In this study, the yield of hydrogen production was investigated under different feedstock pretreatment conditions. The feedstock for dark fermentative hydrogen production was textile wastewater which was obtained from the de-sizing process in a textile factory, located in northern Taiwan. The wastewater was pretreated with activated carbon, cation exchange resin or was not pretreated before being fed into the batch bottles. Biohydrogen production was carried out in a batch reactor with the sludge of mixed-culture using the feedstock from the pretreated wastewater. The sludge was obtained from the Taichung municipal wastewater treatment plant. The yield of hydrogen production using the two pretreatment methods or non - treatment were compared. Results showed that the textile wastewater could remove bio-toxic inhibitors by using activated carbon. The best hydrogen yield was 1.37 mol H-2/mol reducing sugar with an initial pH 7.0, as substrate concentration was fixed at 20 g total sugar/L. Using textile wastewater as feedstock for biohydrogen production was successful, in removing bio-toxic inhibitors from wastewater with activated carbon pretreatment. Copyright (C) 2012, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.