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.

Trace elements, dioxins and PCBs in different fish species and marine regions: Importance of the taxon and regional features.

Chemical contaminant concentrations in wild organisms are used to assess environmental status under the European Marine Strategy Framework Directive. However, this approach is challenged by the complex intra- and inter-species variability, and the different regional features. In this study, concentrations in trace elements (As, Cd, Hg and Pb), polychlorinated biphenyls (PCBs), polychlorodibenzo-para-dioxines (PCDDs) and polychlorodibenzofuranes (PCDFs) were monitored in 8 fish species sampled on the continental shelf of three French regions: the Eastern English Channel (EEC) and Bay of Biscay (BoB) in the Northeast Atlantic Ocean, and the Gulf of Lions (GoL) in Western Mediterranean Sea. Our objectives were to identify species or regions more likely to be contaminated and to assess how to take this variability into account in environmental assessment. While concentrations were higher in benthic and demersal piscivores, PCB and PCDD/F concentrations (lipid-weight) were similar in most teleost species. For Cd, Hg and Pb, the trophic group accumulating the highest concentrations depended on the contaminant and region. Concentrations in Hg, PCBs and PCDD/Fs were higher in the EEC and/or GoL than in BoB. Cadmium and Pb concentrations were highest in the BoB. Lipid content accounted for 35%-84% of organic contaminant variability. Lipid normalisation was employed to enhance robustness in the identification of spatial patterns. Contaminant patterns in chondrichthyans clearly differed from that in teleosts. In addition, trophic levels accounted for ≤1% and ≤33% of the contaminant variability in teleost fishes in the EEC and BoB, respectively. Therefore, developing taxa-specific thresholds might be a more practical way forward for environmental assessment than normalisation to trophic levels.

Chromium speciation analysis in raw and cooked milk and meat samples by species-specific isotope dilution and HPLC-ICP-MS.

This study aimed at the assessment of the impact of various culinary processes on the fate of chromium (Cr) species (Cr(III) and Cr(VI)) in infant formula milk, semi-skimmed milk and bovine meat samples. The cooking procedures were boiling at 70°C/100°C (milk samples) and frying without and with oil (95°C and 120°C) (bovine meat). The levels of Cr(III) and Cr(VI) in raw and cooked samples were determined by high-performance liquid chromatography (HPLC) coupled to inductively coupled plasma mass spectrometry (ICP-MS) using double spike species-specific-isotope dilution (SS-ID). The species were extracted by sequential complexation of Cr(III) with ethylenediaminetetraacetic acid and of Cr(VI) with 1,5-diphenylcarbazide in the same analytical run by heating at 70°C for 50 min. Anion exchange chromatography using a Dionex IonPac™ AG7 column and a mobile phase consisting of 10 mM HNO3, 2.5% MeOH and 30 mM EDTA at pH 2 was employed for species separation. The quantification limits were 0.013 and 0.049 µg kg-1 for Cr(III) and Cr(VI), respectively. ANOVA test used to compare the mean Cr species concentrations showed no significant differences between raw and cooked samples. The results obtained in the present study show that oxidation of Cr(III) to Cr(VI) does not occur during thermal cooking of milk and bovine meat samples. A selection of 10 samples of each type were analysed in terms of total Cr (Crtotal) as well as speciation (Cr(III) and Cr(VI)). Cr(VI) was not quantified in any of these samples, whereas Cr(III) levels ranged from 0.22 (infant formula milk) up to 80 µg kg-1 (chorizo sausage). Additionally, Cr(III) and Crtotal levels were comparable hence demonstrating that in the samples analysed in this study, Cr is found exclusively as Cr(III) species.

Development and validation of a single run method based on species specific isotope dilution and HPLC-ICP-MS for simultaneous species interconversion correction and speciation analysis of Cr(III)/Cr(VI) in meat and dairy products.

