TY - JOUR A1 - Cruz-Alonso, M. A1 - Fernandez, B. A1 - Álvarez, L. A1 - González-Iglesias, H. A1 - Traub, Heike A1 - Jakubowski, Norbert A1 - Pereiro, R. T1 - Bioimaging of metallothioneins in ocular tissue sections by laser ablation-ICP-MS using bioconjugated gold nanoclusters as specific tags JF - Microchimica Acta N2 - An immunohistochemical method is described to visualize the distribution of metallothioneins 1/2 (MT 1/2) and metallothionein 3 (MT 3) in human ocular tissue. It is making use of (a) antibodies conjugated to gold nanoclusters (AuNCs) acting as labels, and (b) laser ablation (LA) coupled to inductively coupled plasma – mass spectrometry (ICP-MS).Water-soluble fluorescent AuNCs (with an average size of 2.7 nm) were synthesized and then conjugated to antibody by carbodiimide coupling. The surface of the modified AuNCs was then blocked with hydroxylamine to avoid nonspecific interactions with biological tissue. Immunoassays for MT 1/2 and MT 3 in ocular tissue sections (5 μm thick) from two post mortem human donors were performed. Imaging studies were then performed by fluorescence using confocal microscopy, and LA-ICP-MS was performed in the retina to measure the signal for gold. Signal amplification by the >500 gold atoms in each nanocluster allowed the antigens (MT 1/2 and MT 3) to be imaged by LA-ICP-MS using a laser spot size as small as 4 μm. The image patterns found in retina are in good agreement with those obtained by conventional fluorescence immunohistochemistry which was used as an established reference method. KW - Metal nanoclusters KW - Fluorescence KW - Protein imaging KW - Thin tissue sections KW - Immunohistochemistry KW - Bioconjugation KW - Carbodiimide crosslinking KW - Laser ablation KW - Mass spectrometry PY - 2018 DO - https://doi.org/10.1007/s00604-017-2597-1 VL - 185 IS - 1 SP - 1 EP - 9 PB - Springer AN - OPUS4-44022 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Laux, P. A1 - Tentschert, J. A1 - Riebeling, Ch. A1 - Braeuning, A. A1 - Creutzenberg, O. A1 - Epp, A. A1 - Fessard, V. A1 - Haas, K.-H. A1 - Haase, A. A1 - Hund-Rinke, K. A1 - Jakubowski, Norbert A1 - Kearns, P. A1 - Lampen, A. A1 - Rauscher, H. A1 - Schoonjans, R. A1 - Störmer, A. A1 - Thielmann, A. A1 - Mühle, U. A1 - Luch, A. T1 - Nanomaterials: certain aspects of application, risk assessment and risk communication JF - Archives of Toxicology N2 - Development and market introduction of new nanomaterials trigger the need for an adequate risk assessment of such products alongside suitable risk communication measures. Current application of classical and new nanomaterials is analyzed in context of regulatory requirements and standardization for chemicals, food and consumer products. The challenges of nanomaterial characterization as the main bottleneck of risk assessment and regulation are presented. In some areas, e.g., quantification of nanomaterials within complex matrices, the establishment and adaptation of analytical techniques such as laser ablation inductively coupled plasma mass spectrometry and others are potentially suited to meet the requirements. As an example, we here provide an approach for the reliable characterization of human exposure to nanomaterials resulting from food packaging. Furthermore, results of nanomaterial toxicity and ecotoxicity testing are discussed, with concluding key criteria such as solubility and fiber rigidity as important parameters to be considered in material development and regulation. Although an analysis of the public opinion has revealed a distinguished rating depending on the particular field of application, a rather positive perception of nanotechnology could be ascertained for the German public in general. An improvement of material characterization in both toxicological testing as well as end-product control was concluded as being the main obstacle to ensure not only safe use of materials, but also wide acceptance of this and any novel technology in the general public. KW - Nanomaterials KW - Toxicity KW - Ecotoxicity KW - Standardization KW - Exposure PY - 2018 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-441096 DO - https://doi.org/10.1007/s00204-017-2144-1 VL - 92 IS - 1 SP - 121 EP - 141 PB - Springer AN - OPUS4-44109 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Meyer, S. A1 - López-Serrano, Ana A1 - Mitze, H. A1 - Jakubowski, Norbert A1 - Schwerdtle, T. T1 - Single-cell analysis by ICP-MS/MS as a fast tool