There is a growing worldwide focus on evaluating the risk potential of substances associated with the manufacture and use of plastic, including the intentionally added constituents (additives) that impart specific technical functions as well as their transformation products. To enhance the consistent and effective evaluation of plastic additive uses, the European Centre for Ecotoxicology and Toxicology of Chemicals (ECETOC) is proposing a framework for risk assessment of plastic additives in complex scenarios. Here, the fundamental elements of the framework are presented, including an overview of available databases, tools and methodologies that enable the safety assessment of individual additives in plastic. The framework is comprised of five modules to identify and efficiently gather pertinent information. Data needs are tailored to the specific plastic additive and/or its major known transformation product(s) and to the scope of the risk assessment. As a risk-based approach, the five modules are pursued in an iterative, ‘circular’ manner. The risk assessment is terminated as soon as it is possible to either exclude risk potential or to characterise and manage any identified risks. While the framework for plastic additive risk assessment, as presented here, is applicable to individual plastic additives and/or their major known transformation products, it can be supplemented by further considerations for the assessment of aggregate or cumulative exposures and complex material cycles. Building thereupon, the second article by ECETOC shall present such further elements to consider when applying the framework for risk assessment of plastic additives at diverse recycling stages.

Indole-3-acetic acid (IAA) is a natural hormone found in all plants and can also be used as a plant growth regulator, which is banned in the EU. To differentiate between its natural presence and pesticide usage, the origin and fate of IAA during coffee roasting were studied. Green beans from the Coffea arabica and Coffea canephora varieties were spiked with a stable isotope precursor, 13C11-tryptophan, and roasted to different degrees. Liquid and gas chromatography coupled with mass spectrometry were used to analyze the degradation products. Alkaline hydrolysis determined free and bound IAA proportion in the coffee samples. The study found that IAA levels increased during roasting, forming indole-3-acetaldehyde and further degrading into 3-methylindole. Hydrolysis experiments showed that IAA was released from the coffee matrix, with higher amounts released during longer roasting times. These results indicate that IAA naturally occurs in green coffee and is not likely due to pesticide use.

Indole-3-acetic acid (IAA) is a natural plant hormone which can also be used as a plant growth regulator. However, its usage on crops is not permitted in the EU, with a maximum residue limit (MRL) currently set at 0.1 mg kg-1 for all food commodities. As available data on the occurrence of IAA in food commodities are limited, this work aims at evaluating the amount of IAA in important food commodities such as coffee and cereals, but also selected processed ingredients such as cocoa and malt. In total, 133 samples representing 18 different food commodities were analysed for the occurrence of free IAA using a liquid chromatography high resolution mass spectrometry method. IAA was detected in all samples except malt powders and tomatoes. The analysed crops were either grown under conventional, organic, or strictly controlled (i.e. excluding the use of IAA as pesticide/plant growth regulator) farming conditions. No significant differences in the amounts of IAA were found in crops grown under the three different farming conditions. A high percentage of the samples (63%) showed levels of free IAA above the EU MRL. We therefore conclude that the natural abundance of IAA was not properly assessed prior to the establishment of some current EU MRLs. Based on the natural occurrence of IAA in a wide range of foods; we question the need for an MRL for IAA in plant-based foods.

