Search

Per- and polyfluoroalkyl substances (PFAS) are under regulatory scrutiny since some of them are persistent, bioaccumulative, and toxic. The occurrence of 4777 PFAS was investigated in the Danube River Basin (DRB; 11 countries) using target and suspect screening. Target screening involved investigation of PFAS with 56 commercially available reference standards. Suspect screening covered 4777 PFAS retrieved from the NORMAN Substance Database, including all individual PFAS lists submitted to the NORMAN Suspect List Exchange Database. Mass spectrometry fragmentation patterns and retention time index predictions of the studied PFAS were established for their screening by liquid chromatography - high resolution mass spectrometry using NORMAN Digital Sample Freezing Platform (DSFP). In total, 82 PFAS were detected in the studied 95 samples of river water, wastewater, groundwater, biota and sediments. Suspect screening detected 72 PFAS that were missed by target screening. Predicted no effect concentrations (PNECs) were derived for each PFAS via a quantitative structure-toxicity relationship (QSTR)-based approach and used for assessment of their environmental risk. Risk characterization revealed 18 PFAS of environmental concern in at least one matrix. The presence of PFAS in all studied environmental compartments across the DRB indicates a potentially large-scale migration of PFAS in Europe, which might require their further systematic regulatory monitoring.

Siverskyi Donets is the fourth longest river in Ukraine and its ecosystem is heavily affected by numerous agricultural and industrial activities. An impact of the on-going armed military conflicts in the Eastern Ukraine to the overall pollution by the chemicals has been studied. Considering the uncontrolled activities in the catchment due to the conflict, there is a high demand to assess the contamination status of the Siverskyi Donets basin. In this study, the occurrence of the EU Water Framework Directive priority substances, selected physicochemical parameters and wide-range emerging contaminants were investigated in surface water, groundwater, biota and river sediments samples from 13 sampling sites in the river basin. The study included metals, inorganic, non-polar and polar organic contaminants. The wide-scope target screening of 2316 substances and suspect screening of 2219 substances revealed occurrence of 83 compounds in the studied samples. A few industrial chemicals such as plasticizers bisphenol A and DEHP, as well as flame retardant brominated diphenylethers were found to be potentially hazardous to the ecosystem, exceeding the established legacy environmental quality standards (EQS) or the provisional no-effect concentration (PNEC) values. River sediment samples contained traces of long-term banned chemicals such as polychlorinated biphenyls (PCBs) and degradation products of DDT (p,p'-DDD and p,p'-DDE). A simplified risk assessment based on comparison of measured concentration of the detected compounds against their (eco)toxicity threshold values from the NORMAN Ecotoxicology Database has been performed to aid their prioritization in future monitoring and, eventually, establishing the list of Siverskyi Donets River Basin Specific Pollutants. A comparison with the recent similar studies in the Dniester and Dnieper river basins in Ukraine has shown that the overall pollution by chemicals in the Siverskyi Donets basin is significantly lower.

Advances in Matrix-assisted Laser Desorption/Ionization -Time-Of-Flight Mass Spectrometry (MALDI-TOF-MS) have led to its supremacy for complex assessment of food authenticity studies, like dairy products fraud, holding promise for the discovery of potential authenticity (bio)markers. In this study, an integrated untargeted protein-based workflow in combination with advanced chemometrics is presented, to address authenticity challenges in PDO feta cheese which is legally manufactured by the mixture of sheep/goat milk. Potential markers attributed to specific animal origin were found from protein profiles acquired for authentic feta and white cheeses (prepared from cow milk), belonging to 4 kDa–18.5 kDa mass area. Rapid detection of feta cheese adulteration from cow milk was also achieved down to 1% adulteration level. The discriminative models showed high predictive ability for feta cheese authenticity (Q2 = 0.920, RMSEE = 0.053) and its adulteration (Q2 = 0.835, RMSEE = 0.121), introducing a reliable approach in routine analysis. The methodology was successfully applied in detection of cow milk in sheep yoghurt.

Stevia rebaudiana Bertoni is a plant cultivated worldwide due to its use as a sweetener. The sweet taste of stevia is attributed to its numerous steviol glycosides, however, their use is still limited, due to their bitter aftertaste. The transglycosylation of steviol glycosides, aiming at the improvement of their taste, has been reported for many enzymes, however, glycosyl hydrolases are not extensively studied in this respect. In the present study, a β-glucosidase, MtBgl3a, and a β-galactosidase, TtbGal1, have been applied in the transglycosylation of two steviol glycosides, stevioside and rebaudioside A. The maximum conversion yields were 34.6 and 33.1% for stevioside, while 25.6 and 37.6% were obtained for rebaudioside A conversion by MtBgl3a and TtbGal1, respectively. Low-cost industrial byproducts were employed as sugar donors, such as cellulose hydrolyzate and acid whey for TtbGal1- and MtBgl3a- mediated bioconversion, respectively. LC-HRMS analysis identified the formation of mono- and di- glycosylated products from stevioside and rebaudioside A. Overall, the results of the present work indicate that both biocatalysts can be exploited for the design of a cost-effective process for the modification of steviol glycosides.

Use of high-resolution mass spectrometry (HRMS) including a MS calibration method has enabled simultaneous identification and quantification of knowns/unknowns. This has expanded our knowledge about the existing sample relevant chemical space in a way beyond reconciliation with a quantification task. This is largely due to fact that reference standards are not always available to achieve quantitative analysis. In this scenario, a semi-quantitative approach can fill the gap and provide a rough estimation of concentration. This research aimed to develop and compare several semi-quantification approaches based on chemical similarity or properties. The ionization efficiency scale was created for several groups of natural products. Advanced modeling approach based on a support vector machine was conducted to learn from the experimental ionization efficiency and apply it to unknowns or suspected compounds to predict their ionization efficiency in electrospray ionization mode. The developed semi-quantification workflows could be useful in most HRMS based "omics" areas, especially in natural products discovery.

INTRODUCTION Pelargonium sidoides is a member of the Geraniaceae family and it originates from the coastal regions of South Africa. In the last decades, Pelargonium sidoides root has been subjected to several surveys due to the assertion of its health benefits, such as the relief of symptoms of acute bronchitis, common cold and acute rhinosinusitis. Many studies have been conducted to reveal its naturally occurring bioactive chemicals, yet no wide-scope chemical characterisation strategies have been done using mass spectrometry. OBJECTIVE This research aimed to comprehensively characterise the chemical profile of Pelargonium sidoides root via high-resolution mass spectrometry. METHODOLOGY The Pelargonium sidoides root was extracted by a mixture of methanol: water in the proportion of 80:20. The extraction procedure included vortexing, shaking as well as the use of an ultrasound sonication bath under 40°C. After centrifugation, the supernatant was evaporated to dryness. The dry residue was reconstituted with a mixture of methanol/water (50:50, v/v), filtered and injected into an ultra-high-pressure liquid chromatography-quadruple time-of-flight mass spectrometer. RESULTS Overall, 33 compounds were identified in the root using suspect and non-target screening. These compounds were originated from different classes of compounds such as amino acids, phenolic acids, α-hydroxy-acids, vitamins, polyphenols, flavonoids, coumarins, coumarins glucosides, coumarin sulphates and nucleotides. Quantitative results were provided for the identified compounds, where their reference standards were available. CONCLUSION Some important compounds were elucidated, belonging to different classes of compounds such as antioxidants (coumarins and phenolic compounds), amino acids, nucleotides and vitamins revealing the importance of the bioactive content of this root.

