AbstractAlfalfa stems and ground aspen were exposed to peracetic acid (0.5 to 9% on biomass) at temperatures ranging from 40 to 100° C and reaction times from 1 to 5 hours. Glucose release as a percentage of total cellulose content was determined using subsequent standard enzymatic hydrolysis. Statistical analysis confirmed that aspen showed a strong response to peracetic acid addition rate. 9% peracetic acid removed 14% of the original lignin and increased the rate of glucose release from 23% to 44%. Temperature and reaction time played a less significant role. For alfalfa stems, low levels of peractic acid (0.5%) increased glucose release from 30 to 47%. The addition of larger doses of peracetic acid did not show any significant improvement; this effect appears to be closely related to rate of lignin removal. While peracetic acid effectively removed lignin from aspen, 98% of the original lignin was still present in alfalfa after higher level peracetic acid treatments; the yield loss observed during pretreatment of alfalfa stems originates from other biomass components.