This study presents the development, validation and application of a new analytical approach for the simultaneous speciation analysis of Cr(III) and Cr(VI) in meat and dairy products by high-performance liquid chromatography (HPLC) coupled to inductively coupled plasma mass spectrometry (ICP-MS) and double spike species specific-isotope dilution (SS-ID). The species extraction was achieved by sequential complexation of Cr(III) with ethylenediaminetetraacetic acid (EDTA) and of Cr(VI) with 1,5-diphenylcarbazide (DPC) in the same analytical run. The HPLC separation of complexed species was carried out using a short (5 cm) microbore anion-exchange HPLC column and a mobile phase consisting of 0.01 mol L-1 HNO3 + 2.5% (v/v) MeOH + 0.30 mol L-1 EDTA (pH = 2) in isocratic elution mode with excellent baseline separation achieved in less than 3 min. The method was validated by means of the accuracy profile approach by carrying out 6 measurement series in duplicate on (six) different days over a timespan of two months. The quantification limit was 0.013 µg kg-1 for Cr(III) and 0.049 µg kg-1 for Cr(VI), respectively. The measurement bias corresponding to the validity domain ranged from 0.01 to 0.11%, whereas the coefficient of variation in terms of repeatability (CVr) varied from 2.9 to 11.6% (depending on the analyte level) for Cr(III) and from 6.7 to 11.8% for Cr(VI). Similarly, the coefficient of variation in terms of intermediate reproducibility (CVR) ranged from 6.8 to 13% for Cr(III) and from 6.8 to 25.9% for Cr(VI), respectively. The method was successfully applied to the analysis of a selection of food samples such as baby and semi-skimmed milk and steak beef samples. Cr(VI) was not quantified in any of these samples while Cr(III) levels ranged between 2.7 and 4.7 µg kg-1, which were comparable with the levels of total chromium analysed in the same samples by ICP-MS (accredited method). The method presented here with combined use of species specific isotope dilution and sequential species complexation is a powerful analytical tool for accurate and precise quantification of Cr(III) and Cr(VI) at trace levels and allows for correction of any species interconversion during sample preparation.

Seasonal and Spatial Variability of Trace Elements in Livers and Muscles of Three Fish Species from the Eastern Mediterranean.

Levels of 20 trace elements (Al, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Ag, Cd, Sn, Sb, Ba, Hg, Pb, and U) were assessed in livers and muscles of two demersal fish species (Siganus rivulatus and Lithognathus mormyrus) and one pelagic species (Etrumeus teres) from the Lebanese coast located in the Eastern Mediterranean. The samples were collected from three sites along the Lebanese coast during the wet and dry seasons in 2017. The trace elements were more concentrated in livers than in muscles and interspecific differences were also found. The herbivorous species S. rivulatus showed the highest levels for most trace elements, while the carnivorous species L. mormyrus showed the least contamination. Elemental seasonal differences were species dependent and were observed for Ti, Cr, Mn, Fe, Ni, As, and Hg, with higher values during the wet season. Multivariate analysis showed spatial differences mainly during the wet season, while being closely related to species that reflected different accumulation patterns in each site. Levels of most trace metals in livers were higher than those reported in other Mediterranean regions (up to 2 to 3 folds). Nevertheless, the levels of Cd, Pb, and Hg in fish muscle were below the maximum levels set by the European Commission indicating that the consumption of these fish species is not likely to have adverse effect on human health. However, exposure depends on dietary habits of the population and a continuous exposure to these elements may result in adverse effects.

Mercury in foods from the first French total diet study on infants and toddlers.

A very sensitive method using a direct mercury analyser was developed and validated according to the accuracy profile procedure to determine mercury levels in foods mainly consumed by infants and toddlers. Total mercury was not detected (LOD of 0.30µg.kg-1 fresh weight) in 92.4% of the 291 food samples analysed or at relatively low concentrations, lesser than or equal to LB/UB 0.5/1.0µg.kg-1 in all samples, except in fish samples (mean LB/UB 25µg.kg-1 with a maximum of 53µg.kg-1). Levels of total mercury in these foods were in all cases within permitted Regulatory limits of 500 or 1000µg.kg-1 in fishery products and muscle meat of fish.

Levels of lead in foods from the first French total diet study on infants and toddlers.