Honey consumption is attributed to potentially advantageous effects on human health due to its antioxidant capacity as well as anti-inflammatory and antimicrobial activity, which are mainly related to phenolic compound content. Phenolic compounds are secondary metabolites of plants, and their content in honey is primarily affected by the botanical and geographical origin. In this study, a high-resolution mass spectrometry (HRMS) method was applied to determine the phenolic profile of various honey matrices and investigate authenticity markers. A fruitful sample set was collected, including honey from 10 different botanical sources (n = 51) originating from Greece and Poland. Generic liquid–liquid extraction using ethyl acetate as the extractant was used to apply targeted and non-targeted workflows simultaneously. The method was fully validated according to the Eurachem guidelines, and it demonstrated high accuracy, precision, and sensitivity resulting in the detection of 11 target analytes in the samples. Suspect screening identified 16 bioactive compounds in at least one sample, with abscisic acid isomers being the most abundant in arbutus honey. Importantly, 10 markers related to honey geographical origin were revealed through non-targeted screening and the application of advanced chemometric tools. In conclusion, authenticity markers and discrimination patterns were emerged using targeted and non-targeted workflows, indicating the impact of this study on food authenticity and metabolomic fields.

In this study, the oxidative degradation of losartan (LOS), a widely administered medicine for high blood pressure by heat-activated persulfate was investigated. Increased temperature and persulfate concentration, as well as acidic conditions enhance the degradation efficiency of LOS, whose rate follows pseudo-first order kinetics. From the respective apparent rate constants in the range 40–60 °C, an apparent activation energy of 112.70 kJ/mol was computed. Radical scavenging tests demonstrated that both HO• and S O 4 • − contribute towards LOS degradation. LOS degradation was suppressed in real water matrices including bottled water (BW) and secondary wastewater effluent (WW), while other experiments indicated that the presence of bicarbonates and humic acid negatively affected its oxidation. Instead, the addition of chloride ions at 250 mg/L resulted in a positive effect on LOS removal. The combination of heat-activated PS with low-frequency ultrasound exhibited a synergistic effect, with the ratio S being 2.29 in BW and 1.52 in WW. Five transformation products of LOS were identified through HRMS suspect and non-target screening approaches, among which two are reported for the first time. Using the in-house risk assessment program, ToxTrAMs was revealed that most of the identified TPs present higher toxicity than LOS against Daphnia magna.

During ozonation in wastewater treatment plants, ozone reacts with emerging pollutants, which are partially removed through the secondary treatment, as long as, with their biotransformation products, triggering the formation of ozonation transformation products (TPs). Although the transformation of parent compounds (PCs) and their metabolites has been reported in the literature, the probable transformation of biotransformation products has not been investigated so far. This study evaluates the fate of citalopram (CTR) and four of its biotransformation products (DESCTR, CTRAM, CTRAC and CTROXO) during ozonation experiments. A Gaussian curve-based trend analysis was performed for the first time for the automated detection of TPs in ozone concentrations ranging from 0.06 to 12 mg/L. In total 46 ozonation TPs were detected; 7 TPs of CTR, 10 of DESCTR, 9 of CTRAM, 12 of CTRAC and 8 of CTROXO and were structurally elucidated based on their high resolution tandem mass spectra interpretation and tandem mass spectra similarity with the respective PC. Results have demonstrated that the examined compounds follow common transformation pathways in reaction with ozone and that common TPs were formed through the ozonation of different structurally-alike compounds. Moreover, the toxicity of the identified TPs was predicted with an in-house risk assessment program.

Since 2019, Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), impaired public health with considerable morbidity and mortality due to the lack of vaccines and effective treatment. The severe disease mainly harmed adults with predisposing medical comorbidities (such as heart disease, hypertension, chronic lung disease), while it can occur in healthy individuals that may be asymptomatic. Wastewater-based Epidemiology (WBE), a non-invasive, objective, chemical tool was used to monitor and estimate the changes in drug's consumption and prescription patterns under normal conditions (2019) and under COVID-19 pandemic conditions (2020). NSAIDs, antihypertensives, diuretics, antiepileptics, antilipidemics, antibiotics, analgesics, antivirals, anticancer drugs, contrast iodinated drugs, antidiabetics, antiallergic drugs, antiulcers and other pharmaceuticals were studied in wastewater and revealed the application of various treatments during the first wave of the pandemic in Athens, Greece. Data were correlated with COVID-19 infection therapeutical plans. The result of the analysis revealed a remarkable increase for antiviral drugs (170%), hydroxychloroquine (387%), and antibiotics (57%), which were the most applied treatments against COVID-19 during the first wave in Greece. In agreement with related authorities urge, NSAIDs presented decrease (27%) during the first lockdown, while paracetamol demonstrated a remarkable increase (198%). The use levels for Angiotensin II receptor blockers such as valsartan, and co-administrated diuretics, such as hydrochlorothiazide, were reduced during 2020, by 32% and 26% respectively., Graphical abstract Unlabelled Image

Between June and November 2020, we assessed plasma antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid protein in 4996 participants (aged 18–82 years, 34.5% men) from the National and Kapodistrian University of Athens. The weighted overall prevalence was 1.6% and monthly prevalence correlated with viral RNA-confirmed SARS-CoV-2 infections in Greece, in the same period. Notably, 49% of seropositive cases reported no history of SARS-CoV-2 infection-related clinical symptoms and 33% were unsuspected of their previous infection. Additionally, levels of anti-SARS-CoV-2 antibodies against the spike-protein receptor-binding domain were similar between symptomatic and asymptomatic individuals, irrespective of age and gender. Using Food and Drug Administration Emergency Use Authorization-approved assays, these results support the need for such studies on pandemic evaluation and highlight the development of robust humoral immune responses even among asymptomatic individuals. The high percentage of unsuspected/asymptomatic active cases, which may contribute to community transmission for more days than that of cases who are aware and self-isolate, underscores the necessity of measures across the population for the efficient control of the pandemic.

The intensity and duration of hot weather and the number of extreme weather events, such as heatwaves, are increasing, leading to a growing need for space cooling energy demand. Together with the building stock’s low energy performance, this phenomenon may also increase households’ energy consumption. On the other hand, the low level of ownership of cooling equipment can cause low energy consumption, leading to a lack of indoor thermal comfort and several health-related problems, yet increasing the risk of energy poverty in summer. Understanding future temperature variations and the associated impacts on building cooling demand will allow mitigating future issues related to a warmer climate. In this respect, this paper analyses the effects of change in temperatures in the residential sector cooling demand in 2050 for a case study of nineteen cities across seven countries: Cyprus, Finland, Greece, Israel, Portugal, Slovakia, and Spain, by estimating cooling degree days and hours (CDD and CDH). CDD and CDH are calculated using both fixed and adaptive thermal comfort temperature thresholds for 2020 and 2050, understanding their strengths and weaknesses to assess the effects of warmer temperatures. Results suggest a noticeable average increase in CDD and CDH values, up to double, by using both thresholds for 2050, with a particular interest in northern countries where structural modifications in the building stock and occupants’ behavior should be anticipated. Furthermore, the use of the adaptive thermal comfort threshold shows that the projected temperature increases for 2050 might affect people’s capability to adapt their comfort band (i.e., indoor habitability) as temperatures would be higher than the maximum admissible values for people’s comfort and health., The paper stems from collaborative work within COST Action ‘European Energy Poverty: Agenda Co-Creation and Knowledge Innovation’ (ENGAGER 2017–2021, CA16232) funded by European Cooperation in Science and Technology—www.cost.eu (5 December 2020). João Pedro Gouveia and Pedro Palma acknowledge and are thankful for the support provided to CENSE by the Portuguese Foundation for Science and Technology (FCT) through the strategic project UIDB/04085/2020 and through the scholarship SFRH/BD/146732/2019 provided to Pedro). Ricardo Barbosa acknowledges the support for this work, which was partly financed by FCT/MCTES through national funds (PIDDAC) under the R&D Unit Institute for Sustainability and Innovation in Structural Engineering (ISISE), under reference UIDB/04029/2020.

