Adsorption of Fluoxetine and Venlafaxine onto the Marine Seaweed Bifurcaria bifurcata

Marine macroalga Bifurcaria bifurcata was investigated as sorbent for two environmental emerging pollutants, the pharmaceuticals venlafaxine (VLF) and fluoxetine (FLX), from aqueous solutions, both in mono and bicomponent batch systems. The alga was characterized by means of IR spectroscopy, allowing to identify the most important groups for biosorption, carboxylic, sulfonic and hidroxile. The point of zero charge was determined, being 6.4. Adsorption of FLX follows a pseudo first order kinetics and kinetic constants are higher for FLX than for VLF. Its adsorption better represented by the pseudo second order model. For both pharmaceuticals, the equilibrium was reached within *100 min. Adsorption studies reveal that the process was more efficient at lower pH range and followed the Langmuir–Freundlich’s model for VLF and Langmuir’s model for FLX. Maximum adsorption capacities reached a level of 12 – 3 and 22 – 4 lmol/g for VLF and FLX, respectively, in the mono-component system. The difference was attributed to the presence of various functional groups of varying polarity within the adsorbates’ molecules that affected their interactions with the adsorbent surface. The Langmuir–Freundlich’s extended model was applied to the adsorption data of the bicomponent system and no changes in the maximum adsorption capacities were found (14 – 2 and 20 – 3 lmol/g for VLF and FLX, respectively), there is no evidence of competition between the adsorbates.Acknowledgments are due to Fundac¸a˜o para a Cieˆncia e a Tecnologia (FCT), FEDER under Programme PT2020 (Project UID/QUI/50006/2013—POCI/01/0145/FEDER/ 007265) and Programme FCT–UT Austin, Emerging Technologies (Project UTAP-ICDT/CTM-NAN/0025/2014) for the financial funding. The authors would like to thank the EU and FCT/UEFISCDI/FORMAS for funding, in the frame of the collaborative international consortium REWATER financed under the ERA-NET Cofund WaterWorks2015 Call. This ERA-NET is an integral part of the 2016 Joint Activities developed by the Water Challenges for a Changing World Joint Programme Initiative (Water JPI).W.S. also thanks FCT/ MEC for his grant (Laborato´rio Associado para Quı´mica Verde–Tecnologia e Processos Limpos–UID/QUI/50006, POCI-01-0145-FEDER-007265).info:eu-repo/semantics/publishedVersio

Ecotoxicological evaluation of chemical indicator substances present as micropollutants in laboratory wastewaters

Laboratories produce a large volume of wastewaters containing different chemical indicators, organic species for which there is no complete knowledge about their effects in the aquatic environment. The aim of this work was to evaluate the ecotoxicity of four chemical indicator substances commonly used in titrations (sodium diphenylamine-4-sulfonate, phenolphthalein, methyl orange, and eriochrome black T) by applying two distinct bioassays that evaluated the growth inhibition of the microalga Chlorella vulgaris and the acute immobilization of the microcrustacean Daphnia magna. All the indicators showed growth inhibition rates in the chronic test performed with the alga C. vulgaris. Only phenolphthalein and eriochrome black-T showed high immobilization rates on the acute test for D. magna. C. vulgaris showed higher sensitivity to the chemical indicators tested than D. magna. Eriochrome black T was the most toxic for both test organisms and, according to the effective concentration that causes inhibition on 50% of C. vulgaris population, it can be considered as “highly toxic to aquatic organisms”. Phenolphthalein and methyl orange may be classified as “toxic to aquatic organisms” and sodium diphenylamine-4-sulfonate is the least toxic, only being considered as “harmful”. This work increases the awareness of the hazardous effects of these chemical indicators and reinforces the need of improved solutions to manage and treat laboratory effluents.info:eu-repo/semantics/publishedVersio

Individual and mixture toxicity evaluation of three pharmaceuticals to the germination and growth of Lactuca sativa seeds

This work aims to assess, individually and in mixtures, possible phytotoxic effects of three pharmaceuticals (paracetamol, ibuprofen and amoxicillin) on germination and early growth of Lactuca sativa seeds. Pharmaceuticals are an important group of emerging contaminants, whose presence has been described in several environmental compartments, including soils. However, knowledge on their possible impact in terrestrial organisms is still sparse and even more when mixtures are considered. Germination tests are important to evaluate the quality of soil and the toxic effects that contaminants can pose to plants. The acute effects of individual pharmaceuticals as well as binary and ternary mixtures were assessed using different endpoints, namely: percentage of seed germination, root elongation, shoot and leaf length, after an exposure time of five days. Overall, in the exposure of L. sativa seeds to individual pharmaceuticals there are indications of acute toxicity in the early plant growth. However, this inhibitory effect tends to be cancelled in the acute exposure to mixtures. This study shows the importance of evaluating the toxicity of mixtures of pharmaceuticals, since they might have distinct toxic effects when compared to the single compounds, and also because, probably, it is the closest scenario to the reality that can be found in the environment.This work received financial support from the European Regional Development Fund (ERDF) through COMPETE – Operational Competitiveness Programme and national funds provided by FCT – Foundation for Science and Technology under the projects UID/QUI/50006/2013.info:eu-repo/semantics/publishedVersio

