The effects of nitrate on fermentative hydrogen production and soluble metabolites from mixed cultures were investigated by varying nitrate concentrations from 0 to 10g N/L at 35°C with an initial pH of 7.0. The results showed that the substrate degradation rate, hydrogen production potential, hydrogen yield, and average hydrogen production rate initially increased with increasing nitrate concentrations from 0 to 0.1g N/L, while they decreased with increasing nitrate concentrations from 0.1 to 10g N/L. The maximum hydrogen production potential of 305.0mL, maximum hydrogen yield of 313.1mL/g glucose, and maximum average hydrogen production rate of 13.3mL/h were obtained at a nitrate concentration of 0.1g N/L. The soluble metabolites produced by the mixed cultures contained only ethanol and acetic acid (HAc) without propionic acid (HPr) and butyric acid (HBu). This study used the Modified Logistic model to describe the progress of cumulative hydrogen production in batch tests. A concise model was proposed to describe the effects of nitrate concentration on average hydrogen production rate.