Honey is a high-value, globally consumed, food product featuring a high market price strictly related to its origin. Moreover, honey origin has to be clearly stated on the label, and quality schemes are prescribed based on its geographical and botanical origin. Therefore, to enhance food quality, it is of utmost importance to develop analytical methods able to accurately and precisely discriminate honey origin. In this study, an all-time scientometric evaluation of the field is provided for the first time using a structured keyword on the Scopus database. The bibliometric analysis pinpoints that the botanical origin discrimination was the most studied authenticity issue, and chromatographic methods were the most frequently used for its assessment. Based on these results, we comprehensively reviewed analytical techniques that have been used in honey authenticity studies. Analytical breakthroughs and bottlenecks on methodologies to assess honey quality parameters using separation, bioanalytical, spectroscopic, elemental and isotopic techniques are presented. Emphasis is given to authenticity markers, and the necessity to apply chemometric tools to reveal them. Altogether, honey authenticity is an ever-growing field, and more advances are expected that will further secure honey quality.

We describe herein the synthesis and laccase mediated oxidation of six novel 1,4-dihydropyridine (DHP)-based hexahydroquinolines (DHP1-DHP3) and decahydroacridines (DHP4-DHP6). We employed different laccase enzymes with varying redox potential to convert DHP1-DHP3 and DHP4-DHP6 to the corresponding pyridine-containing tetrahydroquinoline and octahydroacridine derivatives, respectively. Intensively coloured products were detected in all biocatalytic reactions using laccase from Trametes versicolor (TvLacc), possibly due to the presence of conjugated chromophores formed in products after oxidation. The NMR assessment confirmed that the oxidation product of DHP1 was its corresponding pyridine-bearing tetrahydroquinoline derivative. Laccase from Bacillus subtillis (BacillusLacc) was the most efficient enzyme for this group of substrates using HPLC assessment. Overall, it could be concluded that DHP2 and DHP5, bearing catecholic structures, were easily oxidized by all tested laccases, while DHP3 and DHP6 containing electron-withdrawing nitro-groups are not readily oxidized by laccases. DHP4 with decahydroacridine moiety consisting of three condensed six-membered rings that contribute not only to the volume but also to the higher redox potential of the substrate rendered this compound not to be biotransformed with any of the mentioned enzymes. Overall, we showed that multiple analytical approaches are needed in order to assess biocatalytical reactions. Supplementary material: [https://cherry.chem.bg.ac.rs/handle/123456789/4568]

The scarcity of freshwater has led to a considerable increase of the reuse of reclaimed wastewater for irrigation of field crops [1,2]. This practice potentially exposes agricultural produce to a large variety of xenobiotic compounds including contaminants of emerging concern (CECs) which have been widely recognized to be present in wastewater [3]. Common approaches for the extraction of CECs from crops rely on solid-liquid extraction [4], assisted solvent extraction [5], ultra-sound solvent extraction [6] and recently QuEChERS (QUick, Easy, CHeap, Effective, Rugged and Safe) [[7], [8]–9]. Here, eight QuEChERS-based methodologies were compared for their suitability to determine 45 CECs in roots and leaves of soil-grown radish. The key points of the method development were: • The development of two single-step analytical methods specific for radish root and leaves, after testing eight different approaches using QuEChERS extraction for the quantitation of 45 CECs. The analytical methodology selected requires minimal time and solvent, making it cost-effective. • Methods validation were performed at five concentrations levels (2, 5, 10, 50 and 200 ng g−1), with low limits of quantification between 0.01 and 0.32 ng g−1. • The two optimized methodologies may be applied to identify large number of compounds of different families in radish crop. However, validation will be needed to quantify compounds different from the target compounds of this paper., This study has been financially supported through the Spanish national project TRAPPER (CICLIC, RTI2018-097158-B-C33). This work was supported by the Spanish Ministry of Science and Innovation (Project CEX2018-000794-S-19-2). Authors acknowledge SCIEX for providing the loan instrument LC/HRMS QTOF X500R. Cristina Labad (www.cristinalabad.com) was acknowledged for the graphical support.

The presence of SARS-CoV-2 RNA in wastewater was first reported in March 2020. Over the subsequent months, the potential for wastewater surveillance to contribute to COVID-19 mitigation programmes has been the focus of intense national and international research activities, gaining the attention of policy makers and the public. As a new application of an established methodology, focused collaboration between public health practitioners and wastewater researchers is essential to developing a common understanding on how, when and where the outputs of this non-invasive community-level approach can deliver actionable outcomes for public health authorities. Within this context, the NORMAN SCORE “SARS-CoV-2 in sewage” database provides a platform for rapid, open access data sharing, validated by the uploading of 276 data sets from nine countries to-date. Through offering direct access to underpinning meta-data sets (and describing its use in data interpretation), the NORMAN SCORE database is a resource for the development of recommendations on minimum data requirements for wastewater pathogen surveillance. It is also a tool to engage public health practitioners in discussions on use of the approach, providing an opportunity to build mutual understanding of the demand and supply for data and facilitate the translation of this promising research application into public health practice. Validerad;2021;Nivå 2;2021-05-21 (alebob);Finansiär: För finansiärsinformation se Acknowledgements https://www.sciencedirect.com/science/article/pii/S0043135421003651?via%3Dihub#ack0001 DRIZZLE

International audience; Toxicology has been an active research field for many decades, with academic, industrial and government involvement. Modern omics and computational approaches are changing the field, from merely disease-specific observational models into target-specific predictive models. Traditionally, toxicology has strong links with other fields such as biology, chemistry, pharmacology and medicine. With the rise of synthetic and new engineered materials, alongside ongoing prioritisation needs in chemical risk assessment for existing chemicals, early predictive evaluations are becoming of utmost importance to both scientific and regulatory purposes. ELIXIR is an intergovernmental organisation that brings together life science resources from across Europe. To coordinate the linkage of various life science efforts around modern predictive toxicology, the establishment of a new ELIXIR Community is seen as instrumental. In the past few years, joint efforts, building on incidental overlap, have been piloted in the context of ELIXIR. For example, the EU-ToxRisk, diXa, HeCaToS, transQST, and the nanotoxicology community have worked with the ELIXIR TeSS, Bioschemas, and Compute Platforms and activities. In 2018, a core group of interested parties wrote a proposal, outlining a sketch of what this new ELIXIR Toxicology Community would look like. A recent workshop (held September 30th to October 1st, 2020) extended this into an ELIXIR Toxicology roadmap and a shortlist of limited investment-high gain collaborations to give body to this new community. This Whitepaper outlines the results of these efforts and defines our vision of the ELIXIR Toxicology Community and how it complements other ELIXIR activities.

