Cumulative Dietary Risk Assessment of Benzophenone-Type Photoinitiators from Packaged Foodstuffs

Photoinitiators used in ultraviolet-cured ink may migrate from food packaging materials into food products. Therefore, we conducted a dietary risk assessment of exposure to benzophenone (BP)-type photoinitiators by quantifying and reducing uncertainties associated with the risk characterization. A total of 362 food packaging samples including 180 cereals, 136 fruit and vegetable juices, and 46 milk samples were subjected to fast pesticides extraction to determine photoinitiator residues. The average daily dose (ADD) of BP was the highest in the age group of zero to three years, with a P97.5 ADD of 2.56 × 10−4 mg/kg bw/day. The ADD of 2-hydroxybenzophenone (2-OHBP) was the highest in the age group of three to six years, with a P97.5 UB ADD of 3.52 × 10−5 mg/kg bw/day. The estimated UB P97.5 ADD for each age group was below the toxicological concern threshold of 0.0015 mg/kg bw/day. The cumulative toxicity of all BPs, evaluated using the MOET value, was at an acceptable level. Although the MOET value of BPs was above the safety limit in the foodstuffs studied herein, this result may be different if Taiwan were to follow regulation guidelines for BP-type photoinitiators based on the specific migration limit for the unmeasured BP residues in other foodstuffs.

Application of Microgel as a Sorbent for Bisphenol Analysis in Liquid Food Samples

Bisphenols are well-known endocrine disruptors that can easily migrate from plastic and can containers to food. Due to the complicated matrix and ultra-low concentrations of bisphenols in food, samples require extensive preparation before instrumental analysis. In this paper, an environmental sensitive microgel was employed as a sorbent for the preconcentration of four bisphenols, bisphenol A (BPA), bisphenol B (BPB), bisphenol E (BPE) and bisphenol F (BPF), from liquid food samples. Liquid chromatography with fluorescence detection (LC-FLD) was used for the quantification of bisphenols. By applying microgel solid-phase extraction procedure, the limits of detections achieved in liquid food samples can be lowered to 0.9 µg·L−1 for BPF and BPA, 2.3 µg·L−1 for BPE and 2.9 µg·L−1 for BPB. Only 5 mg of microgel was sufficient to achieve good recoveries (70.5–109%) with precision (RSD 0.21–5.01%, n = 3) for different analyzed liquid food samples spiked at concentration levels of 50 µg·L−1. In five out of twelve of the analyzed samples (pineapple, mandarin, peach, mushroom and pickles), they were contaminated with BPA, and the determined concentration was in the range of 6.2–22 µg·L−1; however, these results are below the specific migration limit (SML) set for BPA (50 µg·kg−1).