for cellular bioavailability studies of arsenite JF - Metallomics N2 - Single-cell inductively coupled plasma mass spectrometry (SC-ICP-MS) has become a powerful and fast tool to evaluate the elemental composition at a single-cell level. In this study, the cellular bioavailability of arsenite (incubation of 25 and 50 μM for 0-48 h) has been successfully assessed by SC-ICP-MS/MS for the first time directly after re-suspending the cells in water. This procedure avoids the normally arising cell membrane permeabilization caused by cell fixation methods (e.g. methanol fixation). The reliability and feasibility of this SC-ICP-MS/MS approach with a limit of detection of 0.35 fg per cell was validated by conventional bulk ICP-MS/MS analysis after cell digestion and parallel measurement of sulfur and phosphorus. KW - Single-cell analysis KW - ICP-MS/MS KW - Arsenite PY - 2018 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-441105 DO - https://doi.org/10.1039/c7mt00285h SN - 1756-5901 VL - 10 IS - 1 SP - 73 EP - 76 PB - RSC Publ. CY - Cambridge AN - OPUS4-44110 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Pisonero, J. A1 - Bouzas-Ramos, D. A1 - Traub, Heike A1 - Cappella, Brunero A1 - Alvarez-Llamas, C. A1 - Richter, Silke A1 - Mayo, J. C. A1 - Costa-Fernandez, J. M. A1 - Bordel, N. A1 - Jakubowski, Norbert T1 - Critical evaluation of fast and highly resolved elemental distribution in single cells using LA-ICP-SFMS JF - Journal of Analytical Atomic Spectrometry N2 - The analytical potential of a nanosecond laser ablation inductively coupled plasma mass spectrometer (ns-LA-ICP-SFMS) system, equipped with an ultra-fast wash-out ablation chamber, is critically investigated for fast and highly spatially resolved (∼μm) qualitative elemental distribution within single cells. Initially, a low surface roughness (< 10 nm) thin In–SnO2 layer (total coating thickness ∼200 nm) deposited on glass is employed to investigate the size, morphology and overlapping of laser-induced craters obtained at different laser repetition rates, making use of Atomic Force Microscopy (AFM). Conical craters with a surface diameter of about 2 µm and depths of about 100 nm were measured after a single laser shot. Furthermore, the influence of the sampling distance (i.e. distance between the sample surface and the inner sniffer of the ablation chamber) on the LA-ICP-MS ion signal wash-out time is evaluated. A significant decrease of the transient 120Sn+ ion signal is noticed after slight variations (±200 μm) around the optimum sampling position. Ultra-fast wash-outs (< 10 ms) are achieved reducing the aerosol mixing from consecutive laser shots even when operating the laser at high repetition rates (25 – 100 Hz). Fast and highly spatially resolved images of elemental distribution within mouse embryonic fibroblast cells (NIH/3T3 fibroblast cells) and human cervical carcinoma cells (HeLa cells), incubated with gold nanoparticles (Au NPs) and Cd-based quantum dots (QDs), respectively, are determined at the optimized operating conditions. Elemental distribution of Au and Cd in single cells is achieved using a high scanning speed (50 µm/s) and high repetition rate (100 Hz). The results obtained for the distribution of fluorescent Cd-based QDs within the HeLa cells are in good agreement with those obtained by confocal microscopy. The size, morphology and overlapping of laser-induced craters in the fixed cells are also investigated using AFM, observing conical craters with a surface diameter of about 2.5 µm and depths of about 800 nm after a single laser shot. KW - Laser ablation KW - ICP-MS KW - Nanoparticle KW - Atomic Force Microscopy KW - Cell PY - 2018 DO - https://doi.org/10.1039/c8ja00096d SP - 1 EP - 9 PB - Royal Society of Chemistry CY - Cambridge AN - OPUS4-45564 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Moraleja, I. A1 - Mena, M. L. A1 - Lázaro, A. A1 - Neumann, B. A1 - Tejedor, A. A1 - Jakubowski, Norbert A1 - Gómez-Gómez, M. M. A1 - Esteban-Fernández, D. T1 - An approach for quantification of platinum distribution in tissues by LA-ICP-MS imaging using isotope dilution analysis JF - Talanta N2 - Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) has been revealed as a convenient technique for trace elemental imaging in tissue sections, providing elemental 2D distribution at a quantitative level. For quantification purposes, in the last years several approaches have been proposed in the