The aim of the present work was to investigate whether a detoxification method already in use during waste water treatment could be functional also for ethanol production based on lignocellulosic substrates. Chemical conditioning of spruce hydrolysate with hydrogen peroxide (H2O2) and ferrous sulfate (FeSO4) was shown to be an efficient strategy to remove significant amounts of inhibitory compounds and, simultaneously, to enhance the enzymatic hydrolysis and fermentability of the substrates. Without treatment, the hydrolysates were hardly fermentable with maximum ethanol concentration below 0.4 g/l. In contrast, treatment by 2.5 mM FeSO4 and 150 mM H2O2 yielded a maximum ethanol concentration of 8.3 g/l. (C) 2014 Elsevier Ltd. All rights reserved.