Migration Studies and Endocrine Disrupting Activities: Chemical Safety of Cosmetic Plastic Packaging

International audienceThe endocrine activity and endocrine disruptor (ED) chemical profiles of eleven plastic packaging materials covering five major polymer types (3PET, 1HDPE, 4LDPE, 2 PP, and 1SAN) were investigated using in vitro cell-based reporter-gene assays and a non-targeted chemical analysis using gas chromatography coupled to mass spectrometry (GC-MS). To mimic cosmetic contact, six simulants (acidic, alkaline, neutral water, ethanol 30%, glycerin, and paraffin) were used in migration assays performed by filling the packaging with simulant. After 1 month at 50 °C, simulants were concentrated by Solid Phase Extraction (SPE) or Liquid-Liquid Extraction (LLE). The migration profiles of seven major endocrine disrupting chemicals detected from GC-MS in the different materials and simulants were compared with Estrogen Receptor (ER) and Androgen Receptor (AR) activities. With low extraction of ED chemicals in aqueous simulants, no endocrine activities were recorded in the leachates. Paraffin was shown to be the most extracting simulant of antiandrogenic chemicals, while glycerin has estrogenic activities. Overall, ED chemical migration in paraffin was correlated with hormonal activity. The NIAS 2,4-di-tert-butyl phenol and 7,9-di-tert-butyl1-oxaspiro (4,5) deca-6,9-diene-2,8-dione were two major ED chemicals present in all polymers (principally in PP and PE) and in the highest quantity in paraffin simulant. The use of glycerin and liquid paraffin as cosmetic product simulants was demonstrated to be relevant and complementary for the safety assessment of released compounds with endocrine activities in this integrated strategy combining bioassays and analytical chemistry approaches

Risk assessment of endocrine-disrupting cocktail effect from multiexposure to plasticizers of medical devices in the neonatal population

International audienc

Use of bioassays to assess hazard of food contact material extracts: State of the art

IF 3.778International audienceThis review focuses on the use of in vitro bioassays for the hazard assessment of food contact materials (FCM) as a relevant strategy, in complement to analytical methods. FCM may transfer constituents to foods, not always detected by analytical chemistry, resulting in low but measurable human exposures. Testing FCM extracts with bioassays represents the biological response of a combination of substances, able to be released from the finished materials. Furthermore, this approach is particularly useful regarding the current risk assessment challenges with unpredicted/unidentified non-intentionally added substances (NIAS) that can be leached from the FCM in the food. Bioassays applied to assess hazard of different FCM types are described for, to date, the toxicological endpoints able to be expressed at low levels; cytotoxicity, genotoxicity and endocrine disruption potential. The bioassay strengths and relative key points needed to correctly use and improve the performance of bioassays for an additional FCM risk assessment is developed. This review compiles studies showing that combining both chemical and toxicological analyses presents a very promising and pragmatic tool for identifying new undesirable NIAS (not predicted) which can represent a great part of the migrating substances and/or "cocktail effect"

Evaluation of the sensitivity of three sublethal cytotoxicity assays in human HepG2 cell line using water contaminants

The in vitro toxicological index IC50 (the millimolar concentration of compound which inhibits response assay by 50% compared to the solvent control) of 11 water contaminants (acrylamide, atrazine, B[a]P, BPA, 2,4-DAT, 17-alphaEE, H(2)O(2), 4-OP, sodium bromate, sodium chlorate, sodium nitrate) was evaluated on the human hepatoma (HepG2) cells using three short-term bioassays related to their morbidity status [radiometric RNA synthesis assay (RNA), luminometric ATP assay (ATP), fluorometric Alamar blue assay (AB)]. Among all substances, we were not able to determine atrazine IC50 value whatever the test used. Furthermore, B[a]P was not cytotoxic in the ATP and AB assays. Statistical analysis revealed a correlation between the IC50 values obtained in the three assays. Except with 4-OP, RNA assay was always inhibited at lower concentrations than those required in the other assays, suggesting that this assay is a very sensitive indicator of the presence of toxic compounds. ATP and AB assays responded to a similar pattern. Due to its higher sensitivity and its reliability, RNA synthesis assay using HepG2 cell line provides the most suitable tool for the screening of water contaminants

Food contact materials: Strategy for quality and food safety of packaging systems

National audienc

In vitro genotoxicity assessment of MTES, GPTES and TEOS, three precursors intended for use in food contact coatings

