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NanoSafetyCluster - Compendium 2012 Figure 4: AF4 fractogram of 100 nm SiO 2 with light scattering detection with different injected volumes (blue 100µL, red 50µL, green 10µL), and respective rms radius fit (triangles). The retention time of the 50 and 10 µL were artificially shifted for better readability. 4.4 WP4: Coupled separation / characterisation methods for organic and functionalised ENPs OBJECTIVES In the case that engineered organic nanoparticles are present in foods their identity and quantity needs to be determined, e.g. for proper exposure assessments or the testing for any (future) legal limits. The aim of WP4 is to develop respective methods for the detection and characterisation of organic nanoparticles in food, including sampling, sample preparation, analytical separation and instrumental detection. • Sampling and sample preparation methods for organic and surface functionalised engineered nanoparticles in food matrices • Validated combined separation and detection methods for organic/functionalised engineered nanoparticles in food matrices based on flow separation techniques and mass spectrometry ACTIVITIES The detection and identification of organic, and functionalized, engineered nanoparticles (ENPs) in food items is difficult since the shell of organic engineered nanoparticles is often of a similar nature as that of many food constituents (e.g. proteins, lipids, carbohydrates). A first necessity is therefore the availability of a technique capable of discriminating between organic engineered nanoparticles and residues of matrix constituents. Techniques with that potential will be selected and sampling, sample preparation and separation/fractionation methods for organic engineered nanoparticles from food will be developed from there. Finally, a separation and detection technique will be combined and validated as a complete method for the detection and characterization of organic engineered nanoparticles in food. RESULTS A liquid chromatography method with mass spectrometric detection (LC-MS/MS) has been established for fullerenes in food samples. A differential mobility analyser (DMA) to separate nanoparticles on size is operational. Nanoparticles in suspension are nebulised and ionized with a system that is comparable with electron spray ionisation in LC/MS analysis. The resolution of the system appears to be good, also for small (>20 nm) particles. At the same time a nanoparticle database for MALDI-TOF is being prepared and additional particles are tested to add them to the database. Suspensions of these particles, and later on sample extracts, are separated using hydrodynamic chromatography to isolate the fraction containing the particles of interest. These are then off-line analysed with MALDI-TOF. In addition to that, if the MALDI-TOF is not specific enough, or if compounds are not ionised in MALDI-TOF, mass fingerprinting using TOFMS is being studied. First results with casein as a model particle show that it can be characterised by MALDI-TOF, but much more in detail (type of casein) by fingerprinting/TOFMS. 138 Compendium of Projects in the European NanoSafety Cluster Vitamin E -CH2-CH2-O- Oxyethylene Δ m/z 44 Figure 5 : MALDI-ToF spectrum of a polysorbate micelle loaded with vitamin E 4.5 WP5: Dissemination and training OBJECTIVES The NanoLyse project intends to exploit the knowledge which is generated within the project in the most beneficial way in various aspects. This includes consumer food safety, competitiveness of the European economy as well as scientific progress. Main goal of WP5 is to ensure that the knowledge and methods which are developed within NanoLyse are distributed to stakeholders and exploited in a proper way. • Active dissemination of results to stakeholders and scientists via website, newsletter, publications and presentations at stakeholder and scientific events • Technology transfer and training to consortium members and external end-users (governmental, education, industry) ACTIVITIES WP5 addresses the key knowledge transfer activities: transfer to the scientific community, to risk assessors and policy makers (e.g. EFSA, DG Sanco, national Food Safety Authorities) and potential users of the developed analytical tools (statutory laboratories, food analysis contract laboratories, food industry, SMEs), but also within the project consortium.
