Determination of emerging pollutants in sewage sludge with LC-QTOF-MS

Οι σύγχρονες υγειονομικές διαδικασίες έχουν ως αποτέλεσμα μεγάλους όγκους ανθρώπινων αποβλήτων, καθώς και αστικών και βιομηχανικών αποβλήτων, τα οποία συλλέγονται και επεξεργάζονται σε κοινά σημεία συλλογής, τα Κέντρα Επεξεργασίας Λυμάτων. Ένα μεγάλο εύρος οργανικών ενώσεων χρησιμοποιείται στις σύγχρονες κοινωνίες οι οποίες καταλήγουν στα ΚΕΛ και ως συνέπεια στα υγρά απόβλητα και ενεργό ιλύ. Για αυτό το λόγο, είναι κρίσιμο να αξιολογηθεί η παρουσία και η επικινδυνότητα των αναδυόμενων ρύπων σε τέτοιου είδους δείγματα. Σε αυτή την διπλωματική εργασία περιγράφεται η στοχευμένη σάρωση αναδυόμενων ρύπων σε δείγματα ενεργού ιλύος, τα οποία συλλέχθηκαν το 2015 από τρία ΚΕΛ από την Νιγηρία (Iponri, Alausa, Ijaiye). Χρησιμοποιήθηκε εκχύλιση με τη βοήθεια υπερήχων για την προκατεργασία δείγματος και τα εκχυλίσματα αναλύθηκαν με υγροχρωματογραφία αντίστροφης φάσης συζευγμένη με φαρματομετρία μάζας τετραπόλου-χρόνου πτήσης (RP)LC-QTOF-MS και η λήψη των δεδομένων πραγματοποιήθηκε με τη λειτουργία bbCID η οποία παρείχε πληροφορίες για τα πρόδρομα ιόντα και τα θραύσματα, χωρίς προεπιλογή των αναλυτών και με μία ανάλυση. Πραγματοποιήθηκε επικύρωση χρησιμοποιώντας αντιπροσωπευτικές ενώσεις της εφαρμοζόμενης ευρείας μεθόδου σάρωσης. Μια βάση δεδομένων περίπου 2500 ενώσεων χρησιμοποιήθηκε για τη στοχευμένη ανάλυση και η ανίχνευση βασίστηκε στο χρόνο ανάκτησης, στην ακρίβεια μάζας, στο ισοτοπικό προφίλ και στα προϊόντα θραυσματοποίησης. Σύμφωνα με τις προαναφερθείσες διαδικασίες, 182 ενώσεις ταυτοποιήθηκαν συμπεριλαμβανομένων φαρμακευτικών ενώσεων, φυτοπροστατευτικών φαρμάκων, ναρκωτικά, χημικά που χρησιμοποιούνται στη βιομηχανία και άλλες κλάσεις ενώσεων. Η στοχευμένη ανάλυση αποκάλυψε το προφίλ για κάθε ΚΕΛ και τις διαφορές μεταξύ των αναλυτών μετά τα στάδια της χώνευσης και της δευτερογενούς επεξεργασίας.Modern sanitary processes result in large volumes of human waste, as well as domestic and industrial sewage, being collected and treated at common collection points, wastewater treatment plants. A wide range of organic compounds is used in modern societies which end up to the WWTPs and consequently to the liquid effluent and sewage sludge. For this reason, it is critical to evaluate the presence and risk of emerging pollutants in such samples. This thesis reports the target screening of emerging pollutants in sewage sludge samples collected in 2015 from three WWTPs from Nigeria (Iponri, Alausa, Ijaiye). An ultrasound assisted extraction was used for the sample preparation and the exracts were analyzed with reversed-phase liquid-chromatography coupled to quadrupole-time-of-flight mass spectrometry (RP)LC-QTOF-MS and the data were acquired through broad-band Collision Induced Dissociation (bbCID) mode, which provided information on parent and fragment ions without pre-selection of analytes in one run. Validation was performed based on representative compounds of the wide-scope screening method. A database of approximately 2500 compounds was used for target screening and the detection was based on retention time, mass accuracy, isotopic pattern and fragmentation products. Following the aforementioned procedures, 182 compounds were identified including pharmaceutical drugs, pesticides, illicit drugs, industrial chemicals and other classes of compounds. The target screening revealed the profile of each WWTP and the differences between the analytes after the stage of digestion and the secondary treatment

