Reference Materials and labelled nanoparticlesReference Materials and labelled nanoparticlesRi G b 1 A i k D dki i 5 S l L 2 A l L h 6 Th P J Li i 1 G ibi M 3Ringo Grombe1, Agneieszka Dudkiewizcs5, Samuel Legros2, Angela Lehner6, Thomas P.J. Linsinger1 , Guibin Ma3 ,

Jonathan G.C. Veinot3, Stephan Wagner2, Stefan Weigel4, p g , g1 EC-JRC Institute for Reference Materials and Measurements (IRMM) Geel (BE) 2 Universitaet Wien Department of Environmental Geosciences Vienna (AT)1 EC-JRC, Institute for Reference Materials and Measurements (IRMM), Geel (BE) 2 Universitaet Wien, Department of Environmental Geosciences, Vienna (AT)3 University of Alberta, Department of Chemistry, Edmonton (CA) 4 Stichting Dienst Landbouwkundig Onderzoek (RIKILT), Wageningen (NL)3 University of Alberta, Department of Chemistry, Edmonton (CA) 4 Stichting Dienst Landbouwkundig Onderzoek (RIKILT), Wageningen (NL)5 FERA, Sand Hutton, Yor (UK) 6 TU Vienna, Department of Analytical Chemistry, Vienna (AT), , ( ) , p y y, ( )

The project foresees two different kinds of reference materials namely Matrix reference materialsThe project foresees two different kinds of reference materials, namelysuspensions of particles in aqueous media/pure solvents and matrix

Matrix reference materialsFor the first time well characterised nanoparticles were spiked into foodsuspensions of particles in aqueous media/pure solvents and matrix

materialsFor the first time, well-characterised nanoparticles were spiked into foodmatrices This allows checking for recovery as well as for potentialmaterials. matrices. This allows checking for recovery as well as for potentialchanges of particle size during sample preparationchanges of particle size during sample preparation.S i d i ll t id d l ti bl

Labelled nanoparticlesSuspensions were used in all cases to avoid deagglomeration problems

Labelled nanoparticlesA labelled Ag and Ge labelled SiO can be sed as internal standards A NP ik d hi k tAu labelled Ag and Ge labelled SiO2 can be used as internal standardst i t bilit d t l i h th

Ag NP spiked chicken meatt t f t t k it f hi k tto improve repeatability and control recovery, in very much the same

i t i ll l b ll d i t l t d d d i th• tests of water-uptake capacity of chicken meat

ill d hi k ik d i h PVP bili d A NP (Fi 3 )way as isotopically labelled internal standards are used in thed t i ti f i d i i t i t i f d

• cryo-milled chicken meat spiked with PVP stabilised Ag NPs (Fig. 3a)determination of organic and inorganic contaminants in food. • Ag mass fractions: 0.1 and 0.5 g/kg

Au labelled Ag (2 % Au) • rapid freezing over liquid Nitrogen to avoid agglomeration (Fig. 3b)Au labelled Ag (2 % Au)• TEM image: shows roughly spherical particles with particle sizes • storage at -130 °CTEM image: shows roughly spherical particles with particle sizes

ranging from 4 to 24 nm (Figure 1a); DLS: polydisperse with a size • homogeneous for total Agranging from 4 to 24 nm (Figure 1a); DLS: polydisperse with a sizemaximum of 73 nm (Figure 1b); • particles are not agglomerated (TEM image Fig. 3b)maximum of 73 nm (Figure 1b); p gg ( g g )

a) b) c)Ge labelled SiO2 (0.8 % Ge) a) b) c)

• SEM: Spherical particles with a diameter of approximately 100 nm(Fig. 4a). DLS: monomodal; mean diameter about 70 nm (Fig. 2b).

• Mesoporoous SiO2 without crystalline products

a) b) c) d) Fig. 3: Ag spiked chicken paste (a), filled vials (b) and TEM image (c)

) ) ) ) g g p p ( ) ( ) g ( )

SiO2 NP spiked tomato soup• tomato soup prepared from fresh ingredients to ensure absence of NPs• fat skimmed off bouillon; skin and seeds removed (Fig. 4a)( g )• spiked with SiO2 NP dispersion and filled in 50 mL jars (Fig. 4b)

Fig. 1: TEM image and size distribution by DLS of the Au labelled Ag p p j ( g )

• SiO2 mass fractions: 10 and 40 g/kgg g y gparticles (1a, b) and Ge labelled SiO2 (1b, c)

