Characterization of Nanomaterials in Complex Matrices Hans Bouwmeester Senior scientist / special Associate Professor Wageningen UR, The Netherlands Hans.bouwmeester@wur.nl
Nanoparticles in the food chain EFSA supporting publication 2014:EN-621
Nanoparticles characterization Misra S et al., 2012 3
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
Outline Analytical methods (for detection of NMs in complex matrices) Selection of biological models Advanced in vitro models: Gut-on-a-chip 5
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
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 2012 7
AF4: Flow field flow fractionation (asymmetric flow field flow fractionation) - separation based on a perpendicular flow. Smaller particles elute earlier. 8
Diameter from MALS (nm) Multi-detector AF4+MALS+ICP-MS Covering size range 10-500 nm with good resolution Size measurement on-line (MALS), element specific mass measurement on-line (ICP-MS), e.g. mass-based particle size distribution 0.03 0.025 500 450 400 28 Si Signal intensity 0.02 0.015 0.01 0.005 350 300 250 200 150 100 50 0 0 10 20 30 40 50 60 70 80 90 time in min 0
Single particle ICP-MS: Counting and sizing
cps prt 3 ms Single particle ICP-MS 200000 180000 160000 140000 120000 100000 80000 60000 40000 20000 0 0 20 40 rt [sec]
Transforming mass-based to number-based distributions Mass-based size distribution Number-based size distribution
HDC coupled with spicp-ms - aggregation, agglomeration and dissolution studies D HDC D spicpms 14
Organic nanoparticles in food: HDC with MALDI-TOFMS
Organic nanoparticles: Measure shell or active ingredient
Collected HDC fractions are analysed with MALDI-TOFMS
Coatsome nanoparticles in orange juice HDC separation of particle and matrix MALDI-TOFMS identification of particle
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
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
Application in biology and risk assessment 21
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
Uptake of silver by cells: ICP-MS and confocal Caco-2 MCF-7 control ions 20 nm 60 nm 100 nm
in vitro intestinal epithelium on a microfluidics chip
~3 µm ~6 µm ~18 µm Caco-2 in CHIP ~15 µm ~18 µm 25
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
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
Funding and (inter)national collaborations Hans.bouwmeester@wur.nl