Durian (Durio zibethinus Murr.) peel, as agricultural waste, is a potential under-utilized lignocellulosic biomass that is sufficiently available in Thailand. In this study, durian peel from monthong (D.zibethinus Murr. cv. Monthong) and chanee (D.zibethinus Murr. cv. Chanee) were subjected to pretreatment with sodium hydroxide (NaOH) under autoclaving conditions to improve glucose recovery. The effect of NaOH concentration (1%, 2%, 3%, and 4%) and autoclave temperature (110°C, 120 °C, and 130 °C) was investigated based on the amount of glucose recovered. The optimal NaOH concentration and autoclave temperature were determined to be 2% and 110 °C, respectively, under which maximum glucose (36% and 35% in monthong and chanee peels, respectively) was recovered. Glucose recovery was improved by about 6-fold at the optimal pretreatment condition for both pretreated monthong and chanee when compared to the untreated durian peels. Scanning electron microscopy (SEM) showed great changes to the surface morphology of pretreated durian peel from the two cultivars. X-ray diffraction (XRD) analysis also revealed a rise in cellulose crystallinity index (CrIs) after pretreatment. A combination of mild NaOH concentration and autoclaving is a very effective pretreatment technique for maximum glucose recovery from durian peel.