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This review describes a wide variety of analytical approaches for the assessment of human exposure to organic chemicals associated with plastic additives, focusing on works published in the last decade on plasticizers, bisphenols, flame retardants and antioxidants. Physiologically based extraction tests serve as preliminary in-vitro assays to determine the bioaccessibility of these compounds from micro/nanoplastics in body fluids of the gastrointestinal tract, skin, or lung. Whenever plastic-laden compounds become bioavailable, human metabolism is to be monitored through the assessment of phase I and II metabolites. In this regard, analytical methods based on chromatography and mass spectrometry for human biomonitoring of parent compounds and their metabolites in biological samples (mostly urine and plasma) are discussed in depth. This review also covers the role of wastewater-based epidemiology in determining the overall human exposure of a given population to plastic-related species.

Tris (1-chloro-2-propyl) phosphate (TCIPP) is one of the major organophosphate flame retardants present in the indoor and outdoor environment. Knowledge of biotransformation pathways is important to elucidate potential bioavailability and toxicity of TCIPP and to identify relevant biomarkers. This study aimed to identify TCIPP metabolites through in vitro human metabolism assays and finally to confirm these findings in urine samples from an occupationally exposed population to propose new biomarkers to accurately monitor exposure to TCIPP. TCIPP was incubated with human liver microsomes and human liver cytosol to identify Phase I and Phase II metabolites, by liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (LC-QTOF-MS). Using a suspect-screening approach, the established biomarkers bis (1-chloro-2-propyl) hydrogen phosphate (BCIPP) and 1-hydroxy-2-propyl bis (1-chloro-2-propyl) phosphate (BCIPHIPP) were identified. In addition, carboxyethyl bis (1-chloro-2-propyl) phosphate (TCIPP-M1), bis (1-chloropropan-2-yl) (-oxopropan-2-yl) phosphate (TCIPP-M2) and 1-chloro-3-hydroxypropan-2-yl bis (1-chloropropan-2-yl) phosphate (TCIPP-M3) were identified. TCIPP-M2, an intermediate product, was not reported before in literature. In urine samples, apart from BCIPP and BCIPHIPP, TCIPP-M1 and TCIPP-M3 were identified for the first time. Interestingly, BCIPP showed the lowest detection frequency, likely due to the poor sensitivity for this compound. Therefore, TCIPP-M1 and TCIPP-M3 could serve as potential additional biomarkers to more efficiently monitor TCIPP exposure in humans.

This work investigated, during one year, the occurrence and fate of 52 contaminants of emerging concern (CECs) in transnational river basins and coastal areas of the North of Portugal and Galicia (NW Spain) and the wastewater treatment plants (WWTPs) discharging on these environments. The different CECs investigated included pharmaceuticals, personal care products, industrial chemicals, among others, of which ca. 90 % would fulfill the persistence, mobility and toxicity criteria proposed by the German Environmental Agency. The results showed the ubiquitous presence of these CECs and an incomplete removal of over 60 % of them with current conventional WWTPs. These findings highlight the requirement of a prominent and coordinated upgrade of WWTP treatments in order to meet the future European Union regulations on urban wastewater treatment and surface water quality. In fact, even some compounds exhibiting high removals, such as caffeine or xylene sulfonate, were frequently detected in river and estuarine waters at the high ng L−1 level. Thus, our preliminary risk assessment study concluded that 18 of the CECs presented a potential risk for the environment, being caffeine, sulpiride, perfluorooctanoic acid (PFOA), diclofenac, fipronil and perfluorobutanoic acid (PFBA) the most concerning ones. Yet, additional toxicity data as well as a more robust information on persistence and mobility of CECs are necessary for better estimating the magnitude of the problem and improve risk assessment. As an example, in the case of the antidiabetic metformin, recent research has revealed toxicity for model fish species at concentration levels below those found in 40 % of the river water samples analyzed in this work.