Determination of psychopharmaceutical adulterants in plant food supplements using UHPLC-MS/MS

In the last decades, medicinal plants and derived products have become increasingly available on the EU market as components of formulations sold as plant food supplements (PFS). Such products are legally considered as foods under the Directive 2002/46/EC [l] and do not require any kind of authorization to be placed on the market. The legal responsibility for its safety relies on the business operators. Among the several issues that may aíFect the safety of PFS, adulterations including the addition of illegal substances represent an increasing public health concern. In this context, the adulteration by the addition of psychopharmaceuticals should be investigated. Chromatographic methods, especially LC, coupled to various detectors are commonly used for the detection of fraudulent addition of pharmaceuticals in complex mixtures such as PFS. Among them, LC-MS/MS provides accurate and sensitive detectíon ofthe target compounds [2]. The purpose of this study was to assess the presence of psychopharmaceutical adulterants namely, fluoxetine, sertraline, citalopram, venlafaxine, paroxetine, trazodone, and diazepam in St. Johns wort (Hypericum perforatum) based PFS since this plant and/or extracts are traditionally used for its antidepressive properties. Analysis was performed in a Nexera Ultra-High Performance Liquid Chromatograph coupled to a triple-quadrupole mass spectrometer (LCMS-8030 Shimadzu) with an electrospray ionization source (ESI), operating in positive ion mode, using a Kinetex C18 fused core column (150 x 2.10 mm i.d.; 1.7 [im) (Phenomenex). Multiple reaction monitoring mode (MRM) was selected and pharmaceuticals were quantified by internai standard calibration method. Prior to analysis PFS samples were extracted with methanol. The methodology was applied to a set of20 St. Johns wort based PFS commercially available in the Portuguese market.This work received financiai support from the European Union (FEDER funds through COMPETE) and National Funds (FCT, Fundação para a Ciência e Tecnologia) through project EXPL/DTPSAP/1438/2013 and Pest-C/EQB/LA0006/2013.info:eu-repo/semantics/publishedVersio

Assessment of plant food supplements adulteration with psychopharmaceutical drugs

The purpose of this study was to compare three different extraction methods (two based on ethanol extraction and one on the Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERs) method) to assess the possible addition of psychopharmaceutical drugs (fluoxetine, sertraline, citalopram, venlafaxine, paroxetine, trazodone, and diazepam) as adulterants in St. John's wort (Hypericum perforatum) based plant food supplements (PFS). Methodology: Analysis was performed in a Nexera Ultra-High Performance Liquid Chromatograph (UHPLC) coupled to a triple-quadrupole mass spectrometer (LCMS-8030 Shimadzu) with an electrospray ionization source (ESI), operating in positive ion mode, using a Kinetex C18 fused core column (150 × 2.10 mm i.d.; 1.7 m) (Phenomenex). Multiple reaction monitoring mode (MRM) was selected and pharmaceuticals were quantified by internal standard calibration method. Calibration curves were constructed in the range 10 – 1000 g/L. The three different extraction methods were compared based on the analysis of spiked samples.This work received financial support from the European Union (FEDER funds through COMPETE) and National Funds (FCT, Fundação para a Ciência e Tecnologia) through project EXPL/DTP-SAP/1438/2013 and Pest- C/EQB/LA0006/2013. To all financing sources the authors are greatly indebted.info:eu-repo/semantics/publishedVersio

Effects and bioaccumulation of gold nanoparticles in the gilthead seabream (Sparus aurata) – Single and combined exposures with gemfibrozil