Ανάπτυξη μεθόδων για την προχωρημένη παρακολούθηση ρύπων προτεραιότητας και αναδυόμενων ρύπων σε περιβαλλοντικά δείγματα με μη στοχευμένη σάρωση με LC/GC-QTOF-MS και χημειομετρικές τεχνικές

The thesis is consisted of five studies. The first two studies (Chapters 3 and 4) are about the occurrence of ECs in aquatic environment and development of target, suspect and non-target screening strategies to create list of chemicals for regulatory monitoring. In addition, following up the second study, Chapter 5 provides information about how chemicals can be prioritized for NTS based identification by looking into their exposure pattern in the daily based environmental sampling. Chapter 6 discusses about the development of strategy for q-NTS in GC-HRMS instrument and its application in environmental samples. The last chapter discusses about the use of marine sponges as potential source for bio-magnification studies of ECs. Here, a brief summary of these studies is provided: In the first study, we evaluated possible occurrence of ECs in drinking water reservoirs by means of wide-scope target, suspect and non-target screening. This is the first comprehensive evaluation of quality of drinking water in Attica, Greece. In the second study, occurrence of ECs in Asopos river in northern region of Attica was evaluated. This was done in a daily sampling base in four different seasons. This was an important study to understand how industrial and agricultural activities affect this river. In the third study, a new prioritization method based on exposure pattern of chemicals in Asopos river was developed for NTS screening. Due to human activity throughout the week, chemicals are released into the river in distinct pattern and this chapter discusses the development of a machine learning based method which learns from this pattern to look for unknowns that show distinguishable exposure pattern. The fourth part of the thesis was devoted to the development of a robust technique to understand how chemicals ionize in APCI source in order to create a semi-quantitative workflow for analysis of ECs in GC-HRMS. This is a very important study fixing analytical gap between detection of a compound via GC-HRMS and its quantification in case the reference standard is not available. Finally, the last study was to propose marine sponge as a new frontier for evaluation of ECs in marine environment. To this end, samples from sediment, seawater, sponge and its porous water were gathered (to understand spatial distribution of ECs in surrounding environment of sponge) mainly from Ionian Sea. Wide-scope HRMS based screening workflows were used based on LC/GC-ESI/APCI-QTOF-MS to discover how chemicals accumulate in different marine sponges' species.