Photo-Fenton (PF) is a promising process for degrading a wide range of contaminants of emerging concern (CECs) present in urban wastewater (UWW) after secondary treatment, mitigating their spread in aquatic systems. However, the near-neutral pH of UWW poses a challenge to PF performance. In this research, a tubular membrane photoreactor, designed for continuous “titration” of Fe2+, is explored to promote the PF process for the tertiary treatment of UWW. The application of persulfate (PF-like) as an alternative oxidant and the impact of phosphate (PO43−) content in UWW were also assessed. Process efficiency was evaluated in continuous mode, applying low residence times (RT: 6.1, 36.6, and 73.2 s) and low Fe2+ doses (1, 2, and 5 mg L−1), for the oxidation of 19 CECs (each spiked at 10 µg/L), in demineralized water (DW) and secondary-treated UWW. Despite the persistence of certain short-chain perfluoroalkyl substances (PFAS) and saccharine across all conditions tested, the PF-like process exhibited superior performance when compared with PF, attaining higher removals for most target CECs, especially for melamine. In UWW, for an RT = 73.2 s, [Fe2+] = 5 mg L−1, and [oxidant] = 1.2 mM, PF process removed 7 CECs > 60 % and PF-like 10 CECs > 60 %. Moreover, higher residual concentrations of Fe2+ and lower precipitation of PO43− were observed for PF-like treatments, evidencing its advantages for tertiary treatment. These results emphasize the importance of photoreactor design to achieve efficient PF/PF-like at neutral pH, avoiding the use of chelating agents while managing iron concentrations compatible with UWW discharge or reuse limits.

This article critically reviews analytical method validation and quality control applied to the environmental chemistry field. The review focuses on the determination of organic micropollutants (OMPs), specifically emerging contaminants and pesticides, in the aquatic environment. The analytical technique considered is (gas and liquid) chromatography coupled to mass spectrometry (MS), including high-resolution MS for wide-scope screening purposes. An analysis of current research practices outlined in the literature has been performed, and key issues and analytical challenges are identified and critically discussed. It is worth emphasizing the lack of specific guidelines applied to environmental analytical chemistry and the minimal regulation of OMPs in waters, which greatly affect method development and performance, requirements for method validation, and the subsequent application to samples. Finally, a proposal is made for method validation and data reporting, which can be understood as starting points for further discussion with specialists in environmental analytical chemistry.

The occurrence of pharmaceuticals in influent wastewater samples (IWW) is a recurrent issue. The monitoring of their presence is not only valuable from an environmental point of view, but also as a tool to analyze patterns of human use by the so-called wastewater-based epidemiology. The development of an analytical method based on solid-phase extraction followed by liquid chromatography coupled to tandem mass spectrometry to monitor the occurrence of a group of seventeen pharmaceuticals including the most representative for various therapeutic families in IWW samples is described in this work. The samples were collected during a monitoring week in six wastewater treatment plants located in different cities and towns across Spain. The developed method provides acceptable figures of merit with apparent recoveries in IWW ranging from 42% to 139%, and low matrix effect (in general lower than ± 30%), and method quantification limits (MQL) between 1 ng/L and 24 ng/L for all compounds, except atenolol (58 ng/L). All the studied pharmaceuticals were found in all samples with concentrations ranging from < MQL to 10,393 ng/L, being the highest concentrations for tramadol. The population normalized daily loads revealed that the use of pharmaceuticals follows, in general, a similar pattern in all cities monitored.

