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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.

LC-MS based metabolomics provide a new perspective in wine authentication enabling a thorough investigation of its chemical composition. In this study, 97 monovarietal white wines derived from four indigenous Greek grape varieties (Assyrtiko, Moschofilero, Malazouzia and Savatiano) and produced in two PDO and one PGI winemaking regions were analyzed with the use of ultra-high-performance liquid chromatography–quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS). A targeted metabolomics method was developed, validated and applied to the analyzed wine samples for the identification and quantification of 22 metabolites. Another 79 compounds were tentatively identified using a “smart” suspect screening workflow based on an in-house developed database. Then, the random forest algorithm was applied on the normalized peak areas of both target and suspect data for supervised statistical analysis. A robust classification model was built enabling the classification and prediction of the varietal origin of the wine samples in acceptable levels. Specific biomarkers, contributing significantly to the discrimination of wine variety, were recognized.

Major, minor and trace elements in wines from Greece were determined by inductively coupled plasma–mass spectrometry (ICP–MS). The concentrations of 44 elements (Na, Mg, P, K, Ca, Cu, Co, Cr, Zn, Sn, Fe, Mn, Li, Be, B, V, Sr, Ba, Al, Ag, Ni, As, Sn, Hg, Pb, Sb, Cd, Ti, Ga, Zr, Nb, Pd, Te, La, Sm, Ho, Tm, Yb, W, Os, Au, Tl, Th, U) in 90 white and red wines from six different regions in Greece for two consecutive vinification years, 2017 and 2018, were determined. Results for the elements aforementioned were evaluated by multivariate statistical methods, such as discriminant analysis and cluster analysis, and the wines were discriminated according to wine variety and geographical origin. Due to the specific choice of the analytes for multivariate statistical investigation, a prediction rate by cross-validation of 98% could be achieved. The aim of this study was not only to reveal specific relationships between the wine samples or between the chemical variables in order to classify the wines from different regions and varieties according to their elemental profile (wine authentication), but also to observe the annual fluctuation in the mineral content of the studied wine samples.

The aim of this work was to study the chromatographic behavior of polar artificial sweeteners on three hydrophilic interaction liquid chromatography (HILIC) columns: Kinetex HILIC (unbounded silica, core shell), ZIC HILIC (sulfoalkylbetain) and TSKgel Amide-80 (amide). The studied analytes were dipeptides (aspartame, neotame, advantame, alitame), sulfamates (acesulfame, saccharin, cyclamate), and sugars (sucralose, neohesperidin dihydrochalcone). Critical parameters such as the mobile phase pH, buffer concentration, column temperature, and organic modifier were studied for the investigation of the retention mechanism. Partition and adsorption models were also used. The retention mechanism of dipeptides seems to be partition for the aqueous layer and weak electrostatic interactions or hydrogen bonds. For the sulfamates, on Kinetex and ZIC HILIC columns, it is likely that partitioning to the aqueous layer is combined with the ERLIC phenomena at pH 5.5. For the TSKgel Amide, the partition and the hyd...

This study investigated the use of liquid chromatography–atmospheric pressure photoionization tandem mass spectrometry (LC-APPI-MS/MS) for the determination of six polybrominated diphenyl ethers (PBDEs) coded as brominated diphenyl ether (BDE) 28, 47, 99, 100, 153, and 154 that are used as flame retardants. It was shown that diphenyl ethers with four to six bromine atoms can be ionized in negative polarity with parent ions corresponding to [M-Br +O]−. On the contrary, the ionization of the diphenyl ether with three bromine atoms (BDE 28), required the use of positive polarity in order to generate precursor ions [M]+. PBDEs were separated by a C18 column (50 × 4.6 mm, 1.8 µm) with a mixture of methanol–water as the mobile phase following gradient elution. Instrumental limits of detection ranged from 41 to 119 pg on the column. The method was applied to surface water samples after liquid–liquid extraction with dichloromethane. Method limits of quantification ranged from 4.1 to 73 ng L−1 and recoveri...

The repowering of already installed wind farms is considered one of the most promising and cost-effective short-term strategies to scale-up wind capacity. In this study, we apply Markowitz's mean-variance (MV) portfolio optimization theory to explore alternative repowering actions in Spain. The efficient portfolios – a direct outcome of the MV optimization – offer optimal repowering alternatives to current wind farm generation mixes. They deliver the highest possible average power output (yield) for a given level of supply risk. Different repowering scenarios are considered in this paper that range from a full restructuring of the existing wind generation mix to restricting by certain amounts the percentage of down-/uprating of each reference region. Results show that, depending on the configuration of the MV portfolio optimization problem, hourly fluctuations in the aggregate power supply can be reduced as much as 12–31%, while retaining the current level of energy productivity. In addition, for the level of energy supply risk experienced with the existing portfolio of Spanish wind farms; we can derive more efficient mixes that boost-up productivity by 16–55%. This work aims at providing valuable insight for energy policy-making in the direction of optimally repowering future renewable generation.

