We investigated the use of low concentrations of butanol (< 40%, all v/v) as an organosolv pretreatment to fractionate lignocellulosic biomass into cellulose, hemicellulose, and lignin. The pretreatment conditions were optimized for sorghum bagasse by focusing on four parameters: butanol concentration, sulfuric acid concentration, pretreatment temperature, and pretreatment time. A butanol concentration of 25% or higher together with 0.5% or higher acid was effective for removing lignin while retaining most of the cellulose in the solid fraction. The highest cellulose (84.9%) and low lignin (15.3%) content were obtained after pretreatment at 200 degrees C for 60 min. Thus, pretreatment comprising 25% butanol, 0.5% acid, 200 degrees C, and 60 min process time was considered optimal. Enzymatic saccharification and fermentation by Saccharomyces cerevisiae produced 61.9 g/L ethanol from 200 g/L solid fraction obtained following pretreatment, and 10.2 g/L ethanol was obtained from the liquid fraction by xylose-utilizing S. cerevisiae following membrane nanofiltration to remove butanol.