Abstract: This study investigates microwave application to assist hydrolysis of Oil Palm’s Empty Fruit Bunch (OPEFB) for the production of bioethanol. Hydrolysis was carried out by two steps microwave irradiation to maximize yield of glucose. On the 1st hydrolysis, microwave energy irradiates OPEFB fibre at various concentrations of H2SO4 (0.5 and 1%) and level of microwave power (medium, medium high and high). Residue from 1st hydrolysis was irradiated on 2nd hydrolysis process. Temperature profile and moisture loss during hydrolysis process were observed to study heat generation inside the reactor. Glucose resulted from each process was determined by DNS method using UV-VIS spectrophotometric. Overall hydrolysis of OPEFB yielded glucose ranged between 2-47%.