The discrimination of organic and conventional production has been a critical topic of public discussion and constitutes a scientific issue. It remains a challenge to establish a correlation between the agronomical practices and their effects on the composition of olive oils, especially the phenolic composition, since it defines their organoleptic and nutritional value. Thus, a liquid chromatography-electrospray ionization-quadrupole time of flight tandem mass spectrometric method was developed, using target and suspect screening workflows, coupled with advanced chemometrics for the identification of phenolic compounds and the discrimination between organic and conventional extra virgin olive oils. The method was optimized by one-factor design and response surface methodology to derive the optimal conditions of extraction (methanol/water (80:20, v/v), pure methanol, or acetonitrile) and to select the most appropriate internal standard (caffeic acid or syringaldehyde). The results revealed that extraction with methanol/water (80:20, v/v) was the optimum solvent system and syringaldehyde 1.30 mg L-1 was the appropriate internal standard. The proposed method demonstrated low limits of detection in the range of 0.002 (luteolin) to 0.028 (tyrosol) mg kg-1. Then, it was successfully applied in 52 olive oils of Kolovi variety. In total, 13 target and 24 suspect phenolic compounds were identified. Target compounds were quantified with commercially available standards. A novel semi-quantitation strategy, based on chemical similarity, was introduced for the semi-quantification of the identified suspects. Finally, ant colony optimization-random forest model selected luteolin as the only marker responsible for the discrimination, during a 2-year study. Graphical abstract Investigation of the organic and conventional production type of olive oil by LC-QTOF-MS.

The fate of four antimicrobials (cefadroxil, CFD; metronidazole, METRO; trimethoprim, TRI; sulfamethoxazole, SMX) was studied in Lemna minor systems and the role of different mechanisms on their removal was evaluated. All micropollutants were significantly removed in batch experiments with active Lemna minor; the highest removal was observed for CFD (100% in 14 d), followed by METRO (96%), SMX (73%) and TRI (59%) during 24 d of the experiment. Calculation of kinetic constants for hydrolysis, photodegradation, sorption to biomass and plant uptake revealed significant differences depending on the compound and the studied mechanism. For METRO, TRI and SMX the kinetic constants of plant uptake were by far higher comparing to those of the other mechanisms. The transformation products of antimicrobials were identified using UHPLC-QToF-MS. Two were the main degradation pathways for TRI; hydroxylation takes place during both phyto- and photodegradation, while demethylation occurs only in absence of Lemna minor. The operation of a continuous-flow duckweed system showed METRO and TRI removal equal to 71 ± 11% and 61 ± 8%, respectively. The application of mass balance and the use of published biodegradation constants showed that plant uptake and biodegradation were the major mechanisms governing METRO removal; the most important mechanism for TRI was plant uptake.

The intramolecular excitation energy transfer (EET) processes in a series of fluorescent-unquenched, self-assembled metallocycles consisting of spatially fixed-separated and parallel-aligned Bodipy chromophores, are investigated here by steady-state and femtosecond-fluorescence upconversion measurements in the solution phase. These multi-Bodipy macrocycles, namely, the rhomboid (A1), the tetragon (A2) and the hexagon (A3), are formed via temperature-regulated Pt(II)–pyridyl coordination and consist, respectively, of two, four, and six Bodipy subunits, which are locked at the corners and aligned with their long molecular axes perpendicular to the rigid polygonal frame formed by the alternating B···Pt(II) connectivities. Extensive simulations and fits to the experimental fluorescence anisotropy decays r(t) show that EET within the cyclic scaffolds is quite uniform and much faster than the intrinsic decay rate of the Bodipy’s. The equalization of the excitation survival probabilities over time of all chromop...

