Ammonia fiber expansion (AFEX) pretreatment is an ammonia-based process for improving the susceptibility of lignocellulosic biomass to enzymatic attack. Four parameters - ammonia loading, water loading, reaction temperature, and residence time - can be varied in order to optimize AFEX pretreatment. The effect of these parameters on process economics of ethanol production was studied using a leading biorefinery model. Ammonia loading and residence time had the greatest impact on the economics of ethanol production, primarily due to processing costs for the chilled water condenser and the capital cost of the AFEX reactor. Water loading and reaction temperature had only modest impact on process economics. In addition, the impact of pretreatment conditions on makeup ammonia requirements was explored experimentally, which ranged from 15 to 25 g ammonia/kg biomass. Overall, pretreatment conditions can change the costs of ethanol production by up to 35 cents per gallon ethanol in an 850 ton/day refinery. By linking the results obtained from this Aspen model to experimental results for ethanol production and makeup ammonia recovery, this study can be used to create an economic optimum for AFEX pretreatment in contrast with simply maximizing fermentable sugar production. (C) 2010 Elsevier Ltd. All rights reserved.