Abstract
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.
References
Young M, Santra S. Copper (Cu)-silica nanocomposite containing valence-engineered Cu: a new strategy for improving the antimicrobial efficacy of Cu biocides. J Agric Food Chem. 2014;62:6043–52. https://doi.org/10.1021/jf502350w.
Kah M, Kookana RS, Gogos A, Bucheli TD. A critical evaluation of nanopesticides and nanofertilizers against their conventional analogues. Nat Nanotechnol. 2018;13:677–84. https://doi.org/10.1038/s41565-018-0131-1.
Liu R, Lal R. Potentials of engineered nanoparticles as fertilizers for increasing agronomic productions. Sci Total Environ. 2015;514:131–9. https://doi.org/10.1016/j.scitotenv.2015.01.104.
Pradhan S, Mailapalli D. Interaction of engineered nanoparticles with agri-environment. J Agric Food Chem. 2017. https://doi.org/10.1021/acs.jafc.7b02528.
Rodrigues SMM, Trindade T, Duarte ACC, Pereira E, Koopmans GFF, Römkens PFAM. A framework to measure the availability of engineered nanoparticles in soils: trends in soil tests and analytical tools. TrAC Trends Anal Chem. 2016;75:129–40.
Dimkpa CO, McLean JE, Britt DW, Anderson AJ. Bioactivity and biomodification of Ag, ZnO, and CuO nanoparticles with relevance to plant performance in agriculture. Ind Biotechnol. 2012;8:344–57. https://doi.org/10.1089/ind.2012.0028.
Ruttkay-Nedecky B, Krystofova O, Nejdl L, Adam V. Nanoparticles based on essential metals and their phytotoxicity. J Nanobiotechnol. 2017;15:33. https://doi.org/10.1186/s12951-017-0268-3.
Ma C, White JC, Zhao J, Zhao Q, Xing B. Uptake of engineered nanoparticles by food crops: characterization, mechanisms, and implications. Annu Rev Food Sci Technol. 2018;9:129–53. https://doi.org/10.1146/annurev-food-030117-012657.
de la Rosa G, García-Castañeda C, Vázquez-Núñez E, Alonso-Castro ÁJ, Basurto-Islas G, Mendoza Á, et al. Physiological and biochemical response of plants to engineered NMs: implications on future design. Plant Physiol Biochem. 2017;110:226–35. https://doi.org/10.1016/j.plaphy.2016.06.014.
Ruotolo R, Maestri E, Pagano L, Marmiroli M, White JC, Marmiroli N. Plant response to metal-containing engineered nanomaterials: an omics-based perspective. Environ Sci Technol. 2018;52:2451–67. https://doi.org/10.1021/acs.est.7b04121.
Gao X, Lowry GV. Progress towards standardized and validated characterizations for measuring physicochemical properties of manufactured nanomaterials relevant to nano health and safety risks. NanoImpact. 2018;9:14–30. https://doi.org/10.1016/j.impact.2017.09.002.
Vencalek BE, Laughton SN, Spielman-sun ER, Rodrigues SMSM, Jason M, Lowry GV, et al. In situ measurement of CuO and Cu (OH)2 nanoparticle dissolution rates in quiescent freshwater mesocosms. Environ Sci Technol Lett. 2016;3:375–80. https://doi.org/10.1021/acs.estlett.6b00252.
Rajasekaran P, Kannan H, Das S, Young M, Santra S. Comparative analysis of copper and zinc based agrichemical biocide products: materials characteristics, phytotoxicity and in vitro antimicrobial efficacy. AIMS Environ Sci. 2016;3:439–55. https://doi.org/10.3934/environsci.2016.3.439.
Wang N, Tong T, Xie M, Gaillard J. Lifetime and dissolution kinetics of zinc oxide nanoparticles in aqueous media. Nanotechnology. 2016;27:1–10. https://doi.org/10.1088/0957-4484/27/32/324001.
Yin H, Casey PS. Effects of iron or manganese doping of ZnO nanoparticles on their dissolution, ROS generation and cytotoxicity. RSC Adv. 2014;4:26149–9. https://doi.org/10.1039/c4ra02481h.
Adeleye AS, Conway JR, Perez T, Rutten P, Keller AA. Influence of extracellular polymeric substances on the long-term fate, dissolution, and speciation of copper-based nanoparticles. Environ Sci Technol. 2014;48:12561–8. https://doi.org/10.1021/es5033426.
