Quantification Approaches in Non-Target LC/ESI/HRMS Analysis: An Interlaboratory Comparison.

Nontargeted screening (NTS) utilizing liquid chromatography electrospray ionization high-resolution mass spectrometry (LC/ESI/HRMS) is increasingly used to identify environmental contaminants. Major differences in the ionization efficiency of compounds in ESI/HRMS result in widely varying responses and complicate quantitative analysis. Despite an increasing number of methods for quantification without authentic standards in NTS, the approaches are evaluated on limited and diverse data sets with varying chemical coverage collected on different instruments, complicating an unbiased comparison. In this interlaboratory comparison, organized by the NORMAN Network, we evaluated the accuracy and performance variability of five quantification approaches across 41 NTS methods from 37 laboratories. Three approaches are based on surrogate standard quantification (parent-transformation product, structurally similar or close eluting) and two on predicted ionization efficiencies (RandFor-IE and MLR-IE). Shortly, HPLC grade water, tap water, and surface water spiked with 45 compounds at 2 concentration levels were analyzed together with 41 calibrants at 6 known concentrations by the laboratories using in-house NTS workflows. The accuracy of the approaches was evaluated by comparing the estimated and spiked concentrations across quantification approaches, instrumentation, and laboratories. The RandFor-IE approach performed best with a reported mean prediction error of 15× and over 83% of compounds quantified within 10× error. Despite different instrumentation and workflows, the performance was stable across laboratories and did not depend on the complexity of water matrices.

Zürich II Statement on Per- and Polyfluoroalkyl Substances (PFASs): Scientific and Regulatory Needs.

Per- and polyfluoroalkyl substances (PFASs) are a class of synthetic organic chemicals of global concern. A group of 36 scientists and regulators from 18 countries held a hybrid workshop in 2022 in Zürich, Switzerland. The workshop, a sequel to a previous Zürich workshop held in 2017, deliberated on progress in the last five years and discussed further needs for cooperative scientific research and regulatory action on PFASs. This review reflects discussion and insights gained during and after this workshop and summarizes key signs of progress in science and policy, ongoing critical issues to be addressed, and possible ways forward. Some key take home messages include: 1) understanding of human health effects continues to develop dramatically, 2) regulatory guidelines continue to drop, 3) better understanding of emissions and contamination levels is needed in more parts of the world, 4) analytical methods, while improving, still only cover around 50 PFASs, and 5) discussions of how to group PFASs for regulation (including subgroupings) have gathered momentum with several jurisdictions proposing restricting a large proportion of PFAS uses. It was concluded that more multi-group exchanges are needed in the future and that there should be a greater diversity of participants at future workshops.

Determination of perfluorinated compounds in aquatic organisms: a review.

Bioaccumulation of PFAS in aquatic organisms is an environmental problem of growing concern around the world. This problem has been tackled by regulatory bodies by proposing EQS for biota in EU water bodies and tolerable daily intake for food. The introduction of regulatory limits requires the availability of harmonised and validated analytical methods of sufficient sensitivity. This paper reviews recent advances in analytical methods for analysis of PFAS in aquatic organisms. The methods available for biota analysis are mostly based on three extraction procedures: ion-pair extraction, solvent liquid extraction, and alkaline digestion. The resulting extracts are then subjected to different clean-up or enrichment steps on solid adsorbents, for example graphitized carbon black, C(18), and WAX phases. All methods reviewed in this work give reliable results but are partially validated only, because of the lack of certified reference materials and regular interlaboratory exercises. The few interlaboratory exercises performed on real unspiked samples did not afford satisfactory results for PFAS other than PFOS, especially for matrices with high lipid content, for example mussels. The reasons for those partially negative results have been identified, and can mainly be attributed to calibration procedures and availability and purity of standards. The urgent need for certified reference materials for this type of analysis is emphasized.

Occurrence and risk characterization of per- and polyfluoroalkyl substances in seafood from the Persian Gulf.

