Analysis of perfluorinated compounds in sewage sludge and hydrochar by UHPLC LTQ/Orbitrap MS and removal assessment during hydrothermal carbonization treatment.

Wastewater treatment plants have been recognized as important sinks for per- and polyfluoroalkyl substances (PFAS) because of their ineffectiveness in removing them reflecting both water and sewage sludge discharge routes. Hydrothermal treatment represents an alternative technology for treating sludge to recover energy and other valuable products. In this study, 15 PFAS were determined in sludge and hydrochar substrates using sonication-solid phase extraction procedure and analyzed using LC-Orbitrap-High Resolution-MS/MS. The method was fully validated, exhibiting very good linearity, recoveries in the range of 48 to 126 %, low detection and quantification limits with expanded uncertainty and precision below 32 % and 21.9 %, respectively. The method was applied to sludge samples from the WWTP of Ioannina city (Greece), as well as to hydrothermally treated samples under various conditions. The most abundant PFAS were PFHxA (0.5-38.3 ng g-1) and PFOS (4.4-22.1 ng g-1). Finally, the hydrothermally treated sludge samples spiked with PFAS presented removal efficiencies for total PFAS of 86.9 %, 91.8 % and 95.7 % at three spiking levels namely 10, 50 and 200 ng g-1, respectively. Results indicated that PFCAs were almost completely removed, except for PFOA, while the concentrations of PFSAs increased in the produced hydrochar with the formation of several intermediates, as detected by HR-LC-MS/MS. The results of this study demonstrate the effect of hydrothermal treatment to the fate of PFAS in sewage sludge and contribute for further studies on design and scale up of hydrothermal carbonization technology as a management option for safer disposal of municipal wastewater sludge.

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.

Target and suspect screening approaches for the identification of emerging and other contaminants in fish feeds using high resolution mass spectrometry.

Fish feed is essential in aquaculture fish production because, along with beneficial nutrients and components, many suspected compounds can be transferred to fish and ultimately to humans. In this context, a comprehensive analysis was conducted to monitor various pesticides and pharmaceutical compounds in aquaculture fish feed through target analysis and many other groups of chemicals via suspect screening approaches. In this study, the QuEChERS extraction method was optimized, validated, and applied to fifty-four fish feed samples collected from different production batches. This was followed by liquid chromatography-high-resolution linear ion trap/Orbitrap mass spectrometry (LC-HR-IT/Orbitrap-MS) for targeted and suspect screening purposes. In general, pesticides provided satisfactory recoveries (70-105.5 %), with quantification limits lower than 5 ng g-1, whereas pharmaceuticals displayed recoveries ranging from 70.5 to 120.2 %, with quantification limits below 25 ng g-1. In addition, the matrix effects and measurement uncertainty were assessed to provide more accurate and high-confidence results. Pirimiphos-methyl was detected and quantified in 20 of 54 fish feed samples (37 %) at concentrations <77 ng g-1. Finally, suspect screening revealed the occurrence of 10 mycotoxins (e.g., citrinin, aflatoxin G2, zearalenone, and alternariol), two pesticides excluding the target pesticides (tebuconazole and fenazaquin), perfluorooctane sulfonic acid (PFOS) in almost 2 % of the samples, and ethoxyquin (antioxidant), with 12 of its Transformation Products (TPs). Finally, suspect analysis incorporated in routine analyses have proven to have great potential for complete monitoring.

Aqueous fate of furaltadone: Kinetics, high-resolution mass spectrometry - based elucidation and toxicity assessment of photoproducts.

Furaltadone (FTD) is an antibiotic belonging to the nitrofurans group. It has been broadly used in livestock and aquaculture for therapeutic purposes, as well as for stimulating promotion. Although the European Union has imposed restrictions on the use of FTD since 1995 due to concerns regarding its toxicity, in many cases FTD has been excessively and/or illegally applied in productive animals in developing countries, because of its high efficacy and low-cost. Unlike other nitrofuran compounds, the hydrolytic and photolytic behavior of FTD in natural aquatic systems has not been thoroughly investigated. To this end, hydrolysis in different pH values and photolysis in aquatic environment, including lake, river and sea water have been both examined. Hydrolysis was found to have an insignificant impact on degradation of FTD in the aquatic environment relevant pH values, whereas indirect photolysis proved to be the main route of its elimination. The identification of tentative photoproducts (PPs) was performed using ultra high performance liquid chromatography coupled to hybrid LTQ/Orbitrap high resolution mass spectrometry. A possible pathway for photolytic transformation of FTD was proposed. Additionally, in silico simulations were used to evaluate the toxicity such as the mutagenicity of FTD and PPs. Complementary to the low-cost and time-limited simulations, an in vitro method (Vibrio Fischeri bioluminescence) was also used to assess ecotoxicity.

Elucidation of 4-Hydroxybenzoic Acid Catabolic Pathways in Pseudarthrobacter phenanthrenivorans Sphe3.

4-hydroxybenzoic acid (4-HBA) is an aromatic compound with high chemical stability, being extensively used in food, pharmaceutical and cosmetic industries and therefore widely distributed in various environments. Bioremediation constitutes the most sustainable approach for the removal of 4-hydroxybenzoate and its derivatives (parabens) from polluted environments. Pseudarthrobacter phenanthrenivorans Sphe3, a strain capable of degrading several aromatic compounds, is able to grow on 4-HBA as the sole carbon and energy source. Here, an attempt is made to clarify the catabolic pathways that are involved in the biodegradation of 4-hydroxybenzoate by Sphe3, applying a metabolomic and transcriptomic analysis of cells grown on 4-HBA. It seems that in Sphe3, 4-hydroxybenzoate is hydroxylated to form protocatechuate, which subsequently is either cleaved in ortho- and/or meta-positions or decarboxylated to form catechol. Protocatechuate and catechol are funneled into the TCA cycle following either the ß-ketoadipate or protocatechuate meta-cleavage branches. Our results also suggest the involvement of the oxidative decarboxylation of the protocatechuate peripheral pathway to form hydroxyquinol. As a conclusion, P. phenanthrenivorans Sphe3 seems to be a rather versatile strain considering the 4-hydroxybenzoate biodegradation, as it has the advantage to carry it out effectively following different catabolic pathways concurrently.

Beneficial properties of Drimia numidica leaf methanolic extract against the cytogenotoxic effects of mitomycin C on human lymphocytes.

This study investigated the phytochemical profile of Drimia numidica leaf methanolic extract, as well as its cyto-genotoxic and cyto/genoprotective potential against mitomycin C (MMC) mediated effects on healthy human lymphocytes. Photosynthetic pigments, trace elements, and secondary metabolites were estimated and/or identified in methanolic extract of mature leaves, and the latter was further used for assessing its in vitro biological effects on MMC-free and/or MMC-treated human lymphocytes (at low, non-toxic concentrations of 0.001 and 0.01% v/v). The results showed that D. numidica leaf methanolic extract, being rich in carotenoids, phenolics, flavonoids, organic acids and bufadienolides, could be protective against MMC mediated cyto/genotoxic potential in healthy human lymphocytes. Biomolecules possessing antioxidant and antitumor potential, such as beta-carotene and lutein among others, chlorogenic acid, caffeic acid and their derivatives, minerals such as Si, as well as apigenin- and luteolin-derived glycosides, either individual or in a mixture, could be beneficial rather than harmful, at least at the extract concentrations tested. Although further in vitro and in vivo studies are still needed for elucidating the beneficial (individual and/or additive/synergistic) role of those compounds, the results of the present study are quite promising, thus encouraging new challenges for the appropriate utilization of D. numidica leaf extract.

Local Anesthesia Thoracoscopy with versus without Midazolam: A Randomized Controlled Trial.

BACKGROUND: Medical thoracoscopy is the gold standard for the diagnosis of pleural diseases. To date, no consensus exists regarding the choice of sedative and analgesic agents in patients undergoing local anesthetic thoracoscopy (LAT), and questions are raised as to whether sedatives may add to respiratory side effects. OBJECTIVE: The aim of the study was to test the hypothesis that administration of midazolam associated with lidocaine versus lidocaine alone in patients with LAT adds to respiratory side effects. METHODS: We randomly assigned 80 patients to a 1:1 study to 2 groups: local anesthesia by lidocaine (n = 40) versus lidocaine and midazolam (n = 40), with the primary end point being the mean lowest oxygen saturation. The secondary end points were cardiovascular parameters, complications, days of drainage, hospital stay, and patients' quality of life (QoL) as assessed by a visual analog scale (VAS). RESULTS: The mean age of all patients was 66.6 ± 13.1 years. The study comprised 50 males (62.5%). No difference was observed in the demographics between the 2 groups. No significant difference was observed between the 2 groups in oxygen saturation (primary end point). A significant difference was observed in favor of the midazolam group regarding the QoL assessed by VAS. CONCLUSION: Midazolam does not add to respiratory side effects when it is used with lidocaine for LAT, while patients' QoL is actually improved in this group. Therefore, in our department, we changed our startegy in favor of the association of lidocaine and midazolam.

Assessment of multiclass pharmaceutical active compounds (PhACs) in hospital WWTP influent and effluent samples by UHPLC-Orbitrap MS: Temporal variation, removals and environmental risk assessment.

Nowadays the occurrence and associated risks of Pharmaceutical Active Compounds (PhACs) in the aquatic environment comprises a major issue. In the present study, a comprehensive survey on contamination profiles, occurrence, removals, temporal variation and ecological risk of multiclass multiresidue PhACs, such as antibiotics, non-steroidal anti-inflammatories, lipid regulators and phsychiatrics, (including past and newly monitored PhACs as well as some of their metabolites) was performed in wastewaters from the WWTP of Ioannina University hospital along one year period on a monthly sampling basis. WWTP influent and effluent samples were analyzed for physicochemical quality parameters and PhACs concentration levels using Ultra High Performance Liquid Chromatography-Orbitrap-Mass Spectrometry (UHPLC-Orbitrap-MS), after Solid Phase Extraction (SPE) through Oasis HLB cartridges. Influent concentrations ranged between < LOQ (Limit of Quantification) for diclofenac and tolfenamic acid and 48586 ng/L for caffeine, while effluent concentrations between < LOQ for tolfenamic acid and simvastatin and 3361 ng/L for caffeine. Removal efficiencies ranged between -132.6% for venlafaxine and 100% for caffeine. Environmental risk assessment by means of Risk Quotient (RQ) for maximum and minimum concentration levels as well as optimized by the frequency of exceeding toxicity threshold values, RQf, was applied revealing that up to 12 PhACs posed acute toxicity (clofibric acid, fenofibrate, sulfadiazine, sulfamethoxazole, trimethoprim, amitryptiline, fluoxetine, fluvoxamine, norfluoxetine, sertraline, venlafaxine, caffeine) while up to 4 compounds exerted long-term toxicity (sulfamethoxazole, fluoxetine, sertraline, caffeine) at least for one of the studied organisms. Furthermore, mixture RQMEC/PNEC and RQSTU effect of multiple compounds showed high potential risks of the target groups in some cases, although some contaminants were not included due to lack of available data. Results can be used to prioritization of PhACs and their metabolites for surveillance in receiving water bodies as well as development of knowledge on toxicity and mechanism(s) of action.

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.

Psychiatrics and selected metabolites in hospital and urban wastewaters: Occurrence, removal, mass loading, seasonal influence and risk assessment.

The occurrence, removal, mass loading, seasonal influence and environmental risk assessment of nine psychiatric pharmaceuticals and four of their selected metabolites, were studied in one hospital and one urban wastewater treatment plant (WWTP) in Ioannina city, in northwestern Greece, providing information about the efficiency of the plants and their contribution into the final receiver's flow. Samples were collected from the influents and the effluents of the plants in different sampling campaigns, from July to December 2016. Analytical methodology was based on ultra-high performance liquid chromatography-Orbitrap high-resolution mass spectrometry, after solid-phase extraction through Oasis HLB cartridges. Concentrations in both WWTPs ranged between

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.

A modified QuEChERS approach for the analysis of pharmaceuticals in sediments by LC-Orbitrap HRMS.

This work reports on a rapid and sensitive analytical method intended to be used for investigation of 25 multiclass pharmaceuticals in sediments. The method comprises a modified quick, easy, cheap, effective, rugged and safe (QuEChERS) extraction, followed by ultra-high-performance liquid chromatography-high-resolution linear ion trap/Orbitrap mass spectrometry, operated in positive ionization mode. The crucial parameters of both extraction and cleanup as well as those related to liquid chromatography and mass spectrometry were optimized and the method was validated in terms of accuracy, method detection and quantification limits, precision, linearity, and matrix effects. A matrix-matched calibration approach and isotopically labeled internal standards were employed for quantification and correction of matrix effects. In the optimized method, recoveries were calculated to be between 64 and 101% in the intermediate spiking level, with a relative standard deviation below 14%. Method quantification limits ranged from 1.3 to 47 ng g-1. The linearity, expressed as a correlation coefficient, was over 0.998 in all cases, within a range from each compound's method quantification limit to 250 ng g-1. The matrix effect study revealed slight signal suppression that decreased after the cleanup step for 50% of the pharmaceuticals, proving its adequacy to limit matrix interferences, without causing any important loss of relevant analytes. The applicability of the method was successfully tested in sediments from two Greek rivers in northwestern Greece, revealing the presence of paracetamol, risperidone, venlafaxine, citalopram, and carbamazepine and estimating for the first time the potential occurrence of pharmaceuticals in these two aquatic systems.

Trace analysis of pesticide residues in sediments using liquid chromatography-high-resolution Orbitrap mass spectrometry.

The present study describes the optimization and validation of an analytical method based on quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction and purification with dispersive solid-phase extraction (dSPE) before analysis, followed by ultrahigh-performance liquid chromatography-high-resolution linear ion trap/Orbitrap (LTQ Orbitrap) mass spectrometry for the determination of 18 pesticides in sediment. To optimize process efficiency, parameters such as pH, extraction salts, sediment amount, and cleanup sorbents were evaluated. Identification was based on both accurate mass and retention time, and further confirmation was achieved by mass spectrometry fragmentation. The optimized analytical method demonstrated good validation characteristics, such as accuracy (recoveries from 70.8% to 106.2%), method quantification limits (below 10 ng g-1 for 89% of the pesticides selected), linearity (coefficient of determination greater than 0.9921 in all cases), precision (repeatability and reproducibility with standard deviations below 18% and 21%, respectively), and matrix effect (signal suppression was exhibited for almost all analytes). The overall method performance, expressed as process efficiency, ranged from 58.8% to 102.1%. The validated method was successfully applied to real samples collected along two rivers in northwestern Greece, revealing the presence of three selected pesticides but at levels below the method quantification limit.

Halogenated and organophosphorus flame retardants in European aquaculture samples.

This work monitors flame retardants in sediment, mussel and water samples from European fish farms. Polybrominated diphenyl ethers (PBDEs) were detected in 95% of the sediment and mussel samples with mean levels of 8.60±22.6ngg-1 dw in sediments and 0.07±0.18ngg-1 dw in mussels. BDE-209 was the main contributor for the sediments and BDE-47 was found in about 60% of the samples of both matrices. Pentabromoethylbenzene (PBEB) and hexabromobenzene (HBB) were detected in 42% of the sediments, but not in mussels. Decabromodiphenyl ethane (DBDPE) was found in about 55% of the samples of both matrices. The same happened for dechloranes in mussels, but they were detected in 92% of the sediments. Syn-DP and anti-DP were always the main contributors. Methoxylated PBDEs (MeO-PBDEs) were detected in all mussels and some sediments, mainly 6-MeO-BDE-47 and 2'-MeO-BDE-68. Organophosphorus flame retardants (OPFRs) were found in all matrices with concentrations of 0.04-92.8ngg-1 dw in sediment, 0.50-102ngg-1 dw in mussel and 0.43-867ngl-1 in water. Only OPFRs were analysed in water samples as halogenated flame retardants and MeO-PBDEs are highly unlikely to be detected in water due to their physicochemical properties. Flame retardants have no application in fish farming so results should reflect the impact of human activity on the farm locations. A large majority of the most contaminated samples were collected from sampling spots that were at urban shores or in enclosed water bodies not completely open to the sea.

Bioaccumulation of persistent and emerging pollutants in wild sea urchin Paracentrotus lividus.

Marine pollution has been increasing as a consequence of anthropogenic activities. The preservation of marine ecosystems, as well as the safety of harvested seafood, are nowadays a global concern. Here, we report for the first time the contamination levels of a large set of 99 emerging and persistent organic contaminants (butyltins (BTs), polycyclic aromatic hydrocarbons (PAHs), pesticides including pyrethroids, pharmaceuticals and personal care products (PCPs) and flame retardants) in roe/gonads of sea urchin Paracentrotus lividus. Sea urchins are a highly prized worldwide delicacy, and the harvesting of this seafood has increased over the last decades, particularly in South West Atlantic coast, where this organism is harvested mainly for exportation. Sampling was performed in three harvesting sites of the NW Portuguese coast subjected to distinct anthropogenic pressures: Carreço, Praia Norte and Vila Chã, with sea urchins being collected in the north and south areas of each site. Butyltins and pharmaceuticals were not found at measurable levels. Several PAHs, four pyrethroids insecticides, four PCPs and eleven flame retardants were found in roe/gonads of sea urchins, though in general at low levels. Differences among harvesting sites and between areas within each site were found, the lowest levels of contaminants being registered in Carreço. The accumulation of contaminants in sea urchins' roe/gonads seemed to reflect the low anthropogenic pressure felt in the sampling sites. Nevertheless, taking into account the low accumulated levels of chemicals, results indicate that sea urchins collected in South West Atlantic coast are safe for human consumption.

An experimental design approach employing artificial neural networks for the determination of potential endocrine disruptors in food using matrix solid-phase dispersion.

Matrix solid-phase dispersion (MSPD) as a sample preparation method for the determination of two potential endocrine disruptors, linuron and diuron and their common metabolites, 1-(3,4-dichlorophenyl)-3-methylurea (DCPMU), 1-(3,4-dichlorophenyl) urea (DCPU) and 3,4-dichloroaniline (3,4-DCA) in food commodities has been developed. The influence of the main factors on the extraction process yield was thoroughly evaluated. For that purpose, a 3 fractional factorial design in further combination with artificial neural networks (ANNs) was employed. The optimal networks found were afterwards used to identify the optimum region corresponding to the highest average recovery displaying at the same time the lowest standard deviation for all analytes. Under final optimal conditions, potato samples (0.5 g) were mixed and dispersed on the same amount of Florisil. The blend was transferred on a polypropylene cartridge and analytes were eluted using 10 ml of methanol. The extract was concentrated to 50 microl of acetonitrile/water (50:50) and injected in a high performance liquid chromatography coupled to UV-diode array detector system (HPLC/UV-DAD). Recoveries ranging from 55 to 96% and quantification limits between 5.3 and 15.2 ng/g were achieved. The method was also applied to other selected food commodities such as apple, carrot, cereals/wheat flour and orange juice demonstrating very good overall performance.