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The effect of ammonia concentration ranging from 0 to 10g N/L on fermentative hydrogen production by mixed cultures was investigated in batch tests using glucose as substrate at 35°C and initial pH 7.0. The experimental results showed that during the fermentative hydrogen production, the substrate degradation efficiency increased with increasing ammonia concentration from 0 to 0.01g N/L. The hydrogen production potential, hydrogen yield and average hydrogen production rate increased with increasing ammonia concentration from 0 to 0.1g N/L. The maximum hydrogen production potential of 291.4mL, maximum hydrogen yield of 298.8mL/g glucose and maximum average hydrogen production rate of 8.5mL/h were all obtained at the ammonia concentration of 0.1g N/L. [Copyright &y& Elsevier]