Η διατριβή αποτελείται από πέντε μελέτες. Οι δύο πρώτες μελέτες (Κεφάλαια 3 και 4) αφορούν την εμφάνιση αναδυόμενων ρύπων στο υδάτινο περιβάλλον και την ανάπτυξη στοχευμένων και μη-στοχευμένων μεθόδων με σκοπό τη δημιουργία λίστας χημικών ουσιών για μελλοντική παρακολούθηση. Σε συνέχεια της δεύτερης μελέτης, το Κεφάλαιο 5 παρέχει πληροφορίες σχετικά με τον τρόπο με τον οποίο οι χημικές ουσίες μπορούν να τεθούν σε προτεραιότητα για ταυτοποίηση βάσει NTS (μη-στοχευμένης ανάλυσης), εξετάζοντας την εμφάνισή τους σε δειγματοληψία συνεχόμενων ημερών. Στο Κεφάλαιο 6 συζητείται η ανάπτυξη στρατηγικής για q-NTS (ποσοτική μη-στοχευμένη ανάλυση) χρησιμοποιώντας το όργανο GC-HRMS και η εφαρμογή της σε περιβαλλοντικά δείγματα. Το τελευταίο κεφάλαιο εξετάζει τη χρήση θαλάσσιων σπόγγων ως πιθανή λύση για μελέτες βιο-μεγέθυνσης των αναδυόμενων ρύπων. Εδώ, παρέχεται μια σύντομη περίληψη αυτών των μελετών: Στην πρώτη μελέτη, αξιολογήσαμε την πιθανή εμφάνιση αναδυόμενων ρύπων σε δεξαμενές πόσιμου νερού μέσω στοχευμένης και μη-στοχευμένης ανάλυσης. Αυτή είναι η πρώτη ολοκληρωμένη αξιολόγηση της ποιότητας, όσον αφορά τους αναδυόμενους ρύπος, του πόσιμου νερού στην Αττική, Ελλάδα. Στη δεύτερη μελέτη, αξιολογήθηκε η εμφάνιση αναδυόμενων ρύπων στον ποταμό Ασωπό στη βόρεια περιοχή της Αττικής. Πραγματοποιήθηκαν δειγματοληψίες σε καθημερινή βάση, για τέσσερις διαφορετικές εποχές. Αυτή ήταν μια σημαντική μελέτη για να κατανοήσουμε πώς οι βιομηχανικές και γεωργικές δραστηριότητες επηρεάζουν την ποιότητα των νερών του ποταμού. Στην τρίτη μελέτη, αναπτύχθηκε μια νέα μέθοδος ιεράρχησης με βάση το μοτίβο εμφάνισης χημικών ουσιών στον ποταμό Ασωπό για την ταυτοποίηση μέσω NTS. Λόγω της ανθρώπινης δραστηριότητας καθ' όλη τη διάρκεια της εβδομάδας, χημικές ουσίες απελευθερώνονται στον ποταμό με ξεχωριστό μοτίβο και αυτό το κεφάλαιο συζητά την ανάπτυξη της μεθόδου που βασίζεται στη μηχανική μάθηση, η οποία μαθαίνει από αυτό το μοτίβο και να αναζητά άγνωστες ουσίες που δείχνουν διακριτό μοτίβο εμφάνισης. Το τέταρτο μέρος της διατριβής αφιερώθηκε στην ανάπτυξη μιας ισχυρής τεχνικής για την κατανόηση του τρόπου με τον οποίο ιονίζονται τα χημικά στην πηγή APCI προκειμένου να δημιουργηθεί μια ημι-ποσοτική πορεία εργασίας για την ανάλυση των αναδυόμενων ρύπων στο GC-HRMS. Αυτή είναι μια πολύ σημαντική μελέτη που καθορίζει το αναλυτικό χάσμα μεταξύ της ανίχνευσης μιας ένωσης μέσω GC-HRMS και της ποσοτικοποίησής της σε περίπτωση που το πρότυπο αναφοράς δεν είναι διαθέσιμο.Τέλος, η τελευταία μελέτη προτείνει το θαλάσσιο σπόγγο ως νέο βιοδείκτη για την αξιολόγηση των αναδυόμενων ρύπων στο θαλάσσιο περιβάλλον. Για το σκοπό αυτό, συλλέχθηκαν δείγματα από ίζημα, θαλασσινό νερό, σπόγγο και το νερό των πόρων του (για την κατανόηση της χωρικής κατανομής των αναδυόμενων ρύπων στο περιβάλλον του σφουγγαριού) κυρίως από το Ιόνιο Πέλαγος. Πορείες εργασιών ανάλυσης ευρείας εμβέλειας χρησιμοποιήθηκαν για τις αναλύσεις με βάση LC/GC-ESI/APCI-QTOF-MS για να ανακαλύψουν πώς συσσωρεύονται χημικές ουσίες σε διαφορετικά είδη θαλάσσιων σπόγγων

The Correlation between the Temperature, Precipitation, and Referrals for Physical Therapy for Patients with Chronic Musculoskeletal Problems