International audienceOrganoalkoxysilanes are precursors that are used increasingly in the synthesis of food contact coatings. To comply with the EU regulation, their potential toxicity must be assessed, and very little information is known. The genotoxicity of three common precursors was studied, namely, tetraethylorthosilicate (TEOS), methyltriethoxysilane (MTES) and 3-glycidyloxypropyltriethoxysilane (GPTES). By the Ames test, MTES and TEOS were not mutagenic for bacteria. A significant positive response was observed with GPTES in the TA100 and TA1535 strains. The mutagenic effect was more pronounced in the presence of the exogenous metabolic activation system with an increase of the induction factor (ten-fold higher for the TA1535 strain). In the micronucleus assay performed with a human hepatoma cell line (HepG2 cells), GPTES gave negative results even in the presence of an exogenous activation system. To ascertain the possibility of using this precursor in food contact material, its migration must be monitored according to the coating formulation because migration might result in hazardous human exposure

Reliable bioassays to detect potential hazard of paperboard food contact material extracts

Food contact materials (FCM) represent a major economic issue and a large field of innovation. Food packaging production must be in compliance with the European Regulation 1935/2004: the third article specifies that FCM must not transfer their constituents to food in quantities which could endanger human health under normal or foreseeable conditions of use. Indeed, these materials are not inert and, in addition to started substances, Non-Intentionally Added Substances (NIAS) are able to be released into the food. NIAS can be contaminants from recycled materials, impurities, synthetic residues, new substances formed along the packaging production chain… Then, NIAS could represent a larget part of migrating substances which are not risk assessed, some being unpredictabled (Grob et al., 2006 ; Skejevrak et al., 2005). The European Regulation 10/2011 on plastic materials, multilayers and articles intended to come into contact with food regulates NIAS. It requires that the notion of the risk engendered by a substance concerns the substance itself, the impurities of this substance and any reaction or degradation products that are foreseeable in the context of the intended use. However there are currently no available guidelines to assess the risk of NIAS. They may also be present in other FCM than plastics, such as paper and boards... Because of the complex nature of FCM extract, it is pertinent to evaluate the potential toxicity of FCM as a whole taking into account any « cocktail effect ». The objectives of this study are to provide to packaging and food companies scientific relevant tools combining physicochemical and toxicological strategies based on bioassays applied to FCM extracts, especially paper and boards. Four toxicological endpoints were checked as relevant for low dose exposure: cytotoxicity, genotoxicity , oxidative stress and endocrine disruption activities

In vitro toxicity assessment of extracts derived from sol–gel coatings on polycarbonate intended to be used in food contact applications

IF 3.584International audiencePolycarbonate is a widely used polymer in food contact applications all around the world. However, due to the potential release of Bisphenol A (BPA) during repeated washing cycles, its use becomes compromised as BPA is known for being an endocrine disruptor for rodents. In order to tackle this issue, sol-gel coatings based on organoalkoxysiloxane were developed on PC, to act as a physical barrier. To this end, two sol-gel systems based on tetraethylorthosilicate (TEOS), methyltriethoxysilane (MTES) and 3-glycidyloxypropyltriethoxysilane (GPTES), three common sol-gel precursors, were prepared. The coatings derived from the latter two systems were then studied with regards to their potential toxicity in vitro. Migration tests were performed in food simulants, and the maximal migration was obtained in ethanol 10% (v/v) for one system and in isooctane for the other one. In vitro genotoxicity was assessed with the Ames test (OECD 471) and the micronucleus assay (OECD 487), and no genotoxic effect was observed. Moreover, the estrogenic activity of the extracts was studied with a transcriptional activation assay using transient transfection in human cells; none of the extracts was found estrogenic. These negative in vitro results are highly promising for the future use of these new barrier coating formulations onto food contact materials. (C) 2016 Elsevier Ltd. All rights reserved

Consequences of physical post treatments (Microwave and Electron Beam) on the food / packaging interactions: a physicochemical and toxicological approach

The safety of microwave and electron-beam treatments has been demonstrated, in regards to the formation of reaction products that could endanger human health. An integrated approach was used combining the potential toxicity of all the substances likely to migrate to their chemical characterizations. This approach was applied to polypropylene (PP) films prepared with a selection of additives. Components were identified by liquid and gas chromatography using a mass selective detector system. Their potential toxicity was assessed using three in vitro short-term bioassays and their migrations were carried out using a standards-based approach. After the electron-beam treatment some additives decomposed and there was a significant increase in the polyolefin oligomeric saturated hydrocarbons concentration. PP prepared with Irgafos 168 led to a significantly strong cytotoxic effect and PP prepared with Irganox 1076 induced a dose-dependant estrogenic effect in vitro. Migration values were low and below the detection limit of the analytical method applied