Chemicals are part of our daily lives, and we are exposed to numerous chemicals through multiple pathways. Relevant scientific evidence contributing to the regulation of hazardous chemicals require a holistic approach to assess simultaneous exposure to multiple compounds. Biomonitoring provides an accurate estimation of exposure to chemicals through very complex and costly sampling campaigns. Finding efficient proxies to predict the risk of chemical exposure in humans is an urgent need to cover large areas and populations at a reasonable cost. We conducted an exploratory study to characterize the human chemical exposome in maternal blood and placenta samples of a population-based birth cohort in Barcelona (2018-2021). Ultimate HRMS-based approaches were applied including wide-scope target, suspect, and nontarget screening. Forty-two chemicals were identified including pesticides, personal care products, or industrial compounds, among others, in the range of ng/mL and ng/g. In parallel, sewage sludge from the wastewater treatment plants serving the residence areas of the studied population were also screened, showing correlations with the type and concentrations of chemicals found in humans. Our findings were suggestive for the potential use of sewage sludge as a proxy of the human exposure and its application in early warning systems to prevent bioaccumulation of hazardous chemicals., This work received support from the “La Caixa” Foundation (ID 100010434), fellowship code LCF/BQ/PR20/11770013, and Barcelona Council (Expo-Bar). The BiSC cohort study is funded by the European Research Council (ERC) under Grant Agreement No. 785994 (AIR-NB), and the Health Effects Institute (HEI) with Grant Agreement No. 4959-RFPA15-1/ 18-1 (FRONTIER). IDAEA-CSIC and ISGlobal are Centres of Excellence Severo Ochoa (Spanish Ministry of Science and Innovation).

Little information exists on the effects of shipping and handling on per- and polyfluoroalkyl substances (PFASs) in environmental samples. Thus, we evaluated the integrity of dried wastewater extracts and the sensitivity of our high-resolution mass spectrometry (HRMS) instrument to perform such analyses by monitoring 13 representative PFASs in samples extracted, evaporated, and stored at room temperature up to one month. Relative to zero-day recoveries of six detected PFASs ranged between 94 and 124% (RSD 38%) for influents, between 88 and 126% (RSD 18%) for effluents after 28 days. Larger variabilities are tentatively associated with the lack of specific mass-labeled standards and the interactions between analytes and remaining matrix components over time. In a second stage, a mix of local and international dry-shipped wastewater samples were analyzed and the same PFASs were quantified. Up to six PFASs were identified, with median concentrations ranging from 1.3 (perfluoro butyl sulfonate (PFBS)) to 7.7 ng/L (perfluoro hexanoic acid (PFHxA)) and from 1.5 (PFBS) to 13.8 ng/L (PFHxA) in local influents and effluents respectively; and from 0.7 (perfluoro hexyl sulfonate (PFHxS)) to 52.8 ng/L (PFHxA) and from 0.5 (PFHxS) to 21.4 ng/L (PFHxA) in Greek influents and effluents, respectively. The importance of this study lies on the need to consider the wider recovery shifts and expanded variability ranges of PFASs derived from the transport and storage times of dried extracts, particularly when applied to HRMS and wide-scope screening approaches.

Background/Purpose Anatomically, hemodynamically relevant and validated animal models for aortic valve stenosis are of great need. Drolet rabbit model with tricuspid anatomy produced conflicting results for unclear reasons. We hypothesized that limitations concentrate in the regimen administration. We sought to evaluate multiple doses, ways of administration and time periods. Methods We included New Zealand rabbits in 4 groups: Group A (Drolet): was fed with normal chaw (nc)+0.5% cholesterol (chol)+3500 IUs Vitamin D2/kg (VD2, ergocalciferol, Sigma) in water daily for 12 weeks (wks), Group B: nc+0.5%chol+3500 IUs/kg VD2 in oil incorporated in a bisquit daily for 8 wks, Group C: nc+0.5%chol+8750 IUs/kg VD2 in oil-biscuit for 8 wks, Group D: nc+0.5%chol+17500 IUs VD2 in oil-biscuit for 8 wks vs controls (fed only with nc). After 12 and 8 wks the rabbits were sacrificed. Aortic valve area (AVA) and mean gradient (meanGr) were assessed with echocardiography (Vivid 7, M3S transducer, GE) and serum obtained, at baseline and before sacrifice. VD2 levels were evaluated through Chemiluminescent Microparticle Immuno Assay (CMIA, Abbott) and liquid chromatography – tandem mass spectrometry (LC-APCI-MS/MS). Animals received i.v. 18F-NaF one hour before sacrifice and valve was ex-vivo imaged with microPET/CT (Mediso nanoScan). Aortic cusps were analyzed with Fourier-Transformed Infrared Spectroscopy (FT-IR, Nicolet 6700 spectrometer, OMNIC 7.3 software). Valves from surgical patients with severe stenosis served for comparison purposes. Results In Group A at 12 wks AVA and meanGr remained unchanged but biomineralization was detected with FT-IR with vibrations in the region of 1800–800 cm–1 demonstrating the deposition of CaCO3 and non-hydroxyapatite Ca3(PO4)2 identical to human lesion. Calcification was detected on cusps with 18F-NaF. VD2 levels were out of upper detection range with CMIA due to cross reaction, whereas all samples measured through LC-MS/MS were below the detection limit of the method ( Conclusions The modified diet produces aortic valve stenosis and biomineralization detectable with 18F-NaF, chemically identical to human lesion. Very high doses of Vitamin D2 directly produce LVH, possibly leading to arrythmiogenesis. The modified high-fat-vitamin D2 rabbit model proved suitable for translational research of aortic valve stenosis disease. Funding Acknowledgement Type of funding source: Public Institution(s). Main funding source(s): National and Kapodistrian University of Athens

In March 2020 the World Health Organization announced a pandemic outbreak. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative pathogen for the coronavirus disease-19 (COVID-19) pandemic. The authorities worldwide use clinical science to identify infected people, but this approach is not able to track all symptomatic and asymptomatic cases due to limited sampling capacity of the testing laboratories. This drawback is eliminated by the Wastewater-Based Epidemiology (WBE) approach. In this review, we summarized the peer-reviewed published literature (available as of September 28, 2020), in the field of WBE. The commonly used steps (sampling, storage, concentration, isolation, detection) of the analytical protocols were identified. The potential limitations of each stage of the protocols and good practices were discussed. Finally, new methods for the efficient detection of SARS-CoV-2 were proposed., Highlights • Review of the methods for the detection of SARS-CoV-2 in wastewater. • Critical discussion of all steps of the analytical protocols. • Proposal for improvement of SARS-CoV-2 methods in wastewater based epidemiology. • Provide techniques from clinical science that are applicable in WBE. • WBE could be used as a tool to detect pandemics and track government interventions.

The huge interest in the health-related properties of foods to improve health has brought about the development of sensitive analytical methods for the characterization of natural products with functional ingredients. Greek olive leaves and drupes constitute a valuable source of biophenols with functional properties. A novel ultra-high-performance liquid chromatography&ndash, quadrupole time of flight tandem mass spectrometry (UHPLC-QTOF-MS) analytical method was developed to identify biophenols through target and suspect screening in Greek olive leaves and drupes of the varieties: Koroneiki, Throumbolia, Konservolia, Koutsourelia, Kalamon, Petrolia, Amigdalolia, Megaritiki, Mastoeidis, Agouromanakolia, Agrilia, Adramitiani and Kolovi. The method&rsquo, s performance was evaluated using the target compounds: oleuropein, tyrosol and hydroxytyrosol. The analytes demonstrated satisfactory recovery efficiency for both leaves (85.9&ndash, 90.5%) and drupes (89.7&ndash, 92.5%). Limits of detection (LODs) were relatively low over the range 0.038 (oleuropein)&ndash, 0.046 (hydroxytyrosol) and 0.037 (oleuropein)&ndash, 0.048 (hydroxytyrosol) for leaves and drupes, respectively For leaves, the precision limit ranged between 4.7 and 5.8% for intra-day and between 5.8 and 6.5% for inter-day experiments, and for drupes, it ranged between 3.8 and 5.2% for intra-day and between 5.1 and 6.2% for inter-day experiments, establishing the good precision of the method. The regression coefficient (r2) was above 0.99 in all cases. Furthermore, the preparation of herbal tea from olive leaves is suggested after investigating the optimum infusion time of dried leaves in boiling water. Overall, 10 target and 36 suspect compounds were determined in leaves, while seven targets and thirty-three suspects were identified in drupes, respectively.

