Identification and quantification of per- and polyfluorinated alkyl substances (PFAS) migrating from food contact materials (FCM).

Per- and polyfluorinated alkyl substances (PFAS) can be added to food contact materials (FCM) to increase their water and/or grease repellent properties. Some well-known PFAS are perfluoroalkyl carboxylic acids (PFCA), perfluoroalkyl sulfonic acids (PFSA), and polyfluorinated telomer alcohols (FTOH). Due to the strength of the carbon-fluorine bond, PFAS are chemically very stable and highly resistant to biological degradation, posing a risk to human health and the environment. To examine the presence of PFAS in paper-based FCM, various samples were collected, including popcorn bags, muffin cups, and pizza boxes with high total organic fluorine (TOF) content from the Danish and Spanish markets. The FCM composition was characterised by FTIR. Quantification of some well-known PFAS such as PFCA, PFSA, and FTOH was performed in food simulants using LC-MS/MS, and in addition a non-targeted screening approach was performed by LC-Orbitrap-HRMS. Among analysed samples, the highest concentrations of PFAS were found in a muffin cup made of cellulose (PFCA âˆ¼ 1.41 µg kg-1 food, FTOH âˆ¼ 11.5 µg kg-1 food), and the results were used to estimate dietary exposures to PFAS migrated from this FCM. Compared to measured TOF value in this sample, the fluorine from all quantified PFAS accounted for only 0.6%. Thus, a more powerful analytical approach was used to further investigate PFAS occurrence in this sample. Using non-targeted screening, an additional twenty compounds were identified, among them five with confidence level 1 and ten with confidence level 2. Many of them were either fluorotelomer carboxylic acids or sulfonic acids or ether-containing compounds.

Per- and poly-fluoroalkyl substances in commercial organic eggs via fishmeal in feed.

Chicken eggs can be a significant source of human PFAS exposure. A survey of PFAS in commercial eggs from larger farms across Denmark showed the absence or low contents of PFAS in free-range and barn eggs. However, organic eggs from eight farms collected in September 2022 had a similar profile of nine PFASs with a predominance of odd over even carbon length PFCAs. Farm 11-13 e.g. had egg yolk ng/g concentrations of PFOA 0.07 ± 0.02; PFNA 0.37 ± 0.04; PFDA 0.13 ± 0.00; PFUnDA 0.22 ± 0.04; PFDoDA 0.06 ± 0.02; PFTrDA 0.15 ± 0.04; PFTeDA 0.02 ± 0.02; PFHxS 0.10 ± 0.04; PFOS 2.62 ± 0.11. Normalised to PFOS, the relative sum of other PFAS showed no difference between the eight organic egg samples, but significant differences between mean individual PFASs (p = 1.4E-25), reflecting a similar profile. The PFAS found in two fishmeal samples with the same origin as the fishmeal used for the organic feed production, could account for the contents in the eggs via estimated transfer from the feed. Furthermore, the estimated transfer from concentration in feed to concentration in egg increased with the carbon length of the PFCA. Exposure (95th percentile) of ∑4PFAS (PFOA, PFNA, PFHxS, PFOS) solely from consumption of 311 g âˆ¼ 5-6 organic eggs/week was for children 4-9 years 10.4 ng/kg bw, i.e. a significant exceedance of the tolerable weekly intake of 4.4 ng/kg bw established by the European Food Safety Authority. Based on the PFAS exposures from organic egg consumption, the organic egg producers decided voluntarily to cease adding fishmeal to the feed. Since the feed-to-egg half-lives are ≤1 week for PFOA, PFOS, and PFHxS, the removal of fishmeal as a feed ingredient should eliminate PFAS after 1-2 months. This was demonstrated in analyses of ten organic egg samples collected by the authorities without PFAS in eight and with 0.1 and 0.4 ng/g ∑4PFAS in two samples.

Antibiotic induced restructuring of the gut microbiota does not affect oral uptake and accumulation of perfluorooctane sulfonic acid (PFOS) in rats.