The aim of this study was to investigate how temperature and precipitation are associated with the number of referrals for physical therapy. Material and Methods: Weather data and monthly referrals were collected retrospectively for three years (2020–2022). A total of 937 referrals were studied. Trend analysis was conducted to identify patterns and trends in data over time, including correlation analysis to discover relationships and multivariate linear regression analyses to model the direction and strength of this. Results: Monthly referrals were found to be strongly associated with weather indicators (rs = −0.744, p s = 0.894, p p p p p < 0.001). Conclusion: Low temperatures and high precipitation rates were found to be associated with an increased number of referrals. Females were more vulnerable to precipitation increases

Can Exercise Affect the Pain Characteristics in Patients with Fibromyalgia? A Randomized Controlled Trial

Exercise is often recommended for fibromyalgia. The aim of this study was to investigate the possible influence and change in the pain characteristics of fibromyalgia patients when breathing exercises were added to their exercise program. A total of 106 patients were included and randomly divided into two groups. &Tau;he first group of patients followed a program of active exercises up to the limits of pain, lasting 30 min with a repetition of two times a week. Patients of the second group followed the same program with the addition of diaphragmatic breaths when they reached the pain limit. The patients completed three questionnaires: the Fibromyalgia Rapid Screening Tool (FiRST), the Brief Pain Inventory (BPI), and the Pain Quality Assessment Scale (PQAS)&mdash;once at the beginning, once again after three weeks of exercise, and again 3 months since the beginning of the program. Independent t-tests for the mean total change scores in pain scales demonstrated that for the second group there was a greater improvement in all pain scales, except for the PQAS Deep Pain subscale (p = 0.38). In conclusion, both groups showed significant improvement in all characteristics of the pain scales; however, the improvement of the second group was significantly higher

Screening of legacy and emerging substances in surface water, sediment, biota and groundwater samples collected in the Siverskyi Donets River Basin employing wide-scope target and suspect screening

Siverskyi Donets is the fourth longest river in Ukraine and its ecosystem is heavily affected by numerous agricultural and industrial activities. An impact of the on-going armed military conflicts in the Eastern Ukraine to the overall pollution by the chemicals has been studied. Considering the uncontrolled activities in the catchment due to the conflict, there is a high demand to assess the contamination status of the Siverskyi Donets basin. In this study, the occurrence of the EU Water Framework Directive priority substances, selected physicochemical parameters and wide-range emerging contaminants were investigated in surface water, groundwater, biota and river sediments samples from 13 sampling sites in the river basin. The study included metals, inorganic, non-polar and polar organic contaminants. The wide-scope target screening of 2316 substances and suspect screening of 2219 substances revealed occurrence of 83 compounds in the studied samples. A few industrial chemicals such as plasticizers bisphenol A and DEHP, as well as flame retardant brominated diphenylethers were found to be potentially hazardous to the ecosystem, exceeding the established legacy environmental quality standards (EQS) or the provisional no-effect concentration (PNEC) values. River sediment samples contained traces of long-term banned chemicals such as polychlorinated biphenyls (PCBs) and degradation products of DDT (p,p&apos;-DDD and p,p&apos;-DDE). A simplified risk assessment based on comparison of measured concentration of the detected compounds against their (eco)toxicity threshold values from the NORMAN Ecotoxicology Database has been performed to aid their prioritization in future monitoring and, eventually, establishing the list of Siverskyi Donets River Basin Specific Pollutants. A comparison with the recent similar studies in the Dniester and Dnieper river basins in Ukraine has shown that the overall pollution by chemicals in the Siverskyi Donets basin is significantly lower. (C) 2021 Elsevier B.V. All rights reserved

Determination of 56 per- and polyfluoroalkyl substances in top predators and their prey from Northern Europe by LC-MS/MS