literature such as the use of CRMs or matrix matched standards. The use of Isotope Dilution (ID) for quantification by LA-ICP-MS has been also described, being mainly useful for bulk analysis but not feasible for spatial measurements so far. In this work, a quantification method based on ID analysis was developed by printing isotope-enriched inks onto kidney slices from rats treated with antitumoral Pt-based drugs using a commercial ink-jet device, in order to perform an elemental quantification in different areas from bio-images. For the ID experiments ¹⁹⁴Pt enriched platinum was used. The methodology was validated by deposition of natural Pt standard droplets with a known amount of Pt onto the surface of a control tissue, where could be quantified even 50 pg of Pt, with recoveries higher than 90%. The amount of Pt present in the whole kidney slices was quantified for cisplatin, carboplatin and oxaliplatin-treated rats. The results obtained were in accordance with those previously reported. The amount of Pt distributed between the medullar and cortical areas was also quantified, observing different behavior for the three drugs. KW - Isotopic dilution KW - LA-ICP-MS KW - Quantification KW - Imaging KW - Kidney KW - Pt-based drugs PY - 2018 DO - https://doi.org/10.1016/j.talanta.2017.09.031 SN - 0039-9140 VL - 178 SP - 166 EP - 171 PB - Elsevier AN - OPUS4-43627 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Grunert, B. A1 - Saatz, Jessica A1 - Hoffmann, Katrin A1 - Appler, F. A1 - Lubjuhn, Dominik A1 - Jakubowski, Norbert A1 - Resch-Genger, Ute A1 - Emmerling, Franziska A1 - Briel, A. T1 - Multifunctional rare-earth element nanocrystals for cell labeling and multimodal imaging JF - ACS Biomaterials Science & Engineering N2 - In this work, we describe a simple solvothermal route for the synthesis of Eu3+-doped gadolinium orthovanadate nanocrystals (Eu:GdVO4−PAA) functionalized with poly(acrylic)acid (PAA), that are applicable as cell labeling probes for multimodal cellular imaging. The Eu3+ doping of the vanadate matrix provides optical functionality, due to red photoluminescence after illumination with UV light. The Gd3+ ions of the nanocrystals reduce the T1 relaxation time of surrounding water protons, allowing these nanocrystals to act as a positive MRI contrast agent with a r1 relaxivity of 1.97 mM−1 s−1. Low background levels of Eu3+, Gd3+, and V5+ in biological systems make them an excellent label for elemental microscopy by Laser Ablation (LA)-ICP-MS. Synthesis resulted in polycrystalline nanocrystals with a hydrodynamic diameter of 55 nm and a crystal size of 36.7 nm, which were further characterized by X-ray diffraction (XRD), photoluminescence spectroscopy (PL) and transmission electron microscopy (TEM). The multifunctional nanocrystals were subsequently used for intracellular labeling of both human adipose-derived stem cells (MSCs) and A549 (adenocarcinomic human alveolar basal epithelial) cells. KW - Bioimaging KW - Nanoparticle KW - Multimodal KW - Lanthanide PY - 2018 DO - https://doi.org/10.1021/acsbiomaterials.8b00495 SN - 2373-9878 VL - 4 IS - 10 SP - 3578 EP - 3587 PB - ACS Publications CY - Washington, USA AN - OPUS4-46244 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Bernevic, Bogdan A1 - El-Khatib, Ahmed H. A1 - Jakubowski, Norbert A1 - Weller, Michael G. T1 - Online immunocapture ICP‑MS for the determination of the metalloprotein ceruloplasmin in human serum JF - BMC Research Notes N2 - The human copper-protein ceruloplasmin (Cp) is the major copper-containing protein in the human body. The accurate determination of Cp is mandatory for the reliable diagnosis of several diseases. However, the analysis of Cp has proven to be difficult. The aim of our work was a proof of concept for the determination of a metalloprotein-based on online immunocapture ICP-MS. The immuno-affinity step is responsible for the enrichment and isolation of the analyte from serum, whereas the compound-independent quantitation with ICP-MS delivers the sensitivity, precision, and large dynamic range. Off-line ELISA (enzyme-linked immunosorbent assay) was used in parallel to confirm the elution profile of the analyte with a structure-selective method. The total protein elution was observed with the 32S mass trace. The ICP-MS signals were normalized on a 59Co signal. The human copper-protein Cp could be selectively determined. This was shown with pure Cp and with a sample of human