Assessment of the analytical methods used for the determination of potential endocrine disruptors and their main metabolites in sediments, natural waters and several food commodities: chemometrics application

This PhD study was primarily focused on the validation, optimization and application of three different preconcentration methods for the simultaneous determination of the target Endocrine Disrupting Compounds, linuron and diuron as well as their common metabolites (DCPMU, DCPU and 3, 4-DCA) in various natural waters, sediments as well as several food commodities. Whenever possible, chemometric methods such as experimental designs - both fractional factorial and fully nested- in combination with artificial neural networks were applied and/or compared to classical approaches. A high-performance liquid chromatography system coupled to UV-diode array detector (HPLC/UV-DAD) was used for the target analytes quantification. Generally, the study comprises of three parts each one dealing respectively with the analysis of sediments, natural waters and selected food commodities. In the first part the validation of an analytical method based on ultrasonic extraction of the target analytes from various freshwater sediments is presented. In the second part, a solid phase extraction (SPE) preconcentration method for the simultaneous determination of the target analytes present in natural waters was optimized and validated. The influence of several experimental variables affecting the extraction efficiency of the target analytes was studied in order to maximize extraction efficiency, including the sorbent material, elution solvents, pH and breakthrough volume, as well as some solution parameters that is, ionic strength and organic matter content. Then, matrix solid-phase dispersion (MSPD) as a sample preparation method for the determination of the two potential EDCs and their metabolites in food commodities was developed. The influence of the main factors on the extraction process yield was thoroughly evaluated. For that purpose, both classical approach and the multivariable one using a 3(4-1) fractional factorial design in further combination with artificial neural networks (ANNs) were employed. A fully nested experimental design was applied to study the measurement uncertainty arising from trueness as well as to examine the usability of the three analytical methods used for different sediments, water types and food commodities. In order to verify the real applicability of the proposed procedure and to assess the contamination extend, representative market baskets were sampled in 10 European countries in 3 sampling campaigns (Jan-Feb, Apr-May, Sept-Oct 2003). It was ensured for the samples to represent different geographical areas and preferred nutrition modes of their inhabitants. Finally a risk assessment originating from food consumption was attempted. Despite the violations of MRLs, in no case linuron residues exceeded the ADIs. However, the latter refer to single substance intake whereas aliment contains a complex mixture of chemical residues. Furthermore, current safety regulation for consumer health protection does not consider endocrine effects.Η εκπόνηση της παρούσας Διδακτορικής Διατριβής πραγματεύεται τη βελτιστοποίηση και εφαρμογή αναλυτικών μεθόδων για τον προσδιορισμό δύο επιλεγμένων φυτοφαρμάκων καθώς και των κοινών τους μεταβολιτών σε διάφορους τύπους υδάτων, σε ιζήματα ποικίλης προέλευσης και σε επιλεγμένα φρούτα και λαχανικά. Το ενδιαφέρον της εστιάζεται επιπλέον στην υπολειμματικότητα των παραπάνω ενώσεων σε ορισμένα είδη τροφίμων που συμπεριλαμβάνονται στο κοινώς λεγόμενο "καλάθι της νοικοκυράς". Η χρήση χημειομετρικών μεθόδων διαπερνά όλο τον κορμό της ερευνητικής εργασίας σε μια προσπάθεια εφαρμογής τους, τόσο για τη βελτιστοποίηση των αναλυτικών μεθόδων όσο και για την εκτίμηση του βαθμού της - προερχόμενης από την ανάκτηση-αβεβαιότητας αυτών. Έτσι γίνεται χρήση δύο διαφορετικών πειραματικών σχεδιασμών και επιπλέον επιχειρείται συνδυασμός με υπολογιστικά μοντέλα που ανήκουν στον κλάδο της Τεχνητής Νοημοσύνης, τα Τεχνητά Νευρωνικά Δίκτυα. Επιπλέον, γίνεται προσπάθεια αξιολόγησης του κινδύνου των καταναλωτών των επιλεγμένων τροφίμων λόγω της υπολειμματικότητας των δύο φυτοφαρμάκων και των μεταβολιτών τους. Επιλέχθηκαν τα ευρέως χρησιμοποιούμενα σε γεωργικές καλλιέργειες ζιζανιοκτόνα linuron και diuron ως ύποπτες ενώσεις για ενδοκρινική διαταρακτική δράση. Η συνεκτίμηση των υπολειμμάτων τριών κοινών μεταβολιτών τους (DCPMU, DCPU και 3,4-DCA) κρίθηκε επίσης απαραίτητη αφού σε μερικές περιπτώσεις τα υπολείμματα των προϊόντων διάσπασης μπορεί να υπερβαίνουν αυτά των αρχικών ενώσεων. Τρεις αναλυτικές μέθοδοι επιλέχθηκαν να μελετηθούν. Η εκχύλιση με τη χρήση υπερήχων για τα ιζήματα, η εκχύλιση δια της στερεάς φάσης (SPE) για τα φυσικά νερά και η μέθοδος εκχύλισης με διασπορά του υποστρώματος στη στερεά φάση (MSPD) για τα τρόφιμα. Σκοπό της παρούσας ερευνητικής εργασίας αποτέλεσε και ο έλεγχος των κυριότερων παραμέτρων που είναι γνωστό ότι επηρεάζουν την απόδοση τόσο της μεθόδου SPE όσο και της MSPD ώστε να προσδιοριστούν οι βέλτιστες συνθήκες εκχύλισης για το μίγμα των δύο EDCs και των μεταβολιτών τους σε κάθε περίπτωση. Περαιτέρω, η βελτιστοποίηση της μεθόδου MSPD έγινε εκτός της κλασσικής (ένας παράγοντας τη φορά) προσέγγισης με τη χρήση πειραματικού σχεδιασμού σε συνδυασμό με τη χρήση μοντέλων όπως τα τεχνητά νευρωνικά δίκτυα με στόχο τη σύγκριση των δύο μεθόδων αλλά και την ουσιαστικότερη χαρτογράφηση και τελικά προσέγγιση των βέλτιστων συνθηκών. Επιπλέον έγινε μια προσπάθεια προσδιορισμού της προερχόμενης από την ανάκτηση αβεβαιότητας των τριών μεθόδων εκχύλισης που χρησιμοποιήθηκαν. Για την επίτευξη του στόχου χρησιμοποιήθηκε ένα πλήρως ένθετο πειραματικό σχέδιο. Τέλος, η μέθοδος MSPD εφαρμόστηκε για τον προσδιορισμό υπολειμμάτων των επιλεγμένων ενώσεων και των μεταβολιτών τους σε δείγματα φρούτων και λαχανικών αντιπροσωπευτικών του «καλαθιού της νοικοκυράς» που περιλάμβαναν μήλα, καρότα, πατάτες, σιτάλευρα και χυμούς πορτοκαλιών. Ο τελικός στόχος ήταν η εκτίμηση της επικινδυνότητας από την ημερήσια πρόσληψη των παραπάνω τροφίμων για τους καταναλωτές