Aquaculture production has globally increased and its environmental impact is not well understood and assessed yet. Therefore, in this work nine metals and metalloids (Cu, Cd, Pb, Hg, Ni, Fe, Mn, Zn and As) and three nutrients (P, N and C) that seem to accumulate in marine sediments, were determined under the fish cages (zero distance) and about 50 and 100 m away from them, in three aquacultures in Greece. The analysis of these data is crucial due to the negative impact of the intensive aquaculture activities on fish population, human health and marine environment. This study investigated the environmental impact associated with aquaculture cages on marine sediments, using Supervised Artificial Neural Networks (ANNs) in parallel with Classification Trees (CTs). Optimised models were constructed in order to detect the significance of each variable, predict the origin of the sediment samples and successfully visualise their results. Three popular ANN architectures, as multi-layer perceptrons (MLPs), radial basis function (RBF) and counter propagation artificial neural networks (CP-ANNs) were used to assess the impact of the intensive aquaculture activities on marine sediments. In addition, more traditional multivariate chemometric techniques like CTs were applied to the same data set for comparison purposes. The modelling study showed that P, N, Cu, Cd were the most critical (and polluting) factors of those metals studied. Moreover, single-element models achieved elevated predictive percentages. The results were justified due to the usual practices used for fish feeding or cages maintenance.

Due to early implementation of public health measures, Greece had low number of SARS-CoV-2 infections and COVID-19 severe incidents in hospitalized patients. The National and Kapodistrian University of Athens (ΝΚUA), especially its health-care/medical personnel, has been actively involved in the first line of state responses to COVID-19. To estimate the prevalence of antibodies (Igs) against SARS-CoV-2 among NKUA members, we designed a five consecutive monthly serosurvey among randomly selected NKUA consenting volunteers. Here, we present the results from the first 2500 plasma samples collected during June–July 2020. Twenty-five donors were tested positive for anti-SARS-CoV-2 Igs; thus, the overall seroprevalence was 1.00%. The weighted overall seroprevalence was 0.93% (95% CI: 0.27, 2.09) and varied between males [1.05% (95% CI: 0.18, 2.92)] and females [0.84% (95% CI: 0.13, 2.49)], age-groups and different categories (higher in participants from the School of Health Sciences and in scientific affiliates/faculty members/laboratory assistants), but no statistical differences were detected. Although focused on the specific population of NKUA members, our study shows that the prevalence of anti-SARS-CoV-2 Igs for the period June–July 2020 remained low and provides knowledge of public health importance for the NKUA members. Given that approximately one in three infections was asymptomatic, continuous monitoring of the progression of the pandemic by assessing Ig seroprevalence is needed.

Food science continually requires the development of novel analytical methods to prevent fraudulent actions and guarantee food authenticity. Greek table olives, one of the most emblematic and valuable Greek national products, are often subjected to economically motivated fraud. In this work, a novel ultra-high-performance liquid chromatography&ndash, quadrupole time of flight tandem mass spectrometry (UHPLC-QTOF-MS) analytical method was developed to detect the mislabeling of Greek PDO Kalamata table olives, and thereby establish their authenticity. A non-targeted screening workflow was applied, coupled to advanced chemometric techniques such as Principal Component Analysis (PCA) and Partial Least Square Discriminant Analysis (PLS-DA) in order to fingerprint and accurately discriminate PDO Greek Kalamata olives from Kalamata (or Kalamon) type olives from Egypt and Chile. The method performance was evaluated using a target set of phenolic compounds and several validation parameters were calculated. Overall, 65 table olive samples from Greece, Egypt, and Chile were analyzed and processed for the model development and its accuracy was validated. The robustness of the chemometric model was tested using 11 Greek Kalamon olive samples that were produced during the following crop year, 2018, and they were successfully classified as Greek Kalamon olives from Kalamata. Twenty-six characteristic authenticity markers were indicated to be responsible for the discrimination of Kalamon olives of different geographical origins.

Food science continually requires the development of novel analytical methods to prevent fraudulent actions and guarantee food authenticity. Greek table olives, one of the most emblematic and valuable Greek national products, are often subjected to economically motivated fraud. In this work, a novel ultra-high-performance liquid chromatography–quadrupole time of flight tandem mass spectrometry (UHPLC-QTOF-MS) analytical method was developed to detect the mislabeling of Greek PDO Kalamata table olives, and thereby establish their authenticity. A non-targeted screening workflow was applied, coupled to advanced chemometric techniques such as Principal Component Analysis (PCA) and Partial Least Square Discriminant Analysis (PLS-DA) in order to fingerprint and accurately discriminate PDO Greek Kalamata olives from Kalamata (or Kalamon) type olives from Egypt and Chile. The method performance was evaluated using a target set of phenolic compounds and several validation parameters were calculated. Overall, 65 table olive samples from Greece, Egypt, and Chile were analyzed and processed for the model development and its accuracy was validated. The robustness of the chemometric model was tested using 11 Greek Kalamon olive samples that were produced during the following crop year, 2018, and they were successfully classified as Greek Kalamon olives from Kalamata. Twenty-six characteristic authenticity markers were indicated to be responsible for the discrimination of Kalamon olives of different geographical origins.

Monitoring of biogenic amines in food is important for quality control, in terms of freshness evaluation and even more for food safety. A novel and cost-effective method was developed and validated for the determination of the main biogenic amines: histamine, putrescine, cadaverine, spermidine and spermine in fish tissues. The method includes extraction of amines with perchloric acid, pre-column derivatization with Pyrene Sulfonyl Chloride (PSCl), extraction of derivatives with toluene, back-dissolution in ACN after evaporation and determination by reversed phase high performance liquid chromatography with UV and intramolecular excimer fluorescence detection. The structure of the pyrene-derivatives was confirmed by liquid chromatography&ndash, mass spectrometry with electrospray ionization. The standard addition technique was applied for the quantitation due to significant matrix effect, while the use of 1,7-diaminoheptane as internal standard offered an additional confirmation tool for the identification of the analytes. Method repeatability expressed as %RSD ranged between 7.4&ndash, 14% for the different amines and recovery ranged from 67% for histamine up to 114% for spermine. The limits of detection ranged between 0.1&ndash, 1.4 mg kg&minus, 1 and the limits of quantification between 0.3&ndash, 4.2 mg kg&minus, 1. The method was applied to canned fish samples and the concentrations of the individual biogenic amines were below the detection limit up to 40.1 mg kg&minus, 1, while their sum was within the range 4.1&ndash, 49.6 mg kg&minus, 1.

The sonochemical degradation of trimethoprim (TMP), a widely used antibiotic, in various water matrices was investigated. The effect of several parameters, such as initial TMP concentration (0.5–3 mg/L), actual power density (20–60 W/L), initial solution pH (3–10), inorganic ions, humic acid and water matrix on degradation kinetics was examined. The pseudo-first order degradation rate of TMP was found to increase with increasing power density and decreasing pH, water complexity (ultrapure water > bottled water > secondary wastewater) and initial TMP concentration. TMP degradation is accompanied by the formation of several transformation products (TPs) as evidenced by LC-QToF-MS analysis. Nine such TPs were successfully identified and their time-trend profiles during degradation were followed. An in silico toxicity evaluation was performed showing that several TPs could potentially be more toxic than the parent compound towards Daphnia magna, Pimephales promelas and Pseudokirchneriella subcapitata.

