Characterisation of titanium dioxide (nano)particles in foodstuffs and E171 additives by single particle inductively coupled plasma-tandem mass spectrometry using a highly efficient sample introduction system.

This study addressed primarily the characterisation and quantification of titanium dioxide (TiO2) (nano)particles (NPs) in a large variety of commercial foodstuffs. The samples were purchased from local markets in Spain before the ban of TiO2 food additive (E171) in the EU. The analyses were carried out by single particle inductively coupled plasma-tandem mass spectrometry (spICP-MS/MS) in mass shift mode (oxidation of 48Ti to 48Ti16O (m/z = 64)) and using a highly efficient sample introduction system (APEX™ Ω). This novel analytical approach allowed accurate characterisation of a large panel of TiO2 NPs sizes ranging from ∼12 to ∼800 nm without isobaric interferences from 48Ca isotope, which is highly abundant in most of the analysed foodstuffs. TiO2 NPs were extracted from foodstuffs using sodium dodecyl sulphate (0.1%, w/v) and diluted with ultra-pure water to reach ∼ 1000 particles signals per acquisition. All the analysed samples contained TiO2 NPs with concentrations ranging from 1010 to 1014 particles kg-1, but with significant low recoveries compared to the total Ti determination. A selection of samples was also analysed using a similar spICP-MS/MS approach with a conventional sample introduction system. The comparison of results highlighted the improvement of the limit of detection in size (12 nm) by the APEX™ Ω system, providing nanoparticulate fractions ranging from ∼4% (cheddar sauce) up to ∼87% (chewing gum), which is among the highest nanoparticulate fractions reported in literature using a spICP-MS approach. In addition, two commercially available E171 additives were analysed using the previous approaches and other techniques in different European laboratories with the aim of methods inter-comparison. This study provides occurrence data related to TiO2 NPs in common commercial foodstuffs but it also demonstrates the potential of the novel analytical approach based on APEX™-ICP-MS/MS to characterise nano-size TiO2 particles in complex matrices such as foodstuffs.

Assessment of TiO2 (nano)particles migration from food packaging materials to food simulants by single particle ICP-MS/MS using a high efficiency sample introduction system.

TiO2 is the most widely used white pigment in plastics and food packaging industry, thus the question of its migration towards food and hence the impact on consumers is raised. Since recent research indicate its potential toxicity, it is necessary to study TiO2 contamination as a consequence of food storage. For this purpose, plastic containers from commercially-available dairy products and custom-made TiO2-spiked polypropylene materials were put in contact with 50% (v/v) ethanol and 3% (w/v) acetic acid, which were used here as food simulants. The migration assays were carried out under standard contact conditions of packaging use (as recommended by Commission Regulation (EU) N° 10/2011 for food contact migration testing), and under conditions of extreme mechanical degradation of the packaging. The TiO2 (nano)particles released in the food simulants were analysed by single particle inductively coupled plasma-tandem mass spectrometry in mass-shift mode and using a high efficiency sample introduction system (APEX™ Ω) to avoid matrix effects from food simulants. For the dairy product containers and for the spiked polypropylene, results showed release of TiO2 particles of rather large sizes (average size: 164 and 175 nm, respectively) under mechanical degradation conditions, i.e. when the polymeric structure is damaged. The highest amounts of TiO2 were observed in 50% ethanol after 10 days of storage at 50 °C (0.62 ng cm-2) for the dairy product containers and after 1 day of storage at 50 °C (0.68 ng cm-2) for the spiked polypropylene. However, the level of Ti released in particle form was very small compared to the total Ti content in the packaging and far below the acceptable migration limits set by European legislation. Release under standard contact conditions of use of the container was not measurable, thus the migration of TiO2 particles from this packaging to dairy products among storage is expected to be negligible.

Traceable characterization of hollow organosilica beads as potential reference materials for extracellular vesicle measurements with optical techniques.

The concentration of cell-type specific extracellular vesicles (EVs) is a promising biomarker for various diseases. However, concentrations of EVs measured by optical techniques such as flow cytometry (FCM) or particle tracking analysis (PTA)  in clinical practice are incomparable. To allow reliable and comparable concentration measurements suitable reference materials (RMs) and SI-traceable (SI-International system of units) methods are required. Hollow organosilica beads (HOBs) are promising RM candidates for concentration measurements of EVs based on light scattering, as the shape, low refractive index, and number concentration of HOBs are comparable to EVs of the respective size range that can be detected with current optical instrumentation. Here, we present traceable methods for measuring the particle size distribution of four HOB types in the size range between 200 and 500 nm by small-angle X-ray scattering (SAXS) and atomic force microscopy (AFM), as well as the number concentration by single-particle inductively coupled plasma mass spectrometry (spICP-MS). Based on the size and shape results, traceable reference values were obtained to additionally determine the refractive index of the shell of the HOB samples by FCM. Furthermore, the estimated refractive indexes of the HOBs plausibly agree with the refractive indexes of EVs of corresponding size. Due to their narrow size distribution and their similar shape, and low refractive index, all HOB samples studied are suitable RM candidates for calibration of the measured sample volume by optical methods within the photon wavelength range used, and thus for calibration of number concentration measurements of EVs in the size range indicated. This was confirmed as the number concentration values obtained by PTA and two independent flow cytometric measurements agreed with the concentration reference values obtained by two independent spICP-MS measurements within the calculated uncertainty limits.

The food additive titanium dioxide hinders intestinal production of TGF-ß and IL-10 in mice, and long-term exposure in adults or from perinatal life blocks oral tolerance to ovalbumin.

Food hypersensitivities are increasing in industrialized countries, and foodborne nanoparticles (NPs) are suspected as co-factors in their aetiology. Food-grade titanium dioxide (fg-TiO2), a food colouring agent, is composed of NPs with immunomodulatory properties. We investigated whether fg-TiO2 may compromise the establishment of oral tolerance (OT) to food proteins using a model of OT induction to ovalbumin (OVA) in mice, and whether a perinatal exposure could trigger this effect. In pregnant mice fed a TiO2-enriched diet, ICP-MS and TEM-EDX analyses showed passage of TiO2 NPs into the foetus. When their weaned offspring were fed the same diet, a breakdown in OT to OVA was observed at adulthood, characterized by a high anti-OVA IgG production compared to controls. However, adult mice directly exposed to fg-TiO2 did not induce OT to OVA either, ruling out a developmental origin for these effects. When these mice were orally challenged with OVA, intestinal inflammation demonstrated hypersensitivity to OVA. In OVA-naïve mice, fg-TiO2 exposure impaired intestinal TGF-ß and IL-10 production, of key role in OT induction and maintenance. These findings showed that long-term exposure to TiO2 as food additive alters anti-inflammatory cytokine profile, and leads to OT failure regardless of the timing of TiO2 exposure throughout life.