Assessment of Four Studies on Developmental Neurotoxicity of Bisphenol A

The Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) has on the request from the Norwegian Food Safety Authority (Mattilsynet) assessed four studies on developmental neurotoxicity following low dose exposure to bisphenol A (BPA) (Adriani et al., 2003; Carr et al., 2003; Negishi et al., 2004; Ryan and Vandenbergh, 2006). The background for the request is uncertainties related to developmental neurotoxcity of BPA raised by the Nordic environmental agencies in Norway, Sweden and Denmark. VKM was asked to consider if these studies provide sufficient evidence to set a lower no observed adverse effect level (NOAEL) in the hazard characterisation of BPA. Further, a Norwegian exposure scenario based on available exposure data should be performed. The task has been assessed by the Scientific Panel on Food Additives, Flavourings, Processing Aids, Materials in Contact with Food and Cosmetics (Panel 4). Bisphenol A (CAS number 80-05-7) is primarily used as a monomer in the production of polycarbonate, which is used to make food containers, such as beverage bottles, baby bottles, tableware and storage containers. It is also used as a precursor of certain epoxy resins used for protective coatings for food and beverage cans. BPA is permitted for use in food contact materials in the European Union (EU) with a specific migration limit (SML) of 0.6 mg/kg food. The migration limit in the EU regulations has yet to be modified according to an opinion from the European Food Safety Authority (EFSA) from 2006 where a new established tolerable daily intake (TDI) of 0.05 mg BPA/kg body weight (bw) was derived from a NOAEL of 5 mg/kg bw/day. A European Union Risk Assessment Report (RAR) of BPA produced in accordance with Council Regulation (EEC) 793/93 has recently been updated (April 2008) reviewing a previously requested 2-generation study in mice (Tyl et al., 2007) and new data on human exposure and effects of BPA. A NOAEL of 50 mg/kg bw/day was suggested in this report. The Nordic environmental agencies (Norway, Sweden and Denmark) have participated in the discussions on this updated EU RAR of BPA and they strongly disagreed that this NOAEL also covers developmental neurotoxicity. According to the Nordic environmental agencies, the four above mentioned studies indicate a possible risk for developmental neurotoxicity of BPA at very low exposure levels (0.1-0.25 mg/kg bw/day). The position of the Nordic environmental agencies has been included as a footnote in the revised EU RAR. Recently, in April 2008, the U.S. National Toxicology Program (NTP), Health Canada and Environment Canada have published draft reports on effects of BPA, including developmental effects (neural and behavioural effects) and expressed some concern for neural and behavioural effects in fetuses, infants and children at current human exposures. The European Commission has therefore asked EFSA to further assess possible age dependent toxicokinetics of BPA in animals and humans and their implications for hazard and risk assessment of BPA taken into account the most recent information and data available. The present opinion from VKM Panel 4 is based on an evaluation of the design, conduct (or accomplishment) and the results in the four above mentioned studies. The study design has been evaluated in light of international recommendations given in relevant guidelines dealing with developmental neurotoxicity testing in animals. The recent international developments on BPA in the U.S. and Canada are not addressed in this opinion. The report by Tyl and co-workers was central in the EFSA opinion from 2006 and the updated EU RAR from 2008. The Tyl study is a GLP compliant 2-generation reproductive toxicity evaluation in mice performed according to a modified OECD 416 guideline. However, the study did not include functional tests for developmental neurotoxicity. VKM has reviewed the four above mentioned studies on neurodevelopmental toxicity of BPA as requested by the Norwegian Food Safety Authority. Although the design and reporting of these studies suffer from major and serious shortcomings, the overall findings may raise some concern. It is the opinion of the VKM Panel 4 that the four studies do not provide sufficient evidence for setting a robust lower NOAEL for BPA than the current EFSA NOAEL of 5 mg/kg bw/day. The Panel is aware that the EU Commission recently has requested EFSA to re-evaluate the information available on BPA. In order to eliminate any uncertainty regarding potential developmental effects of BPA at low doses, it is recommended that a GLP compliant study is carried out according to OECD guideline 426. Such a study should utilize a broad concentration range from the very low doses up to those with known maternal effects. A Norwegian exposure scenario based on available data on exposure to BPA from food and beverages and via the environment was performed. In general, exposure levels of BPA in Norway are low. The estimated exposure of infants and children is in the range of 3.5 – 13.2 μg/kg bw/day, whereas the estimated aggregated exposure of adults is 1.5 μg/kg bw/day. As a result of the current use of BPA in food contact materials and other consumer products, infants and children are exposed to higher levels of BPA per kg body weight than the rest of the population.

Migration and Degradation in Composting Environment of Active Polylactic Acid Bilayer Nanocomposites Films: Combined Role of Umbelliferone, Lignin and Cellulose Nanostructures

This study was dedicated to the functional characterization of innovative poly(lactic acid) (PLA)-based bilayer films containing lignocellulosic nanostructures (cellulose nanocrystals (CNCs) or lignin nanoparticles (LNPs)) and umbelliferone (UMB) as active ingredients (AIs), prepared to be used as active food packaging. Materials proved to have active properties associated with the antioxidant action of UMB and LNPs, as the combination of both ingredients in the bilayer formulations produced a positive synergic effect inducing the highest antioxidant capacity. The results of overall migration for the PLA bilayer systems combining CNCs or LNPs and UMB revealed that none of these samples exceeded the overall migration limit required by the current normative for food packaging materials in both non-polar and polar simulants. Finally, all the hydrophobic monolayer and bilayer films were completely disintegrated in composting conditions in less than 18 days of incubation, providing a good insight on the potential use of these materials for application as active and compostable food packaging.