Perfluorooctane sulfonic acid (PFOS) is a manmade legacy compound belonging to the group of persistent per- and polyfluorinated substances (PFAS). While many adverse health effects of PFOS have been identified, knowledge about its effect on the intestinal microbiota is scarce. The microbial community inhabiting the gut of mammals plays an important role in health, for instance by affecting the uptake, excretion, and bioavailability of some xenobiotic toxicants. Here, we investigated (i) the effect of vancomycin-mediated microbiota modulation on the uptake of PFOS in adult Sprague-Dawley rats, and (ii) the effects of PFOS exposure on the rat microbiota composition. Four groups of twelve rats were exposed daily for 7 days with either 3 mg/kg PFOS plus 8 mg/kg vancomycin, only PFOS, only vancomycin, or a corn oil control. Vancomycin-induced modulation of the gut microbiota composition did not affect uptake of branched and linear PFOS over a period of 7 days, measured in serum samples. 16S rRNA amplicon sequencing of faecal and intestinal samples revealed that vancomycin treatment lowered microbial alpha-diversity, while PFOS increased the microbial diversity in vancomycin-treated as well as in non-antibiotic treated animals, possibly because an observed decrease in the Enterobacteriaceae abundance allows other microbial species to propagate. Colonic short-chain fatty acids were significantly lower in vancomycin-treated animals but remained unaffected by PFOS. Our results suggest that PFOS exposure may disturb the intestinal microbiota, but that antibiotic-induced modulation of the intestinal ecosystem does not affect systemic uptake of PFOS in rats.

Levels of nitrate, nitrite and nitrosamines in model sausages during heat treatment and in vitro digestion - The impact of adding nitrite and spinach (Spinacia oleracea L.).

Nitrite derivatives react with endogenous precursors forming N-nitrosamines associated with development of colorectal cancer. The present study aims to investigate the formation of N-nitrosamines in sausage during processing and in vitro gastrointestinal digestion after adding sodium nitrite and/or spinach emulsion. The INFOGEST digestion protocol was used to simulate the oral, gastric, and small intestinal phases of digestion, and sodium nitrite was added in the oral phase to mimic the input of nitrite from saliva as it has shown to affect the endogenous formation of N-nitrosamines. The results show that the addition of spinach emulsion, in spite of it being a source of nitrate, did not affect the nitrite content in either batter, sausage, or roasted sausage. The levels of N-nitrosamines increased with the added amount of sodium nitrite, and further formation of some volatile N-nitrosamines was observed during roasting and in vitro digestion. In general, N-nitrosamine levels in the intestinal phase followed the same trend as in the undigested products. The results further indicate that nitrite present in saliva may cause a significant increase in N-nitrosamine levels in the gastrointestinal tract and that bioactive components in spinach may protect against the formation of volatile N-nitrosamines both during roasting and digestion.

Is the use of paper food contact materials treated with per- and polyfluorinated alkyl substances safe for high-temperature applications? - Migration study in real food and food simulants.

Migration of per- and polyfluorinated alkyl substances (PFAS) from paper food contact materials (FCMs) can pose a consumer risk. However, risk assessment procedures typically do not consider PFAS contribution from FCMs. Moreover, migration studies are often limited to one subclass of PFAS or simplified by using food simulants (FS). To assess the risk comprehensively, migration of three PFAS subclasses (perfluorinated carboxylic acids/ sulfonic acids (PFCAs/PFSAs), polyfluoroalkyl phosphate esters (PAPs), and fluorotelomer alcohols (FTOHs)) from six FCMs were investigated to FS (50% and 20% ethanol) and food (oatmeal porridge, muffins, and tomato soup) under high-temperature conditions. Migration of PFCAs and FTOHs to all food samples was observed. Migration of PFCAs and FTOHs to 50% ethanol was significantly higher than migration to real food whilst FTOHs did not migrate into 20% ethanol. Estimated dietary PFAS exposure for children (1.06 - 5.67 ng/kgbw/day) exceeded EFSA's proposed safety threshold (0.63 ng/kgbw/day), risking consumer health.