This study describes the extension of a gas chromatography mass spectrometry (GC-MS) method, initially devoted to the analysis of ethylene oxide (EO) in ice cream, to a larger range of food items including herbs, spices, vegetables, inorganic salts, food supplements, thickeners, etc. Results are reported as EOTotal according to EC 2015/868 definition (expressed as EO equivalents as the sum of native EO and 2-chloroethanol (2-CE) after acidic hydrolysis) with a limit of quantification at 0.01 mg/kg regardless of the food item. Its ruggedness was demonstrated through fortification experiments on hundreds of samples. Re-analysis of 146 positive food samples without hydrolysis demonstrated that not EO but 2-CE is the predominant analyte detected in the different processed ingredients suspected to have been previously treated with EO. A series of eight contaminated dried herbs and spices were also re-analysed by four ISO 17025 accredited commercial laboratories making use of different analytical strategies for EO determination in foods. Each laboratory reported EOTotal levels within the same concentration range, but the resulting reproducibility ranged from 23% to 41% depending on the sample. Additionally, we show that results of free EO from methods based on conversion to 2-iodoethanol may lead to artefactual detection of native EO (false positive). An official method of analysis applicable for different food matrices would be useful to avoid discrepancies of results. Altogether, these data re-enforce the fact that in absence of native EO in food items, risk assessment of EO in foodstuffs should consider the predominance of 2-CE. A toxicological risk assessment using the food additive xanthan gum as a case study is discussed.

The idea that previously unknown hazards can be readily revealed in complex mixtures such as foods is a seductive one, giving rise to the hope that data from effect-based assays of food products collected in market surveys is of suitable quality to be the basis for data-driven decision-making. To study this, we undertook a comparative study of the oestrogenicity of blinded cereal samples, both in a number of external testing laboratories and in our own facility. The results clearly showed little variance in the activities of 9 samples when using a single method, but great differences between the activities from each method. Further exploration of these findings suggest that the oestrogenic activity is likely an inherent part of the natural food matrix which the varying sample preparation methods are able to release and extract to differing degrees. These issues indicate the current poor suitability of these types of datasets to be used as the basis for consumer advice or food decision-making. Data quality must be improved before such testing is used in practice.

Background and objectives The presence of endocrine disruptors in food is creating concern, and consequently, efficient methods for their identification are needed. In vitro bioassays have been developed to study endocrine properties of pure chemicals as well as mixtures. Because of their relative ease of execution and low cost, application of such techniques to whole foods is tempting. This raises a number of technical challenges. To address them, a study was commissioned in a contract laboratory offering in vitro estrogenicity testing of food products. Findings Four different commercial and regulatory compliant cereal products were sent in blinded triplicates. Some samples were reported to be active by the laboratory. However, the results were not reproducible and very close to the 0% measurable activity. Furthermore, a statistical analysis of the raw data indicated an absence of activity. This was surprising since these products were expected to naturally contain estrogenic compounds such as the phytoestrogens lignans and flavones. However, phytoestrogens in cereals are known to occur mainly in bound, inactive, forms, suggesting that the sample preparation used in the present study was not suitable to release active estrogenic substances. This hypothesis was supported by preliminary data using a high-resolution mass spectrometry method which did not detect any significant levels of phytoestrogens in the tested cereal extracts. Conclusions Using a yeast-based method, no estrogenic activity was found in breakfast cereals. However, benchmarking the bioassay data against quality criteria raised the question of the qualification of the sample preparation and bioassay used to adequately address estrogenic activity of whole foods. Significance and novelty The quality requirements for testing whole food samples were formally addressed. The application of validated and harmonized methods for both sample preparation and bioassay testing is strongly advocated.

Residues of ethylene oxide (EO), a banned fumigant in the EU, were found at amounts above the maximum residue limit (MRL) in carob (locust) bean gum (additive E410). The pesticide entered the food chain via stabiliser blends that are used as minor ingredients in the manufacture of ice cream. Consequently, all products that contained the non-compliant ingredient were withdrawn or recalled in several countries across the EU, in most cases irrespective of whether the pesticide residue was detectable or not in the final product. This is the first report of a reliable method to determine EO and its metabolite/marker compound 2-chloroethanol (2-CE), either together or independently in ice cream, with a limit of quantification at 0.01 mg EO/kg and recovery in the range of 87-104% across the levels investigated (0.01, 0.02 and 0.06 mg EO/kg). The method applies QuEChERS extraction and isotope dilution gas chromatography coupled with tandem mass spectrometry (GC-MS/MS). High resolution mass spectrometry (HRMS) confirmed the specificity of low mass ions. Data on the stability of EO and 2-CE under thermal conditions revealed that 2-CE is relatively stable in an ice cream matrix (ca. 80% recovery of spiked material). Importantly, this study also demonstrates that not EO, but 2-CE is the predominant analyte detected in the contaminated samples, which is new information of significance in terms of the overall risk assessment of EO in foodstuffs.