Two-Layer Functional Coatings of Chitosan Particles with Embedded Catechin and Pomegranate Extracts for Potential Active Packaging

Two-layer functional coatings for polyethylene (PE) and polypropylene (PP) films were developed for the active packaging concept. Prior to coating, the polymer films were activated by O2 and NH3 plasma to increase their surface free energy and to improve the binding capacity and stability of the coatings. The first layer was prepared from a macromolecular chitosan solution, while the second (upper) layer contained chitosan particles with embedded catechin or pomegranate extract. Functionalized films were analyzed physico-chemically to elemental composition using ATR-FTIR spectroscopy and XPS. Further, oxygen permeability and wettability (Contact Angle) were examined. The antimicrobial properties were analyzed by the standard ISO 22196 method, while the antioxidative properties were determined with an ABTS assay. Functionalized films show excellent antioxidative and antimicrobial efficacy. A huge decrease in oxygen permeability was achieved in addition. Moreover, a desorption experiment was also performed, confirming that the migration profile of a compound from the surfaces was in accordance with the required overall migration limit. All these properties indicate the great potential of the developed active films/foils for end-uses in food packaging.

Oleamide, a Bioactive Compound, Unwittingly Introduced into the Human Body through Some Plastic Food/Beverages and Medicine Containers

The purpose of the study was to investigate the migration of oleamide, a polymer lubricant, and a bioactive compound, from various plastic, marketed containers for food/beverages and medicines into polymer contact liquid. Methanol, food/medicine simulants or real samples were used to extract polymer leachables and extractables. Migrated oleamide into polymer contact liquids was determined by ultra-high performance liquid chromatography coupled to mass spectrometry (UHPLC-MS). The concentration of oleamide in the extracts of medicinal and insulin syringes was 7351 ng mL−1 and 21,984 ng mL−1, respectively. The leachates of intravenous (i.v.) infusion bottle, medicinal and insulin syringes contained 17 ng mL−1, 12 ng mL−1 and 152 ng mL−1, respectively. Oleamide in the extracts of dummies ranged from 30 to 39 ng mL−1, while in the leachates of baby bottles, from 12 to 23 ng mL−1. Leachates of soft drink bottles contained from 6 to 15 ng mL−1 oleamide, milk bottles from 3 to 9 ng mL−1, liquid yogurt bottles 17 ng mL−1 and water bottles from 11 to 18 ng mL−1. Bottled real matrices of oil and milk contained oleamide in the range from 217 to 293 ng mL−1. Moreover, the source of migrated oleamide (e.g., containers, caps, other parts) was identified. Oleamide is listed in the current EU regulations without a specific migration limit. Accordingly, these values are considered of no concern, unless future toxicological studies prove the opposite.

Dialkylketones in Paperboard Food Contact Materials—Method of Analysis in Fatty Foods and Comparative Migration into Liquid Simulants Versus Foodstuffs

