Hans Bouwmeester Senior scientist / special Associate Professor Wageningen UR, The Netherlands

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1 Characterization of Nanomaterials in Complex Matrices Hans Bouwmeester Senior scientist / special Associate Professor Wageningen UR, The Netherlands Hans.bouwmeester@wur.nl

2 Nanoparticles in the food chain EFSA supporting publication 2014:EN-621

3 Nanoparticles characterization Misra S et al.,

4 Definition Commission Recommendation of 18 October 2011 on the definition of nanomaterial (2011/696/EU) Nanomaterial means a natural, incidental or manufactured material containing particles, in an unbound state or as an aggregate or as an agglomerate and where, for 50 % or more of the particles in the number size distribution, one or more external dimensions is in the size range 1 nm-100 nm. 4

5 Outline Analytical methods (for detection of NMs in complex matrices) Selection of biological models Advanced in vitro models: Gut-on-a-chip 5

$Separation Hydrodynamic chromatography Field flow fractionation Counting and sizing Single particle ICPMS$

6 Analytical techniques for nanoparticles in complex matrices Imaging Light scattering Electron microscopy Separation Hydrodynamic chromatography Field flow fractionation Counting and sizing Single particle ICPMS Mobility analysis

7 Hydrodynamic Chromatography (HDC) Separation by size dependent exclusion from wall, diffusion in laminar flow wide dynamic range (<10 to 1000 nm) robust however... (far) lower resolution than AF4 Striegel & Brewer

$AF4: Flow field flow fractionation (asymmetric flow field flow fractionation)$

8 AF4: Flow field flow fractionation (asymmetric flow field flow fractionation) - separation based on a perpendicular flow. Smaller particles elute earlier. 8

10 Diameter from MALS (nm) Multi-detector AF4+MALS+ICP-MS Covering size range nm with good resolution Size measurement on-line (MALS), element specific mass measurement on-line (ICP-MS), e.g. mass-based particle size distribution Si Signal intensity time in min 0

11 Single particle ICP-MS: Counting and sizing

12 cps prt 3 ms Single particle ICP-MS rt [sec]

13 Transforming mass-based to number-based distributions Mass-based size distribution Number-based size distribution

14 HDC coupled with spicp-ms - aggregation, agglomeration and dissolution studies D HDC D spicpms 14

15 Organic nanoparticles in food: HDC with MALDI-TOFMS

16 Organic nanoparticles: Measure shell or active ingredient

17 Collected HDC fractions are analysed with MALDI-TOFMS

18 Coatsome nanoparticles in orange juice HDC separation of particle and matrix MALDI-TOFMS identification of particle

19 Nanoparticles: detection and characterization Huge range of technics to detect and/ or characterize particles One of the most promising, hyphenated methods e.g. HDC-spICP-MS or AF4-spICP-MS HDC-spICP-MS low resolution separation but relatively fast without NP-column interactions AF4-spICP-MS high resolution separation, however, long analysis times, possible NP and matrix interactions with the membrane 19

20 Application in models for the oral exposure route Food and food ingredients Realistic architecture in microfluidic chips Advanced analytics Healthy life, gut health Gut epithelium 20

21 Application in biology and risk assessment 21

22 Key descriptors: linking phys-chem and biology Nanomaterial Dissolution of NMs in biological matrix methods based on ICP-MS, elementary detection Ultrafiltration (UF) ICP-MS 22

23 Uptake of silver by cells: ICP-MS and confocal Caco-2 MCF-7 control ions 20 nm 60 nm 100 nm

24 in vitro intestinal epithelium on a microfluidics chip

25 ~3 µm ~6 µm ~18 µm Caco-2 in CHIP ~15 µm ~18 µm 25

26 Gut-on-a-chip Personalized (stem) cell biology Microbiome ips organoids human biopts microbiome On chip, on line testing Chip sensor technology 3D extracellular environment Advanced microchips

27 Acknowledgements Research staff Post docs dr ir H (Hans) Bouwmeester AK (Anna) Undas, MSc S (Steffi) Böhme, PhD dr HJP (Hans) Marvin Y (Yamine) Bouzembrak, PhD dr RJB (Ruud) Peters C (Claudia) Cascio, PhD dr S (Stefan) Weigel PhD A (Ashraf) Abdelkhaliq KN (Karrar) Mahdi dr M (Meike) van der Zande K (Kornphimol) Kulthong Technicians dr ing MK (Martijn) van der Lee ing MEM (Greet) van Bemmel dr ing SPJ (Stefan) van Leeuwen ing SM (Sandra) Munniks JR (Richard) Helsdingen 27

28 Funding and (inter)national collaborations

Nanoparticles in food and non food Recent methods and measurements

Nanoparticles in food and non food Recent methods and measurements Ruud Peters, Stefan Weigel, Hans Marvin and Hans Bouwmeester RIKILT Institute of Food Safety, Wageningen, The Netherlands ruudj.peters@wur.nl,