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We compared the toxicity of the neonicotinoids imidacloprid, thiacloprid, thiamethoxam, acetamiprid, and clothianidin in terms of the survival and reproduction of 2 species of soil invertebrates, Folsomia candida and Eisenia andrei. Tests were performed using LUFA 2.2 natural soil, following standard protocols aimed at answering 2 questions: 1) Is there a difference in the toxicity between pure compound and its formulation? and 2) Is there a difference in the sensitivity of the species exposed to the same compound? For E. andrei, formulations and pure compounds had similar toxicity to both endpoints tested. For F. candida, acetamiprid and imidacloprid had different toxicities, with acetamiprid being 4 times more toxic to survival (median lethal concentration [LC50] 0.12 mg active substance [a.s.]/kg dry soil) and imidacloprid being 4 times more toxic to reproduction of the springtail (median effect concentration [EC50] 0.25 mg a.s./kg dry soil) than their commercial formulations. The most toxic compound to E. andrei was acetamiprid (LC50 0.80 and EC50 0.35–0.40 mg a.s./kg), and the most toxic to F. candida was clothianidin (LC50 0.07 and EC50 0.05 mg a.s./kg). Estimated risk ratios indicated that only one application/yr of clothianidin in the formulation Poncho® may pose a threat to the populations of springtails and earthworms. Environ Toxicol Chem 2020;39:548–555. © 2019 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.