Potential exposure to 14 per- and polyfluoroalkyl substances (PFAS) through seafood consumption was investigated in widely consumed seafood (Platycephalus indicus, Lethrinus nebulosus, and Penaeus semisulcatus) from the Persian Gulf. A total of 61 samples of fish and prawns were purchased from local fishers at Bushehr port (Persian Gulf, South-West of Iran) and were analyzed for PFAS compounds. In addition, potential factors influencing factor of PFAS bioaccumulation in fish and invertebrates such as age, sex, and habitat, were investigated. Æ©PFAS concentrations were in the range of 2.3- 6.1 ng/g-d.w (mean = 3.9 ± 1.9) in studied species which are equal to 0.46-1.2 ng/g-w.w according to their conversion factor. Perfluorooctane sulfonic acid (PFOS) was the most abundant perfluorinated compound in studied organisms and tissues. The results of correlation analysis showed that the bioaccumulation of PFAS in aquatic organisms is significantly correlated to the length of the compound's carbon chain, the identity of anionic group, and organism's age, sex, and habitant. The risk assessment using hazard index calculation and Monte-Carlo simulation indicated that weekly consumption of prawn and fish fillets does not pose a health risk to adults but might threaten children's health. However, the risk posed by PFAS exposure via entire fish or fish liver intake is an important issue for wild marine mammals (i.e., dolphins). So, accurate and routine monitoring of PFAS in aquatic environments seems mandatory to preserve wildlife and human health in the Persian Gulf.

Study of antibiotic resistance in freshwater ecosystems with low anthropogenic impact.

This study aimed to investigate the bacterial diversity and the background level of antibiotic resistance in two freshwater ecosystems with low anthropogenic impact in order to evaluate the presence of natural antimicrobial resistance in these areas and its potential to spread downstream. Water samples from a pre-Alpine and an Apennine river (Variola and Tiber, respectively) were collected in three different sampling campaigns and bacterial diversity was assessed by 16S sequencing, while the presence of bacteria resistant to five antibiotics was screened using a culturable approach. Overall bacterial load was higher in the Tiber River compared with the Variola River. Furthermore, the study revealed the presence of resistant bacteria, especially the Tiber River showed, for each sampling, the presence of resistance to all antibiotics tested, while for the Variola River, the detected resistance was variable, comprising two or more antibiotics. Screening of two resistance genes on a total of one hundred eighteen bacterial isolates from the two rivers showed that blaTEM, conferring resistance to ß-lactam antibiotics, was dominant and present in ~58 % of isolates compared to only ~9 % for mefA/E conferring resistance to macrolides. Moreover, ß-lactam resistance was detected in various isolates showing also resistance to additional antibiotics such as macrolides, aminoglycosides and tetracyclines. These observations would suggest the presence of co-resistant bacteria even in non-anthropogenic environments and this resistance may spread from the environment to humans and/or animals.

What is in the fish? Collaborative trial in suspect and non-target screening of organic micropollutants using LC- and GC-HRMS.

A collaborative trial involving 16 participants from nine European countries was conducted within the NORMAN network in efforts to harmonise suspect and non-target screening of environmental contaminants in whole fish samples of bream (Abramis brama). Participants were provided with freeze-dried, homogenised fish samples from a contaminated and a reference site, extracts (spiked and non-spiked) and reference sample preparation protocols for liquid chromatography (LC) and gas chromatography (GC) coupled to high resolution mass spectrometry (HRMS). Participants extracted fish samples using their in-house sample preparation method and/or the protocol provided. Participants correctly identified 9-69 % of spiked compounds using LC-HRMS and 20-60 % of spiked compounds using GC-HRMS. From the contaminated site, suspect screening with participants' own suspect lists led to putative identification of on average ∼145 and ∼20 unique features per participant using LC-HRMS and GC-HRMS, respectively, while non-target screening identified on average ∼42 and ∼56 unique features per participant using LC-HRMS and GC-HRMS, respectively. Within the same sub-group of sample preparation method, only a few features were identified by at least two participants in suspect screening (16 features using LC-HRMS, 0 features using GC-HRMS) and non-target screening (0 features using LC-HRMS, 2 features using GC-HRMS). The compounds identified had log octanol/water partition coefficient (KOW) values from -9.9 to 16 and mass-to-charge ratios (m/z) of 68 to 761 (LC-HRMS and GC-HRMS). A significant linear trend was found between log KOW and m/z for the GC-HRMS data. Overall, these findings indicate that differences in screening results are mainly due to the data analysis workflows used by different participants. Further work is needed to harmonise the results obtained when applying suspect and non-target screening approaches to environmental biota samples.