The information obtained from the chemical analysis of specific human excretion products (biomarkers) in urban wastewater can be used to estimate the exposure or consumption of the population under investigation to a defined substance. A proper biomarker can provide relevant information about lifestyle habits, health and wellbeing, but its selection is not an easy task as it should fulfil several specific requirements in order to be successfully employed. This paper aims to summarize the current knowledge related to the most relevant biomarkers used so far. In addition, some potential wastewater biomarkers that could be used for future applications were evaluated. For this purpose, representative chemical classes have been chosen and grouped in four main categories: (i) those that provide estimates of lifestyle factors and substance use, (ii) those used to estimate the exposure to toxicants present in the environment and food, (iii) those that have the potential to provide information about public health and illness and (iv) those used to estimate the population size. To facilitate the evaluation of the eligibility of a compound as a biomarker, information, when available, on stability in urine and wastewater and pharmacokinetic data (i.e. metabolism and urinary excretion profile) has been reviewed. Finally, several needs and recommendations for future research are proposed. This work was supported by the COST Action ES1307 “SCORE – Sewage biomarker analysis for community health assessment”. Emma Gracia-Lor is very grateful to Generalitat Valenciana, Conselleria d'Educació, Investigació, Cultura i Esport (APOSTD/2015, Programa VALi + d) for her post-doctoral contract. Lubertus Bijlsma acknowledges NPS-Euronet (HOME/2014/JDRUG/AG/DRUG/7086), co-funded by the European Union, for his post-doctoral fellowship. Erika Castrignanò, Richard Bade, Juliet Kinyua, Pedram Ramin, Nikolaos I. Rousis, Yeonsuk Ryu would like to thank the SEWPROF MC ITN project, ‘A new paradigm in drug use and human health risk assessment: Sewage profiling at the community level’ [grant agreement 317205] supported by the European Union's Seventh Framework Programme for research, technological development and demonstration for the financial support. Iria González-Mariño extends her gratitude to the Galician Council of Culture, Education and Universities for her postdoctoral contract (Plan Galego de Investigación, Innovación e Crecemento 2011–2015). Foon Yin Lai acknowledges her postdoctoral fellowship from the University of Antwerp. Luigi Lopardo, Axel Rydevik and Barbara Kasprzyk-Hordern would like to acknowledge Leverhulme Trust for funding ‘TOX-EDC, Wastewater profiling for community-wide human exposure assessment from environmental endocrine disrupting chemicals in personal care and consumer products’ (Project No: RPG-2013-297). Frederic Been would like to thank the Swiss National Science Foundation (SNF, P2LAP2_164892) for his post-doctoral grant. This publication reflects the views only of the authors, and the European Commission cannot be held responsible for any use which may be made of the information contained therein.

The complexing capacity for Cu ions ( L T ) and the apparent stability constant values (log K app ) were measured in throughfall (TF) and open field bulk deposition (BD), at two Mediterranean forest ecosystems of western Greece. L T was measured for the first time in the TF of natural forests. Concentrations of L T were three-to-four-fold higher in TF (mean ± st.dev: 2014 ± 769 nM for Varetada and 1565 ± 595 nM for Karpenissi) compared to those in BD (531 ± 517 nM and 468 ± 321 nM, respectively). In all TF and BD samples, L T concentrations were significantly higher than the corresponding total Cu concentrations, indicating that Cu was fully complexed. The L T /TOC ratios in TF were found comparable between the two study sites (235 ± 149 nM mg −1 C for Varetada and 256 ± 233 nM mg −1 C for Karpenissi) and with those of BD (226 ± 257 and 163 ± 163 nM mg −1 C, respectively). The determined mean log K app values were almost identical in TF (6.8 ± 0.7 at Varetada; 6.8 ± 0.6 at Karpenissi) and BD (6.5 ± 0.6 at Varetada; 6.8 ± 0.3 at Karpenissi), pointing to the fact that regardless of the enrichment of TF in ligands ( L T (TF) > L T (BD) ) the type of binding sites remain the same. The log K app values which were obtained herewith are similar to those obtained for humic-like substances (HULIS), indicating them as the most widespread type of ligands in BD and TF.

An investigation into the occurrence of priority substances regulated by 2000/60/EC Water Framework Directive and 2008/105/EC Directive was conducted for a period of one year in the surface water sources supplying the water treatment plants (WTPs) of Athens and in the raw water of WTPs. Samples from four reservoirs and four water treatment plants of Athens were taken seasonally. The substances are divided into seven specific groups, including eight volatile organic compounds (VOCs), diethylhexylphthalate, four organochlorine pesticides (OCPs), three organophosphorus/organonitrogen pesticides (OPPs/ONPs), four triazines and phenylurea herbicides, pentachlorophenol, and four metals. The aforementioned substances belong to different chemical categories, and different analytical methods were performed for their determination. The results showed that the surface waters that feed the WTPs of Athens are not burdened with significant levels of toxic substances identified as European Union (EU) priority substances. Atrazine, hexachlorocyclohexane, endosulfan, trifluralin, anthracene and 4-nonylphenol were occasionally observed at very low concentrations. Their presence in a limited number of cases could be attributed to waste disposal, agricultural activities, and to a limited industrial activity in the area nearby the water bodies.