Screening of Commonly Used Antibiotics in Fresh and Saltwater Samples Impacted by Aquacultures: Analytical Methodology, Occurrence and Environmental Risk Assessment

Traditionally, antibiotics have been used to treat human and animal diseases caused by pathogenic bacteria. The aquaculture industry, which is massively expanding currently, also makes use of several antibiotic classes, resulting in potential antibiotic residues in the surrounding aquatic environment, as well as the cultured products raising bacterial resistance. The aim of this study was the optimization, validation, and application of a solid-phase extraction (SPE) method in combination with liquid chromatography (LC)-LTQ/Orbitrap mass spectrometry in order to determine the most commonly used antibiotics in waters sampled from fish farms, both saltwater and freshwater, located in Greece. Under optimum conditions, the method was validated, achieving recoveries in the range of 57.7% (for sulfamethoxazole in river water) to 95.8% (for florfenicol in river water). The method quantification limits were within the range of 0.25 and 10 ng·L−1 in all cases, with relative standard deviations (RSDs) −6 < RQ < 0.496) or the mixture RQ (0.005 < RQ < 0.682), proving that in all cases, the risk was medium to low

Photocatalytic Degradation of Psychiatric Pharmaceuticals in Hospital WWTP Secondary Effluents Using g-C₃N₄ and g-C₃N₄/MoS₂ Catalysts in Laboratory-Scale Pilot