Holistic approach to chemical and microbiological quality of aquatic ecosystems impacted by wastewater effluent discharges.

Wastewater treatment plants (WWTPs) collect wastewater from various sources and use different treatment processes to reduce the load of pollutants in the environment. Since the removal of many chemical pollutants and bacteria by WWTPs is incomplete, they constitute a potential source of contaminants. The continuous release of contaminants through WWTP effluents can compromise the health of the aquatic ecosystems, even if they occur at very low concentrations. The main objective of this work was to characterize, over a period of four months, the treatment steps starting from income to the effluent and 5 km downstream to the receiving river. In this context, the efficiency removal of chemical pollutants (e.g. hormones and pharmaceuticals, including antibiotics) and bacteria was assessed in a WWTP case study by using a holistic approach. It embraces different chemical and biological-based methods, such as pharmaceutical analysis by HPLC-MSMS, growth rate inhibition in algae, ligand binding estrogen receptor assay, microbial community study by 16S and shotgun sequencing along with relative quantification of resistance genes by quantitative polymerase chain reaction. Although both, chemical and biological-based methods showed a significant reduction of the pollutant burden in effluent and surface waters compared to the influent of the WWTP, no complete removal of pollutants, pathogens and antibiotic resistance genes was observed.

Suspect screening of wastewaters to trace anti-COVID-19 drugs: Potential adverse effects on aquatic environment.

During the first period of the SARS-CoV-2 pandemic, the lack of specific therapeutic treatments led to the provisional use of a number of drugs, with a continuous review of health protocols when new scientific evidence emerged. The management of this emergency sanitary situation could not take care of the possible indirect adverse effects on the environment, such as the release of a large amount of pharmaceuticals from wastewater treatment plants. The massive use of drugs, which were never used so widely until then, implied new risks for the aquatic environment. In this study, a suspect screening approach using Liquid Chromatography-High Resolution Mass Spectrometry techniques, allowed us to survey the presence of pharmaceuticals used for COVID-19 treatment in three WWTPs of Lombardy region, where the first European cluster of SARS-CoV-2 cases was detected. Starting from a list of sixty-three suspect compounds used against COVID-19 (including some metabolites and transformation products), six compounds were fully identified and monitored together with other target analytes, mainly pharmaceuticals of common use. A monthly monitoring campaign was conducted in a WWTP from April to December 2020 and the temporal trends of some anti-COVID-19 drugs were positively correlated with those of COVID-19 cases and deaths. The comparison of the average emission loads among the three WWTPs evidenced that the highest loads of hydroxychloroquine, azithromycin and ciprofloxacin were measured in the WWTP which received the sewages from a hospital specializing in the treatment of COVID-19 patients. The monitoring of the receiving water bodies evidenced the presence of eight compounds of high ecological concern, whose risk was assessed in terms of toxicity and the possibility of inducing antibiotic and viral resistance. The results clearly showed that the enhanced, but not completely justified, use of ciprofloxacin and azithromycin represented a risk for antibiotic resistance in the aquatic ecosystems.

Integrated Exposure and Algal Ecotoxicological Assessments of Effluents from Secondary and Advanced-Tertiary Wastewater-Treatment Plants.