Core elements of the dissemination strategy are (i) The public NanoLyse website (www.NanoLyse.eu) The website is a main portal for the dissemination of the results dedicated to general public, official authorities, food and feed sectors and scientists. (ii) The e-Newsletter Bi-annual e-newsletter is distributed actively to stakeholders and other interested parties. The newsletter informs on the scientific progress in the analysis of engineered nanoparticles in food and beverages within NanoLyse, as well as on upcoming events relevant for this field. (iii) Presentation of results The scientific outcome of the project will be published in peer reviewed international journals and presented at international scientific conferences, after careful consideration of IPR issues. In addition, two “NanoLyse Open Days” are organised to present the approach and the outcome of the project to a wider public, focusing on potential stakeholders of the developed methods. (iv) Training workshops will be organised at the end of the project. Goal of these hands-on workshops will be the technology transfer of the developed tools to laboratories which have or will have the need to analyse food and beverages for presence and levels of engineered nanoparticles. In the first place laboratories involved in the risk assessment and in the (future) monitoring of food for engineered nanoparticles will be addressed, but interested parties from food industry and private contract food laboratories will also be invited. 5 NanoLyse Project outcomes 5.1 Scientific publications Thomas P.J. Linsinger, Gert Roebben, Conxita Solans, Roland Ramsch (2011), Reference materials for measuring the size of nanoparticles, Trends in Analytical Chemistry 30:18-27 Agnieszka Dudkiewicz, Karen Tiede, Alistair B.A. Boxall, Katrin Loeschner, Louise Helene Soegaard Jensen, Eric Jensen, Rafal Wierzbicki, Kristian Molhave (2011), Characterisation of nanomaterials in food by electron microscopy, Trends in Analytical Chemistry 30:28-43 Ruud Peters, Guillaime ten Dam, Hans Bouwmeester, Hans Helsper, Günter Allmaier, Frank vd Kammer, Roland Ramsch, Conxita Solans, Monika Tomaniová, Jana Hajslova, Stefan Weigel 6 Directory Table 1 Directory of people involved in this project. NanoSafetyCluster - Compendium 2012 (2011), Identification and characterization of organic nanoparticles in food, Trends in Analytical Chemistry 30:100-112 Frank Von der Kammer, Samuel Legros (2011), Separation and characterization of nanoparticles in complex samples (food/environment) by FFF, Trends in Analytical Chemistry 30:425- 436 Günter Allmaier, Anne Maißer, Christian Laschober, Paul Messner, Wladyslaw W. Szymanski (2011), Parallel differential mobility analysis for electrostatic characterization and manipulation of nanoparticles and viruses, Trends in Analytical Chemistry 30:123-132 5.2 Open days The first NanoLyse Open Day had been organised on the 2 nd November 2011, during the 5 th International Symposium on Recent Advances in Food Analysis (RAFA 2011), held from the 1 st to the 4 th November 2011 in Prague, Czech Republic. The Open day was attended by more than 50 participants from various countries and different sectors as well as from national and European authorities. They actively engaged in discussions with the present NanoLyse scientists. The level of interaction was remarkably high and there was an intense exchange not only on the presented goals and results of the project, but also about future needs and possible collaborations. Figure 6 : Impression of the NanoLyse Open Day 5.3 Training workshops Training workshops are scheduled for the second half of 2013 for the technology transfer of the developed methods to potential end-users of the new methods for the analysis of engineered nanoparticles in food and other complex matrices. First Name Last Name Affiliation Address e-mail Stefan Weigel (Coordinator) RIKILT – Institute of Food Safety (part of Wageningen University and Research Centre / Stichting DLO) Akkermaalsbos 2 6708 WB Wageningen The Netherlands stefan.weigel@wur.nl Compendium of Projects in the European NanoSafety Cluster 139
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Compendium of Projects in the Europ
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EDITED BY Michael Riediker, PD Dr.s
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CONTENTS NanoSafetyCluster - Compen
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Project Acronym ENNSATOX ENPRA EURO
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Foreword NanoSafetyCluster - Compen
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ENNSATOX ENgineered Nanoparticle Im
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The biological membrane and its dep
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distinguishes between the interacti
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Figure 10. Schematic representation
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WP5: • Permeability of NPs in hyd
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NanoSafetyCluster - Compendium 2012
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ENPRA RISK ASSESSMENT OF ENGINEERED
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vitro findings with in vivo models;
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protocols with real biological samp
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and it was organised in the frame o
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First Name Last Name Affiliation Ad
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EURO-NanoTox European Center for Na
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silver nanoparticles, iron nanopart
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The core functions of the Center, h
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8 Directory Table 1 Directory of pe
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HINAMOX Health Impact of Engineered
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High Resolution Electron Microscopy
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integrated risk assessment. Integra
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powders: What is the difference in
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InLiveTox Intestinal, Liver and End
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- to develop and validate a novel m
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environments, IEEE Industrial Elect
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INSTANT Innovative Sensor for the f
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complementary research which will l
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7 Copyright NanoSafetyCluster - Com
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ITS-NANO Intelligent testing strate
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approach to conveying the appropria
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in the discussion. The integration
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MARINA MANAGING RISKS of NANOMATERI
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for risk analysis, these tools need
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environmental ENM toxicity, but tox
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propose it as a Method Validation F
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and SiO2-NM200 and NM203, and a SiO
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ModNanoTox Modelling nanoparticle t
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3) To model environmental exposure
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Nanodetector European Network on th
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measurement interval. Measurements
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5 Directory Table 1 Directory of pe
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NANODEVICE Novel Concepts, Methods,
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Timing, hosts and locations of (grouped) events of NanoImpactNet

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