Sekine R, Marzouk ER, Khaksar M, Scheckel KG, Stegemeier JP, Lowry GV, et al. Aging of dissolved copper and copper-based nanoparticles in five different soils: short-term kinetics vs. long-term fate. J Environ Qual. 2017;46:1198–205. https://doi.org/10.2134/jeq2016.12.0485.
Gao X, Spielman-Sun E, Rodrigues SM, Casman EA, Lowry GV. Time and nanoparticle concentration affect the extractability of cu from CuO NP-amended soil. Environ Sci Technol. 2017;51:2226–34. https://doi.org/10.1021/acs.est.6b04705.
Kent RD, Vikesland PJ. Dissolution and persistence of copper-based nanomaterials in undersaturated solutions with respect to cupric solid phases. Environ Sci Technol. 2016;50:6772–81. https://doi.org/10.1021/acs.est.5b04719.
Gao X, Rodrigues SSMS, Spielman-Sun E, Lopes S, Rodrigues SSMS, Zhang Y, et al. Effect of soil organic matter, soil pH, and moisture content on solubility and dissolution rate of CuO NPs in soil. Environ Sci Technol. 2019;53:4959–67. https://doi.org/10.1021/acs.est.8b07243.
Rippner DA, Green PG, Young TM, Parikh SJ. Dissolved organic matter reduces CuO nanoparticle toxicity to duckweed in simulated natural systems. Environ Pollut. 2018;234:692–8. https://doi.org/10.1016/j.envpol.2017.12.014.
Larue C, Castillo-Michel H, Sobanska S, Cécillon L, Bureau S, Barthès V, et al. Foliar exposure of the crop Lactuca sativa to silver nanoparticles: evidence for internalization and changes in Ag speciation. J Hazard Mater. 2014;264:98–106. https://doi.org/10.1016/j.jhazmat.2013.10.053.
Kranjc E, Mazej D, Regvar M, Drobne D, Remškar M. Foliar surface free energy affects platinum nanoparticle adhesion, uptake, and translocation from leaves to roots in arugula and escarole. Environ Sci Nano. 2018;5:520–32. https://doi.org/10.1039/C7EN00887B.
Keller AA, Huang Y, Nelson J. Detection of nanoparticles in edible plant tissues exposed to nano-copper using single-particle ICP-MS. J Nanopart Res. 2018;20:101. https://doi.org/10.1007/s11051-018-4192-8.
De la Calle I, Menta M, Séby F. Current trends and challenges in sample preparation for metallic nanoparticles analysis in daily products and environmental samples: a review. Spectrochim Acta B At Spectrosc. 2016;125:66–96. https://doi.org/10.1016/j.sab.2016.09.007.
Laborda F, Bolea E, Cepriá G, Gómez MT, Jiménez MS, Pérez-Arantegui J, et al. Detection, characterization and quantification of inorganic engineered nanomaterials: a review of techniques and methodological approaches for the analysis of complex samples. Anal Chim Acta. 2016;904:10–32. https://doi.org/10.1016/j.aca.2015.11.008.
Singh G, Stephan C, Westerhoff P, Carlander D, Duncan TV. Measurement methods to detect, characterize, and quantify engineered nanomaterials in foods. Compr Rev Food Sci Food Saf. 2014;13:693–704. https://doi.org/10.1111/1541-4337.12078.
von der Kammer F, Legros S, Hofmann T, Larsen EH, Loeschner K. Separation and characterization of nanoparticles in complex food and environmental samples by field-flow fractionation. TrAC Trends Anal Chem. 2011;30:425–36. https://doi.org/10.1016/j.trac.2010.11.012.
Meisterjahn B, Neubauer E, Von der Kammer F, Hennecke D, Hofmann T. Asymmetrical flow-field-flow fractionation coupled with inductively coupled plasma mass spectrometry for the analysis of gold nanoparticles in the presence of natural nanoparticles. J Chromatogr A. 2014;1372C:204–11. https://doi.org/10.1016/j.chroma.2014.10.093.
Montaño MD, Olesik JW, Barber AG, Challis K, Ranville JF. Single particle ICP-MS: advances toward routine analysis of nanomaterials. Anal Bioanal Chem. 2016;408:5053–5074. https://doi.org/10.1007/s00216-016-9676-8.
Pace HE, Rogers NJ, Jarolimek C, Coleman VA, Higgins CP, Ranville JF. Determining transport efficiency for the purpose of counting and sizing nanoparticles via single particle inductively coupled plasma mass spectrometry. Anal Chem. 2011;83:9361–9. https://doi.org/10.1021/ac201952t.
