Study of the Stability, Uptake and Transformations of Zero Valent Iron Nanoparticles in a Model Plant by Means of an Optimised Single Particle ICP-MS/MS Method.

In the context of the widespread distribution of zero valent iron nanoparticles (nZVI) in the environment and its possible exposure to many aquatic and terrestrial organisms, this study investigates the effects, uptake, bioaccumulation, localisation and possible transformations of nZVI in two different forms (aqueous dispersion-Nanofer 25S and air-stable powder-Nanofer STAR) in a model plant-Arabidopsis thaliana. Seedlings exposed to Nanofer STAR displayed symptoms of toxicity, including chlorosis and reduced growth. At the tissue and cellular level, the exposure to Nanofer STAR induced a strong accumulation of Fe in the root intercellular spaces and in Fe-rich granules in pollen grains. Nanofer STAR did not undergo any transformations during 7 days of incubation, while in Nanofer 25S, three different behaviours were observed: (i) stability, (ii) partial dissolution and (iii) the agglomeration process. The size distributions obtained by SP-ICP-MS/MS demonstrated that regardless of the type of nZVI used, iron was taken up and accumulated in the plant, mainly in the form of intact nanoparticles. The agglomerates created in the growth medium in the case of Nanofer 25S were not taken up by the plant. Taken together, the results indicate that Arabidopsis plants do take up, transport and accumulate nZVI in all parts of the plants, including the seeds, which will provide a better understanding of the behaviour and transformations of nZVI once released into the environment, a critical issue from the point of view of food safety.

To-Do and Not-To-Do in Model Studies of the Uptake, Fate and Metabolism of Metal-Containing Nanoparticles in Plants.

Due to the increasing release of metal-containing nanoparticles into the environment, the investigation of their interactions with plants has become a hot topic for many research fields. However, the obtention of reliable data requires a careful design of experimental model studies. The behavior of nanoparticles has to be comprehensively investigated; their stability in growth media, bioaccumulation and characterization of their physicochemical forms taken-up by plants, identification of the species created following their dissolution/oxidation, and finally, their localization within plant tissues. On the basis of their strong expertise, the authors present guidelines for studies of interactions between metal-containing nanoparticles and plants.

Analysis of cerium oxide and copper oxide nanoparticles bioaccessibility from radish using SP-ICP-MS.

BACKGROUND: The transformation of nanoparticles (NPs) internalized in plant tissues is the human digestive system that can provide a better understanding of the impact of NPs on the human system. The presented methodology was developed to study the bioaccessibility of cerium oxide (CeO2 ) and copper oxide (CuO) NPs from radish after the in vitro simulation of gastrointestinal digestion using single-particle inductively coupled plasma mass spectrometry (SP-ICP-MS). RESULTS: Radish plants were cultivated hydroponically in a growth medium containing: (i) CeO2 NPs and (ii) CuO NPs. Both cerium (Ce) and copper (Cu) were found in all organs of the radish plants after analysis by standalone ICP-MS. This confirms the bioaccumulation of CeO2 and CuO NPs and the translocation of their Ce and Cu to the aerial parts of the plant. Less Ce (4.095 µg g-1 ) has been detected in leaves than in roots (1.156 mg g-1 ) while Cu content in leaves was 5.245 µg g-1 and in roots was 10.41 µg g-1 . Analysis of the digestive extracts obtained after the in vitro simulation of gastro (pepsin) and gastrointestinal (pancreatin) digestion showed that Ce has easy access to human system at least by 73%. CONCLUSION: The size of CeO2 NPs in digestive extracts showed no significant changes. However, the results obtained for CuO NPs digestion were variable and suggested that CuO NPs dissolved during the digestion process. The CuO NPs were observed in roots after the gastrointestinal digestion concluding that CuO NPs recovered after the initial dissolution. © 2020 Society of Chemical Industry.

Uptake, translocation, size characterization and localization of cerium oxide nanoparticles in radish (Raphanus sativus L.).