Gold nanoparticles (AuNPs) are found in a wide range of applications and therefore expected to present increasing levels in the environment. There is however limited knowledge concerning the potential toxicity of AuNPs as well as their combined effects with other pollutants. Hence, the present study aimed to investigate the effects of AuNPs alone and combined with the pharmaceutical gemfibrozil (GEM) on different biological responses (behaviour, neurotransmission, biotransformation and oxidative stress) in one of the most consumed fish in southern Europe, the seabream Sparus aurata. Fish were exposed for 96 h to waterborne 40 nm AuNPs with two coatings - citrate and polyvinylpyrrolidone (PVP), alone or combined with GEM. Antioxidant defences were induced in liver and gills upon both AuNPs exposure. Decreased swimming performance (1600 μg.L-1) and oxidative damage in gills (4 and 80 μg.L-1) were observed following exposure to polyvinylpyrrolidone coated gold nanoparticles (PVP-AuNPs). Generally, accumulation of gold in fish tissues and deleterious effects in S. aurata were higher for PVP-AuNPs than for cAuNPs exposures. Although AuNPs and GEM combined effects in gills were generally low, in liver, they were higher than the predicted. The accumulation and effects of AuNPs showed to be dependent on the size, coating, surface charge and aggregation/agglomeration state of nanoparticles. Additionally, it was tissue' specific and dependent on the presence of other contaminants. Although, gold intake by humans is expected to not exceed the estimated tolerable daily intake, it is highly recommended to keep it on track due to the increasing use of AuNPs.This research was supported through the COMPETE – Operational Competitiveness Program and national funds through FCT – Foundation for Science and Technology, under the project “NANOAu – Effects of Gold Nanoparticles to Aquatic Organisms” (FCT PTDC/MAR-EST/3399/2012) (FCOMP-01-0124-FEDER-029435), through FCT/MCTES through national funds (PIDDAC) and the cofounding by FEDER, within the PT2020 Partnership Agreement and Compete 2020 to CESAM (UID/AMB/50017 – POCI-01-0145-FEDER-007638). A. Barreto has a doctoral fellowship from FCT (SFRH/BD/97624/2013); L. G. Luis had a fellowship from FCT (BI/UI88/6881/2014). MO has financial support of the program Investigator FCT, co-funded by the Human Potential Operational Programme and European Social Fund (IF/00335(2015).info:eu-repo/semantics/publishedVersio

Microplastics as vectors of pharmaceuticals in aquatic organisms – An overview of their environmental implications

Microplastics (MPs) have been recognized as an environmental threat due to their persistence, ubiquity, and toxicity potential. Due to their small size, MPs are endowed with a large specific surface area, having the ability to sorb and accumulate other contaminants that co-exist with them in the aquatic environment as it is the case of pharmaceuticals. In this way, MPs can act as vectors, facilitating the contact of pharmaceuticals with aquatic organisms. Once ingested, MPs may desorb pharmaceuticals, increasing their bioavailability, which could promote their bioaccumulation and biomagnification through the food web as well as modulate their toxic effects. Microplastics may differently impact the toxicity of pharmaceuticals by potentiating (synergism) or decreasing (antagonism) it. This review aims at highlighting the role of MPs as vectors of pharmaceuticals to aquatic organisms and how the interaction of these emerging contaminants may influence the bioaccumulation and toxicity of pharmaceuticals on biota. Examples of different scenarios resulting from the exposure of aquatic life to MPs and pharmaceuticals are presented. Future studies should cover a broader range of polymer types and environmental realistic concentrations to better understand the impact of MPs on the bioaccumulation and toxicity of pharmaceuticals.This work was funded by the Spanish State Research Agency of the Spanish Ministry of Science, Innovation and Universities and European Fund for Regional Development (project PLAS-MED; FEDER-MCIU-AEI/CTM2017-89701-C3-2-R). Authors thank Generalitat de Catalunya through Consolidated Research Group (ICRA-ENV 2017 SGR 1124 & 2017 SGR 1404). ICRA researchers thank funding from CERCA program. Lúcia H.M.L.M. Santos thanks the Juan de la Cierva program (IJCI-2017-32747) and Sara Rodríguez-Mozaz thanks the Ramon y Cajal program (RYC-2014-16707) from the Spanish State Research Agency of the Spanish Ministry of Science, Innovation and Universities (AEI-MCIU).Peer reviewe

Analysis of microplastics in the environment: Identification and quantification of trace levels of common types of plastic polymers using pyrolysis-GC/MS

This work describes the development of analytical workflows based on pyrolysis coupled with gas chromatography-mass spectrometry (Pyr-GC/MS) for the qualitative and quantitative analysis of 12 of the most common plastic polymers in environmental samples. The most suitable characteristic pyrolyzate compounds and respective indicator ions were selected for each polymer in order to obtain the most appropriate response for analytical purposes. Additionally, commercial pyrolyzates and polymers libraries were used to confirm the identity of the detected microplastics. The method was validated, showing a good linearity for all the plastic polymers (R2 > 0.97) and limits of detection between 0.1 (polyurethane) to 9.1 µg (polyethylene). The developed methodology was successfully applied for the analysis of plastic polymers in environmental microplastic samples collected in three Mediterranean beaches (NE Spain). • Fast and reproducible Pyr-GC/MS method for the analysis of the 12 most common plastic polymers in a single GC/MS run • Straightforward analytical workflows using pyrolyzates and polymers libraries enable a fast identification and quantification of microplastics in environmental sample