Peters RJB, Rivera ZH, Van Bemmel G, Marvin HJP, Weigel S, Bouwmeester H. Development and validation of single particle ICP-MS for sizing and quantitative determination of nano-silver in chicken meat characterisation of nanomaterials in biological samples. Anal Bioanal Chem. 2014;406:3875–85. https://doi.org/10.1007/s00216-013-7571-0.
Dan Y, Zhang W, Xue R, Ma X, Stephan C, Shi H. Characterization of gold nanoparticle uptake by tomato plants using enzymatic extraction followed by single-particle inductively coupled plasma–mass spectrometry analysis. Environ Sci Technol. 2015;49:3007–14. https://doi.org/10.1021/es506179e.
Dan Y, Ma X, Zhang W, Liu K, Stephan C, Shi H. Single particle ICP-MS method development for the determination of plant uptake and accumulation of CeO2 nanoparticles. Anal Bioanal Chem. 2016;408:5157–67. https://doi.org/10.1007/s00216-016-9565-1.
Zhang W, Dan Y, Shi H, Ma X. Elucidating the mechanisms for plant uptake and in-planta speciation of cerium in radish (Raphanus sativus L.) treated with cerium oxide nanoparticles. J Environ Chem Eng. 2017;5:572–7. https://doi.org/10.1016/j.jece.2016.12.036.
Bao D, Oh ZG, Chen Z. Characterization of silver nanoparticles internalized by Arabidopsis plants using single particle ICP-MS analysis. Front Plant Sci. 2016;7:32. https://doi.org/10.3389/fpls.2016.00032.
Deng Y, Petersen EJ, Challis KE, Rabb SA, Holbrook RD, Ranville JF, et al. Multiple method analysis of TiO 2 nanoparticle uptake in rice ( Oryza sativa L.) plants. Environ Sci Technol. 2017;51:10615–23. https://doi.org/10.1021/acs.est.7b01364.
Gray EP, Coleman JG, Bednar AJ, Kennedy AJ, Ranville JF, Higgins CP. Extraction and analysis of silver and gold nanoparticles from biological tissues using single particle inductively coupled plasma mass spectrometry. Environ Sci Technol. 2013;47:14315–23. https://doi.org/10.1021/es403558c.
Loeschner K, Brabrand MSJ, Sloth JJ, Larsen EH. Use of alkaline or enzymatic sample pretreatment prior to characterization of gold nanoparticles in animal tissue by single-particle ICPMS characterisation of nanomaterials in biological samples. Anal Bioanal Chem. 2014;406:3845–51. https://doi.org/10.1007/s00216-013-7431-y.
Palmer DA, Bénézeth P (2008) Solubility of copper oxides in water and steam. In: 14th International Conference on the Properties of Water and Steam in Kyoto. pp 491–496
Bradfield SJ, Kumar P, White JC, Ebbs SD. Zinc, copper, or cerium accumulation from metal oxide nanoparticles or ions in sweet potato: yield effects and projected dietary intake from consumption. Plant Physiol Biochem. 2017;110:128–37. https://doi.org/10.1016/j.plaphy.2016.04.008.
Cakmak I, Kalayci M, Kaya Y, Torun AA, Aydin N, Wang Y, et al. Biofortification and localization of zinc in wheat grain. J Agric Food Chem. 2010;58:9092–102. https://doi.org/10.1021/jf101197h.
Vidmar J, Buerki-Thurnherr T, Loeschner K. Comparison of the suitability of alkaline or enzymatic sample pre-treatment for characterization of silver nanoparticles in human tissue by single particle ICP-MS. J Anal At Spectrom. 2018;33:752–61. https://doi.org/10.1039/c7ja00402h.
Dettmer K, Nürnberger N, Kaspar H, Gruber MA, Almstetter MF, Oefner PJ. Metabolite extraction from adherently growing mammalian cells for metabolomics studies: optimization of harvesting and extraction protocols. Anal Bioanal Chem. 2011;399:1127–39. https://doi.org/10.1007/s00216-010-4425-x.
Wu H, Southam AD, Hines A, Viant MR. High-throughput tissue extraction protocol for NMR- and MS-based metabolomics. Anal Biochem. 2008;372:204–12. https://doi.org/10.1016/j.ab.2007.10.002.
Zhang H, Lin Q, Ponnusamy S, Kothandaraman N, Teck KL, Zhao C, et al. Differential recovery of membrane proteins after extraction by aqueous methanol and trifluoroethanol. Proteomics. 2007;7:1654–63. https://doi.org/10.1002/pmic.200600579.