Formation and mitigation of acrylamide in oven baked vegetable fries.

Investigation into oven baked sweet potato and carrot fries at various temperatures and times demonstrated the in situ formation of acrylamide in an exponential manner. High levels of acrylamide were found in these food items: up to 327 µg/kg for sweet potato baked at 190 °C for 14 min, and 99 µg/kg for carrot baked at 190 °C for 13 min. Risk assessment via Margin of Exposures estimation showed that consumption of these fries might pose adverse health effects to consumers from toddlers to adults, especially when the fries were prepared at high temperatures above 175 °C and for a long time. Raw ingredient blanching and immersion in acetic acid prior to preparation have been proven to greatly reduce acrylamide formation, up to 99%. It is recommendable to apply these techniques either at industrial or domestic cooking scales to ensure minimal health risk from dietary exposure to acrylamide.

Increasing Seaweed Consumption in the Netherlands and Portugal and the Consequences for the Intake of Iodine, Sodium, and Exposure to Chemical Contaminants: A Risk-Benefit Study.

Background: Seaweed has a high potential for nourishing the future planet. However, besides being beneficial, it also contains adverse components; this poses the question whether consumption of seaweed foods overall contributes beneficially or detrimentally to human health, and hence if their consumption should be promoted or restricted. Methods: This study evaluated the impact of substituting regular foods with seaweed foods in the diet, both in terms of nutritional quality (via iodine and sodium) and food safety (via arsenic, cadmium, lead, and mercury). Food consumption data from the Netherlands and Portugal (adults aged >18 years) were used, in which 10% of the amounts of pasta, bacon, and lettuce consumed were replaced by seaweed-derived products made from kelp (Saccharina latissima). Using Monte Carlo Risk Assessment software (MCRA), long-term nutrient intake and exposure to contaminants were assessed. The results obtained for the Netherlands and Portugal were compared with data from Japan, a country that has a high natural consumption of seaweed. Results: This low-tier risk-benefit study reveals that an increased seaweed consumption (as assessed by the 10% replacement with seaweed products) has no consequences in terms of intake of sodium and exposure to cadmium, lead, and mercury, and the associated (absence of) adverse health aspects. The alternative scenario almost doubled the mean iodine intake in the Netherlands (to 300 µg/day) and Portugal (to 208 µg/day) and increased the average exposure to arsenic levels in the Netherlands (to 1.02 µg/kg bw/day) and Portugal (to 1.67 µg/kg bw/day). Conclusion: The intake of iodine and exposure to arsenic in the Netherland and Portugal were certainly higher due to the modeled increase of seaweed foods. If seaweed consumption increases close to the 10% substitution, the public health consequences thereof may trigger further research.

Growth performance, bioavailability of toxic and essential elements and nutrients, and biofortification of iodine of rainbow trout (Onchorynchus mykiss) fed blends with sugar kelp (Saccharina latissima).

Aquaculture production is demanding novel feed ingredients that reflect natural marine nutrient levels, that are also essential to humans. In this regard, biofortification through addition of iodine-rich sugar kelp in feed formulations was assessed in a 12 week rainbow trout trial. Yttrium inclusion in feed allowed determinations of apparent absorption coefficients of essential and potentially toxic elements and apparent digestibility coefficient of nutrients. E.g. apparent absorption coefficients in trouts fortified feed with 1-4% dw kelp were 67-61% As, 32-40% Cd, <5% Fe; 80-83% I; 66-58% Se. Iodine concentrations in feed up to 239 mg/kg (~4% kelp) was proportional to iodine accumulation in trout fillets (R2 = 1.00) with 0.5% transfer ratio. Feed iodine concentrations up to 117 mg/kg (~2% kelp) did not affect growth performance negatively, but increased significantly protein efficiency ratio after eight weeks feeding. However, 4% kelp meal inclusion affected final growth and hepato somatic index, and caused histomorphological changes in the intestine. All fillets had low toxic element concentrations (As, Cd, Hg, Pb). The potential applicability of Saccharina latissima as feed ingredient to tailor iodine concentration in farmed fish is evident. Consuming of a 160 g fillet (2% kelp) contributes ~60% of recommended daily iodine intake for adults.

