Prairie cordgrass (PCG) was pretreated by sequential extrusion and clean fractionation (CF) processing. Following CF, PCG was fractionated into cellulose, hemicellulose and lignin-rich fractions. Cellulose pulp was then enzymatically hydrolyzed, producing glucose. The main purpose of this study was to produce the highest glucose yield as possible. The effects of time, temperature, catalyst concentration and solvent mixture composition on the fractionation were tested. Different proportions of methyl isobutyl ketone (MIBK), ethanol and water with sulfuric acid as a catalyst were evaluated. Optimal conditions for sequential extrusion and clean fractionation (39 min, 129 C, 0.69% catalyst, and 28% MIBK) resulted in higher glucose yield (92%), and more lignin (87%) and xylan (95%) removal than for clean fractionation alone. Pairwise comparison of raw PCG with extruded PCG clean fractionation revealed no difference in glucose yields, but xylan and AIL removal were higher in the case of clean fractionation of the pre-extruded PCG. (C) 2012 Elsevier Ltd. All rights reserved.