This research involved optimizing acid hydrolysis in the development of ethanol, a promising alternative energy source for restricted crude oil, from lignocellulosic materials (bamboo). The conversion of bamboo to ethanol can mainly be accomplished through three process steps: pretreatment of bamboo wood for the removal of lignin and hemicellulose, acid hydrolysis of pretreated bamboo for the conversion of cellulose into sugar reduction (glucose) and fermentation of sugars into ethanol using anaerobic Saccharomyces cerevisiae. The effects of parameters (factors) in the hydrolysis step were investigated and the optimum combination of parameters values (temperature, time and acid concentration) was set by experimentation. Factorial design of three-factors-at-two-level with a replica of two (23 = 8, 8•2 = 16) was applied to the hydrolysis step to investigate the effect of hydrolysis parameters on the response variable (ethanol yield) using Design-Expert® 7 software.