A new facile and fast method was developed in this study for the determination of pigments (chlorophylls and carotenoids), tocopherols (&alpha, sum of (&beta, + &gamma, ), and &delta, ), and squalene in olive oil. This method consisted of a dilution of olive oil in 2-propanol, followed by reversed phase-high-pressure liquid chromatography equipped with a diode array detector (RP-HPLC-DAD). Chromatographic separation was performed using a C18 column, while the mobile phase consisted of acetonitrile and methanol using a gradient elution program. The methodology was optimized, validated, and applied to the analysis of 452 samples of Extra Virgin Olive Oil (EVOOs) and Virgin Olive Oil (VOOs) originated from five islands of the Northeastern Aegean Region, in Greece. From the obtained results, it was indicated that the carotenoid, tocopherol, and squalene content was relatively high, while the chlorophyll content was low. Furthermore, the acquired results were studied and compared in order to obtain useful information about the correlation of the concentration levels of these compounds in olive oil to different cultivation and olive oil production parameters. Several parameters were found to play a significant role on the pigment and antioxidant content of olive oil, such as the olive tree variety, geographical origin, fruit maturation stage during harvesting, and addition of water during malaxation, while other parameters such as the altitude of cultivation, the type of farming (organic or conventional), and the type of olive mill did not seem to affect the levels of these compounds.

Background The Max Weaver Dye Library (MWDL) from North Carolina State University is a repository of around 98,000 synthetic dyes. Historically, the uses for these dyes included the coloration of textiles, paper, packaging, cosmetic and household products. However, little is reported about their ecotoxicological properties. It is anticipated that prediction models could be used to help provide this type information. Thus, the purpose of this work was to determine whether a recently developed QSAR (quantitative structure–activity relationships) model, based on ACO-SVM techniques, would be suitable for this purpose. Results We selected a representative subset of the MWDL, composed of 15 dyes, for testing under controlled conditions. First, the molecular structure and purity of each dye was confirmed, followed by predictions of their solubility and pKa to set up the appropriate test conditions. Only ten of the 15 dyes showed acute toxicity in Daphnia, with EC50 values ranging from 0.35 to 2.95 mg L−1. These values were then used to determine the ability of the ACO-SVM model to predict the aquatic toxicity. In this regard, we observed a good prediction capacity for the 10 dyes, with 90% of deviations within one order of magnitude. The reasons for this outcome were probably the high quality of the experimental data, the consideration of solubility limitations, as well as the high purity and confirmed chemical structures of the tested dyes. We were not able to verify the ability of the model to predict the toxicity of the remaining 5 dyes, because it was not possible to determine their EC50. Conclusions We observed a good prediction capacity for the 10 of the 15 tested dyes of the MWDL, but more dyes should be tested to extend the existing training set with similar dyes, to obtain a reliable prediction model that is applicable to the full MWDL.

Electricity prices are known to have special statistical properties, where long-memory effects, varying-level periodicities, cross-dependencies and other exogenous influences often work together to form a complex data-generating process. The need for accurately representing all these features typically leads us to complex parametrizations, which are prone to numerical instabilities, overfitting and poor transparency. A natural counterpart to an overparameterized model is accomplished by the use of factor decomposition techniques to simplify the modeling process by implementing a more compact factor structure for the cross-section of electricity prices. In this paper, we exploit multi-level factor modelling techniques to unravel systematic unobserved determinants of the intraday and interzonal price curve dynamics. These techniques make an explicit separation of global drivers from region-specific common factors, thereby facilitating the identification of the actual sources of co-variability. Our findings on the Pennsylvania-New Jersey-Maryland (PJM) interconnection confirm the hypothesis that the common unobserved determinants of power prices in this interconnection obey a block structure, some of which affect different segments of our panel. We argue that a multi-level factor approach offers a more systematic and transparent representation of intertemporal and cross-sectional patterns in PJM electricity prices compared to alternative brute-force VARMAX parametrizations and the ``flat" factor models, which are often put forward in the literature as viable modelling alternatives.

The number of new psychoactive substances (NPS) is constantly increasing. However, although the number might be large, most NPS have a low prevalence of use, so keeping screening libraries updated with the relevant analytical targets becomes a challenge. One way to ensure sufficient screening coverage is to use shared high resolution-mass spectrometry (HR-MS) databases, such as HighResNPS.com: a free, online, spreadsheet-format, crowd-sourced HR-MS database for NPS screening. The aims of this study were (i) to present the database to the scientific community and (ii) to verify that the HighResNPS database can be utilized in suspect screening workflows for LC–HR-MS instruments and software from four different instrument vendors. A sample was spiked with 10 NPS, and participating laboratories then analyzed the sample with their respective HR-MS vendor platforms and the HighResNPS database. The HighResNPS data were obtained via a spreadsheet converted to fit the import specifications of the different vendor platforms. Suspect screening was performed using LC–HR-MS vendor platforms from Thermo Fisher, Waters, Bruker and Agilent. All 10 NPS were identified in at least three workflows used for the four different vendor platforms. Multiple users have submitted data to HighResNPS for the same NPS, which resulted in multiple true-positive identifications for these NPS. Suspect screening with LC–HR-MS can be based on diagnostic fragment ions reported by users of different vendor platforms and can support NPS identification in biological samples and/or seizure analyses when no reference standard is available in-house. The present work clearly demonstrates that HighResNPS data is compatible with instruments and screening software from at least four different vendor platforms. The database can thus serve as a useful add-on in LC–HR-MS screening workflows.