Per- and polyfluoroalkyl substances (PFAS) are a group of emerging substances that have proved to be persistent and highly bioaccumulative. They are broadly used in various applications and are known for their long-distance migration and toxicity. In this study, 65 recent specimens of a terrestrial apex predator (Common buzzard), freshwater and marine apex predators (Eurasian otter, harbour porpoise, grey seal, harbour seal) and their potential prey (bream, roach, herring, eelpout) from northern Europe (United Kingdom, Germany, the Netherlands and Sweden) were analyzed for the presence of legacy and emerging PFAS, employing a highly sensitive liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) method. 56 compounds from 14 classes were measured; 13 perfluoroalkyl carboxylic acids (PFCAs), 7 perfluoroalkyl sulphonic acids (PFSAs), 3 perfluorooctane sulfonamides (FOSAs), 4 perfluoroalkylphosphonic acids (PFAPAs), 3 perfluoroalkylphosphinic acids (PFPi's), 5 telomer alcohols (FTOHs), 2 mono-substituted polyfluorinated phosphate esters (PAPs), 2 di-substituted polyfluorinated phosphate esters (diPAPs), 6 saturated fluorotelomer acids (FTAS), 3 unsaturated fluorotelomer acids (FTUAs), 2 N-Alkyl perfluorooctane sulfonamidoethanols (FOSEs), 3 fluorotelomer sulphonic acids (FTSAs), 2 perfluoroether carboxylic acids (PFECAs) and 1 chlorinated perfluoroether sulphonic acid (Cl-PFESA). All samples were lyophilized before analysis, in order to enhance extraction efficiency, improve the precision and achieve lower detection limits. The analytes were extracted from the dry matrices through generic methods of extraction, using an accelerated solvent extraction (ASE), followed by clean-up through solid phase extraction (SPE). Method detection limits and method quantification limits ranged from 0.02 to 1.25 ng/g wet weight (ww) and from 0.05 to 3.79 ng/g (ww), respectively. Recovery ranged from 40 to 137%. Method precision ranged from 3 to 20 %RSD. The sum of PFAS concentration in apex predators livers ranged from 0.2 to 20.2 μg/g (ww), whereas in the fish species muscle tissues it ranged from 16 to 325 ng/g (ww). All analyzed specimens were primarily contaminated with PFOS, while the three PFPi's included in this study exhibited frequency of appearance (FoA) 100 %. C9 to C13 PFCAs were found at high concentrations in apex predator livers, while the overall PFAS levels in fish fillets also exceeded ecotoxicological thresholds. The findings of our study show a clear association between the PFAS concentrations in apex predators and the geographical origin of the specimens, with samples that were collected in urban and agricultural zones being highly contaminated compared to samples from pristine or semi-pristine areas. The high variety of PFAS and the different PFAS composition in the apex predators and their prey (AP&P) samples is alarming and strengthens the importance of PFAS monitoring across the food chain

Assessment of contaminants of emerging concern in European apex predators and their prey by LC-QToF MS wide-scope target analysis

Apex predators are good indicators of environmental pollution since they are relatively long-lived and their high trophic position and spatiotemporal exposure to chemicals provides insights into the persistent, bioaccumulative and toxic (PBT) properties of chemicals. Although monitoring data from apex predators can considerably support chemicals’ management, there is a lack of pan-European studies, and longer-term monitoring of chemicals in organisms from higher trophic levels. The present study investigated the occurrence of contaminants of emerging concern (CECs) in 67 freshwater, marine and terrestrial apex predators and in freshwater and marine prey, gathered from four European countries. Generic sample preparation protocols for the extraction of CECs with a broad range of physicochemical properties and the purification of the extracts were used. The analysis was performed utilizing liquid (LC) chromatography coupled to high resolution mass spectrometry (HRMS), while the acquired chromatograms were screened for the presence of more than 2,200 CECs through wide-scope target analysis. In total, 145 CECs were determined in the apex predator and their prey samples belonging in different categories, such as pharmaceuticals, plant protection products, per- and polyfluoroalkyl substances, their metabolites and transformation products. Higher concentration levels were measured in predators compared to prey, suggesting that biomagnification of chemicals through the food chain occurs. The compounds were prioritized for further regulatory risk assessment based on their frequency of detection and their concentration levels. The majority of the prioritized CECs were lipophilic, although the presence of more polar contaminants should not be neglected. This indicates that holistic analytical approaches are required to fully characterize the chemical universe of biota samples. Therefore, the present survey is an attempt to systematically investigate the presence of thousands of chemicals at a European level, aiming to use these data for better chemicals management and contribute to EU Zero Pollution Ambition