The great concern over the environmental impact of wastewaters has led to the designing of advanced treatment processes to upgrade conventional treatment plants and achieve a significant reduction of contaminants in receiving waters. In the present study we combined chemical and ecotoxicological analyses, aiming to evaluate the reduction of toxicity effects associated with the removal of micropollutants and to define the contribution of the detected compounds to the overall toxicity of the mixtures in a series of wastewater effluents collected from a secondary treatment (OUT 2) and from a tertiary activated carbon treatment (OUT 3) plant. The target compounds were selected after a screening procedure among pharmaceuticals, musk fragrances, and trace metals. The classical algal growth inhibition test was conducted on the original effluent samples and on different fractions obtained by solid-phase extraction (SPE) treatment. A good accordance was found between the removal of toxicity (30%-80%) and organic compounds (70%-80%) after the tertiary treatment, suggesting its high efficiency to improve the wastewater quality. The discrepancy between the contribution to the overall toxicity of the nonadsorbable compounds (i.e., inorganic or very polar organic compounds) as experimentally measured by the SPE bioassays (18%-76%) and calculated by the concentration addition approach (>97%) could be mitigated by including the bioavailability correction in metal-toxicity modeling of wastewater mixtures. For the organic compounds, the toxic equivalency method enabled us to quantify the portion of toxicity explained by the detected chemicals in both OUT 2 (82%-104%) and OUT 3 (5%-57%), validating the selection of the target molecules. The applied integrating approach could be implemented by the inclusion of both additional target chemicals and toxicity endpoints. Environ Toxicol Chem 2022;41:2404-2419. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Determination of perfluoroalkyl acids in different tissues of graminaceous plants.

A method for the determination of 12 perfluoroalkyl acids (PFAA) in vegetal samples was proposed. The analytical procedure was developed to optimize the detection of short-chain PFAA (C < 8) due to their higher potential to be translocated and bioaccumulated in plants than long-chain congeners. The method, based on ultrasonic extraction, clean-up and HPLC-MS/MS analysis, determined PFAA in different plant tissues allowing the PFAA distribution and partition in vegetal compartments to be studied. The performance of this analytical procedure was validated by analysing samples (root, stem and leaf) of reed grass. The validated method was then applied to graminaceous plants from an agricultural area impacted by a fluorochemical plant discharge (Northern Italy). The PFAA congeners were detected in most of the samples with ΣPFAA concentrations in the whole plant ranging from

Sediment quality assessment framework for per- and polyfluoroalkyl substances: Results from a preparatory study and regulatory implications.

A preparatory study was performed to develop a sediment quality and risk assessment strategy for Switzerland, addressing the following questions: the sediment fraction to be analyzed chemically (< 63 µm, or alternatively < 2 mm); the suitability of using perfluorooctanesulfonic acid (PFOS) as an indicator of per- and polyfluoroalkyl substances (PFAS) contamination in sediments; the availability of data for the derivation of sediment quality guidelines; and the suitability of normalization to total organic carbon (TOC). The results confirmed PFOS as a suitable indicator of PFAS contamination in sediments from small streams, being the most detected and on average with the highest concentrations among the analyzed PFAS. The fine fraction (< 63 µm) was more appropriate to screening for possible sources and studying the compound profiles at the study sites, but the analysis of the < 2 mm fraction and the normalization to a sample consisting of 100% of the < 63 µm fraction was, in principle, feasible for PFOS. Sediment quality guidelines for PFOS aiming to protect benthic invertebrates from generic adverse effects could be derived, but the available toxicity database is still too sparse. It was only possible to derive preliminary values. Sediment quality guidelines to protect wildlife and human health from secondary poisoning, using the equilibrium partitioning approach and simple trophic web models, were also derived. The use of food web models to derive sediment quality guidelines has not been validated, and the available database of Biota-Sediment Accumulation Factors and Trophic Magnification Factors for PFOS remains limited. However, it is still larger than the effect concentrations database based on sediment studies using benthic invertebrates. Normalization to TOC content in the field (0.2%-12.9%) was decisive in the quality assessment outcome. This is a widely accepted practice for hydrophobic organic contaminants, but its endorsement for PFOS would also benefit from validation. Integr Environ Assess Manag 2021;17:716-725. © 2021 SETAC.

Exposure assessment of PFAS-contaminated sites using avian eggs as a biomonitoring tool: A frame of reference and a case study in the Po River valley (Northern Italy).