Untargeted analysis of a composite house dust sample has been performed as part of a collaborative effort to evaluate the progress in the field of suspect and nontarget screening and build an extensive database of organic indoor environment contaminants. Twenty-one participants reported results that were curated by the organizers of the collaborative trial. In total, nearly 2350 compounds were identified (18%) or tentatively identified (25% at confidence level 2 and 58% at confidence level 3), making the collaborative trial a success. However, a relatively small share (37%) of all compounds were reported by more than one participant, which shows that there is plenty of room for improvement in the field of suspect and nontarget screening. An even a smaller share (5%) of the total number of compounds were detected using both liquid chromatography–mass spectrometry (LC-MS) and gas chromatography–mass spectrometry (GC-MS). Thus, the two MS techniques are highly complementary. Most of the compounds were detected using LC with electrospray ionization (ESI) MS and comprehensive 2D GC (GC×GC) with atmospheric pressure chemical ionization (APCI) and electron ionization (EI), respectively. Collectively, the three techniques accounted for more than 75% of the reported compounds. Glycols, pharmaceuticals, pesticides, and various biogenic compounds dominated among the compounds reported by LC-MS participants, while hydrocarbons, hydrocarbon derivatives, and chlorinated paraffins and chlorinated biphenyls were primarily reported by GC-MS participants. Plastics additives, flavor and fragrances, and personal care products were reported by both LC-MS and GC-MS participants. It was concluded that the use of multiple analytical techniques was required for a comprehensive characterization of house dust contaminants. Further, several recommendations are given for improved suspect and nontarget screening of house dust and other indoor environment samples, including the use of open-source data processing tools. One of the tools allowed provisional identification of almost 500 compounds that had not been reported by participants. [Figure not available: see fulltext.].

Paola Movalli et al., European Raptor Biomonitoring Facility COST Action (CA16224) is supported by COST (European Cooperation in Science and Technology) and funded by the Horizon 2020 Framework Programme of the European Union. Life APEX is financed by the European Union through the program LIFE17ENV/SK/000355‘Systematic use ofcontaminant data from apex predators and their prey in chemicals management’.

Averaged 7-day composite effluent wastewater samples from twelve wastewater treatment plants (WWTPs) in nine countries (Romania, Serbia, Hungary, Slovenia, Croatia, Slovakia, Czechia, Austria, Germany) in the Danube River Basin were collected. WWTPs' selection was based on countries' dominant technology and a number of served population with the aim to get a representative holistic view of the pollution status. Samples were analyzed for 2248 chemicals of emerging concern (CECs) by wide-scope target screening employing LC-ESI-QTOF-MS. 280 compounds were detected at least in one sample and quantified. Spatial differences in the concentrations and distribution of the compounds classes were discussed. Additionally, samples were analyzed for the possible agonistic/antagonistic potencies using a panel of in vitro transactivation reporter gene CALUX® bioassays including ERα (estrogenics), anti-AR (anti-androgens), GR (glucocorticoids), anti-PR (anti-progestins), PPARα and PPARγ (peroxisome proliferators) and PAH assays. The potency of the wastewater samples to cause oxidative stress and induce xenobiotic metabolism was determined using the Nrf2 and PXR CALUX® bioassays, respectively. The signals from each of the bioassays were compared with the recently developed effect-based trigger values (EBTs) and thus allowed for allocating the wastewater effluents into four categories based on their measured toxicity, proposing a putative action plan for wastewater operators. Moreover, samples were analyzed for antibiotics and 13 antibiotic-resistant genes (ARGs) and one mobile genetic element (intl1) with the aim to assess the potential for antibiotic resistance. All data collected from these various types of analysis were stored in an on-line database and can be viewed via interactive map at https://norman-data.eu/EWW_DANUBE. Keywords: Emerging substances, Wide-scope target screening, Effluent wastewater, Bioassays, Antibiotic resistant genes, Danube River Basin

