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Indonesia is the largest producer of palm oil commodity in the world. Around on fifth of the palm oil processing weight end up as oil palm empty fruit bunches (OPEFB). OPEFB is lignocellulosic biomass which contains cellulose, hemicellulose, and lignin. The utilizations of cellulose and hemicellulose require pretreatment to open the lignin bond that covers those components. In this study, pretreatment of OPEFB using microwave assisted dilute ammonia method was carried out. OPEFB (30 - 40 mesh) were pretreated by ammonia solution with solid-liquid ratio of 1:10 under variated parameters such as ammonia concentration (7.5; 10; 12.5 %), microwave power (280; 560; 840 Watt) and reaction time (3; 6; 9 minutes).The hemicellulose contents of all samples were then characterized according to SNI 01-1561-1989. The results obtained were finally optimized using response surface methodology with the application of Box-Behnken model. The model showed that highest hemicellulose content of 27.3% can be reached at pretreatment condition of 665 Watt microwave power at 7.5 % ammonia concentration for 3 minutes.