This study examines detrimental effects of acid mine drainage (AMD) on stream invertebrate communities and tests for a direct relationship between trophic position and accumulation of three metals (Fe, Cu, Zn) by stream invertebrates in situ. On two dates in each of seven stream sites, we measured food chain length, mean trophic level, taxa richness, and trophic position of stream macroinvertebrates comprising the food webs using stable nitrogen isotope ratios. Metals in tissue of representatives of 35 taxa were measured by ICP-OES. Our results are the first direct comparison of uptake of these metals in stream invertebrate taxa according to trophic position as identified by delta15N. As predicted, metal concentrations were generally greater in water and insects from sites adjacent to mining activity and invertebrate taxa richness was significantly lower. Taxa richness increased with distance away from contaminated headwaters. Despite reductions in diversity at sites nearest AMD, food chain length and mean trophic level did not differ between streams. The relationship between trophic position and metal accumulation differed considerably among metals. Specifically, Fe declined (biodilution) and Zn increased (biomagnification) with trophic level, but trophic position had no effect on Cu levels in these insects. Our results highlight fundamental differences in trophic transfer of specific metals through aquatic food webs and identify ecologically important impacts of AMD on stream invertebrates.