Almost all licit and illicit drugs consumed by the society end up either unchanged or as a mixture of metabolites in the sewage systems. The analysis of influent wastewater samples and the estimation of drug consumption is the field of wastewater-based epidemiology (WBE). A new trend of WBE is the estimation of the consumption of New Psychoactive Substances (NPS), which are legal replacements of established narcotic and psychotropic drugs with slightly modified chemical structures and similar or new effects. To investigate the occurrence of NPS, 30 composite daily influent wastewater samples from the wastewater treatment plant (WWTP) of Athens (Greece) were collected in a four-year sampling campaign (2015–2018). A generic four-sorbent solid-phase extraction (SPE) sample preparation protocol able to retain compounds with wide physicochemical properties was used. Extracts were analyzed by liquid-chromatography coupled to quadrupole-time-of-flight mass spectrometry (LC-QTOF-MS) using target screening for 278 NPS and suspect screening for 451 NPS. Target screening method was validated for a subset of 49 representative NPS and illicit drugs with similar structures with the NPS. 24 NPS and related compounds were detected by target screening and two compounds were tentatively identified based on mass accuracy, prediction of retention time using in-house QSRR prediction models, isotopic pattern and HRMS/MS fragmentation, whereas the excreted mass loads were also calculated. The results indicated an occasional and low occurrence of NPS in wastewater during the week and over the years, whereas the estimation of the exact sources and the evaluation of the patterns in wastewater were critically discussed.

We apply linear and nonlinear panel models to investigate fundamental pricing laws in the Greek day-ahead electricity market. We link the persistence behaviour and other dynamic properties of electricity price trajectories to an extended array of fundamental variables reflecting market conditions and the price course of energy commodities in global markets. Assuming that the dynamics of the intraday power pricing curve is adequately specified within a panel modelling framework with long memory and common unobserved stochastic effects, we are able to provide consistent estimates of price responsiveness to the main fundamental drivers and thus avoid biases induced by other popular modelling approaches. We also device a panel variant of the standard smooth-transition regression model to investigate regime switching effects in the price responsiveness mechanism. Empirical results suggest that Greek day-ahead electricity market prices respond significantly to hour-specific (load, marginal generation capacity) and global (Brent price, emission allowance rights cost) fundamental variables. In some intraday power delivery zones, these covariates are in a fractional cointegrating relationship with electricity prices being responsible for their drifting behaviour. The strength and type of cointegration changes across intraday trading periods in response to market conditions. The results of the nonlinear model demonstrate the ineffectiveness of linear model designs (either univariate or panel) to accurately estimate the levels of price risk and associate it with changes in the market climate.

Wine metabolomics constitutes a powerful discipline towards wine authenticity assessment through the simultaneous exploration of multiple classes of compounds in the wine matrix. Over the last decades, wines from autochthonous Greek grape varieties have become increasingly popular among wine connoisseurs, attracting great interest for their authentication and chemical characterization. In this work, 46 red wine samples from Agiorgitiko and Xinomavro grape varieties were collected from wineries in two important winemaking regions of Greece during two consecutive vintages and analyzed using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QToF-MS). A targeted metabolomics methodology was developed, including the determination and quantification of 28 phenolic compounds from different classes (hydroxycinnamic acids, hydroxybenzoic acids, stilbenes and flavonoids). Moreover, 86 compounds were detected and tentatively identified via a robust suspect screening workflow using an in-house database of 420 wine related compounds. Supervised chemometric techniques were employed to build an accurate and robust model to discriminate between two varieties.

The aim of this study was to determine lead, mercury, cadmium, arsenic and selenium in muscle and cephalothorax of giant red Mediterranean shrimp (Aristaeomorpha foliacea). Arsenic was also determined in the lipid fraction of the shrimp to provide an estimate of the arsenolipids. The results indicated that Pb and Cd were higher in cephalothorax than in muscle (p

Two liquid chromatographic systems, one coupled to atmospheric pressure chemical ionization and tandem mass spectrometric methods (UHPLC-APCI-MS/MS) and the other a high-performance liquid chromatographic coupled to diode array detector (HPLC-DAD) were used to develop and validate methods for the simultaneous determination of fat-soluble vitamins A, D3 and E in rice cereal baby foods. The chromatographic separation was performed on C18 columns with a mixture of methanol-acetonitrile as mobile phase for all methods. The extraction of fat-soluble vitamins included enzymatic hydrolysis with α-amylase, saponification, extraction with petroleum ether or n-hexane and purification with silica cartridge (only for vitamin D3). Quantification of vitamin D3 and E through UHPLC-APCI-MS/MS was performed by the use of internal standards (IS) D3-d3 and E-d6, respectively, while IS was not used for vitamin A. The methods were optimized and validated in terms of linearity, precision, trueness, limits of detection and quantification. The recoveries were in the range of 85.0–107% for retinol, 92.0–105% for α-tocopherol and 95.2–106% for cholecalciferol and the %RSD (Relative Standard Deviation) values ranged from 6.4% to 15%. The evaluation of the methods was also conducted through the estimation of uncertainties, the application in commercial samples and the participation in a proficiency test