Due to their unique physical and chemical properties, the production and use of cerium oxide nanoparticles (CeO2 NPs) in different areas, especially in automotive industry, is rapidly increasing, causing their presence in the environment. Released CeO2 NPs can undergo different transformations and interact with the soil and hence with plants, providing a potential pathway for human exposure and leading to serious concerns about their impact on the ecosystem and human organism. This study investigates the uptake, bioaccumulation, possible translocation and localization of CeO2 NPs in a model plant (Raphanus sativus L.), whose edible part is in direct contact with the soil where contamination is more likely to happen. The stability of CeO2 NPs in plant growth medium as well as after applying a standard enzymatic digestion procedure was tested by single particle ICP-MS (SP-ICP-MS) showing that CeO2 NPs can remain intact after enzymatic digestion; however, an agglomeration process was observed in the growth medium already after one day of cultivation. An enzymatic digestion method was next used in order to extract intact nanoparticles from the tissues of plants cultivated from the stage of seeds, followed by size characterization by SP-ICP-MS. The results obtained by SP-ICP-MS showed a narrower size distribution in the case of roots suggesting preferential uptake of smaller nanoparticles which led to the conclusion that plants do not take up the CeO2 NPs agglomerates present in the medium. However, nanoparticles at higher diameters were observed after analysis of leaves plus stems. Additionally, a small degree of dissolution was observed in the case of roots. Finally, after CeO2 NPs treatment of adult plants, the spatial distribution of intact CeO2 NPs in the radish roots was studied by laser ablation ICP-MS (LA-ICP-MS) and the ability of NPs to enter and be accumulated in root tissues was confirmed.

Speciation analysis and bioaccessibility evaluation of trace elements in goji berries (Lycium Barbarum, L.).

Goji berries (Lycium Barbarum, L.) are known for their nutritional potential as a great source of trace metals (e.g., copper, zinc and manganese) which are present in the form of highly bioaccessible compounds. In order to assess the bioaccessibility of trace elements and to identify compounds responsible for better bioaccessibility of copper and zinc, an in vitro simulation of gastrointestinal digestion was used in this study. The total content of trace metals was evaluated using sample digestion followed by inductively coupled plasma mass spectrometry. Bioaccessibility of trace elements was estimated by size exclusion chromatography coupled to inductively coupled plasma mass spectrometry. These analytical methods were used to analyse samples of goji berries to determine the highest amount of elements. For total trace metal content in goji berries, Zn had the highest level of the three studied (10.6µgg-1), while the total content of manganese and copper was 9.9µgg-1 and 6.1µgg-1, respectively. Additionally, the analysed metals were found to be highly bioaccessible to the human body (about 56% for Mn, 72% for Cu and 64% for Zn in the gastric extract and approximately 35% for Mn, 23% for Cu and 31% for Zn in the case of gastrointestinal extract). To obtain information about metal complexes present in goji berries, extraction treatment using different solutions (ionic liquid, HEPES, SDS, Tris-HCl, ammonium acetate, water) was performed. Enzymatic treatment using pectinase and hemicellulase was also checked. Extracts of berries were analysed by SEC-ICP-MS and µHPLC-ESI-MS/MS techniques. The ionic liquid and pectinase extraction helped efficiently extract copper (seven compounds) and zinc (four compounds) complexes. Compounds identified in goji berries are most likely to be responsible for better bioaccessibility of those elements to the human organism.

In vitro digestion method for estimation of copper bioaccessibility in Açaí berry.

ABSTRACT: Copper is an essential trace element for humans and its deficiency can lead to numerous diseases. A lot of mineral supplements are available to increase intake of copper. Unfortunately, only a part of the total concentration of elements is available for human body. Thus, the aim of the study was to determine bioaccessibility of copper in Açai berry, known as a "superfood" because of its antioxidant qualities. An analytical methodology was based on size exclusion chromatography (SEC) coupled to a mass spectrometer with inductively coupled plasma (ICP MS) and on capillary liquid chromatography coupled to tandem mass spectrometer with electrospray ionization (µ-HPLC-ESI MS/MS). To extract various copper compounds, berries were treated with the following buffers: ammonium acetate, Tris-HCl, and sodium dodecyl sulfate (SDS). The best extraction efficiency of copper was obtained for SDS extract (88 %), while results obtained for Tris-HCl and ammonium acetate were very similar (47 and 48 %, respectively). After SEC-ICP-MS analysis, main signal was obtained for all extracts in the region of molecular mass about 17 kDa. A two-step model simulated gastric (pepsin) and gastrointestinal (pancreatin) digestion was used to obtain the knowledge about copper bioaccessibility. Copper compounds present in Açai berry were found to be highly bioaccessible. The structures of five copper complexes with amino acids such as aspartic acid, tyrosine, phenylalanine, were proposed after µ-HPLC-ESI MS/MS analysis. Obtained results show that copper in enzymatic extracts is bound by amino acids and peptides what leads to better bioavailability of copper for human body.

Ionic liquids as a key medium for efficient extraction of copper complexes from chia seeds (Salvia hispanica L.).