Diets supplemented with Saccharina latissima influence the expression of genes related to lipid metabolism and oxidative stress modulating rainbow trout (Oncorhynchus mykiss) fillet composition.

This study aimed to evaluate the impact of diets including increasing amounts (1, 2 and 4%) of an iodine-rich macroalgae, Saccharina latissima, on gene expression and fillet composition of commercial-sized rainbow trout. Liver and muscle expression of genes related to growth, iodine, oxidative stress, and lipid metabolism, and the fillet content of fatty acids, cholesterol, and vitamin D3 were assessed. The highest kelp inclusion led to lower final body weight and HSI, without significant differences in mRNA transcription of genes involved in growth (ghr1, ghr2 and igf1) or iodine metabolism (dio1, thra, and thrb). A significant downregulation of an oxidative stress marker, gpx1b2, was observed in fish fed 2% S. latissima, which might suggest the need for less endogenous antioxidants. Dietary inclusion of kelp impacted lipid metabolism, with a downregulation of fatty acid synthase, accompanied by a general decrease of fatty acids in fillet. The present study demonstrated that supplementation of diets with 1 or 2% S. latissima can be achieved without detrimental effects on rainbow trout final weight. Evidence suggest a lipid-lowering effect of diets that did not compromise fillet EPA and DHA concentrations, being 3.7 times above the recommended levels for human consumption.

Dietary exposure to selected chemical contaminants in fish for the Danish population.

The exposure to selected chemical contaminants from fish has been calculated for the Danish population, both for adults (15-75 years of age) and children (4-14 years of age). The Danish mean consumption of fish is 21 g person-1 day-1 for adults and 12 g person-1 day-1 for children. Fish consumption is the main food group contributor for exposure to mercury and dioxins and dioxin-like polychlorinated biphenyls (PCDD/F and DL-PCB) for the Danish population. Comparison of the mean exposure with the TDI or TWI values shows for these substances as well as for perfluorooctane sulphuric acid (PFOS) that the exposure is below the TDI/TWI values. However, even without taking other food groups into account, PCDD/Fs and DL-PCB exposure is close to the actual TWI-value. Calculation of the Margin of Exposure (MOE) for the sum of hexabromocyclododecanes (HBCDD) and polycyclic aromatic hydrocarbons (PAHs) revealed fish consumption to be of low concern for the consumer health regarding these contaminants.

Pharmaceuticals and endocrine disruptors in raw and cooked seafood from European market: Concentrations and human exposure levels.

Pharmaceuticals (PhACs) and endocrine disrupting compounds (EDCs) are chemicals of emerging concern that can accumulate in seafood sold in markets. These compounds may represent a risk to consumers through effects on the human reproductive system, metabolic disorders, pathogenesis of breast cancer or development of microbial resistance. Measuring their levels in highly consumed seafood is important to assess the potential risks to human health. Besides, the effect of cooking on contaminant levels is relevant to investigate. Therefore, the objectives of this research were to study the presence and levels of PhACs and EDCs in commercially available seafood in the European Union market, to investigate the effect of cooking on contaminant levels, and to evaluate the dietary exposure of humans to these compounds through seafood consumption. A sampling survey of seafood from 11 European countries was undertaken. Twelve highly consumed seafood types were analysed raw and cooked with 3 analytical methods (65 samples, 195 analysis). PhACs were mostly not detectable or below quantification limits in seafood whereas EDCs were a recurrent group of contaminants quantified in the majority of the samples. Besides, cooking by steaming significantly increased their levels in seafood from 2 to 46-fold increase. Based on occurrence and levels, bisphenol A, methylparaben and triclosan were selected for performing a human exposure assessment and health risk characterisation through seafood consumption. The results indicate that the Spanish population has the highest exposure to the selected EDCs through seafood consumption, although the exposure via seafood remained below the current toxicological reference values.

