Comparison of Three Bacterial Toxicity Assays for Imidazolium-Derived Ionic Liquids

Ionic liquids have become leading candidates for replacing many common organic solvents used in the chemical process industry. There is, however, a general lack of toxicology data relevant to wastewater treatment facility microbes for these compounds. In this study, we performed three bacterial-based toxicity assays on several imidazolium-derived compounds as well as the precursor compound 1-methylimidazole. Two of the assays, the Shk1 and Microtox assays, are used as surrogate assays for toxicity to bacterial respiration in activated sludge wastewater treatment plants. The third assay was a direct measure of the effect of toxicity on mixed bacterial culture respiration, using a commercially available consortium of naturally occurring bacteria to obtain IC50 values for direct comparison to the EC50 values from the surrogate assays. The Shk1 assay is based on a genetically engineered bioluminescent Pseudomonas bacterium and is more highly correlated with the respiration inhibition than the Microtox assay. The Shk1 assay gave EC50 values more similar to IC50 values from the bacterial respiration inhibition assay for the compounds tested in this work. The Shk1 EC50 values were similar to that of 1-butanol, an alcohol with an alkyl chain length similar to that of the cation of the tested compounds, which were 1-butyl-3-methylimidazolium bromide ([bmim][Br]), 1-butyl-3-methylimidazolium hexafluorophosphate ([bmim][PF6]), 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide, ([bmim][Tf2N]), and the precursor compound 1-methylimidazole, and were generally smaller than those typical of aromatic organic solvents.