AbstractResponse surface methodology (RSM) was employed to optimize dilute alkaline pretreatment conditions for the maximum glucose yield of poppy stalk, with respect to NaOH concentration (1.0 to 3.5%, w/v), pretreatment temperature (50 to 100 °C), and pretreatment time (10 to 110 min). Recovery of glucan ranged from 61.02% to 99.14%, based on the initial glucan in the raw material. The highest lignin removal (43.43%) was obtained at the pretreatment conditions of 90 °C and 3.50% NaOH for 90 min. The optimum pretreatment conditions for maximum glucose yield after enzymatic hydrolysis were found to be 2.40% w/v NaOH, 70 min, and 80 °C. Under these conditions, experimental glucose and xylose yields were 499.35 mg/g glucan and 498.66 mg xylose/g xylan, respectively.