Prioritization before risk assessment: The viability of uncertain data on food contact materials.

The shortage of data on non-intentionally added substances (NIAS) present in food contact material (FCM) limits the ability to ensure food safety. Recent strategies in analytical method development permit NIAS investigation by using chemical exploration, but this has not been sufficiently investigated in risk assessment context. Here, exploration is utilized and followed by risk prioritization on chemical compounds that can potentially migrate to food from two paperboard FCM samples. Concentration estimates from exploration are converted to tentative exposure assessment, while predicted chemical structures are assessed using quantitative structure-activity relationships (QSAR) models for carcinogenicity, mutagenicity, and reproductive toxicity. A selection of 60 chemical compounds from two FCMs is assessed by four risk assessors to classify compounds based on probable risk. For almost 60% of cases, the assessors classified compounds as either high priority or low priority. Unclassified compounds are due to disagreements between experts (18%) or due to a perceived lack of data (23%). Among the high priority substances are high-concentration compounds, benzophenone derivatives, and dyes. The low priority compounds contained e.g. oligomers from plasticizers and linear alkane amides. The classification scheme provides valuable information based on tentative data and is able to prioritize discovered chemical compounds for pending risk assessment.

Effects of steaming on contaminants of emerging concern levels in seafood.

Seafood consumption is a major route for human exposure to environmental contaminants of emerging concern (CeCs). However, toxicological information about the presence of CeCs in seafood is still insufficient, especially considering the effect of cooking procedures on contaminant levels. This study is one among a few who evaluated the effect of steaming on the levels of different CeCs (toxic elements, PFCs, PAHs, musk fragrances and UV-filters) in commercially relevant seafood in Europe, and estimate the potential risks associated with its consumption for consumers. In most cases, an increase in contaminant levels was observed after steaming, though varying according to contaminant and seafood species (e.g. iAs, perfluorobutanoate, dibenzo(ah)anthracene in Mytilus edulis, HHCB-Lactone in Solea sp., 2-Ethylhexyl salicylate in Lophius piscatorius). Furthermore, the increase in some CeCs, like Pb, MeHg, iAs, Cd and carcinogenic PAHs, in seafood after steaming reveals that adverse health effects can never be excluded, regardless contaminants concentration. However, the risk of adverse effects can vary. The drastic changes induced by steaming suggest that the effect of cooking should be integrated in food risk assessment, as well as accounted in CeCs regulations and recommendations issued by food safety authorities, in order to avoid over/underestimation of risks for consumer health.

The influence of microplastics and halogenated contaminants in feed on toxicokinetics and gene expression in European seabass (Dicentrarchus labrax).

When microplastics pollute fish habitats, it may be ingested by fish, thereby contaminating fish with sorbed contaminants. The present study investigates how combinations of halogenated contaminants and microplastics associated with feed are able to alter toxicokinetics in European seabass and affect the fish. Microplastic particles (2%) were added to the feed either with sorbed contaminants or as a mixture of clean microplastics and chemical contaminants, and compared to feed containing contaminants without microplastics. For the contaminated microplastic diet, the accumulation of polychlorinated biphenyls (PCBs) and brominated flame retardants (BFRs) in fish was significantly higher, increasing up to 40 days of accumulation and then reversing to values comparable to the other diets at the end of accumulation. The significant gene expression results of liver (cyp1a, il1ß, gstα) after 40 days of exposure indicate that microplastics might indeed exacerbate the toxic effects (liver metabolism, immune system, oxidative stress) of some chemical contaminants sorbed to microplastics. Seabass quickly metabolised BDE99 to BDE47 by debromination, probably mediated by deiodinase enzymes, and unlike other contaminants, this metabolism was unaffected by the presence of microplastics. For the other PCBs and BFRs, the elimination coefficients were significantly lower in fish fed the diet with contaminants sorbed to microplastic compared to the other diets. The results indicate that microplastics affects liver detoxification and lipid distribution, both of which affect the concentration of contaminants.