Raw and treated leachate samples were collected from different landfills in Greece and analyzed for several groups of emerging contaminants using high resolution mass spectrometric workflows to investigate the possible threat from their discharge to the aquatic environment. Fifty-eight compounds were detected; 2−OH-benzothiazole was found at 84 % of the samples and perfluorooctanoic acid at 68 %. Bisphenol A, valsartan and 2−OH-benzothiazole had the highest average concentrations in raw leachates, after biological treatment and after reverse osmosis, respectively. In untreated leachates, Risk Quotients > 1 were calculated for 35 and 18 compounds when maximum and average concentrations were used, indicating an ecological threat for the aquatic environment. Leachates’ biological treatment partially removed COD and NH4+-N, as well as 52.3 % of total emerging contaminants. The application of reverse osmosis resulted in a 98 % removal of major pollutants, 99 % removal of total emerging contaminants and a significant decrease of ecotoxicity to Lemna minor. Beside the decrease of the detected micropollutants during treatment, RQs > 1 were still calculated for 13 and 3 compounds after biological treatment and reverse osmosis, respectively. Among these, special attention should be given to 2−OH-benzothiazole and bisphenol A that had RQ values much higher than 1 for all tested organisms.

Previous reports support that wine contains a mixture of micro-constituents in a proper quality and quantity that possess cardio-protective effect partly through Platelet-Activating Factor (PAF) inhibition. Grape pomace (GP) is a source of wine-like micro-constituents that may be a suitable alternative in food fortification. Limited data exist concerning their effects on thrombosis and inflammation. Therefore, the purpose of this study was to examine a grape pomace extract regarding its anti-platelet and anti-inflammatory properties. GP from four red grape varieties were extracted with 80% ethanol (GP:solvent 1:5w/v). The extract's total phenolic compounds were evaluated and the phenolic profile was performed by Ultrahigh-Performance-Liquid-Chromatography coupled to Mass-Spectrometry and the determination of fatty acids profile was performed by Gas-Chromatography. The extract's anti-platelet properties were tested in healthy volunteers’ platelet rich plasma by the light transmittance method, against three agonists: PAF, ADP and TRAP. The results expressed as IC50 values (μg of extract that cause 50% inhibition of aggregation) and EC50 values (agonist concentration that causes 50% of the maximum aggregation) in the extract's presence and absence. Concerning the extract's anti-inflammatory properties, peripheral blood mononuclear cells from healthy volunteers were pre-incubated with different extract concentrations, which were tested for their effect on cell viability, for 1 h and then stimulated with LPS for 4 h. Secretion of IL-1β and TNF-a was measured and normalized with the total cell protein. Phenolic compounds were calculated at 8.79 ± 1.17 mg gallic acid per g of GP. The most abundant ones were catechin, epicatechin and quercetin at 202.9 ± 6.9, 84.8 ± 1.5 and 83.7 ± 3.5μg per g of GP respectively. Out of the 18 fatty acids detected, the most abundant ones were palmitic, oleic, linoleic, and linolenic acid at 28.7 ± 0.1, 11.4 ± 0.01, 32.5 ± 0.07, 12.7 ± 0.005 g per 100 g of fat. The extract's IC50 was calculated at 162.1 ± 66.9, 181.2 ± 82.3 and 156.3 ± 97.5μg against PAF, ADP and TRAP, respectively. The EC50 values in the presence of 150μg extract were increased (lower platelet aggregation sensitivity) approximately at 100%, 45% and 13% against PAF, ADP and TRAP respectively, compared to EC50 values in the absence of extract. The presence of 500 and 1000μg/mL of extract reduced LPS-induced TNF-a secretion at approximately 38.2% (p = 0.04) and 6.0% (p < 0.000), respectively. Potent anti-platelet and anti-inflammatory properties are combined in a grape pomace extract. The use of its bioactive micro-constituents is likely to lead to the production of functional foods with cardioprotective properties.

This study presents the development and validation of a comprehensive quantitative target methodology for the analysis of 2316 emerging pollutants in water based on Ultra-Performance Liquid Chromatography Quadrupole-Time-Of-Flight Mass Spectrometry (UPLC-Q-ToF-HRMS/MS). Target compounds include pesticides, pharmaceuticals, drugs of abuse, industrial chemicals, doping compounds, surfactants and transformation products, among others. The method was validated for 195 analytes, chosen to be representative of the chemical space of the target list, enabling the assessment of the performance of the method. The method involves a generic sample preparation based on mixed mode solid phase extraction, a UPLC-QTOF-MS/MS screening method using Data Independent Acquisition (DIA) mode, which provides MS and MS/MS spectra simultaneously and an elaborate strong post-acquisition evaluation of the data. The processing method was optimized to provide a successful identification rate >95 % and to minimize the number of false positive results (< 5 %). Decision limit (CCα) and detection capability (CCβ) were also introduced in the validation scheme to provide more realistic metrics on the performance of a HRMS-based wide-scope screening method. A new system of identification points (IPs) based on the one described in the Commission Decision 2002/657/EC was applied to communicate the confidence level in the identification of the analytes. This system considers retention time, mass accuracy, isotopic fit and fragmentation; taking full advantage of the capacities of the HRMS instruments. Finally, 398 contaminants were detected and quantified in real wastewater This project was implemented under the Operational Program “Education and Lifelong Learning” and funded by the European Union (European Social Fund) and Greek National Resources – ARISTEIA 624 (TREMEPOL project). Pablo Gago-Ferrero acknowledges the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement Smart-Workflow No 747698

