Fermentative hydrogen production was carried out using Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564). This work investigates the effects of initial substrate concentration, initial medium pH, and temperature. The hydrogen yield was about 3.1 mol (mol glucose)(-1) when starting with an initial glucose concentration of 10 gl(-1) and initial a pH of 6.0 +/- 0.2 at a temperature of 37 degrees C. The volume of hydrogen produced decreased when higher initial glucose concentrations were applied. The most suitable conditions for hydrogen production in a batch reactor were observed at initial PH 6.0 +/- 0.2 and 37 degrees C. (C) 2008 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights reserved.