Combined effects of microplastics and chemical contaminants on the organ toxicity of zebrafish (Danio rerio).

Microplastics contamination of the aquatic environment is considered a growing problem. The ingestion of microplastics has been documented for a variety of aquatic animals. Studies have shown the potential of microplastics to affect the bioavailability and uptake route of sorbed co-contaminants of different nature in living organisms. Persistent organic pollutants and metals have been the co-contaminants majorly investigated in this field. The combined effect of microplastics and sorbed co-contaminants in aquatic organisms still needs to be properly understood. To address this, we have subjected zebrafish to four different feeds: A) untreated feed; B) feed supplemented with microplastics (LD-PE 125-250µm of diameter); C) feed supplemented with 2% microplastics to which a mixture of PCBs, BFRs, PFCs and methylmercury were sorbed; and D) feed supplemented with the mixture of contaminants only. After 3 weeks of exposure fish were dissected and liver, intestine, muscular tissue and brain were extracted. After visual observation, evaluation of differential gene expression of some selected biomarker genes in liver, intestine and brain were carried out. Additionally, quantification of perfluorinated compounds in liver, brain, muscular tissue and intestine of some selected samples were performed. The feed supplemented with microplastics with sorbed contaminants produced the most evident effects especially on the liver. The results indicate that microplastics alone does not produce relevant effects on zebrafish in the experimental conditions tested; on the contrary, the combined effect of microplastics and sorbed contaminants altered significantly their organs homeostasis in a greater manner than the contaminants alone.

Exploring the chemistry of complex samples by tentative identification and semiquantification: A food contact material case.

In fields such as food safety and environmental chemistry, ensuring safety is greatly challenged by large numbers of unknown substances occurring. Even with current state-of-the-art mass spectrometers, dealing with nonidentified substances is a very laborious process as it includes structure elucidation of a vast number of unknowns, of which only a fraction may be relevant. Here, we present an exploration and prioritization approach based on high-resolution mass spectrometry. The method uses algorithm-based precursor/product-ion correlations on quadrupole time-of-flight tandem mass spectrometry data to retrieve the most likely chemical match from a structure database. In addition, time-of-flight-only data are used to estimate analyte concentration via semiquantification. The method is demonstrated in recycled paper food contact material. Here, 585 chromatographic peaks were discovered, of which 117 were unique to the sample and could be tentatively elucidated via accurate mass, isotopic pattern, and precursor/product-ion correlations. Nearly 85% of these 117 peaks were matched with database entries, which provided varying certainty of information about the analyte structure. Semiquantitative concentration ranges of investigated compounds were between 0.7 and 1600 µg dm-2 . With these data, a subgroup of chemicals was risk-categorized and prioritized by using the most likely candidate structure(s) obtained. Prioritization based on expected health impact was possible by using the tentatively assigned data. Overall, the described method not only is a valuable chemical exploration tool for nonidentified substances but may also be used as a preliminary prioritization tool for substances expected to have the highest health impact, for example, in food contact materials.

Occurrence of cyclic imines in European commercial seafood and consumers risk assessment.

