Water quality influences the bioavailability and toxicity of copper to aquatic organisms. Understanding the relationships between water-quality parameters and copper toxicity may facilitate the development of site-specific criteria for water quality and result in better protection of aquatic biota. Many studies have examined the influence of a single water-quality parameter on copper toxicity, but the interactions of several characteristics have not been well studied in low-hardness water. The goal of the present research was to examine the interactions among water-quality characteristics and their effects on copper toxicity to larval fathead minnows (Pimephales promelas). The effects of dissolved organic carbon (DOC) concentration, DOC source, pH, and hardness on acute copper toxicity were determined using a complete factorially designed experiment. Hardness, pH, DOC, and interaction of pH and DOC all significantly affected copper toxicity. A predictive model based on these data described 88% of the variability in copper toxicity. This model also explained 58% of the variability in copper toxicity for an independent dataset of South Carolina (USA) waters. The biotic ligand model underpredicted the acute copper toxicity to fathead minnows when compared with observed values.