The activity of individual biotransformation products cannot be measured in multicomponent mixtures by the current status-quo assays. A prior separation and tedious isolation of compounds is required, and often in addition, a concentration step into a solvent suitable for the cell-/enzyme-based assay. Hence, the metabolizing S9 system, mimicking the complex biotransformation reactions in the liver, was aimed to be integrated into two orthogonal methods for analysis of the acetylcholinesterase (AChE) inhibition. For the microtiter plate assay method, the evaluation of the generated fluorescence signal was impaired by the incorporated S9 system. In contrast, the metabolic activator (S9 mixture) was successfully incorporated into the high-performance thin-layer chromatography (HPTLC) method. As proof of principle, four reference AChE inhibitors were studied in complex samples with and without metabolic activation. In addition to the neurotoxic carbamate eserine and the organophosphate insecticides chlorpyrifos, quinalphos and parathion, the tris(nonylphenyl) phosphite and nonylphenol, both originating from food contact materials, were tested in isolation but also in food packaging migrate and extract. A method comparison and benchmarking pointed to multifold advantages of using this newly developed bioanalytical tool for assessment of individual neurotoxins in complex samples. The sensitive HPTLC-S9-AChE assay allowed the detection of neurotoxic chemicals with and without metabolic activation, at levels consistent with the threshold of toxicological concern of organophosphates and carbamates. This new on surface metabolism system can be applied to other toxicities and samples.

Many food innovations rely on the introduction and use of new or modified proteins. New or modified food proteins may lead to major health risks due to their inherent potential to cause food allergy. Currently, the pre-market allergenicity assessment for new or modified food proteins and protein sources relies on methods for identifying allergenic hazards based on characteristics of known allergens. However, there is no general consensus on the allergenicity parameters to use and the criteria that should apply for the evaluation and decisions to be made. In this paper, we propose that the strategy for allergenicity risk assessment of new or modified food proteins and the methodologies applied should be governed by the risk management questions to be answered, reflected in the information needed by risk managers to enable their informed decision making. We generated an inventory of health outcome-related assessment parameters and criteria potentially important for risk management decision-making and we discuss the implications of selecting different optional criteria (e.g. cut-off values) for what could be accepted as safe with regards to the health outcomes in the (at risk) population. The impact of these various options on both method development and risk management practices was investigated.

Chemical contaminants can occur at trace levels in foods or food ingredients either as intrinsic toxins produced in plant/animal metabolism or by external contamination due to fungal growth, agricultural practice, environment or food manufacturing processes. Monitoring and managing such unavoidable chemical contaminants in raw materials of foods is important for ensuring safe food for the consumer. The aim of this study was to adapt an existing tool for ranking the chemical contaminants of raw materials used to produce food for infants and young children based on their likelihood to cause harm for the consumer. This tool was built on a risk-assessment based approach. The level of risk was defined according to a decision tree where the overall exposure, the main contributor and the distribution of the occurrence levels are key criteria. For developing this approach, a conservative global food intake scenario for infants and young children was established using several local data publicly available. Two case studies on cadmium and deoxynivalenol are described and showed consistent results with the findings of local total diet studies. In addition, this tool is flexible to adapt to global and specific local populations in adjusting the food intake scenario. It supports risk management in the identification and prioritisation of chemical contaminants.