Background and aims Wastewater‐based epidemiology is an additional indicator of drug use that is gaining reliability to complement the current established panel of indicators. The aims of this study were to: (i) assess spatial and temporal trends of population‐normalized mass loads of benzoylecgonine, amphetamine, methamphetamine and 3,4‐methylenedioxymethamphetamine (MDMA) in raw wastewater over 7 years (2011–17); (ii) address overall drug use by estimating the average number of combined doses consumed per day in each city; and (iii) compare these with existing prevalence and seizure data. Design Analysis of daily raw wastewater composite samples collected over 1 week per year from 2011 to 2017. Setting and Participants Catchment areas of 143 wastewater treatment plants in 120 cities in 37 countries. Measurements Parent substances (amphetamine, methamphetamine and MDMA) and the metabolites of cocaine (benzoylecgonine) and of Δ9‐tetrahydrocannabinol (11‐nor‐9‐carboxy‐Δ9‐tetrahydrocannabinol) were measured in wastewater using liquid chromatography–tandem mass spectrometry. Daily mass loads (mg/day) were normalized to catchment population (mg/1000 people/day) and converted to the number of combined doses consumed per day. Spatial differences were assessed world‐wide, and temporal trends were discerned at European level by comparing 2011–13 drug loads versus 2014–17 loads. Findings Benzoylecgonine was the stimulant metabolite detected at higher loads in southern and western Europe, and amphetamine, MDMA and methamphetamine in East and North–Central Europe. In other continents, methamphetamine showed the highest levels in the United States and Australia and benzoylecgonine in South America. During the reporting period, benzoylecgonine loads increased in general across Europe, amphetamine and methamphetamine levels fluctuated and MDMA underwent an intermittent upsurge. Conclusions The analysis of wastewater to quantify drug loads provides near real‐time drug use estimates that globally correspond to prevalence and seizure data., This study was supported by the following countries, institutions and projects Australia: Thyne Reid foundation; Canada: Natural Sciences and Engineering Research Council of Canada (NSERC); Iceland: The Icelandic Research Fund (Grant number 163049–051); Slovenia: Slovene Research Agency (funding Project L1–9191); Spain: MINECO/AEI projects (CTM2014–56628‐C3–2‐R, CTQ2015–65603‐P, CTM2016–81935‐REDT, CTM2017–84763‐C3–2‐R), Galician Council of Culture, Education and Universities (ED481D 2017/003); UK: Environmental Sustainability Knowledge Transfer Network, Engineering and Physical Sciences Research Council and Thermo Fisher Scientific (CASE industrial scholarship for K. Munro, Ref.: EP/J502029/1). European Monitoring Centre for Drugs and Drug Addiction (EMCDDA); EU International Training Network SEWPROF (Marie Curie‐FP7‐PEOPLE, grant number 317205); COST Action ES1307 supported by COST (European Cooperation in Science and Technology); WATCH (Wastewater Analysis of Traces of illicit drug‐related Chemicals for law enforcement and public Health DG migration and Home affairs ‐ HOME/2015/ISFP/PR/DRUG/0062). Sampling support and provision of catchment information Austria: Klemens Geiger and Michael Schlapp; Belgium: Jonathan Phariseau, Bart Coene, Luc Van Os, Peter Nys, Dieter Lemaire, Jean‐François Mougel, Patrick Vantroyen and Karel Claes; Croatia: Marin Ganjto; Czech Republic: Jiri Stara and Robert Hrich Brněnské; France: Véronique Bremont; Italy: Roberto Mazzini and Francesca Pizza (Milan), Francesco Avolio (Bologna), Fabrizio Moratto and Sergio Ghezzi (Gorizia), Rosanna Brienza and Simona Panariello (Potenza), Pier Paolo Abis and Antonia Attanasio (Bari), Antonella Cicala (Palermo); Norway: Pia Ryrfors; Portugal: João Goulão, José Martins, Pedro Alvaro and Fátima Paixão; Slovakia: Silvia Antalová and Jozef Tichy; Spain: Begoña Martínez López, Cristian Mesa, Santiago Querol Rodriguez, Fernando Llavador, Enrique Albors and Gloria Fayos; the Netherlands: Alex Veltman, Peter Theijssen, Peter van Dijk, Stefan Wijers, Mark Stevens, Ferry de Wilde. Analytical support Australia: Maulik Ghetia; Cyprus: Popi Karaolia; Greece: Nikoilaos Raikos; Spain: Ana Maria Botero‐Coy, Clara Boix, Alberto Celma, Jorge Pitarch, Inés Racamonde and Eddie Fonseca; Italy: Emma Gracia‐Lor, Nikolaos I. Rousis and Noelia Salgueiro‐Gonzalez. Data repository and data screening Eawag: Harald von Waldow; EMCDDA: Renate Hochwieser, Liesbeth Vandam, João Matias and Federica Mathis.

© The Author(s) 2020., The Partnership for Chemicals Risk Assessment (PARC) is currently under development as a joint research and innovation programme to strengthen the scientific basis for chemical risk assessment in the EU. The plan is to bring chemical risk assessors and managers together with scientists to accelerate method development and the production of necessary data and knowledge, and to facilitate the transition to next-generation evidence-based risk assessment, a non-toxic environment and the European Green Deal. The NORMAN Network is an independent, well-established and competent network of more than 80 organisations in the field of emerging substances and has enormous potential to contribute to the implementation of the PARC partnership. NORMAN stands ready to provide expert advice to PARC, drawing on its long experience in the development, harmonisation and testing of advanced tools in relation to chemicals of emerging concern and in support of a European Early Warning System to unravel the risks of contaminants of emerging concern (CECs) and close the gap between research and innovation and regulatory processes. In this commentary we highlight the tools developed by NORMAN that we consider most relevant to supporting the PARC initiative: (i) joint data space and cutting-edge research tools for risk assessment of contaminants of emerging concern; (ii) collaborative European framework to improve data quality and comparability; (iii) advanced data analysis tools for a European early warning system and (iv) support to national and European chemical risk assessment thanks to harnessing, combining and sharing evidence and expertise on CECs. By combining the extensive knowledge and experience of the NORMAN network with the financial and policy-related strengths of the PARC initiative, a large step towards the goal of a non-toxic environment can be taken.

Screening of a large number of chemicals of emerging concern is highly desirable for the control of crops irrigated with reclaimed water since it is considered an alternative water source of great value. This study describes a high resolution mass spectrometry approach for developing methods for quantification in lettuce leaves of 48 different wastewater-borne pollutants (including analgesics and anti-inflammatories, anti-hypertensives, antifungal agents, lipid regulators, psychiatric drugs and stimulants, β-blockers, antibiotics, antimycotics, and sweeteners) frequently found in water resources. In this respect, a simple and fast QuEChERS-based method for the determination of contaminants in lettuce has been developed. During extraction, the use of formic acid was adopted to further improve the results of some problematic compounds (e.g., fenofibrate, furosemide, metronidazole, oxcarbazepine, sulfanilamide). High resolution multiple reaction monitoring (MRM) and SWATH acquisition were compared in term of accuracy, repeatability, sensitivity, linearity and matrix effect. Both methods provided similar recoveries between 80 and 120% in lettuce leaves, although sulfanilamide, ciprofloxacin, and sulfamethazine presenting values of 26.8, 27.8, and 28.4% in MRM and 25, 33.9, and 35% in SWATH, respectively. The effectiveness of a two-step cleanup on analyte recovery was also assessed and matrix effects were also taken into consideration during the method validation. The developed method allows the simultaneous quantitative analysis of 48 compounds (drug residues and metabolites) in lettuce leaves irrigated with treated wastewater for human consumption. Application of the present method to lettuce crops growth in controlled conditions showed the presence of 14 out 48 studied compounds with similar concentrations in both acquisition modes ranging from 3.3 and 1.3 ng g for climbazole (for MRM and SWATH, respectively) to 33.2 and 17.7 ng g for sulfamethazine. Drug residues such as carbamazepine (6.0 and 8.5 ng g ), and its metabolite carbamazepine epoxide (18.1 and 16.5 ng g ), frequently found in wastewater effluents, were also detected., This study has been financially supported by the EU through the WaterJPI-2015 AWARE project (PCIN-2017–067). This work was supported by the Spanish Ministry of Science and Innovation (Project CEX2018–000794-S). The authors thank the Water Challenges for a Changing World Joint Programming Initiative. The authors also acknowledge SCIEX for providing the loan instrument LC/HRMS QTOF X500R and Bekolut GmbH & Co. KG for the contribution with QuEChERS kits extraction. The EU is not liable for any use that may be made of the information contained therein.

Treated wastewater irrigation (TWW) releases antibiotics and antibiotic resistance genes (ARGs) into the environment and might thus promote the dissemination of antibiotic resistance in groundwater (GW). We hypothesized that TWW irrigation increases ARG abundance in GW through two potential mechanisms: the contamination of GW with resistant bacteria and the accumulation of antibiotics in GW. To test this, the GW below a real-scale TWW-irrigated field was sampled for six months. Sampling took place before, during and after high-intensity TWW irrigation. Samples were analysed with 16S rRNA amplicon sequencing, qPCR of six ARGs and the class 1 integron-integrase gene intI1, while liquid chromatography tandem mass spectrometry was performed to detect antibiotic and pharmaceutical residues. Absolute abundance of 16S rRNA in GW decreased rather than increased during long-term irrigation. Also, the relative abundance of TWW-related bacteria did not increase in GW during long-term irrigation. In contrast, long-term TWW irrigation increased the relative abundance of sul1 and intI1 in the GW microbiome. Furthermore, GW contained elevated concentrations of sulfonamide antibiotics, especially sulfamethoxazole, to which sul1 confers resistance. Total sulfonamide concentrations in GW correlated with sul1 relative abundance. Consequently, TWW irrigation promoted sul1 and intI1 dissemination in the GW microbiome, most likely due to the accumulation of drug residues.

