The fermentable sugars in lignocellulosic biomass are derived from cellulose and hemicellulose, which are not readily accessible to enzymatic saccharification because of their recalcitrance. An ethanosolv pretreatment method was applied for the enzymatic saccharification of barley straw with an inorganic acid. The effects of four process variables (temperature, time, catalyst dose, and ethanol concentration) on the barley straw pretreatment were analyzed over a broad range using a small composite design and a response surface methodology. The yield of the residual solid and composition of the solid fraction differed as ethanosolv conditions varied within the experimental range. A glucan recovery, xylan recovery, and delignification were 85%, 14%, and 69% at center point conditions (170°C, 60 min, 1.0% (w/w) H(2)SO(4), and 50% (w/w) ethanol), respectively. Ethanosolv pretreatment removed lignin effectively. Additionally, the highest enzymatic digestibility of 85.3% was obtained after 72 h at center point conditions.