Quantitative risk assessment (QRA) for food allergens has made considerable progress in recent years, yet acceptability of its outcomes remains stymied because of the limited extent to which it has been possible to incorporate severity as a variable. Reaction severity, particularly following accidental exposure, depends on multiple factors, related to the allergen, the host and any treatments, which might be administered. Some of these factors are plausibly still unknown. Quantitative risk assessment shows that limiting exposure through control of dose reduces the rates of reactions in allergic populations, but its impact on the relative frequency of severe reactions at different doses is unclear. Food challenge studies suggest that the relationship between dose of allergenic food and reaction severity is complex even under relatively controlled conditions. Because of these complexities, epidemiological studies provide very limited insight into this aspect of the dose-response relationship. Emerging data from single-dose challenges suggest that graded food challenges may overestimate the rate of severe reactions. It may be necessary to generate new data (such as those from single-dose challenges) to reliably identify the effect of dose on severity for use in QRA. Success will reduce uncertainty in the susceptible population and improve consumer choice.

Vacuum frying (VF) was tested as an alternative technique to produce high-quality fried food. This study investigated the effects of oil temperature on the moisture loss, oil uptake, texture, color, and acrylamide content of vacuum-fried versus atmospheric-fried breaded shrimps. The moisture loss and oil absorption of breaded shrimps during VF were significantly affected (p < 0.05) by oil temperature. Oil absorption was directly proportional to moisture loss during VF. The oil absorption of breaded shrimps during VF increased with increasing temperature for the same frying time. After 10 min of frying, the oil contents of breaded shrimps fried at 80 °C, 100 °C, and 120 °C under vacuum conditions were 0.20 ± 0.013, 0.23 ± 0.012, and 0.25 ± 0.019 goil/gdry solid, respectively. These values were significantly lower than those (0.32 ± 0.019 goil/gdry solid) obtained under atmospheric conditions (170 °C). VF effectively decreased the acrylamide content of breaded shrimps. Furthermore, breaded shrimps with desirable color and low hardness can be produced using VF. These results indicate that VF is a feasible alternative technique to produce high-quality breaded shrimps.

Outbreaks of foodborne illness associated with berries often involve contamination with hepatitis A virus (HAV) and norovirus but also bacteria such as Escherichia coli O157:H7 and parasites such as Cyclospora caytanensis. We evaluated the applicability of UV-C to the inactivation of pathogens on strawberries, raspberries and blueberries. Our three-step approach consisted of assessing the chemical safety of UV-C-irradiated berries, evaluating the sensory quality after UV-C treatment and finally studying the inactivation of the target microorganisms. Treatments lasting up to 9 min (4000 mJ cm−2) did not produce detectable levels of furan (<5 μg/kg), a known photolysis product of fructose with genotoxic activity and thus were assessed to be toxicologically safe. No effect on taste or appearance was observed, unless treatment was excessively long. 20 s of treatment (an average fluence of ~ 212 mJ cm−2) reduced active HAV titer by >1 log10 unit in 95% of cases except on frozen raspberries, while 120 s were required to inactivate murine norovirus to this extent on fresh blueberries. The mean inactivation of HAV and MNV was greater on blueberries (2–3 log10) than on strawberries and raspberries (<2 log10). MNV was more sensitive on fresh than on frozen berries, unlike HAV. Inactivation of Salmonella, E. coli O157:H7 and Listeria monocytogenes was poor on all three berries, no treatment reducing viable counts by >1 log10 unit. In most matrices, prolonging the treatment did not improve the result to any significant degree. The effect was near its plateau after 20 s of treatment. These results provide insight into the effectiveness of UV-C irradiation for inactivating bacterial and viral pathogens and surrogates on fresh and frozen berries having different surface types, under different physical conditions and at different levels of contamination. Overall they show that UV-C as single processing step is unsuitable to inactivate significant numbers of foodborne pathogens on berries.