Dialkyl diketene dimers are used as sizing agents in the manufacture of paper and board for food contact applications to increase wetting stability. Unbound residues can hydrolyze and decarboxylate into dialkylketones. These non-intentionally added substances (NIAS) have potential to migrate to fatty foods in contact with those packaging materials. In Germany, the Federal Institute for Risk Assessment (BfR) established a specific migration limit (SML) of 5 mg/kg for the transfer of these dialkylketones into foodstuffs. In order to investigate the differences between simulants and real foods, an analytical method was optimized for extraction and quantification of dialkylketones in edible oils and fatty foods by gas chromatography coupled with flame ionization detection (GC-FID), and additionally by gas chromatography with mass spectrometry (GC-MS), to confirm their identification and to quantify them in case of interferences. Dialkylketones are separated from the extracted fat by alkaline saponification of the triglycerides. Dialkylketones migration from paper-based food contact articles into organic solvents isooctane and dichloromethane, in olive and sunflower oils, and in fatty foods (croissants, Gouda, cheddar cheese, and salami was studied). As a result, it was found that the simulating tests, including the edible oil extraction tests, gave migration values that exceeded the SML largely, while the migration with the food samples were largely below the SML.

The Impact of Storage on Phthalic Acid Esters Concentrations in Yogurts Packed in Plastic Cups

Phthalic acid esters are used as plastic softeners and also can be found in food packaging materials. European legislation defines specific migration limits of plastic additives for plastic materials that come into contact with food. This study monitors the phthalic acid ester concentrations in yogurts after manufacturing and then after a 3‑week storage. The studied yoghurts were natural yogurt with 1 % of chia flour, natural yogurt with 5 % of chia flour, natural yogurt with 1 % of bamboo fibre, natural yogurt with 5 % of bamboo fibre and natural yogurt. The analysed phthalic acid esters were dibutyl phthalate (DBP) and di‑(2‑ethylhexyl) phthalate (DEHP). The average phthalate concentrations in plastic cups were detected for DBP of 59.5 µg/g and for DEHP of 9.0 µg/g of the plastic material. Higher DBP concentrations than DEHP concentrations were also found in all studied yogurts. The average DBP concentrations in yogurts were detected from 1.8 µg/g up to 5.0 µg/g of the original matter and the average DEHP concentrations were determined from 0.5 µg/g up to 1.0 µg/g of the original matter. No statistically significant difference was found when comparing phthalic acid ester concentrations in yogurts immediately after production and after three weeks of storage. However, in our study in all cases of yogurts, the DBP concentrations were higher than the specific migration limit set by the legislation (0.3 mg/kg) and the DEHP concentrations were in all cases of yogurts lower than the specific migration limit set by the legislation (1.5 mg/kg).

Time course of chemotaxis and chemokinesis of neutrophils following stimulation with IL-8 or FMLP

Polymorphonuclear cells (PMNs) attend to inflammatory sites by chemotactic movement, caused by chemoattractants (CAs) like n-formyl-l-methionyl-l-leucyl-l-phenylalanine (FMLP) and interleukin-8 (IL-8). However, distinct but applicable assays for investigations of PMNs’ migration limit in vitro examination. We integrated CD15-bead-based isolation of PMNs with analysing their chemotaxis in a novel 3D-µ-Slide migration chamber. The PMNs were exposed to different concentrations of FMLP and IL-8 (1, 10 and 100 nM) and observed for 180 min in cell-physiological environment conditions. Moving PMNs’ percentage (median and interquartile range) decreased from 62% (27%) to 36% (31%) without CA, from 88% (30%) to 22% (26%) for 1 nM IL-8, from 70% (22%) to 28% (13%) for 100 nM IL-8, from 30% (23%) to 18% (46%) for 1 nM FMLP and from 76% (20%) to 28% (13%) for 100 nM FMLP. Centres of cell movement turned towards the CAs (negative values) within a single 30-min observation period: 5.37 µm (16.82 µm) without CA, −181.37 µm (132.18 µm) with 10 nM and −239.34 µm (152.19 µm) with 100 nM IL-8; −116.2 µm (69.07 µm) with 10 nM and −71.59 µm (98.58 µm) with 100 nM FMLP. FMLP and IL-8 ensure chemotaxis without increase of chemokinesis. 3D-µ-Slide chemotaxis chambers facilitate time course analyses of PMNs’ migration in stable conditions over a long time with concise distinction of chemotaxis and chemokinesis.