Abstract
Despite the increasing concern about the harmful effects of micro- and nanoplastics (MNPs), there are no harmonized guidelines or protocols yet available for MNP ecotoxicity testing. Current ecotoxicity studies often use commercial spherical particles as models for MNPs, but in nature, MNPs occur in variable shapes, sizes and chemical compositions. Moreover, protocols developed for chemicals that dissolve or form stable dispersions are currently used for assessing the ecotoxicity of MNPs. Plastic particles, however, do not dissolve and also show dynamic behavior in the exposure medium, depending on, for example, MNP physicochemical properties and the medium’s conditions such as pH and ionic strength. Here we describe an exposure protocol that considers the particle-specific properties of MNPs and their dynamic behavior in exposure systems. Procedure 1 describes the top-down production of more realistic MNPs as representative of MNPs in nature and particle characterization (e.g., using thermal extraction desorption-gas chromatography/mass spectrometry). Then, we describe exposure system development for short- and long-term toxicity tests for soil (Procedure 2) and aquatic (Procedure 3) organisms. Procedures 2 and 3 explain how to modify existing ecotoxicity guidelines for chemicals to target testing MNPs in selected exposure systems. We show some examples that were used to develop the protocol to test, for example, MNP toxicity in marine rotifers, freshwater mussels, daphnids and earthworms. The present protocol takes between 24 h and 2 months, depending on the test of interest and can be applied by students, academics, environmental risk assessors and industries.
Key points
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In nature, micro- and nanoplastics (MNPs) occur in various shapes, sizes and chemical compositions. Each of these properties can affect both their dynamic behavior and their toxicology and should be considered when performing ecotoxicology experiments to assess their risk.
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Here, MNPs are generated from plastic waste by using either a ball or centrifugal mill. Their physicochemical properties are measured. Ecotoxicology experiments in soil and aquatic systems are described.
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Acknowledgements
This work received financial support from the UEF Water research program, which is jointly funded by the Saastamoinen Foundation, the Wihuri Foundation and the Olvi Foundation. The study was also partially funded by the European Union’s Horizon 2020 research and innovation program, via the projects PLASTICSFATE (Grant Agreement number 965367) and POLYRISK (Grant Agreement number 964766). N.T. acknowledges the Canada Research Chair program and the Killam Research Fellowship.
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F.A.M. designed the protocol, and conceptualized and supervised the protocol development. All the co-authors contributed to developing, writing, editing and reviewing the protocol.
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Nature Protocols thanks Win Cowger and Xianfei Huang for their contribution to the peer review of this work.
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Key references using this protocol
Jeong, C.-B. et al. Environ. Sci. Technol. 50, 8849–8857 (2016): https://doi.org/10.1021/acs.est.6b01441
Abdelsaleheen, O. et al. Sci. Total Environ. 798, 149196 (2021): https://doi.org/10.1016/j.scitotenv.2021.149196
Abdolahpur Monikh, F. et al. Nano Today 46, 101611 (2022): https://doi.org/10.1016/j.nantod.2022.101611
Xu, E. G. et al. Environ. Sci. Technol. 54, 6859–6868 (2020): https://doi.org/10.1021/acs.est.0c00245
Altmann, K. et al. Appl. Res. e202200078 (2023): https://doi.org/10.1002/appl.202200078
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Abdolahpur Monikh, F., Baun, A., Hartmann, N.B. et al. Exposure protocol for ecotoxicity testing of microplastics and nanoplastics. Nat Protoc 18, 3534–3564 (2023). https://doi.org/10.1038/s41596-023-00886-9
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DOI: https://doi.org/10.1038/s41596-023-00886-9
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