Assessment of contaminants of emerging concern in European apex predators and their prey by LC-QToF MS wide-scope target analysis

Apex predators are good indicators of environmental pollution since they are relatively long-lived and their high trophic position and spatiotemporal exposure to chemicals provides insights into the persistent, bioaccumulative and toxic (PBT) properties of chemicals. Although monitoring data from apex predators can considerably support chemicals’ management, there is a lack of pan-European studies, and longer-term monitoring of chemicals in organisms from higher trophic levels. The present study investigated the occurrence of contaminants of emerging concern (CECs) in 67 freshwater, marine and terrestrial apex predators and in freshwater and marine prey, gathered from four European countries. Generic sample preparation protocols for the extraction of CECs with a broad range of physicochemical properties and the purification of the extracts were used. The analysis was performed utilizing liquid (LC) chromatography coupled to high resolution mass spectrometry (HRMS), while the acquired chromatograms were screened for the presence of more than 2,200 CECs through wide-scope target analysis. In total, 145 CECs were determined in the apex predator and their prey samples belonging in different categories, such as pharmaceuticals, plant protection products, per- and polyfluoroalkyl substances, their metabolites and transformation products. Higher concentration levels were measured in predators compared to prey, suggesting that biomagnification of chemicals through the food chain occurs. The compounds were prioritized for further regulatory risk assessment based on their frequency of detection and their concentration levels. The majority of the prioritized CECs were lipophilic, although the presence of more polar contaminants should not be neglected. This indicates that holistic analytical approaches are required to fully characterize the chemical universe of biota samples. Therefore, the present survey is an attempt to systematically investigate the presence of thousands of chemicals at a European level, aiming to use these data for better chemicals management and contribute to EU Zero Pollution Ambition

Network analysis to reveal the most commonly detected compounds in predator-prey pairs in freshwater and marine mammals and fish in Europe

Marine and freshwater mammalian predators and fish samples, retrieved from environmental specimen banks (ESBs), natural history museum (NHMs) and other scientific collections, were analysed by LIFE APEX partners for a wide range of legacy and emerging contaminants (2545 in total). Network analysis was used to visualize the chemical occurrence data and reveal the predominant chemical mixtures for the freshwater and marine environments. For this purpose, a web tool was created to explore these chemical mixtures in predator-prey pairs. Predominant chemicals, defined as the most prevalent substances detected in prey-predator pairs were identified through this innovative approach. The analysis established the most frequently co-occurring substances in chemical mixtures from AP&P in the marine and freshwater environments. Freshwater and marine environments shared 23 chemicals among their top 25 predominant chemicals. Legacy chemical, including perfluorooctanesulfonic acid (PFOS), brominated diphenyl ethers (BDEs), polychlorinated biphenyls (PCBs), hexachlorobenzene and mercury were dominant chemicals in both environments. Furthermore, N-acetylaminoantipyrine was a predominant pharmaceutical in both environments. The LIFE APEX chemical mixture application (https://norman-data.eu/LIFE_APEX_Mixtures) was proven to be useful to establish most prevalent compounds in terms of number of detected counts in prey-predator pairs. Nonetheless, further research is needed to establish food chain associations of the predominant chemicals

A novel role for natural science collections in European contaminant monitoring

International audienceIntegration of the world’s natural history collections can provide a resource for decision-maker