The environmental profile of hydrogen depends greatly on the nature of the feedstock and the production process. In this Well-to-Wheels (WTW) study, the environmental impacts of hydrogen production from lignocellulosic biomass via pyrolysis and subsequent steam reforming of bio-oil were evaluated and compared to the conventional production of hydrogen from natural gas steam reforming. Hydrogen was assumed to be used as transportation fuel in an internal combustion engine vehicle. Two scenarios for the provision of lignocellulosic biomass were considered: wood waste and dedicated willow cultivation. The WTW analysis showed that the production of bio-hydrogen consumes less fossil energy in the total lifecycle, mainly due to the renewable nature of the fuel that results in zero energy consumption in the combustion step. The total (fossil and renewable) energy demand is however higher compared to fossil hydrogen, due to the higher process energy demands and methanol used to stabilize bio-oil. Improvements could occur if these are sourced from renewable energy sources. The overall benefit of using a CO(2) neutral renewable feedstock for the production of hydrogen is unquestionable. In terms of global warming, production of hydrogen from biomass through pyrolysis and reforming results in major GHG emissions, ranging from 40% to 50%, depending on the biomass source. The use of cultivated biomass aggravates the GHG emissions balance, mainly due to the N(2)O emissions at the cultivation step. Copyright (C) 2011, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.