The effect of sulphate on hydrogen production by fermentation of distillery wastewater was studied in small scale batch experiments at 37.5 °C with an initial pH of 6.5 and adding sulphate varying in concentrations from 0-3000 mg/L. Hydrogen yield and substrate degradation efficiency was estimated at various concentrations of sulphate. The results showed that the hydrogen production and substrate degradation efficiency decreased with increase in sulphate concentrations from 0 to 3000 mg/L. Maximum hydrogen yield of 3.8 ml/g was found when distillery wastewater (having 1892 mg/L sulphate) was used without any further addition of sulphate. Since in the presence of high sulphate additions of 1000- 3000mg/L, substrate degradation and hydrogen production sharply declined, the fermentative hydrogen production technology using distillery effluents, which have high sulphate concentrations may not be practicable and wastewaters with low sulphates should be utilized for fermentative hydrogen production.