serum. The good correlation with off-line ELISA shows that Cp could be captured and eluted selectively from the anti-Cp affinity column and subsequently determined by the copper signal of ICP-MS. KW - ELISA KW - Affinity chromatography KW - Affinity extraction KW - IgY KW - Chicken antibodies KW - Immunoaffinity extraction KW - Copper KW - Diagnostics PY - 2018 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-446157 UR - https://bmcresnotes.biomedcentral.com/articles/10.1186/s13104-018-3324-7 UR - https://static-content.springer.com/esm/art%3A10.1186%2Fs13104-018-3324-7/MediaObjects/13104_2018_3324_MOESM1_ESM.pdf UR - https://static-content.springer.com/esm/art%3A10.1186%2Fs13104-018-3324-7/MediaObjects/13104_2018_3324_MOESM2_ESM.pdf DO - https://doi.org/10.1186/s13104-018-3324-7 SN - 1756-0500 VL - 11 IS - 1 SP - Article 213, 1 EP - 5 PB - Springer Nature CY - Heidelberg AN - OPUS4-44615 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - López-Serrano Oliver, Ana A1 - Baumgart, S. A1 - Bremser, Wolfram A1 - Flemig, Sabine A1 - Wittke, D. A1 - Grützkau, A. A1 - Luch, A. A1 - Haase, A. A1 - Jakubowski, Norbert T1 - Quantification of silver nanoparticles taken up by single cells using inductively coupled plasma mass spectrometry in the single cell measurement mode JF - Journal of Analytical Atomic Spectrometry N2 - The impact of nanoparticles, NPs, at the single cell level has become a major field of toxicological research and different analytical methodologies are being investigated to obtain biological and toxicological information to better understand the mechanisms of cell–NP interactions. Here, inductively coupled plasma mass spectrometry in the single cell measurement mode (SC-ICP-MS) is proposed to study the uptake of silver NPs, AgNPs, with a diameter of 50 nm by human THP-1 monocytes in a proof-ofprinciple experiment. The main operating parameters of SC-ICP-MS have been optimized and applied for subsequent quantitative analysis of AgNPs to determine the number of particles in individual cells using AgNP suspensions for calibration. THP-1 cells were incubated with AgNP suspensions with concentrations of 0.1 and 1 µg/mL for 4 and 24 hours. The results reveal that the AgNP uptake by THP-1 monocytes is minimal at the lower dose of 0.1 µg/mL (roughly 1 AgNP per cell was determined), whereas a large cell-to-cell variance dependent on the exposure time is observed for a 10 times higher concentration (roughly 7 AgNPs per cell). The method was further applied to monitor the AgNP uptake by THP-1 cells differentiated macrophages incubated at the same AgNP concentration levels and exposure times demonstrating a much higher AgNP uptake (roughly from 9 to 45 AgNPs per cell) that was dependent on exposure concentration and remained constant over time. The results have been compared and validated by sample digestion followed by ICP-MS analysis as well as with other alternative promising techniques providing single cell analysis. KW - Silbernanopartikel KW - ICP-MS KW - Einzelzellanalyse PY - 2018 DO - https://doi.org/10.1039/C7JA00395A SN - 0267-9477 VL - 33 IS - 7 SP - 1256 EP - 1263 PB - Royal Society of Chemistry CY - London AN - OPUS4-45473 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Phukphatthanachai, Pranee A1 - Vogl, Jochen A1 - Traub, Heike A1 - Jakubowski, Norbert A1 - Panne, Ulrich T1 - A new approach of using polyethylene frits for the quantification of sulphur in copper metals by isotope dilution LA-ICP-MS and comparison with conventional IDMS techniques JF - Journal of Analytical Atomic Spectrometry N2 - Polyethylene (PE) frits were used to quantify sulphur in copper and its alloys by isotope dilution combined with LA-ICP-MS as an alternative approach to conventional sample preparation: the copper samples were spiked, the spiked samples were dissolved, the resulting solutions were absorbed in the PE frits and finally the PE frits were analysed by LA-ICP-MS. A prerequisite for such a support material is a low sulphur blank and thus PE was selected for this purpose. The absorption efficiency of the PE frits was studied for varying sulphur amounts ranging from 2 mg S to 80 mg S showing that more than 99.5% of the loaded sulphur was absorbed by the frit. The so prepared PE frits were measured by LA-ICP-MS and yielded a good linearity (R2 ¼ 0.999) for the sulphur ion intensities corresponding to sulphur amounts up to 40 mg S; the associated sensitivity is approximately 