Liu S, Beauchemin D. Effect of methanol and sodium dodecylsulfate on radial profiles of ion abundance in inductively coupled plasma mass spectrometry. Spectrochim Acta B At Spectrosc. 2006;61:319–25. https://doi.org/10.1016/j.sab.2006.02.010.
Laughton S, Laycock A, von der Kammer F, Hofmann T, Casman EA, Rodrigues SMSM, et al. Persistence of copper-based nanoparticle-containing foliar sprays in Lactuca sativa ( lettuce ) characterized by spICP-MS. J Nanopart Res. 2019;21:174. https://doi.org/10.1007/s11051-019-4620-4.
Bland GD, Lowry GV. Multi-step method to extract moderately soluble copper oxide nanoparticles from soil for quantification and characterization. Anal Chem. 2020;92:9620–8. https://doi.org/10.1021/acs.analchem.0c00824.
Spielman-Sun E, Lombi E, Donner E, Howard DL, Unrine JM, Lowry GV. Impact of surface charge on cerium oxide nanoparticle uptake and translocation by wheat (Triticum aestivum). Environ Sci Technol. 2017;51:7361–8. https://doi.org/10.1021/acs.est.7b00813.
Hendriks L, Gundlach-Graham A, Hattendorf B, Gunther D. Characterization of a new ICP-TOFMS instrument with continuous and discrete introduction of solutions. J Anal At Spectrom. 2017;32:548–61. https://doi.org/10.1039/c6ja00400h.
Loosli F, Wang J, Rothenberg S, Bizimis M, Winkler C, Borovinskaya O, et al. Sewage spills are a major source of titanium dioxide engineered (nano)-particle release into the environment. Environ Sci Nano. 2019;6:763–77. https://doi.org/10.1039/c8en01376d.
U.S. EPA. Method 3050B: acid digestion of sediments, sludges, and soils. Washington, DC.: Revision 2; 1996.
Shared MS, Angeles L, States U, Li M, Pokhrel S, Jin X, et al. Stability, bioavailability, and bacterial toxicity of ZnO and iron-doped ZnO nanoparticles in aquatic media. Environ Sci Technol. 2011;45:755–61. https://doi.org/10.1021/es102266g.
Jiang C, Castellon BT, Matson CW, Aiken GR, Hsu-Kim H. Relative contributions of copper oxide nanoparticles and dissolved copper to cu uptake kinetics of Gulf killifish (Fundulus grandis) embryos. Environ Sci Technol. 2017;51:1395–404. https://doi.org/10.1021/acs.est.6b04672.
Deng Y, Petersen EJ, Challis KE, Rabb SA, Holbrook RD, Ranville JF, et al. Multiple method analysis of TiO2 nanoparticle uptake in rice (Oryza sativa L.) plants. Environ Sci Technol. 2017;51:10615–23. https://doi.org/10.1021/acs.est.7b01364.
Kińska K, Jiménez-Lamana J, Kowalska J, Krasnodębska-Ostręga B, Szpunar J. Study of the uptake and bioaccumulation of palladium nanoparticles by Sinapis alba using single particle ICP-MS. Sci Total Environ. 2018;615:1078–85. https://doi.org/10.1016/j.scitotenv.2017.09.203.
Jiménez-Lamana J, Wojcieszek J, Jakubiak M, Asztemborska M, Szpunar J. Single particle ICP-MS characterization of platinum nanoparticles uptake and bioaccumulation by Lepidium sativum and Sinapis alba plants. J Anal At Spectrom. 2016;31:2321–9. https://doi.org/10.1039/C6JA00201C.
Spielman-Sun E, Avellan A, Bland GD, Tappero RV, Acerbo AS, Unrine JM, et al. Nanoparticle surface charge influences translocation and leaf distribution in vascular plants with contrasting anatomy. Environ Sci Nano. 2019;6:2508–19. https://doi.org/10.1039/c9en00626e.
Niklas KJ. A mechanical perspective on foliage leaf form and function. New Phytol. 1999;143:19–31. https://doi.org/10.1046/j.1469-8137.1999.00441.x.
Scanlon MJ. Developmental complexities of simple leaves. Curr Opin Plant Biol. 2000;3:31–6. https://doi.org/10.1016/S1369-5266(99)00040-0.
He X, Zhang H, Shi H, Liu W, Sahle-Demessie E. Fates of Au, Ag, ZnO, and CeO 2 nanoparticles in simulated gastric fluid studied using single-particle-inductively coupled plasma-mass spectrometry. J Am Soc Mass Spectrom. 2020. https://doi.org/10.1021/jasms.0c00278.
Lee S, Bi X, Reed RB, Ranville JF, Herckes P, Westerhoff P. Nanoparticle size detection limits by single particle ICP-MS for 40 elements. Environ Sci Technol. 2014;48:10291–300. https://doi.org/10.1021/es502422v.
Navratilova J, Praetorius A, Gondikas A, Fabienke W, von der Kammer F, Hofmann T. Detection of engineered copper nanoparticles in soil using single particle ICP-MS. Int J Environ Res Public Health. 2015;12:15756–68. https://doi.org/10.3390/ijerph121215020.
Naasz S, Weigel S, Borovinskaya O, Serva A, Cascio C, Undas AK, et al. Multi-element analysis of single nanoparticles by ICP-MS using quadrupole and time-of-flight technologies. J Anal At Spectrom. 2018;33:835–45. https://doi.org/10.1039/c7ja00399d.
Peters RJB, Herrera-Rivera Z, Undas A, van der Lee M, Marvin H, Bouwmeester H, et al. Single particle ICP-MS combined with a data evaluation tool as a routine technique for the analysis of nanoparticles in complex matrices. J Anal At Spectrom. 2015;30:1274–85. https://doi.org/10.1039/C4JA00357H.
Erhardt T, Jensen CM, Borovinskaya O, Fischer H. Single particle characterization and total elemental concentration measurements in polar ice using continuous flow analysis-inductively coupled plasma time-of-flight mass spectrometry. Environ Sci Technol. 2019;53:13275–83. https://doi.org/10.1021/acs.est.9b03886.
May TW, Wiedmeyer RH. A table of polyatomic interferences in ICP-MS. At Spectrosc. 1998;19:150–5.
Van Koetsem F, Verstraete S, Wallaert E, Verbeken K, Van der Meeren P, Rinklebe J, et al. Use of filtration techniques to study environmental fate of engineered metallic nanoparticles: factors affecting filter performance. J Hazard Mater. 2017;322:105–17. https://doi.org/10.1016/j.jhazmat.2016.05.098.
Marinsky JA, Wolf A, Bunzl K. The binding of trace amounts of lead (II), copper (II), cadmium (II), zinc (II) and calcium (II) to soil organic matter. Talanta. 1979;27:461–8.
Cheng T, Allen HE. Comparison of zinc complexation properties of dissolved natural organic matter from different surface waters. J Environ Manag. 2006;80:222–9. https://doi.org/10.1016/j.jenvman.2005.09.007.
Bryan SE, Tipping E, Hamilton-Taylor J. Comparison of measured and modelled copper binding by natural organic matter in freshwaters. Comp Biochem Physiol C Toxicol Pharmacol. 2002;133:37–49. https://doi.org/10.1016/S1532-0456(02)00083-2.
Manceau A, Matynia A. The nature of Cu bonding to natural organic matter. Geochim Cosmochim Acta. 2010;74:2556–80. https://doi.org/10.1016/j.gca.2010.01.027.
Wojcieszek J, Jimenez-Lamana J, Bierla K, Asztemborska M, Ruzik L, Jarosz M, et al. Elucidation of the fate of zinc in model plants using single particle ICP-MS and ESI tandem MS. J Anal At Spectrom. 2019;34:683–93. https://doi.org/10.1039/c8ja00390d.
Fragni R, Trifirò A, Nucci A, Seno A, Allodi A, Di Rocco M. Italian tomato-based products authentication by multi-element approach: a mineral elements database to distinguish the domestic provenance. Food Control. 2018;93:211–8. https://doi.org/10.1016/j.foodcont.2018.06.002.
Alvarez AM, Estévez Alvarez JR, do Nascimento CWA, González IP, Rizo OD, Carzola LL, et al. Lead isotope ratios in lichen samples evaluated by ICP-ToF-MS to assess possible atmospheric pollution sources in Havana, Cuba. Environ Monit Assess. 2017;189:28. https://doi.org/10.1007/s10661-016-5739-8.
Saha A, Deb SB, Saxena MK. Determination of trace impurities in advanced metallic nuclear fuels by inductively coupled plasma time-of-flight mass spectrometry (ICP-TOF-MS). J Anal At Spectrom. 2016;31:1480–9. https://doi.org/10.1039/c6ja00138f.
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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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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). https://doi.org/10.1007/s00216-020-03014-8
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DOI: https://doi.org/10.1007/s00216-020-03014-8
Keywords
- Single particle ICP-MS
- Metallic nanoparticle characterization
- Agrichemical characterization