High δ-manganese dioxide’s initial concentration (800 mg/L), temperature (50 °C) and acidic pH (3.0–3.5) were found to be the optimum conditions for the removal of acetaminophen. Additionally, acetaminophen was found to be removed more rapidly at high initial concentrations (70−100 mg/L). The kinetic analysis of the experiments revealed that the two tested kinetic models, the one assuming the electron transfer and the other assuming the complex formation as the rate-limiting step, provided a good fit. The positive value of activation energy estimated by the Arrhenius equation indicated that the reaction is endothermic. The transformation products analysis by means of RPLC-QToF, revealed the formation of acetaminophen’s dimer and trimer in the early stages of the reaction, followed by their complete removal in the latter stages of the reaction. Furthermore, four new compounds were detected. Three isomeric compounds fit the formulas C14H11NO4, attributed to aggregates of 1,4-benzoquinone and N-Acetyl-p-benzoquinone Imine, and one additional transformation product was attributed to C17H8N3O5. The abundance of the isomeric compounds was increasing after 60 min of the reaction, and decreasing during the last stages of the reaction, while the latter’s abundance was increasing during the reaction process.

Optimal siting of wind farms based on a pre-assessment of the spatiotemporal variability of wind resources is considered a suitable method for reducing fluctuations in the delivered output. In this study, we explore the potential for balancing wind energy generation in the Iberian Peninsula using Principal Component Analysis (PCA). This technique permits the discovery of possibly new promising locations for wind power harvesting and an evaluation of the existing wind farm network in terms of reliability in energy generation. Data input to the PCA consists of hourly wind capacity factor in a 5-km spatial resolution grid covering the entire peninsula. These data are derived from an equivalent wind farm power curve fed by modeled wind speed data from 80 m above ground level. PCA reveals three significant balancing patterns prevailing over the IP, where half of the currently operating wind farms in Spain are placed. Hence, among the many constituents of the existing wind farm network, these spots offer the best opportunity for stable power supply. The paper concludes by making proposals on an optimum wind capacity allocation based on the idea of equally distributing installed power between positive/negative dipoles emerging from balancing principal components.

Citalopram (CTR) is a worldwide highly consumed antidepressant which has demonstrated incomplete removal by conventional wastewater treatment. Despite its global ubiquitous presence in different environmental compartments, little is known about its behaviour and transformation processes during wastewater treatment. The present study aims to expand the knowledge on fate and transformation of CTR during the biological treatment process. For this purpose, batch reactors were set up to assess biotic, abiotic and sorption losses of this compound. One of the main objectives of the study was the identification of the formed transformation products (TPs) by applying suspect and non-target strategies based on liquid chromatography quadrupole-time-of-flight mass spectrometry (LC-QTOF-MS). The complementary use of reversed phase liquid chromatography (RPLC) and hydrophilic interaction liquid chromatography (HILIC) for the identification of polar TPs, and the application of in-house developed quantitative structure-retention relationship (QSRR) prediction models, in addition to the comprehensive evaluation of the obtained MS/MS spectra, provided valuable information to support identification. In total, fourteen TPs were detected and thirteen of them were tentatively identified. Four compounds were confirmed (N-desmethylCTR, CTR amide, CTR carboxylic acid and 3-oxo-CTR) through the purchase of the corresponding reference standard. Probable structures based on diagnostic evidence were proposed for the additional nine TPs. Eleven TPs are reported for the first time. A transformation pathway for the biotransformation of CTR was proposed. The presence of the identified TPs was assessed in real wastewater samples through retrospective analysis, resulting in the detection of five compounds. Finally, the potential ecotoxicological risk posed by CTR and its TPs to different trophic levels of aquatic organisms was evaluated by means of risk quotients.

The economic crisis plaguing Greece was expected to impact consumption of pharmaceuticals and illicit drugs - a priori to an unknown extent. We quantified the change of use for various classes of licit and illicit drugs by monitoring Athens' wastewater from 2010 to 2014. A high increase in the use of psychoactive drugs was detected between 2010 and 2014, especially for antipsychotics (35-fold), benzodiazepines (19-fold), and antidepressants (11-fold). This directly reflects the perceived increase of incidences associated with mental illnesses in the population, as a consequence of severe socioeconomic changes. Other therapeutic classes, like antiepileptics, hypertensives, and gastric and ulcer drugs also showed an increase in use (from 2-fold increase for antiepileptics to 13-fold for hypertensives). In contrast, the overall use of antibiotics and NSAIDs decreased. For mefenamic acid, an almost 28-fold decrease was observed. This finding is likely related to the reduction in drug expenditure applied in public health. A 2-fold increase of methamphetamine use was detected, associated with a cheap street drug called ″sisa″ (related to marginal conducts), which is a health concern. MDMA (5-fold) and methadone (7-fold) use showed also an increase, while cocaine and cannabis estimates did not show a clear trend.

