Herein, to research the toxic effect of ionic liquids (ILs) on earthworms and compare their different toxicities in different soils, 1-butyl-3-methyl imidazolium tetrafluoroborate ([Bmim]BF4) was selected as a test substance, Eisenia fetida was selected as the experimental indicator organism, and artificial and fluvo-aquic soils were selected as the media. The acute toxicity, reactive oxygen species (ROS) content, detoxification enzyme (GST) activity, anti-oxidant enzyme activities, lipid peroxidation oxidative and DNA damage in earthworms were all measured to evaluate the toxicity of [Bmim]BF4. The results showed that either in fluvo-aquic soil or artificial soil, [Bmim]BF4 can stimulate the accumulation of ROS in earthworms, inducing activities of antioxidant enzymes and detoxification enzymes, inevitably causing lipid peroxidation and DNA damage in earthworms. The integrated biomarker response (IBR) indicated that the toxicity of [Bmim]BF4 in fluvo-aquic soil was greater than that in artificial soil. This experiment is relevant to the reliability of artificial soil toxicity research, and maybe this paper can provide a more authentic understanding of traditional toxicity experiments.