Today, the pollution caused by a multitude of pharmaceuticals used by humans has been recognized as a major environmental problem. The objective of this study was to evaluate and compare the photocatalytic degradation of ten target psychiatric drugs in hospital wastewater effluents using g-C3N4 and 1%MoS2/g-C3N4 (1MSCN) as photocatalytic materials. The experiments were performed using real wastewater samples collected from hospital wastewater treatment plant (WWTP) secondary effluent in spiked and inherent pharmaceutical concentration levels. The photocatalytic experiments were performed in a laboratory-scale pilot plant composed of a stainless-steel lamp reactor (46 L) equipped with ten UVA lamps and quartz filters connected in series with a polypropylene recirculation tank (55–100 L). In addition, experiments were carried out in a solar simulator apparatus Atlas Suntest XLS+ at a 500 Wm−2 irradiation intensity. The analysis of the samples was accomplished by solid-phase extraction, followed by liquid chromatography-Orbitrap high-resolution mass spectrometry. Results showed that the photocatalytic degradation of pharmaceutical compounds followed first-order kinetics. In all cases, 1MSCN presented higher photocatalytic performance than g-C3N4. The removal rates of the pharmaceutical compounds were determined above 30% and 54% using g-C3N4 and 1MSCN, respectively. Parallel to kinetic studies, the transformation products (TPs) generated during the treatment were investigated

Enrichment of Olive Oils with Natural Bioactive Compounds from Aromatic and Medicinal Herbs: Phytochemical Analysis and Antioxidant Potential

Olive oil and herbs, two key components of the Mediterranean diet, are known for their beneficial effects on humans. In our study, we incorporated aromatic and medicinal herbs into local monovarietal olive oils via maceration procedures for enrichment. We identified the herbal-derived ingredients that migrate to olive oils and contribute positively to their total phenolic content and functional properties, such as radical scavenging activity. Thus, we characterized the essential oil composition of the aromatic herbs (GC-MS), and we determined the phenolic content and antioxidant capacity of the additives and the virgin olive oils before and after enrichment. The herbal phenolic compounds were analyzed by LC-LTQ/Orbitrap HRMS. We found that olive oils infused with Origanum vulgare ssp. hirtum, Rosmarinus officinalis and Salvia triloba obtained an increased phenolic content, by approximately 1.3 to 3.4 times, in comparison with the untreated ones. Infusion with S. triloba led to a significantly higher antioxidant capacity. Rosmarinic acid, as well as phenolic glucosides, identified in the aromatic herbs, were not incorporated into olive oils due to their high polarity. In contrast, phenolic aglycones and diterpenes from R. officinalis and S. triloba migrated to the enriched olive oils, leading to a significant increase in their phenolic content and to an improvement in their free radical scavenging capacity

Exploring the Efficiency of UHPLC-Orbitrap MS for the Determination of 20 Pharmaceuticals and Acesulfame K in Hospital and Urban Wastewaters with the Aid of FPSE

Aside from the classical residues of persistent organic pollutants (POPs), the occurrence of emerging contaminants (ECs) in the environment has become a subject of increasing concern due to their harmful impact on the aquatic environment. Wastewater treatment plant (WWTP) effluents are major sources of environmental pollution. Therefore, data concerning their existence is required. In this study, twenty compounds representative of different drug groups considered ECs and belonging to antibiotics, antipsychotics, anti-inflammatory drugs plus acesulfame K were selected to be accurately detected and quantified with UHPLC&ndash;LTQ-Orbitrap MS in hospital and urban WWTP effluents. Chromatographic parameters (column efficiency, mobile phase, etc.), as well as mass spectrometry conditions concerning ionization mode and Orbitrap analysis (ESI options, mass resolving power, AGC target, tube lens, injection time), were evaluated. Moreover, a novel fabric phase sorptive extraction (FPSE) method based on fiber glass coated with PEG300 was employed as sample preparation process. Experimental parameters affecting extraction and desorption steps such as sample pH, extraction time, ionic strength, elution time and solvent have been optimized. The optimized methodology was validated providing excellent linearity (R2 &gt; 0.99), and low detection and quantification limits up to 3.1 and 9.3 ng/L, for carbamazepine, respectively. Relative recoveries ranged from 81.1% to 114.0%, while a medium matrix effect for most of the target compounds occurred. Applying the above analytical method in effluents of WWTPs from NW Greece, nine compounds were quantified with concentrations that varied from 55.4 to 728.4 ng/L