For many years, eggs of diverse bird species have been used as monitoring tools in studies investigating perfluoroalkyl substances (PFAS) contamination, especially in marine and remote areas. Avian eggs are a suitable monitoring matrix because they are relatively easy to collect and their yolks store diverse maternally transferred PFAS. Moreover, the concentrations of PFAS detected in the eggs are a good proxy for maternal exposure and allow the assessment of the potential risk for birds. These features support the use of avian eggs as a key monitoring tool in exposure assessment of PFAS-contaminated sites. We first review the recent application of avian eggs in PFAS monitoring in environmental risk assessment schemes, highlighting strengths and limitations and suggesting which criteria should be considered when selecting a proper study species and structuring the sampling and analytical protocol. Eventually, we report findings from a field study realized in 2020 near a perfluoropolymer factory site in the upper Po plain (Northern Italy), revealing an unprecedented contamination level of PFOA and C6O4 in three species of wild passerines. In future, long-term monitoring of PFAS contamination using avian eggs should be maintained, to provide crucial information on the temporal trend of fluorochemical production and waste disposal, while facilitating early identification of emerging PFAS as well as the quantification of their biomagnification across the trophic web. Integr Environ Assess Manag 2021;17:733-745. © 2021 SETAC.

Occurrence, distribution and pollution pattern of legacy and emerging organic pollutants in surface water of the Kongsfjorden (Svalbard, Norway): Environmental contamination, seasonal trend and climate change.

This work aimed to investigate the contamination pattern in Kongsfjorden marine environment (Svalbard, 79°N 12°E) and to disentangle primary and secondary emissions. Surface seawater, sampled in two seasons, was analysed by GC-MS and LC-MS/MS to detect polychlorobiphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), nonylphenols (NPs), bisphenol A (BPA) and perfluoroalkyl and polyfluoroalkyl substances (PFASs). In summer, average ΣPAHs, BPA, ΣNPs, ΣPFASs and ΣPCBs concentrations were 17.3 ± 11.1 ng/L, 0.9 ± 0.3 ng/L, 10.0 ± 6.9 ng/L, 0.4 ± 0.7 ng/L and 1.8 ± 1.3 pg/L, respectively; while in winter, they were 13.6 ± 10.1 ng/L, 0.5 ± 0.2 ng/L, 6.8 ± 3.3 ng/L,

Per- and Polyfluoroalkyl Substances (PFAS) in Fish from European Lakes: Current Contamination Status, Sources, and Perspectives for Monitoring.

Concentrations in fish of per- and polyfluoroalkyl substances (PFAS) were reported for 7 deep lakes in the European subalpine area: Lakes Geneva, Lugano, Maggiore, Iseo, Como, Garda, and Mergozzo; one shallow lowland lake (Varese); and 2 high-altitude alpine lakes (>2000 m a.s.l). Fillets and, in selected cases, other body fractions (viscera, liver, and residual carcass) from 8 fish species were analyzed. The possibility of harmonizing the monitoring protocols was tested. Results suggest that the sampling season is not critical for PFASs and the total protein content cannot be used for normalization of tissue concentrations because PFASs bind to specific proteins. Moreover, the polar lipid content could be used to reduce the variability of PFAS concentrations in phospholipid rich fractions of fish such as viscera and carcass. The data comparison and analysis show that the PFAS contamination in lake fish is generally correlated with the degree of urbanization of the lake catchment; however, it is sometimes difficult to compare absolute concentrations in lake fish because the lake hydro-morphological characteristics play a substantial role in determining the chemical concentrations of persistent and mobile contaminants. Environ Toxicol Chem 2021;40:658-676. © 2020 SETAC.

Within- and Among-Clutch Variation of Yolk Perfluoroalkyl Acids in a Seabird from the Northern Adriatic Sea.

Perfluoroalkyl substances (PFAS) are surface-active agents used in diverse industrial and commercial applications. They contaminate both freshwater and marine ecosystems, are highly persistent, and accumulate through trophic transfer. Seabirds are exposed to environmental contaminants due to their high trophic position in food webs and relatively long lifespan. We measured levels of 10 perfluoroalkyl acids (PFAAs) in egg yolks of yellow-legged gulls (Larus michahellis) breeding in the northern Adriatic Sea (Northeast Italy). We examined variations in PFAAs within clutches (between eggs of different laying order) and among clutches. Perfluorooctane sulfonate (PFOS) was the most abundant yolk PFAA (mean = 42.0 ng/g wet wt), followed by perfluorooctanoic acid (PFOA; 3.8 ng/g wet wt) and perfluorododecanoic acid (PFDoDa; 2.8 ng/g wet wt). The ∑PFAAs averaged 57.4 ng/g wet weight, ranging between 26.5 and 115.0 ng/g wet weight. The PFAA levels varied substantially among clutches (0.29-0.79 of the total variation), whereas the effects of laying order were considerably weaker (0.01-0.13). Egg-laying order effects were detected for ∑PFAAs, PFOS, perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluoroundecanoic acid (PFUnA), and PFDoDa, whereby the last-laid eggs exhibited lower PFAA concentrations than early-laid eggs. Our results indicate that seagulls from the northern Adriatic basin deposit measurable amounts of PFAAs in their eggs. The large among-clutches differences in PFAAs suggest that exposure of yellow-legged gull females to these compounds is highly variable. Environ Toxicol Chem 2021;40:744-753. © 2020 SETAC.

