A hydrothermal-chemical process for the simultaneous biomass hydrolysis and bio-hydrogen formation was proposed in this study. Current results revealed that a maximum reducing sugar concentration of 26.01 g/L and the yield of bio-hydrogen of 0.9098 mmol/g-cellulose were attained with the designated experimental parameters. Consequently, this study provided a potential route to produce bio-hydrogen and reducing sugar simultaneously in a one-step reaction process by using the energy crops or the agriculture wastes as the substrate. Copyright (C) 2012, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.