Sertraline accumulation and effects in the estuarine decapod Carcinusmaenas: Importance of the history of exposure to chemical stress

Sertraline is widely prescribed worldwide and frequently detected in aquatic systems. There is, however, a remarkable gap of information on its potential impact on estuarine and coastal invertebrates. This study investigated sertraline accumulation and effects in Carcinus maenas. Crabs from a moderately contaminated (Lima) and a low-impacted (Minho) estuary were exposed to environmental and high levels of sertraline (0.05, 5, 500 μg L−1). A battery of biomarkers related to sertraline mode of action was employed to assess neurotransmission, energy metabolism, biotransformation and oxidative stress pathways. After a seven-day exposure, sertraline accumulation in crabs’ soft tissues was found in Lima (5 μg L−1: 15.3 ng L−1 ww; 500 μg L−1: 1010 ng L−1 ww) and Minho (500 μg L−1: 605 ng L−1 ww) animals. Lima crabs were also more sensitive to sertraline than those from Minho, exhibiting decreased acetylcholinesterase activity, indicative of ventilatory and locomotory dysfunction, inhibition of anti-oxidant enzymes and increased oxidative damage at ≥0.05 μg L−1. The Integrated Biomarker Response (IBR) index indicated their low health status. In addition, Minho crabs showed non-monotonic responses of acetylcholinesterase suggestive of hormesis. The results pointed an influence of the exposure history on differential sensitivity to sertraline and the need to perform evaluations with site-specific ecological receptors to increase relevance of risk estimations when extrapolating from laboratory to field conditions

Insights on the metabolization of the antidepressant venlafaxine by meagre (Argyrosomus regius) using a combined target and suspect screening approach

Bioaccumulation of pharmaceuticals in fish exposed to contaminated water can be shaped by their capability to metabolize these xenobiotics, affecting their toxicity and animal welfare. In this study the in vivo metabolization of the antidepressant venlafaxine by the juvenile marine fish meagre (Argyrosomus regius) was evaluated using a combined target and suspect screening analytical approach. Thirteen venlafaxine metabolites were identified, namely N-desmethylvenlafaxine and N,N-didesmethylvenlafaxine, which were unequivocally identified using analytical standards, and 11 more tentatively identified by suspect screening analysis, including two Phase II metabolites formed by amino acid conjugation. All of them were detected in the liver, while in plasma and brain only 9 and 6 metabolites, respectively, were detected. Based on these findings, for the first time, a tentative metabolization pathway of venlafaxine by A. regius is proposed. Contrarily to what happen in humans, N-demethylation was identified as the main route of metabolization of venlafaxine by fish. Our findings highlight species-specificity in the metabolization of venlafaxine and allow a better understanding of venlafaxine's toxicokinetic in fish. These results emphasize the need to investigate the biotransformation of xenobiotics by non-target organisms to have an integrated overview of their environmental exposure and to improve future evaluations of environmental risk assessment.This work was funded by the 7th EU-FP (ECsafeSEAFOOD project; GA n° 311820) and the European Regional Development Fund (FEDER) and by Spanish Ministry of Economy and Competitiveness (PGE-2010). This work was also funded by the Spanish State Research Agency of the Spanish Ministry of Science, Innovation and Universities and European Fund for Regional Development (project PLAS-MED; FEDER-MCIU-AEI/CTM2017-89701-C3-2-R). Authors thank Generalitat de Catalunya through Consolidated Research Group (ICRA-ENV 2017 SGR 1124 & 2017 SGR 1404). ICRA researchers thank funding from CERCA program. Lúcia H.M.L.M. Santos thanks the Juan de la Cierva program (IJCI-2017-32747) and Sara Rodríguez-Mozaz thanks the Ramon y Cajal program (RYC-2014-16707) from the Spanish State Research Agency of the Spanish Ministry of Science, Innovation and Universities (AEI-MCIU). A. Jaén-Gil thanks the predoctoral grant from the Agency for Management of University and Research Grants (AGAUR) (2019FI_B2_00202) co-financed by the European Social Fund. A. Marques (IF program) and A.L. Maulvault (Project FishBudget PTDC/BIA-BMA/28630/2017) thank the Portuguese Foundation for Science and Technology.Peer reviewe