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Elevated nitrate concentrations in groundwater and surface water supplies can negatively impact the quality of the environment and human health. Recent studies have examined the use of zero-valent iron technology to treat nitrate-contaminated groundwater. Mechanistic aspects of nitrate reduction by zero-valent iron are unresolved. This project investigated the kinetics and mechanism of nitrate reduction by zero-valent iron under anoxic conditions and under oxic conditions. Stirred-batch reactions were studied over environmentally relevant ranges of reactant concentration, pH, and temperature. A complex rate expression was derived with a 1.8 order dependence on nitrate, a 1.4 order dependence on zero-valent iron, and a fractional order (0.8) dependence on proton concentrations under anoxic conditions. An apparent activation energy of 35 kJ mol−1 was observed indicating that nitrate reduction was diffusion controlled under our conditions. Furthermore, the calculated entropy of activation value of −162 J mol−1K−1 indicates that this reaction occurred by an associative mechanism. Under oxic conditions, there was a lag period in nitrate reduction where oxygen was preferentially utilized, leading to a slower rate of nitrate reduction when compared with anoxic conditions. These rate data can be used in predicting nitrate disappearance in nitrate-contaminated groundwater and wastewater treated with zero-valent iron.