New compounds, old problems. The case of C6O4 - a substitute of PFOA - and its effects to the clam Ruditapes philippinarum.

C6O4 (difluoro{[2,2,4,5-tetrafluoro-5-(trifluoromethoxy)-1,3-dioxolan-4-yl]oxy}acetic acid) is a new surfactant and emulsifier used as a substitute of perfluorooctanoic acid (PFOA). Recently, C6O4 has been detected in aquatic environments, but, at present, no information concerning the effects of C6O4 on aquatic species, such as bivalves, are available in the literature. Therefore, in this study we evaluated for the first time the effects of C6O4 (0.1 and 1 µg/L) and PFOA (1 µg/L) to the clam Ruditapes philippinarum. Short-term (7 days) and long-term (21 days) exposures of clams to the two compounds were carried out and numerous biomarkers were measured in haemocytes/haemolymph, as well as in gills and digestive gland. The MANOVA analysis demonstrated statistically significant effects of the independent variables "treatment", "time" and "treatment-time interaction" on the whole dataset of biomarker responses. The two-way ANOVA analysis performed for each biomarker response indicated that the two compounds affected most of the cellular and tissue parameters measured. Despite preliminary, the results obtained suggested that C6O4 - similarly to PFOA - can affect both cellular and biochemical parameters of clams.

Assessing the Ecological Risks of Per- and Polyfluoroalkyl Substances: Current State-of-the Science and a Proposed Path Forward.

Per- and poly-fluoroalkyl substances (PFAS) encompass a large, heterogenous group of chemicals of potential concern to human health and the environment. Based on information for a few relatively well-understood PFAS such as perfluorooctane sulfonate and perfluorooctanoate, there is ample basis to suspect that at least a subset can be considered persistent, bioaccumulative, and/or toxic. However, data suitable for determining risks in either prospective or retrospective assessments are lacking for the majority of PFAS. In August 2019, the Society of Environmental Toxicology and Chemistry sponsored a workshop that focused on the state-of-the-science supporting risk assessment of PFAS. The present review summarizes discussions concerning the ecotoxicology and ecological risks of PFAS. First, we summarize currently available information relevant to problem formulation/prioritization, exposure, and hazard/effects of PFAS in the context of regulatory and ecological risk assessment activities from around the world. We then describe critical gaps and uncertainties relative to ecological risk assessments for PFAS and propose approaches to address these needs. Recommendations include the development of more comprehensive monitoring programs to support exposure assessment, an emphasis on research to support the formulation of predictive models for bioaccumulation, and the development of in silico, in vitro, and in vivo methods to efficiently assess biological effects for potentially sensitive species/endpoints. Addressing needs associated with assessing the ecological risk of PFAS will require cross-disciplinary approaches that employ both conventional and new methods in an integrated, resource-effective manner. Environ Toxicol Chem 2021;40:564-605. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

The new PFAS C6O4 and its effects on marine invertebrates: First evidence of transcriptional and microbiota changes in the Manila clam Ruditapes philippinarum.

