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Methanol-based extraction protocol for insoluble and moderately water-soluble nanoparticles in plants to enable characterization by single particle ICP-MS


The detection and characterization of soluble metal nanoparticles in plant tissues are an analytical challenge, though a scientific necessity for regulating nano-enabled agrichemicals. The efficacy of two extraction methods to prepare plant samples for analysis by single particle ICP-MS, an analytical method enabling both size determination and quantification of nanoparticles (NP), was assessed. A standard enzyme-based extraction was compared to a newly developed methanol-based approach. Au, CuO, and ZnO NPs were extracted from three different plant leaf materials (lettuce, corn, and kale) selected for their agricultural relevance and differing characteristics. The enzyme-based approach was found to be unsuitable because of changes in the recovered NP size distribution of CuO NP. The MeOH-based extraction allowed reproducible extraction of the particle size distribution (PSD) without major alteration caused by the extraction. The type of leaf tissue did not significantly affect the recovered PSD. Total metal losses during the extraction process were largely due to the filtration step prior to analysis by spICP-MS, though this did not significantly affect PSD recovery. The methanol extraction worked with the three different NPs and plants tested and is suitable for studying the fate of labile metal-based nano-enabled agrichemicals.

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Data availability

The datasets generated during and/or analyzed during the current study are included in this manuscript and its Supplemental Information. Raw data is available from the corresponding author upon reasonable request.


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This material is based upon work supported by the US National Science Foundation (NSF) and the Environmental Protection Agency (EPA) under NSF Cooperative Agreement EF-1266252, Center for the Environmental Implications of NanoTechnology (CEINT), from the NSF Integrated Graduate Education and Research Traineeship Nanotechnology Environmental Effects and Policy (IGERT-NEEP) (DGE-0966227), and CBET-1530563 (NanoFARM). This study was financially supported by Austrian FFG in the framework of the ERA-NET SIINN project 849880 (NanoFARM).

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All authors contributed to the study conception and design. Funding and resource acquisition was performed by Gregory Lowry, Frank von der Kammer, and Thilo Hofmann. Material preparation and data collection and analysis were performed by Stephanie Laughton, Adam Laycock, and Garret Bland. The first draft of the manuscript was written by Adam Laycock and Stephanie Laughton and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Gregory V. Lowry.

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Laughton, S., Laycock, A., Bland, G. et al. Methanol-based extraction protocol for insoluble and moderately water-soluble nanoparticles in plants to enable characterization by single particle ICP-MS. Anal Bioanal Chem 413, 299–314 (2021).

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  • Single particle ICP-MS
  • Metallic nanoparticle characterization
  • Agrichemical characterization