2 g g• sterilisation by -irradiationparticles (1a, b) and Ge labelled SiO2 (1b, c) s e sa o by ad a o• homogeneous as determined by total Si determinationhomogeneous as determined by total Si determination• no agglomerates found by acid digestion-FFF-MALLS (Fig. 4c)

Nanoparticle suspensionsno agglomerates found by acid digestion FFF MALLS (Fig. 4c)

Nanoparticle suspensionsPure suspensions were prepared for training as well as for having a) b) c)Pure suspensions were prepared for training as well as for havingcharacterised spikes for matri RMs

a) b) c) characterised spikes for matrix RMs

• Suspensions of SiO2, Ag, cross-linked gelatin and C60 fullerenes wereSuspensions of SiO2, Ag, cross linked gelatin and C60 fullerenes were preparedprepared

• Homogeneity and stabilty were tested according to ISO Guide 34 and 35;Homogeneity and stabilty were tested according to ISO Guide 34 and 35; for uncertainty of homogeneity see Table 1

Fig. 4: removing coarse fractions from raw soup (a); spiked soup in for uncertainty of homogeneity see Table 1

• Characterisation of Ag and SiO2 by several labs; cross linked gelatinjars (b); FFF-MALLS data fractogram of blank, spike and spiked

• Characterisation of Ag and SiO2 by several labs; cross-linked gelatin contains very small and large particles

soupcontains very small and large particles

p

Cross-linked gelatin NP spiked soft drinkCross-linked gelatin NP spiked soft drink• Cross linked gelatin NPs spiked to Aquarius Green Spalsh™ (Fig 5a b);

a) b) c)• Cross-linked gelatin NPs spiked to Aquarius Green Spalsh™ (Fig. 5a, b);

filled into 20 mL ampoulesa) b) c) filled into 20 mL ampoules• Stabilisation by addition of Ethanol (2 suspensions) or ProClin150 (2• Stabilisation by addition of Ethanol (2 suspensions) or ProClin150 (2

suspensions)suspensions)M f ti f 5 d 20 /k• Mass fractions of 5 and 20 g/kgNP di ti i h bl f t i (Fi 5 d) h d t bl

Fig 2: Nanoparticle suspensions (a) DLS of silica (b)and• NPs distinguishable from matrix (Fig. 5 c, d); homogeneous and stable

Fig. 2: Nanoparticle suspensions (a), DLS of silica (b)and TEM of cross linked gelatin (c)

Size Distribution by Intensity

a) b) c) d)TEM of cross-linked gelatin (c)15

20a) b) c) d)

10

ensi

ty (

%)

SiO SiO Ag Ag Gelatin Gelatin5

Inte

SiO2

10 /k

SiO2

40 /k

Ag

0 2 /k

Ag

1 0 /k

Gelatin

10 /k

Gelatin

40 /k

00.1 1 10 100 1000 10000

Size (d.nm)

10 g/kg 40 g/kg 0.2 g/kg 1.0 g/kg 10 g/kg 40 g/kg

ubb [%] 0.27 0.59 1.1 6.7 0.4 3.8Fig 5: Aquarius ® Green Splash (la) mixed with gelatin nanoparticle dispersion touchar [%] 2.2 2.3 7.3 5.8 n.a. n.a. Fig. 5: Aquarius ® Green Splash (la) mixed with gelatin nanoparticle dispersion to obtain doped beverage matrices (b); DLS distribution of blank Aquarius (c) and NPSize by DLS 143 ± 5 143 ± 15 68 ± 15 66 ± 18 184 690 obtain doped beverage matrices (b); DLS distribution of blank Aquarius (c) and NP

ik d A i (d)[nm] spiked Aquarius (d)

Tab 1: homogeneity and characterisation data of silica,Tab 1: homogeneity and characterisation data of silica, Ag and gelatin NPs Fullerenes in rapeseed oilAg and gelatin NPs

• Rapeseed oil spiked with C60 fullerenes• Poor recovery and homogeneity: adsorption on ampoule walls?

The work leading to these results has receivedThomas |Linsinger The work leading to these results has received funding from the European Union Seventh Framework

Thomas |LinsingerInstitute for Reference Materials and Measurements (IRMM)

www NanoLyse eufunding from the European Union Seventh Framework Programme (FP7/2007 2013) nder grant agreement

( )Retieseweg 111, 2440 Geel, Belgium Ph 32 (0) 14 571 956www.NanoLyse.eu Programme (FP7/2007-2013) under grant agreement

° 245162 ll f H lth C dPhone +32 (0) 14 571 956thomas linsinger@ec europa eu

yn° 245162 as well as from Health Canada. thomas.linsinger@ec.europa.eu