The occurrence of chemical and biological contaminants of emerging concern (CECs) was investigated in treated wastewater intended for reuse in agriculture. An agarose hydrogel diffusion-based passive sampler was exposed to the outlet of a wastewater treatment plant (WWTP) located in Cyprus, which is equipped with membrane bioreactor (MBR). Passive samplers in triplicate were exposed according to a time-series exposure plan with maximum exposure duration of 28 days. Composite flow-proportional wastewater samples were collected in parallel with the passive sampling exposure plan and were processed by solid phase extraction using HORIZON SPE-DEX 4790 and the same sorbent material (Oasis HLB) as in the passive sampler. The analysis of passive samplers and wastewater samples enabled (i) the field-scale calibration of the passive sampler prototype by the calculation of in situ sampling rates of target substances, and (ii) the investigation of in silico predicted transformation products of the four most ecotoxicologically hazardous antibiotics (azithromycin, clarithromycin, erythromycin, ofloxacin). Additionally, the wastewater samples were subjected to the analysis of seven preselected antibiotic resistant genes (ARGs) and one mobile resistant element (int1). All extracts were analyzed for chemicals in a single batch using a highly sensitive method for pharmaceuticals, antibiotics and illicit drugs by liquid chromatography tandem MS/MS (LC-QQQ) and for various other target compounds (2316 compounds in total) by liquid chromatography high-resolution mass spectrometry (LC-HRMS). 279 CECs and all investigated ARGs (except for blaCTX-M−32) were detected, highlighting potential chemical and biological hazards related to wastewater reuse practices. 16 CECs were prioritized following ecotoxicological risk assessment, whereas sul1 and the mobile resistant element (int1) showed the highest abundance. Comprehensive monitoring efforts using novel sampling methods such as passive sampling, wide-scope target screening and molecular analysis are required to assure safe application of wastewater reuse and avoid spread and crop uptake of potentially hazardous chemicals. Keywords: Hydrogel-based passive sampler, Wastewater reuse, Contaminants of emerging concern, Transformation products, Antibiotics, Antibiotic resistance genes

The ready biodegradability of twenty food additives, belonging to the classes of artificial sweeteners, natural sweeteners, preservatives and colorings, was investigated using activated sludge as inoculum and OECD 301F respirometric test. According to the results, saccharin, aspartame, sodium cyclamate, xylitol, erythritol, maltitol, potassium sorbate, benzoic acid and sodium ascorbate are characterized as readily biodegradable compounds, partial biodegradation (60% during the test) was noticed for steviol, inulin, alitame, curcumin, ponceau 4R and tartrazine, while no biodegradation was observed for the other five compounds. The duration of lag phase before the start of biodegradation varied between the target compounds, while their ultimate biodegradation half-life values ranged between 0.7 ± 0.1 days (benzoic acid) and 24.6 ± 1.0 days (curcumin). The expected removal of target compounds due to ultimate biodegradation mechanism was estimated for a biological wastewater treatment system operated at a retention time of one day and percentages higher than 40% were calculated for sodium cyclamate, potassium sorbate and benzoic acid. Higher removal percentages are expected in full-scale Sewage Treatment Plants (STPs) due to the contribution of other mechanisms such as sorption to suspended solids, (bio)transformation and co-metabolic phenomena. Further biodegradation experiments should be conducted under different experimental conditions for the food additives that did not fulfill the requirements of the applied protocol. Future studies should also focus on the occurrence and fate of food colorants and natural sweeteners in full-scale STPs.

Bisphenol A (BPA), a known potential endocrine disrupting compound (EDC) is expected to be present in low quantities in canned food due to its migration from the inner surface coating of cans made of epoxy resins. A selective and confirmatory analytical method, based on microwave assisted extraction (MAE), molecularly imprinted solid phase extraction (MISPE) using a polymer prepared by a non-covalent molecular imprinting technique and liquid chromatography coupled with electrospray ionization mass spectrometry (LC-ESI/MS) was developed for the determination of BPA in canned pineapple, tuna and mushrooms. First, the effect of the loading medium of hydro- organic solutions on the binding of BPA and its deuterated analogue on the MISPE sorbent was investigated. Subsequently, the effects of the experimental conditions of the microwave assisted extraction (solvent, sample mass/solvent volume, time and temperature) on the obtained recovery of BPA from canned food were assessed and the parameters were optimized to provide maximum recovery and selectivity. It was demonstrated that the combination of MAE with MISPE permits the use of a selective extraction solvent (methanol/water, 4/6, v/v), simplifying the sample preparation steps and enhancing sample clean-up of complex food matrices. The method was validated in different food matrices, using BPA-d16 as internal standard and quantitative relative recoveries were determined. The precision (RSD %) of the method ranged from 7% to 10% and the limit of detection was at low ng/g level for all food matrices. The determined concentration of BPA in commercial canned samples ranged between 7.3 and 42.3 ng/g.