Infants and toddlers are highly vulnerable to exposure to lead due to its higher absorption in small children than in adults. This study describes the optimisation and validation of a very sensitive method for the determination of low levels of lead in foods mostly consumed by infants and toddlers. This method, based on inductively coupled plasma-mass spectrometry with a programmable temperature cyclonic spray chamber, attained a limit of quantification (LOQ) of 0.6 or 0.9µgPbkg-1 for a liquid or a solid sample, that was improved by a factor 5.6-8.3 compared to the previous method (LOQ: 5µgkg-1). The analytical method was then applied to 291 food samples from the first French total diet study on infants and toddlers. Lead was detected in most samples at relatively low concentrations (range 0.0-16µgkg-1). The highest lead concentrations were mainly found in processed food products (e.g. products containing chocolate).

Optimization and validation of the methods for the total mercury and methylmercury determination in breast milk.

The objective of the work was to develop and validate methods for the total Hg and methylmercury (MeHg) in breast milk that could be further used to obtain first data on chemical contamination of French breast milk. For total Hg determination, the potential of two techniques, namely Advanced Mercury Analyzer (AMA) and ICP MS, was compared. For MeHg determination, ICP MS detection associated to a quantification by isotopic dilution was used and the potential of a preliminary separation by gas or liquid chromatography was evaluated and discussed. The optimization studies have shown that AMA for total Hg determination and HPLC - ID - ICP MS, after a preconcentration step by freeze-drying, for MeHg quantification were the most relevant methods to use for epidemiologic studies. The figures of merit for both methods were evaluated by means of accuracy profiles in terms of limits of quantification (1.82 and 1.35µg Hg/kg dry weight, corresponding to 0.22 and 0.16µg Hg/kg wet basis for total Hg and MeHg, respectively), repeatability (2-11% and 3-8% for total Hg and MeHg respectively), intermediate precision reproducibility (4-12% and 4-8% for total Hg and MeHg respectively) and trueness bias (-0.1-9% and -4-0% for total Hg and MeHg respectively). The methods were then applied to 180 breast milk samples. Total Hg concentrations ranged from

Distribution and relationships of As, Cd, Pb and Hg in freshwater fish from five French fishing areas.

The concentrations of arsenic, cadmium, mercury and lead in 149 muscle samples of eight freshwater fish species (European eel, bream, common carp, European catfish, roach, perch, pike and pikeperch) from five different French fishing areas from contaminated and control sites were measured by inductively coupled plasma mass spectrometry after microwave digestion under pressure. No significant correlation was found between the condition factor (CF), based on the length-mass relationship, and As, Cd and Pb levels in all the samples analysed, but a positive correlation was detected between CF and Hg levels (P<0.0001, R=0.49). Positive correlations with body length were only found for Hg in roach (P<0.05, R=0.32) and Pb in bream (P<0.05, R=0.48) and correlations with both body weight and length were also found for Hg in pike (P<0.05, R=0.90 and 0.86) and Cd in European eel (P<0.01, R=-0.35 and -0.37). The average content and the standard deviation in fish muscle samples was 0.007±0.012, 0.102±0.077, 0.142±0.097 and 0.035±0.053 mg kg(-1) of wet mass for Cd, As, Hg and Pb, respectively. Significant differences were established between groups of predatory fish and non-predatory fish for Hg and Pb, and between control and contaminated sites in the whole selection and also within feeding guilds, i.e. the values of Hg in the benthophagic fish were significantly different between these sites. Finally, these results were also compared for each species with previous French and European studies.

Li, Cr, Mn, Co, Ni, Cu, Zn, Se and Mo levels in foodstuffs from the Second French TDS.