The increasing use of chemicals in the European Union (EU) has resulted in environmental emissions and wildlife exposures. For approving a chemical within the EU, producers need to conduct an environmental risk assessment, which typically relies on data generated under laboratory conditions without considering the ecological and landscape context. To address this gap and add information on emerging contaminants and chemical mixtures, we analysed 30 livers of white-tailed sea eagles (Haliaeetus albicilla) from northern Germany with high resolution-mass spectrometry coupled to liquid and gas chromatography for the identification of >2400 contaminants. We then modelled the influence of trophic position (δ15N), habitat (δ13C) and landscape on chemical residues and screened for persistent, bioaccumulative and toxic (PBT) properties using an in silico model to unravel mismatches between predicted PBT properties and observed exposures. Despite having generally low PBT scores, most detected contaminants were medicinal products with oxfendazole and salicylamide being most frequent. Chemicals of the Stockholm Convention such as 4,4′-DDE and PCBs were present in all samples below toxicity thresholds. Among PFAS, especially PFOS showed elevated concentrations compared to other studies. In contrast, PFCA levels were low and increased with δ15N, which indicated an increase with preying on piscivorous species. Among plant protection products, spiroxamine and simazine were frequently detected with increasing concentrations in agricultural landscapes. The in silico model has proven to be reliable for predicting PBT properties for most chemicals. However, chemical exposures in apex predators are complex and do not solely rely on intrinsic chemical properties but also on other factors such as ecology and landscape. We therefore recommend that ecological contexts, mixture toxicities, and chemical monitoring data should be more frequently considered in regulatory risk assessments, e.g. in a weight of evidence approach, to trigger risk management measures before adverse effects in individuals or populations start to manifest.

The aim of this work is the evaluation of quality indicators for the estimation of the shelf life of opened cans using the migration of specific metals as variables. For this reason the determination of targeted analytes such as: Cd, Pb, As, Cu, Cr, Ni, Fe, Mn and Sn in different canned samples (cardoon, tuna, green and red beans, corn, and fungi) by Electrothermal Atomic Absorption Spectrometry (Perkin Elmer, SIMAA 6000) was developed and validated. Statistical analysis was performed using SPSS version 22. Principal component analysis (PCA) and cluster analysis were performed in order to examine the correlation between the content of metals and metalloids and the storage time of opened cans. The results showed that there is a strong correlation between metal concentration and storage time, especially for Fe and Sn. The storage time ranged from hours to days and the vast number of collected data sets, led to reliable conclusions about the evaluation of a new shelf life indicator. Principal component analysis indicated the appropriate storage time for opened cans, where no significant migration was observed.

At the individual level, exposure to contaminants is generally assessed through the analysis of specific biomarkers m biological matnces. However, these studies are costly and logistically demanding, limiting their applicability to momtor population-wide exposure over time and space. By focusing on a selection of exposure biomarkers to phosphorus flame retardants and plasticizers (PFRs), this study aims to explore the possibility of using wastewater as a complementary source of information about exposure. Wastewater samples were collected from five cities in Europe and analyzed using a previously establised method. Substantial differences in biomarker levels were observed between the investigated catchments, suggesting differences in exposure. Time trends in biomarkers observed between 2013 and 2016 were found to agree with results from human biomonitoring studies and reports about production volumes. Using Monte Carlo simulations, average urinary concentrations were estimated. These were generally higher compared to results from human biomonitoring studies. Various explanations for these differences were formulated (i.e., other excretion routes, external sources and different sampling approaches). Obtained results show that wastewater analysis provides unique information about geographical and temporal differences in exposure, which would be difficult to gather using other monitoring tools.

A robust and sensitive methodology, utilizing GC-MS, for the identification and quantitation of seven β-agonist residues (mabuterol, clenbuterol, brombuterol, mapenterol, clenpenterol, clenproperol, and clencyclohexerol), in liver and meat samples, is described. Different extraction procedures, followed by GC/MS measurement, were tested and compared in order to achieve optimum extraction conditions and eliminate matrix effects. The optimized method consisted of consecutive extractions with hydrochloric tris(hydroxymethyl)aminomethane (TRIS-HCl) and tert-butyl-methyl ether (TBME), defatting with hexane and solid-phase extraction (SPE) with C18 cartridges. Ν,O-bis(trimethylsilyl) trifluoroacetamide (BSTFA) with 1% trimethylchlorosilane (TMCS) was used as a derivatization agent and the GC-EI-MS determination was performed using a MDN-5S capillary column in single-ion monitoring (SIM) acquisition mode. The method was validated according to the Commission Decision 2002/657/EC, fulfilling all the EU criteria. Quantification was performed via internal standard calibration using deuterated analogs of the compounds. Recoveries ranged from 83 to 118% (mabuterol at concentration levels of 2.0 and 1.5 μg kg−1, respectively) and precision, expressed as %relative standard deviation (% RSD), was in every case lower than the % RSDs obtained from Horwitz equation. The obtained decision limit (CCα) and detection capability (CCβ) values varied from 0.21 ng g−1 (clenbuterol) to 0.49 ng g−1 (clenproperol, clencyclohexerol) and from 0.60 (mapenterol) to 0.69 ng g−1 (clenpenterol), respectively.

COVID-19 pandemic spread rapidly worldwide with unanticipated effects on mental health, lifestyle, stability of economies and societies. Although many research groups have already reported SARS-CoV-2 surveillance in untreated wastewater, only few studies evaluated the implications of the pandemic on the use of chemicals by influent wastewater analysis. Wide-scope target and suspect screening were used to monitor the effects of the pandemic on the Greek population through wastewater-based epidemiology. Composite 24 h influent wastewater samples were collected from the wastewater treatment plant of Athens during the first lockdown and analyzed by liquid chromatography mass spectrometry. A wide range of compounds was investigated (11,286), including antipsychotic drugs, illicit drugs, tobacco compounds, food additives, pesticides, biocides, surfactants and industrial chemicals. Mass loads of chemical markers were estimated and compared with the data obtained under non-COVID-19 conditions (campaign 2019). The findings revealed increases in surfactants (+196%), biocides (+152%), cationic quaternary ammonium surfactants (used as surfactants and biocides) (+331%), whereas the most important decreases were estimated for tobacco (−33%) and industrial chemicals (−52%). The introduction of social-restriction measures by the government affected all aspects of life., Graphical abstract Unlabelled Image

Maturity is one of the most important factors associated with the quality of olive products, however the molecular events underlying olive drupe development remain poorly characterized. Using proteomic and metabolomic approaches, this study investigated the changes in the olive drupes (cv. Chondrolia Chalkidikis) across six developmental stages (S1-S6) that characterize the dynamics of fruit growth and color. Primary metabolites, including carbohydrates and organic acids (i.e., xylose, malic acid), showed significant accumulation in the black maturation stage. Temporal changes in various secondary metabolites (e.g., oleuropein, oleacin and tyrosol) were also observed. Proteins involved in oxidation-reduction (i.e., LOX1/5), carbohydrate metabolism (i.e., GLUA, PG) and photosynthesis (i.e., chlorophyll a-b binding proteins) significantly altered in the turning black compared to the green mature stage. By providing the first proteometabolomic study of olive drupe development, this investigation offers a novel framework for further studies on this economically relevant crop.