each lake, log Hg was significantly related to <515N (r2 ranged from 0.47 to 0.91, P < 0.01). For four species, yellow perch, northern pike, lake cisco, and lake trout, log Hg was positively related to δ15N (r2 ranged from 0.37 to 0.47, P ≤; 0.09) across all lakes. We also used δ15N measurements (assuming a 3%c shift between an organism and its diet) and the developed within-lake regression equations to calculate a prey Hg for each individual fish. These food Hg values were then used to predict predator Hg using Norstrom et al’s bioenergetics model. Predicted results were strongly correlated to measured Hg concentrations (r = 0.91, P < 0.001), indicating that δ15N has potential to be used in modeling.