In 2006, the French Food Safety Agency (AFSSA) conducted the Second French Total Diet Study (TDS) to estimate dietary exposures to the main minerals and trace elements from 1319 samples of foods typically consumed by the French population. The foodstuffs were analysed by inductively coupled plasma-mass spectrometry (ICP-MS) after microwave-assisted digestion. Occurrence data for lithium, chromium, manganese, cobalt, nickel, copper, zinc, selenium and molybdenum were reported and compared with results from the previous French TDS. The results indicate that the food groups presenting the highest levels of these essential trace elements were "tofu" (for Li, Mn, Ni, Cu, Zn and Mo),"fish and fish products" particularly "shellfish" (for Li, Co, Cu, Zn, Se and Mo), "sweeteners, honey and confectionery" particularly dark chocolate (for Cr, Mn, Co, Ni and Cu), "cereals and cereal products" (for Mn, Ni and Mo) and "ice cream" (for Cr, Co and Ni).

Optimisation of ICP-MS collision/reaction cell conditions for the determination of elements likely to be interfered (V, Cr, Fe, Co, Ni, As and Se) in foodstuffs.

A strategy for the accurate determination in foodstuffs of seven elements liable to be interfered with (V, Cr, Fe, Co, Ni, As and Se), was successfully applied. Firstly, to reduce spectroscopic interferences, four influential factors (hexapole and quadrupole bias, helium and hydrogen flows) of the collision/reaction cell device were optimised through the experimental design methodology. Secondly, non-spectroscopic interferences, which may severely disturb the analysis of matrices containing large amounts of non-target elements, were significantly reduced by a limited decrease in the flow rate of the optimum initial nebuliser rather than with a specific time-consuming dilution. Finally, the optimised multi-element method was subjected to a full validation that demonstrated its acceptable analytical performance.

Pb, Hg, Cd, As, Sb and Al levels in foodstuffs from the 2nd French total diet study.

In 2006, the French Food Safety Agency (AFSSA) conducted the second French total diet study (TDS) to estimate dietary exposures of main minerals and trace elements from 1319 samples of foods habitually consumed by the French population. The foodstuffs were analysed by ICP-MS after microwave-assisted digestion. Contamination data for lead, mercury, cadmium, arsenic, antimony and aluminium were reported and compared with results from the previous French total diet study. The results are comparable with those from the rest of Europe. "Fish and fish products" and "sweeteners, honey and confectionery" were the food groups showing the highest cumulated contents in Pb, Hg, Cd, As, Al and Sb. However, observed levels remained low and were generally well below the maximum levels set by the current European regulation for lead, cadmium and mercury.

Determination of 20 trace elements in fish and other seafood from the French market.

The levels of 20 essential or toxic trace elements in 159 fish, other seafood and seafood products on the French coastal market collected between January and April 2005 were measured by ICP-MS. The concentration ranges (mg/kg of fresh mass) for the elements determined were compared with previous studies. The contents of Co, Cu, Fe, Li, Mn, Se, Zn and Pb found in fish are close to or often lower than previous studies. For other seafood, comparison is difficult due to the lack of data on a more global scale. However, it should be noted that the contents of Ag were found considerably higher in this study.

Determination of calcium, magnesium, sodium, and potassium in foodstuffs by using a microsampling flame atomic absorption spectrometric method after closed-vessel microwave digestion: method validation.

This paper describes a validation process in compliance with the NFIEN ISO/IEC 17025 standard for the determination of the macrominerals calcium, magnesium, sodium, and potassium in foodstuffs by microsampling with flame atomic absorption spectrometry after closed-vessel microwave digestion. The French Standards Commission (Agence Francaise de Normalisation) standards NF V03-110, NF EN V03-115, and XP T-90-210 were used to evaluate this method. The method was validated in the context of an analysis of the 1322 food samples of the second French Total Diet Study (TDS). Several performance criteria (linearity, LOQ, specificity, trueness, precision under repeatability conditions, and intermediate precision reproducibility) were evaluated. Furthermore, the method was monitored by several internal quality controls. The LOQ values obtained (25, 5, 8.3, and 8.3 mg/kg for Ca, Mg, Na, and K, respectively) were in compliance with the needs of the TDS. The method provided accurate results as demonstrated by a repeatability CV (CVr) of < 7% and a reproducibility CV (CVR) of < 12% for all the elements. Therefore, the results indicated that this method could be used in the laboratory for the routine determination of these four elements in foodstuffs with acceptable analytical performance.