3.4 x 10⁴ cps μg⁻¹ for ³²S. For the validation of the developed procedure the reference materials BAM-M376a, BAM-228 and BAM-227 were applied such that 2 μg S, 5 μg S and 11 μg S were absorbed in the PE frits, respectively. These samples were pre-quantified for the adsorbed sulphur amount by external calibration LA-ICP-MS yielding sulphur amounts of 0.9 μg, 5.1 μg and 8.5 μg (quantified for ³²S only), respectively. Relative Standard deviations of the isotope ratios were below 5% in average (n ¼ 3 lines) in all cases (except for the pure spike solution). These samples were then analysed by LA-ICP-IDMS and the measurement results were validated by comparing them with the results obtained by conventional ICP-IDMS. The obtained relative expanded measurement uncertainties ranged between 10% and 26%. Pearson's coefficient was used to express the correlation between both techniques; the obtained value was 0.999 demonstrating a strong correlation. Contrary to most published LA-ICP-IDMS procedures, the developed procedure enables SI-traceability for the measurement results. The metrological traceability to the SI for the sulphur mass fractions in copper was established by an unbroken chain of comparisons, each accompanied by an uncertainty budget. Thus, the measurement results are considered reliable, acceptable and comparable within the stated measurement uncertainty. The metrological traceability chain from the kg down to mass fraction in the samples obtained by LA-ICP-IDMS is presented as well. KW - Laser ablation KW - IDMS KW - Traceability KW - Uncertainty KW - Purity PY - 2018 DO - https://doi.org/10.1039/c8ja00116b SN - 0267-9477 SN - 1364-5544 VL - 33 IS - 9 SP - 1506 EP - 1517 PB - Royal Society of Chemistry CY - Cambridge AN - OPUS4-45899 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - García Fernández, J. A1 - Sánchez-González, C. A1 - Bettmer, J. A1 - Llopi, J. A1 - Jakubowski, Norbert A1 - Panne, Ulrich A1 - Montes-Bayón, M. T1 - Quantitative assessment of the metabolic products of iron oxide nanoparticles to be used as iron supplements in cell cultures JF - Analytica Chimica Acta N2 - Iron nanoparticles (NPs) metabolism is directly associated to human health due to their use as anemia treatment and should be studied in detail in cells. Here we present a speciation strategy for the determination of the metabolic products of iron oxide nanoparticles coated by tartaric and adipic acids in enterocytes-like cell models (Caco-2 and HT-29). Such methodology is based on the use of SDS-modified reversed phase high performance liquid chromatography (HPLC) separation using inductively coupled plasma-mass spectrometry (ICP-MS) as Fe selective detector. Post-column isotope dilution analysis is used as quantification tool by adding Fe-57 as isotopically enriched standard. To assess the separation capability of the method, two different iron nanostructures: iron sucrose nanoparticles -Venofer®- used as model suspension and iron tartrate/adipate-modified nanoparticles, both of about 4 nm (core size) were evaluated. The two nanostructures were injected into the system showing good peak profiles and quantitative elution recoveries (>80%) in both cases. In addition, both nanoparticulate fractions could be based-line separated from ionic iron species, which needed to be complexed with 1mM citrate to elute from the column. Exposed cells up to 0.5mM of iron tartrate/adipate-modified nanoparticles were specifically treated to extract the internalized NPs and the extracts examined using the proposed strategy. The obtained results revealed the presence of three different fractions corresponding to nanoparticle aggregates, dispersed nanoparticles and soluble iron respectively in a single chromatographic run. Quantitative experiments (column recoveries ranging from 60 to 80%) revealed the presence of the majority of the Fe in the nanoparticulated form (>75%) by summing up the dispersed and aggregate particles. Such experiments point out the high uptake and low solubilization rate of the tartrate/adipate NPs making these structures highly suitable as Fe supplements in oral anemia treatments. KW - Fe nanoparticles metabolism KW - Cells KW - HPLC-ICP-MS KW - Species-unspecific on-line isotope dilution PY - 2018 DO - https://doi.org/10.1016/j.aca.2018.08.003 SN - 0003-2670 VL - 1039 SP - 24 EP - 30 PB - Elsevier CY - Amsterdam AN - OPUS4-46817 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -