Xenia Trier

The Madrid Statement on Poly- and Perfluoroalkyl Substances (PFASs)

As scientists and other professionals from a variety of disciplines, we are concerned about the production and release into the environment of an increasing number of poly- and perfluoroalkyl substances (PFASs) for the following reasons: PFASs are man-made and found everywhere. PFASs are highly persistent, as they contain perfluorinated chains that only degrade very slowly, if at all, under environmental conditions. It is documented that some polyfluorinated chemicals break down to form perfluorinated ones (D’Eon and Mabury 2007). PFASs are found in the indoor and outdoor environments, wildlife, and human tissue and bodily fluids all over the globe. They are emitted via industrial processes and military and firefighting operations (Darwin 2011; Fire Fighting Foam Coalition 2014), and they migrate out of consumer products into air (Shoeib et al. 2011), household dust (Bjorklund et al. 2009), food (Begley et al. 2008; Tittlemier et al. 2007; Trier et al. 2011), soil (Sepulvado et al. 2011; Strynar et al. 2012), ground and surface water, and make their way into drinking water (Eschauzier et al. 2012; Rahman et al. 2014). In animal studies, some long-chain PFASs have been found to cause liver toxicity, disruption of lipid metabolism and the immune and endocrine systems, adverse neurobehavioral effects, neonatal toxicity and death, and tumors in multiple organ systems (Lau et al. 2007; Post et al. 2012). In the growing body of epidemiological evidence, some of these effects are supported by significant or suggestive associations between specific long-chain PFASs and adverse outcomes, including associations with testicular and kidney cancers (Barry et al. 2013; Benbrahim-Tallaa et al. 2014), liver malfunction (Gallo et al. 2012), hypothyroidism (Lopez-Espinosa et al. 2012), high cholesterol (Fitz-Simon et al. 2013; Nelson et al. 2009), ulcerative colitis (Steenland et al. 2013), lower birth weight and size (Fei et al. 2007), obesity (Halldorsson et al. 2012), decreased immune response to vaccines (Grandjean et al. 2012), and reduced hormone levels and delayed puberty (Lopez-Espinosa et al. 2011). Due to their high persistence, global distribution, bioaccumulation potential, and toxicity, some PFASs have been listed under the Stockholm Convention (United Nations Environment Programme 2009) as persistent organic pollutants (POPs). As documented in the Helsingor Statement (Scheringer et al. 2014), Although some of the long-chain PFASs are being regulated or phased out, the most common replacements are short-chain PFASs with similar structures, or compounds with fluorinated segments joined by ether linkages. While some shorter-chain fluorinated alternatives seem to be less bioaccumulative, they are still as environmentally persistent as long-chain substances or have persistent degradation products. Thus, a switch to short-chain and other fluorinated alternatives may not reduce the amounts of PFASs in the environment. In addition, because some of the shorter-chain PFASs are less effective, larger quantities may be needed to provide the same performance. While many fluorinated alternatives are being marketed, little information is publicly available on their chemical structures, properties, uses, and toxicological profiles. Increasing use of fluorinated alternatives will lead to increasing levels of stable perfluorinated degradation products in the environment, and possibly also in biota and humans. This would increase the risks of adverse effects on human health and the environment. Initial efforts to estimate overall emissions of PFASs into the environment have been limited due to uncertainties related to product formulations, quantities of production, production locations, efficiency of emission controls, and long-term trends in production history (Wang et al. 2014). The technical capacity to destroy PFASs is currently insufficient in many parts of the world. Global action through the Montreal Protocol (United Nations Environment Programme 2012) successfully reduced the use of the highly persistent ozone-depleting chlorofluorocarbons (CFCs), thus allowing for the recovery of the ozone layer. However, many of the organofluorine replacements for CFCs are still of concern due to their high global warming potential. It is essential to learn from such past efforts and take measures at the international level to reduce the use of PFASs in products and prevent their replacement with fluorinated alternatives in order to avoid long-term harm to human health and the environment. For these reasons, we call on the international community to cooperate in limiting the production and use of PFASs and in developing safer nonfluorinated alternatives. We therefore urge scientists, governments, chemical and product manufacturers, purchasing organizations, retailers, and consumers to take the following actions:

Helsingør Statement on poly- and perfluorinated alkyl substances (PFASs)

In this discussion paper, the transition from long-chain poly- and perfluorinated alkyl substances (PFASs) to fluorinated alternatives is addressed. Long-chain PFASs include perfluoroalkyl carboxylic acids (PFCAs) with 7 or more perfluorinated carbons, perfluoroalkyl sulfonic acids (PFSAs) with 6 or more perfluorinated carbons, and their precursors. Because long-chain PFASs have been found to be persistent, bioaccumulative and toxic, they are being replaced by a wide range of fluorinated alternatives. We summarize key concerns about the potential impacts of fluorinated alternatives on human health and the environment in order to provide concise information for different stakeholders and the public. These concerns include, amongst others, the likelihood of fluorinated alternatives or their transformation products becoming ubiquitously present in the global environment; the need for more information on uses, properties and effects of fluorinated alternatives; the formation of persistent terminal transformation products including PFCAs and PFSAs; increasing environmental and human exposure and potential of adverse effects as a consequence of the high ultimate persistence and increasing usage of fluorinated alternatives; the high societal costs that would be caused if the uses, environmental fate, and adverse effects of fluorinated alternatives had to be investigated by publicly funded research; and the lack of consideration of non-persistent alternatives to long-chain PFASs.

Fluorochemicals used in food packaging inhibit male sex hormone synthesis

Polyfluoroalkyl phosphate surfactants (PAPS) are widely used in food contact materials (FCMs) of paper and board and have recently been detected in 57% of investigated materials. Human exposure occurs as PAPS have been measured in blood; however knowledge is lacking on the toxicology of PAPS. The aim of this study was to elucidate the effects of six fluorochemicals on sex hormone synthesis and androgen receptor (AR) activation in vitro. Four PAPS and two metabolites, perfluorooctanoic acid (PFOA) and 8:2 fluorotelomer alcohol (8:2 FTOH) were tested. Hormone profiles, including eight steroid hormones, generally showed that 8:2 diPAPS, 8:2 monoPAPS and 8:2 FTOH led to decreases in androgens (testosterone, dehydroepiandrosterone, and androstenedione) in the H295R steroidogenesis assay. Decreases were observed for progesterone and 17-OH-progesterone as well. These observations indicated that a step prior to progestagen and androgen synthesis had been affected. Gene expression analysis of StAR, Bzrp, CYP11A, CYP17, CYP21 and CYP19 mRNA showed a decrease in Bzrp mRNA levels for 8:2 monoPAPS and 8:2 FTOH indicating interference with cholesterol transport to the inner mitochondria. Cortisol, estrone and 17β-estradiol levels were in several cases increased with exposure. In accordance with these data CYP19 gene expression increased with 8:2 diPAPS, 8:2 monoPAPS and 8:2 FTOH exposures indicating that this is a contributing factor to the decreased androgen and the increased estrogen levels. Overall, these results demonstrate that fluorochemicals present in food packaging materials and their metabolites can affect steroidogenesis through decreased Bzrp and increased CYP19 gene expression leading to lower androgen and higher estrogen levels.

Tools to discover anionic and nonionic polyfluorinated alkyl surfactants by liquid chromatography electrospray ionisation mass spectrometry

A tiered approach is proposed for the discovery of unknown anionic and nonionic polyfluorinated alkyl surfactants (PFASs) by reversed phase ultra high performance liquid chromatography (UHPLC)--negative electrospray ionisation--quadrupole time of flight mass spectrometry (UHPLC-ESI(-)-QTOF-MS). The chromatographic separation, ionisation and detection of PFASs mixtures, was achieved at high pH (pH=9.7) with NH(4)OH as additive. To distinguish PFASs from other chemicals we used the characteristic negative mass defects of PFASs, their specific losses of 20 Da (HF) and the presence of series of chromatographic peaks, belonging to homologues series with m/z of n×50 Da (CF(2)) or n×100 Da (CF(2)CF(2)). The elemental composition of the precursor ions were deducted from the accurate m/z values of the deprotonated molecules [M-H](-). In case of in-source fragmentation, the presence of dimers, e.g. [M(2)-H](-) and adduct ions such as [M-H+solvent](-) and [(M-H)(M-H+Na)(n)](-) were used to confirm the identity of the precursor ions. In relation to quantification of PFASs, we discuss how their surfactancy influence the ESI processes, challenge their handling in solution and choices of precursor-to-product ions for MSMS of e.g., structural PFAS isomers. The method has been used to discover PFASs in industrial blends and in extracts from food contact materials.

Structural isomers of polyfluorinated di- and tri-alkylated phosphate ester surfactants present in industrial blends and in microwave popcorn bags

IntroductionIn this study, we provide strategies for detecting and quantifying the structural isomers of polyfluorinated di- and tri-alkyl surfactants (PFAS) by mass spectrometry (MS). We specifically investigate polyfluorinated dialkylated phosphate ester surfactants (x:2/y:2 diPAPS, (F(CF2)xCH2CH2O-P(O)(O)−-OCH2CH2(CF2)yF)) and their thioether analogues (x:2/y:2 S-diPAPS, F(CF2)xCH2CH2SCH2-C[CH2O)2P(O)(O)−]-CH2SCH2CH2(CF2)yF), which are used for industrial applications, such as oil- and water-repellent coatings on paper and board. DiPAPS have been found in human blood and are metabolised to the persistent perfluoroalkyl carboxylic acids (PFCA) in rats.Materials and methodsA microwave popcorn bag extract was analysed by ultrahigh-pressure liquid chromatography coupled to a negative electrospray ionisation-quadrupole time-of-flight MS.Results and discussionThe extract contained S-diPAPS, diPAPS and trialkylated (triPAPS) impurities. TriPAPS were also present in industrial and synthetic diPAPS standards, and were verified with an 8:2/8:2/8:2 triPAPS standard. The eight elemental compositions (m/z’s) of diPAPS in the extract represent 19 precursor ion structures, and the six S-diPAPS m/z’s represent at least 13 structures. The diPAPS had [M-H]− precursor ions of m/z 789, 889,…1,489 and the S-diPAPS of m/z 921, 1,021,…1,421, corresponding to fluorinated chains from C6–18. Each m/z appeared as one to three chromatographic peaks of structural isomers, where, e.g. m/z 1,189 was present as 10:2/10:2, 8:2/12:2 and 6:2/14:2 diPAPS. The isomers formed different products ions, thus only half of the m/z 1,189 diPAPS concentration was measured with one precursor ion > product ion transition.ConclusionIn general, knowledge about structural isomers of poly-alkylated PFAS is needed for the estimation of types and amounts of perfluorinated degradation products, such as PFCA from diPAPS.

Non-targeted screening for contaminants in paper and board food-contact materials using effect-directed analysis and accurate mass spectrometry

ABSTRACT Due to large knowledge gaps in chemical composition and toxicological data for substances involved, paper and board food-contact materials (P&B FCM) have been emerging as a FCM type of particular concern for consumer safety. This study describes the development of a step-by-step strategy, including extraction, high-performance liquid chromatography (HPLC) fractionation, tentative identification of relevant substances and in vitro testing of selected tentatively identified substances. As a case study, we used two fractions from a recycled pizza box sample which exhibited aryl hydrocarbon receptor (AhR) activity. These fractions were analysed by gas chromatography (GC) and ultra-HPLC (UHPLC) coupled to quadrupole time-of-flight mass spectrometers (QTOF MS) in order tentatively to identify substances. The elemental composition was determined for peaks above a threshold, and compared with entries in a commercial mass spectral library for GC-MS (GC-EI-QTOF MS) analysis and an in-house built library of accurate masses for substances known to be used in P&B packaging for UHPLC-QTOF analysis. Of 75 tentatively identified substances, 15 were initially selected for further testing in vitro; however, only seven were commercially available and subsequently tested in vitro and quantified. Of these seven, the identities of three pigments found in printing inks were confirmed by UHPLC tandem mass spectrometry (QqQ MS/MS). Two pigments had entries in the database, meaning that a material relevant accurate mass database can provide a fast tentative identification. Pure standards of the seven tentatively identified substances were tested in vitro but could not explain a significant proportion of the AhR-response in the extract. Targeted analyses of dioxins and PCBs, both well-known AhR agonists, was performed. However, the dioxins could explain approximately 3% of the activity observed in the pizza box extract indicating that some very AhR active substance(s) still remain to be identified in recycled low quality P&B.

Primary aromatic amines (PAAs) in black nylon and other food-contact materials, 2004–2009

Primary aromatic amines (PAAs) were analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in migrates from 234 samples of food-contact materials, including black nylon (polyamide) kitchen utensils (n = 136), coloured plastics (28), and clear/printed multilayer film/laminates (41), from retailers, importers, and food producers. A further 29 utensils in use were obtained from colleagues. Very high PAA migration was found from black nylon kitchen utensils to the food simulant 3% acetic acid: the ‘non-detectable’ limit (20 µg aniline equivalents kg−1 food) was exceeded by up to 2100 times. All the other materials were compliant. The majority of the non-compliant utensils came from China. The predominant PAAs were aniline and 4,4′-methylenedianiline (4,4′-MDA). The frequency of violations decreased from the year 2004 (55%) to the autumn of 2005 (13%), possibly due to increased demands for in-house documentation, but they remained almost constant from 2005 to 2009. The validity of the results was shown by recovery studies, participation in proficiency testing, and comparative testing of utensils by two laboratories. Migration modelling was used to compare how various compliance migration test conditions influenced the final test results. Long-term release of PAAs was fitted by diffusion modelling experiments and long-term release was also seen as expected from used utensils. Toxicologists consider these migration levels of the suspected carcinogenic PAAs as a problem of major concern.

Fluorinated alkyl substances and technical mixtures used in food paper-packaging exhibit endocrine-related activity in vitro

Migration of chemicals from packaging materials to foods may lead to human exposure. Polyfluoroalkyl substances (PFAS) can be used in technical mixtures (TMs) for use in food packaging of paper and board, and PFAS have been detected in human serum and umbilical cord blood. The specific structures of the PFAS in TMs are often unknown, but polyfluorinated alkyl phosphate esters (PAPs) have been characterized in TMs, food packaging, and in food. PAPs can be metabolized into fluorotelomer alcohols (FTOHs) and perfluoroalkyl carboxylic acids (PFCAs). Some PFAS have endocrine activities, highlighting the need to investigate these effects. Herein, we studied the endocrine activity of less characterized PFAS, including short‐chain PFCAs and FTOHs, PAPs, and TMs of unknown chemical composition. Long‐chain PFCAs were also included. We applied seven assays covering effects on estrogen, glucocorticoid, androgen, and peroxisome proliferator‐activated receptor (PPAR) activity, as well as steroidogenesis in vitro and ex vivo. In general, PAPs, FTOHs, TMs, and long‐chain PFCAs showed estrogenic activity through receptor activation and/or increasing 17β‐estradiol levels. Furthermore, short‐ and long‐chain PFCAs activated PPARα and PPARγ. Collectively, this means that (i) PAPs, FTOHs, and PFCAs exhibit endocrine activity through distinct and sometimes different mechanisms, (ii) two out of three tested TMs exhibited estrogenic activity, and (iii) short‐chain FTOHs showed estrogenic activity and short‐chain PFCAs generally activate both PPARα and PPARγ with similar potency and efficacy as long‐chain PFCAs. In conclusion, several new and divergent toxicological targets were identified for different groups of PFAS.

Scientific challenges in the risk assessment of food contact materials

Background: Food contact articles (FCAs) are manufactured from food contact materials (FCMs) that include plastics, paper, metal, glass, and printing inks. Chemicals can migrate from FCAs into food during storage, processing, and transportation. Food contact materials’ safety is evaluated using chemical risk assessment (RA). Several challenges to the RA of FCAs exist. Objectives: We review regulatory requirements for RA of FCMs in the United States and Europe, identify gaps in RA, and highlight opportunities for improving the protection of public health. We intend to initiate a discussion in the wider scientific community to enhance the safety of food contact articles. Discussion: Based on our evaluation of the evidence, we conclude that current regulations are insufficient for addressing chemical exposures from FCAs. RA currently focuses on monomers and additives used in the manufacture of products, but it does not cover all substances formed in the production processes. Several factors hamper effective RA for many FCMs, including a lack of information on chemical identity, inadequate assessment of hazardous properties, and missing exposure data. Companies make decisions about the safety of some food contact chemicals (FCCs) without review by public authorities. Some chemical migration limits cannot be enforced because analytical standards are unavailable. Conclusion: We think that exposures to hazardous substances migrating from FCAs require more attention. We recommend a) limiting the number and types of chemicals authorized for manufacture and b) developing novel approaches for assessing the safety of chemicals in FCAs, including unidentified chemicals that form during or after production. https://doi.org/10.1289/EHP644

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.