The efficacy of levulinic acid (LVA) in combination with sodium dodecyl sulfate (SDS) in removal of foodborne viruses, enteric bacterial pathogens and their surrogates on fresh strawberries was investigated. Inoculated strawberries were treated with potable water, sodium hypochlorite solution (50 ppm), 0.5% LVA plus 0.5% SDS solution, and 5% LVA plus 2% SDS solution respectively for 2 min, followed by spray-rinsing with potable water. Water washing removed at least 1.0-log of the tested viral and bacterial strains from the strawberries' surfaces. The 50 ppm chlorine wash induced 3.4, 1.5 and 2.1-log reductions for hepatitis A virus (HAV), murine norovirus-1 (MNV-1) and MS2 bacteriophage, respectively. In comparison, the tested bacterial strains showed uniform reductions around 1.6-log CFU/ml. The 0.5% LVA plus 0.5% SDS wash induced 2.7, 1.4 and 2.4-log reductions for HAV, MNV-1 and MS2, which were comparable with the reductions induced by chlorine (P > 0.05). For bacteria, over 2.0-log reductions were obtained for Enterococcus faecium, Listeria monocytogenes and Salmonella, while Escherichia coli O157:H7 and Escherichia coli P1 showed reductions of 1.9 and 1.8-log CFU/ml. Higher concentration of LVA plus SDS showed no significantly higher reductions (P > 0.05). Sensory tests of washed strawberries and chemical residue analysis of LVA on strawberries after washing were also performed. In conclusion, this study demonstrates good performance of 0.5% LVA plus 0.5% SDS to reduce the levels of enteric pathogens if present on strawberries without altering taste and introducing chemical safety issues.

The purpose of this study is to develop a globally valid chemical risk assessment tool that provides the user with a priority rating in terms of which chemicals are important to manage in raw materials. The process entails the use of decision trees that enable the determination of risk (or “likelihood to cause harm”), and severity using objective and transparent selection criteria. Taken together, severity and risk are positioned in an HACCP-like matrix informing on the prioritization level of each combination of chemical hazard and raw material. The proposed model is intended to be adequately protective for consumer's health, as it considers a conservative food intake scenario, as well as various sources of contaminant exposure. The model's design is flexible and can easily be adapted to the needs of different food product categories and scenarios. Case studies are presented to illustrate the feasibility of the approach, and the model was tested using several examples, the results of which are consistent with existing data in the literature.

Endocrine activity of drinking water is a matter of growing interest for scientists as well as health authorities. A concentration technique for endocrine activity screening was developed, optimized, and transposed from 200 mL to 10 L water samples. To avoid any contamination during concentration, the method was developed using exclusively glass, Teflon and stainless steel materials. Any potential losses were tracked using three model radiolabeled molecules, namely BPA, DEHP and 4n-NP. The final method allowed 10 L water samples to be concentrated 5000-fold, with good recovery and repeatability. After validation, by concentrating spiked and non-spiked 10 L samples of EVIAN natural mineral water, 14 different drinking water samples were concentrated and screened for endocrine disrupting activity using bioluminescent assays. Samples consisting of bottled water, conditioned in various materials (glass, PET) and subjected to different storage conditions, had no hormone-like activities whereas estrogenic activity was found in the filtered tap water.

DEHP is known to cause reproductive toxicity in rats, particularly during the neonatal period. Pregnant and brood rats were treated by gavage with 750 mg/kg b.w./day DEHP starting on GD14 within PND4. Two hours after 14C-DEHP administration on GD15, GD18, GD21 and PND4, the radioactivity content was measured in the dams blood and in the liver, gonads and carcass of the offspring. The radioactivity concentration recovered in the fetuses was one or two order of magnitude lower than the concentration found in the dam plasma. A low proportion of radioactivity was present in fetal gonads, ca. 2%, 5% and 3.6% on GD18, GD21 and PND4, respectively. The effect on testosterone production of DEHP and its metabolites (MEHP, metabolites VI and IX) was assessed in fetal testis cultures using a dose-range which included the maximal exposure observed in vivo. None of the compounds affected testosterone production. Thus, DEHP and/or its metabolites appear to cross the placental barrier, reach the fetal gonads. In vitro, neither DEHP nor its main metabolites decreased the testosterone production.