There is growing concern for the wide use ofperfluorooctanoic acid (PFOA) because of its toxic effects on the environment and on human health. A new compound - the so called C6O4 (perfluoro ([5-methoxy-1,3-dioxolan-4-yl]oxy) acetic acid) - was recently introduced as one of the alternative to traditional PFOA, however this was done without any scientific evidence of the effects of C6O4 when dispersed into the environment. Recently, the Regional Agency for the Protection of the Environment of Veneto (Italy) detected high levels of C6O4 in groundwater and in the Po river, increasing the alarm for the potential effects of this chemical into the natural environment. The present study investigates for the first time the effects of C6O4 on the Manila clam Ruditapes philippinarum exposed to environmental realistic concentrations of C6O4 (0.1 µg/L and 1 µg/L) for 7 and 21 days. Furthermore, in order to better understand if C6O4 is a valid and less hazardous alternative to its substitute, microbial and transcriptomic alterations were also investigated in clams exposed to 1 µg/L ofPFOA. Results indicate that C6O4 may cause significant perturbations to the digestive gland microbiota, likely determining the impairment of host physiological homeostasis. Despite chemical analyses suggest a 5 times lower accumulation potential of C604 as compared to PFOA in clam soft tissues, transcriptional analyses reveal several alterations of gene expression profile. A large part of the altered pathways, including immune response, apoptosis regulation, nervous system development, lipid metabolism and cell membrane is the same in C6O4 and PFOA exposed clams. In addition, clams exposed to C6O4 showed dose-dependent responses as well as possible narcotic or neurotoxic effects and reduced activation of genes involved in xenobiotic metabolism. Overall, the present study suggests that the potential risks for marine organism following environmental contamination are not reduced by replacing PFOA with C6O4. In addition, the detection of both C6O4 and PFOA into tissues of clams inhabiting the Lagoon of Venice - where there are no point sources of either compounds - recommends a similar capacity to spread throughout the environment. These results prompt the urgent need to re-evaluate the use of C6O4 as it may represent not only an environmental hazard but also a potential risk for human health.

Inter-laboratory mass spectrometry dataset based on passive sampling of drinking water for non-target analysis.

Non-target analysis (NTA) employing high-resolution mass spectrometry is a commonly applied approach for the detection of novel chemicals of emerging concern in complex environmental samples. NTA typically results in large and information-rich datasets that require computer aided (ideally automated) strategies for their processing and interpretation. Such strategies do however raise the challenge of reproducibility between and within different processing workflows. An effective strategy to mitigate such problems is the implementation of inter-laboratory studies (ILS) with the aim to evaluate different workflows and agree on harmonized/standardized quality control procedures. Here we present the data generated during such an ILS. This study was organized through the Norman Network and included 21 participants from 11 countries. A set of samples based on the passive sampling of drinking water pre and post treatment was shipped to all the participating laboratories for analysis, using one pre-defined method and one locally (i.e. in-house) developed method. The data generated represents a valuable resource (i.e. benchmark) for future developments of algorithms and workflows for NTA experiments.

Uptake and translocation of perfluoroalkyl acids (PFAA) in red chicory (Cichorium intybus L.) under various treatments with pre-contaminated soil and irrigation water.

Perfluoroalkyl acids (PFAAs), particularly short-chained ones, have high potential for crop uptake, posing a threat to human health in contaminated areas. There is a scarcity of studies using contaminated water as the medium for PFAAs delivery to crops, and a lack of data on the partitioning of PFAA mixtures in growing media. In this context, a controlled experimental study was carried out in a greenhouse to investigate the uptake of a PFAA mixture into red chicory, a typical crop from a major PFAA contamination hot-spot in northern Italy, under treatments with environmentally relevant concentrations in spiked irrigation water and soil, separately and simultaneously. To our knowledge, this is the first study involving multiple exposure media and laboratory adsorption/desorption batch tests as a way of assessing the decrease in the bioavailability of PFAAs from soil. Exposure concentrations for each of the 9 utilized PFAAs were 0, 1, 10 and 80 µg/L in irrigation water and 0, 100 and 200 ng/gdw in soil, combined into 12 treatments. The highest bioaccumulation was measured for PFBA in roots (maximum of 43 µg/gdw), followed by leaves and heads of the chicory plants in all treatments, with the concentrations exponentially decreasing with an increasing PFAA chain length in all plant compartments. The use of irrigation water as the delivery medium increased the transport of PFAAs to the aerial chicory parts, long-chain substances in particular. Additionally, the distribution of PFAAs in the soil was assessed by depth and compared with laboratory measured soil-water equilibrium partition coefficients, revealing only partial dependency of PFAAs bioavailability on the adsorption in soil.