Ionic liquid (IL) pretreatment of lignocellulosic materials has provided a new technical tool to improve lignocellulosic ethanol production. To evaluate the influence of the residual IL in the fermentable sugars from enzymatic hydrolysis of IL pretreatment of lignocellulosic materials on the subsequent ethanol fermentation, the toxicity of the IL 1-butyl-3-methylimidazolium chloride ([BMIM]Cl) to Saccharomyces cerevisiae AY93161 was investigated. Firstly, the morphological structure, budding and metabolic activity of Saccharomyces cerevisiae AY93161 at different [BMIM]Cl concentrations were observed under an optical microscope. The results show that its single cell morphology remained unchanged at all [BMIM]Cl concentrations, but its reproduction rate by budding and its metabolic activity decreased with the [BMIM]Cl concentration increasing. The half effective concentration (EC50) and the half inhibition concentration (IC50) of [BMIM]Cl to Saccharomyces cerevisiae AY93161 were then measured using solid and liquid suspension culture and their value were 0.53 and 0.39 g.L-1 respectively. Finally, the influence of [BMIM]Cl on ethanol production was investigated. The results indicate that the [BMIM]Cl inhibited the growth and ethanol production of Saccharomyces cerevisiae AY93161. This toxicity study provides useful basic data for further development in lignocellulosic ethanol production by using IL technology and it also enriches the IL toxicity data.