Due to insufficient information, the aim of study was to concern on the optimization of extraction procedure of selected metal complexes with flavonoids from chia seeds. Evaluation of the amount of elements in compound, not only their total concentration content, is highly important due to the fact, that only a part from total content of metal is absorbed by human body. At the beginning the total amount of elements in chia seeds was established as 14.51±0.42 µg g(-1) for copper, 57.44±1.23 µg g(-1) for manganese, 81.12±1.89 µg g(-1) for zinc and 0.35±0.13 µg g(-1) for cobalt. After the most suitable solvent was established, effects of several parameters on the efficiency of metal extraction were studied. Solvent concentration, solid-solvent ratio, extraction method, extraction time and temperature have been investigated as independent variables. The optimal extraction conditions included vortexing during 20 min in 50°C, using an ionic liquid (1-butyl-3-methylimidazolium bromide) as an extractant, with solid-solvent ratio of 1:20. The determination of total and extractable amount of metals in chia seeds was carried out by standalone ICP MS. In addition, a complementary analysis of extracted metal complexes was performed using SEC-ICP MS method. It was confirmed that the ionic liquid is able to extract different copper complexes in comparison with commonly used solvents. The study indicated that extraction by using an ionic liquid has been successfully applied for determination of metals and metal complexes in chia seeds.

Operationally defined species characterization and bioaccessibility evaluation of cobalt, copper and selenium in Cape gooseberry (Physalis Peruviana L.) by SEC-ICP MS.

Physalis peruviana could attract great interest because of its nutritional and industrial properties. It is an excellent source of vitamins, minerals, essential fatty acids and carotenoids. Physalis Peruviana is also known to have a positive impact on human health. Unfortunately, still little is known about trace elements present in Physalis Peruviana and their forms available for the human body. Thus, the aim of this study was to estimate bioaccessibility and characterization of species of cobalt, copper and selenium in Physalis Peruviana fruits. Total and extractable contents of elements were determined by mass spectrometer with inductively coupled plasma (ICP MS). In order to separate the different types of metal complexes Physalis peruviana fruits were treated with the following solvents: Tris-HCl (pH 7.4), sodium dodecyl sulfate (SDS) (pH 7.4) and ammonium acetate (pH 5.5). The best efficiency of extraction of: cobalt was obtained for ammonium acetate (56%) and Tris-HCl (60%); for copper was obtained for SDS (66%), for selenium the best extraction efficiency was obtained after extraction with SDS (48%). To obtain information about bioaccessibility of investigated elements, enzymatic extraction based on in vitro simulation of gastric (pepsin) and intestinal (pancreatin) digestion was performed. For copper and selenium the simulation of gastric digestion leads to the extraction yield above 90%, while both steps of digestion method were necessary to obtain satisfactory extraction yield in the case of cobalt. Size exclusion chromatography (SEC) coupled to on-line ICP MS detection was used to investigate collected metal species. The main fraction of metal compounds was found in the 17 kDa region. Cobalt and copper create complexes mostly with compounds extracted by means of ammonium acetate and SDS, respectively. Cobalt, copper and selenium were found to be highly bioaccessible from Physalis Peruviana. Investigation of available standards of cobalt and selenium allows confirming the presence of vitamin B12 and probably selenomethionine in the fraction bioaccessible by human body (obtained during enzymatic extraction). It should be noted that the presence of small seleno-compounds in Cape gooseberry was performed for the first time. The results show that the combination of SEC and ICP MS could provide a simple method for separating of soluble element species.

Comparison of copper and zinc in vitro bioaccessibility from cyanobacteria rich in proteins and a synthetic supplement containing gluconate complexes: LC-MS mapping of bioaccessible copper complexes.

An analytical procedure was proposed to estimate bioaccessibility of copper and zinc in Spirulina Pacifica tablets with respect to that of copper and zinc in gluconate complexes. Spirulina is the common name for diet supplements produced primarily from two species of cyanobacteria, namely Arthrospira platensis and Arthrospira maxima. Spirulina tablets are an excellent source of proteins, vitamins and minerals. To obtain information about the bioavailability of these elements, an in vitro bioaccessibility test was performed by application of a two-step protocol which simulated the gastric (pepsin) and intestinal (pancreatin) digestion. The species obtained were investigated by size exclusion chromatography on a chromatograph coupled to a mass spectrometer with inductively coupled plasma (SEC-ICP-MS) and an on-capillary liquid chromatograph coupled to an electrospray mass spectrometer (µ-HPLC-ESI-MS). Both copper and zinc were found to be highly bioaccessible in Spirulina tablets (90-111%) and those containing gluconate complexes (103% for Cu and 62% for Zn). In Spirulina tablets, copper was found to form two types of complex: (1) polar ones with glycine and aspartic acid and (2) more hydrophobic ones containing amino acids with cyclic hydrocarbons (phenylalanine, histidine, proline and tyrosine). Zinc and copper were also proved to form complexes during the digestion process with products of pepsin digestion, but the stability of these complexes is lower than that of the complexes formed in Spirulina. The results proving the involvement of proteins in the enhancement of copper and zinc bioaccessibility will be useful for the design of new copper and zinc supplements.