Environmental risks associated with contaminants of legacy and emerging concern at European aquaculture areas

The contamination of marine ecosystems by contaminants of emerging concern such as personal care products or per- and polyfluoroalkyl substances is of increasing concern. This work assessed the concentrations of selected contaminants of emerging concern in water and sediment of European aquaculture areas, to evaluate their co-variation with legacy contaminants (polycyclic aromatic hydrocarbons) and faecal biomarkers, and estimate the risks associated with their occurrence. The 9 study sites were selected in 7 European countries to be representative of the aquaculture activities of their region: 4 sites in the Atlantic Ocean and 5 in the Mediterranean Sea. Musks, UV filters, preservatives, per- and polyfluoroalkyl substances and polycyclic aromatic hydrocarbons were detected in at least one of the sites with regional differences. While personal care products appear to be the main component of the water contamination, polycyclic aromatic hydrocarbons were mostly found in sediments. As expected, generally higher levels of personal care products were found in sewage impacted sites, urbanised coasts and estuaries. The risk assessment for water and sediment revealed a potential risk for the local aquatic environment from contaminants of both legacy and emerging concern, with a significant contribution of the UV filter octocrylene. Despite marginal contributions of per- and polyfluoroalkyl substances to the total concentrations, PFOS (perfluorooctane sulfonate) aqueous concentrations combined to its low ecotoxicity thresholds produced significant hazard quotients indicating a potential risk to the ecosystems

Persistent and emerging pollutants assessment on aquaculture oysters (Crassostrea gigas) from NW Portuguese coast (Ria De Aveiro)

The study aim was to determine a range of relevant persistent and emerging pollutants in oysters produced in an aquaculture facility located in an important production area, to assure their safety for human consumption. Pollutants, including 16 PAHs, 3 butyltins (BTs), 29 flame retardants (FRs, including organophosphate and halogenated FRs), 35 pesticides (including 9 pyrethroid insecticides) and 13 personal care products (PCPs, including musks and UV filters), were determined in oysters' tissues collected during one year in four seasonal sampling surveys. The seasonal environmental pollution on the production site was evaluated by water and sediment analysis. Furthermore, oysters' nutritional quality was also assessed and related with the consumption of healthy seafood, showing that oysters are a rich source of protein with low fat content and with a high quality index all year around. Results showed that most analysed pollutants were not detected either in oyster tissues or in environmental matrixes (water and sediments). The few pollutants detected in oyster tissues, including both regulated and non-legislated pollutants, such as a few PAHs (fluorene, phenanthrene, anthracene, fluoranthene, pyrene and indenopyrene), FRs (TPPO, TDCPP, DCP, BDE-47, BDE-209 and Dec 602) and PCPs (galaxolide, galaxolidone, homosalate and octocrylene), were present at low levels (in the ng/g dw range) and did not represent a significant health risk to humans. The observed seasonal variations related to human activities (e.g. tourism in summer) highlights the need for environmental protection and sustainable resource exploration for safe seafood production. © 2019 Elsevier B.V.This work was supported by the Structured R&D&I Project INNOVMAR – “Innovation and Sustainability in the Management and Exploitation of Marine Resources” (ref. NORTE-01-0145-FEDER-000035) within the research line “INSEAFOOD - Innovation and valorization of seafood products: meeting local challenges and opportunities”, founded by the Northern Regional Operational Programme (NORTE2020) through the European Regional Development Fund (ERDF). This study was also supported by the SEA-on-a-CHIP project, funded from European Union Seventh Framework Programme (FP7-OCEAN-2013) under grant agreement No. 614168. This work has been also financially supported by the Generalitat de Catalunya (Consolidated Research Group 2017 SGR 01404 – Water and Soil Quality Unit). Biotage is acknowledged for providing SPE cartridges. Authors acknowledge Carlos R. Gomes for helping on multivariated analysis.Peer reviewe