Cyclic imines constitute a quite recently discovered group of marine biotoxins that act on neural receptors and that bioaccumulate in seafood. They are grouped together due to the imino group functioning as their common pharmacore, responsible for acute neurotoxicity in mice. Cyclic imines (CIs) have not been linked yet to human poisoning and are not regulated in the European Union (EU), although the European Food Safety Authority (EFSA) requires more data to perform conclusive risk assessment for consumers. Several commercial samples of bivalves including raw and processed samples from eight countries (Italy, Portugal, Slovenia, Spain, Ireland, Norway, The Netherlands and Denmark) were obtained over 2 years. Emerging cyclic imine concentrations in all the samples were analysed on a LC-3200QTRAP and LC-HRMS QExactive mass spectrometer. In shellfish, two CIs, pinnatoxin G (PnTX-G) and 13-desmethylspirolide C (SPX-1) were found at low concentrations (0.1-12µg/kg PnTX-G and 26-66µg/kg SPX-1), while gymnodimines and pteriatoxins were not detected in commercial (raw and processed) samples. In summary, SPX-1 (n: 47) and PnTX-G (n: 96) were detected in 9.4% and 4.2% of the samples, respectively, at concentrations higher than the limit of quantification (LOQ), and in 7.3% and 31.2% of the samples at concentrations lower than the LOQ (25µg/kg for SPX-1 and 3µg/kg for PnTX-G), respectively. For the detected cyclic imines, the average exposure and the 95th percentile were calculated. The results obtained indicate that it is unlikely that a potential health risk exists through the seafood diet for CIs in the EU. However, further information about CIs is necessary in order to perform a conclusive risk assessment.

An effect-directed strategy for characterizing emerging chemicals in food contact materials made from paper and board.

Food contact materials (FCM) are any type of item intended to come into contact with foods and thus represent a potential source for human exposure to chemicals. Regarding FCMs made of paper and board, information pertaining to their chemical constituents and the potential impacts on human health remains scarce, which hampers safety evaluation. We describe an effect-directed strategy to identify and characterize emerging chemicals in paper and board FCMs. Twenty FCMs were tested in eight reporter gene assays, including assays for the AR, ER, AhR, PPARγ, Nrf2 and p53, as well as mutagenicity. All FCMs exhibited activities in at least one assay. As proof-of-principle, FCM samples obtained from a sandwich wrapper and a pizza box were carried through a complete step-by-step multi-tiered approach. The pizza box exhibited ER activity, likely caused by the presence of bisphenol A, dibutyl phthalate, and benzylbutyl phthalate. The sandwich wrapper exhibited AR antagonism, likely caused by abietic acid and dehydroabietic acid. Migration studies confirmed that the active chemicals can transfer from FCMs to food simulants. In conclusion, we report an effect-directed strategy that can identify hazards posed by FCMs made from paper and board, including the identification of the chemical(s) responsible for the observed activity.

A framework to estimate concentrations of potentially unknown substances by semi-quantification in liquid chromatography electrospray ionization mass spectrometry.

Risk assessment of exposure to chemicals from food and other sources rely on quantitative information of the occurrence of these chemicals. As screening analysis is increasingly used, a strategy to semi-quantify unknown or untargeted analytes is required. A proof of concept strategy to semi-quantifying unknown substances in LC-MS was investigated by studying the responses of a chemically diverse marker set of 17 analytes using an experimental design study. Optimal conditions were established using two optimization parameters related to weak-responding compounds and to the overall response. All the 17 selected analytes were semi-quantified using a different analyte to assess the quantification performance under various conditions. It was found that source conditions had strong effects on the responses, with the range of low-response signals varying from -80% to over +300% compared to centerpoints. Positive electrospray (ESI+) was found to have more complex source interactions than negative electrospray (ESI-). Choice of quantification marker resulted in better quantification if the retention time difference was minimized (12 out of 12 cases error factor < 4.0) rather than if the accurate mass difference was minimized (7 out of 12 cases error factor < 4.0). Using optimal conditions and retention time selection, semi-quantification in ESI+ (70% quantified, average prediction error factor 2.08) and ESI- (100% quantified, average prediction error factor 1.74) yielded acceptable results for untargeted screening. The method was successfully applied to an extract of food contact material containing over 300 unknown substances. Without identification and authentic standards, the method was able to estimate the concentration of a virtually unlimited number of compounds thereby providing valuable data to prioritize compounds in risk assessment studies.