Metformin (MET), an anti-diabetic pharmaceutical of large-scale consumption, is increasingly detected in surface waters. However, current knowledge on the long-term effects of MET on non-target organisms is limited. The present study aimed to investigate the effects of MET in the model freshwater teleost Danio rerio, following a full life-cycle exposure to environmentally relevant concentrations (390 to 14 423 ng/L). Considering that the mode of action (MoA) of MET on non-target organisms remains underexplored and that MET may act through similar human pathways, i.e., lipid and energy metabolisms, biochemical markers were used to determine cholesterol and triglycerides levels, as well as mitochondrial complex I activity in zebrafish liver. Also, the hepatosomatic index as an indication of metabolic disruption, and the expression levels of genes involved in MET's putative MoA, i.e. acaca, acadm, cox5aa, idh3a, hmgcra, prkaa1, were determined, the last by qRT-PCR. A screening of mRNA transcripts, associated with lipid and energy metabolisms, and other signaling pathways potentially involved in MET-induced toxicity were also assessed using an exploratory RNA-seq analysis. The findings here reported indicate that MET significantly disrupted critical biochemical and molecular processes involved in zebrafish metabolism, such as cholesterol and fatty acid biosynthesis, mitochondrial electron transport chain and tricarboxylic acid cycle, concomitantly to changes on the hepatosomatic index. Likewise, MET impacted other relevant pathways mainly associated with cell cycle, DNA repair and steroid hormone biosynthesis, here reported for the first time in a non-target aquatic organism. Non-monotonic dose response curves were frequently detected in biochemical and qRT-PCR data, with higher effects observed at 390 and 2 929 ng/L MET treatments. Collectively, the results suggest that environmentally relevant concentrations of MET severely disrupt D. rerio metabolism and other import

A tubular porous borosilicate membrane contactor was investigated for ozone gas/water mass transfer and the removal of contaminants of emerging concern (CECs) in water. Ozone gas/water contact occurs on the membrane shell-side, which is coated with a photocatalyst (TiO2-P25), as the ozone gas stream is fed from the lumen side and permeates through the pores generating micro-sized ozone bubbles uniformly delivered to the annular reaction zone where the contaminated water to be treated flows. Under continuous flow, water pH at 3.0 and temperature at 20 ºC, the volumetric mass transfer coefficient (KLa) ranged from 3.5 to 9.0 min−1 and improved with the increase of gas flow rate (QG, 1.5-fold from 0.15 to 1.0 Ndm3 min−1) and liquid flow rate (QL, 2.0-fold from 20 to 50 L h−1), due to enhanced turbulence on the membrane shell-side and annular zone. The mass transfer efficiency was more pronounced as the QG decreased and the QL increased, which is advantageous for large-scale applications. The main resistances to ozone transfer were in the water phase boundary layer (53–76%) and in the membrane (24–47%; kM = (1.14 ± 0.01) × 10−4 m s−1). For an ozone dose of 12 g m−3 and residence time of 3.9 s, removals ≥ 80% were achieved for 13 of 19 CECs spiked in demineralized water (each 10 µg L−1), demonstrating the applicability of this membrane contactor for ozonation treatment. Photocatalytic ozonation (O3/UVC/TiO2) did not significantly improve the treatment performance due to the low residence time inside the contactor.

The primary focus of environmental toxicological studies is to address the direct effects of chemicals on exposed organisms (parental generation – F0), mostly overlooking effects on subsequent non-exposed generations (F1 and F2 – intergenerational and F3 transgenerational, respectively). Here, we addressed the effects of simvastatin (SIM), one of the most widely prescribed human pharmaceuticals for the primary treatment of hypercholesterolemia, using the keystone crustacean Gammarus locusta. We demonstrate that SIM, at environmentally relevant concentrations, has significant inter and transgenerational (F1 and F3) effects in key signaling pathways involved in crustaceans’ neuroendocrine regulation (Ecdysteroids, Catecholamines, NO/cGMP/PKG, GABAergic and Cholinergic signaling pathways), concomitantly with changes in apical endpoints, such as depressed reproduction and growth. These findings are an essential step to improve hazard and risk assessment of biological active compounds, such as SIM, and highlight the importance of studying the transgenerational effects of environmental chemicals in animals’ neuroendocrine regulation

This study explores the combination of two sampling strategies (polar organic compounds integrative sampler (POCIS) vs. spot sampling) and four chromatographic retention modes (reversed-phase liquid chromatography (RPLC), hydrophilic interaction liquid chromatography (HILIC), mixed-mode liquid chromatography (MMLC) and supercritical fluid chromatography (SFC)) for high-resolution mass spectrometry (HRMS) screening of organic pollutants in water samples. To this end, a suspect screening approach, using iterative data-dependent tandem mass spectrometry (MS/MS) driven by a library of 3227 chemicals (including pharmaceuticals, pesticides, drugs of abuse, human metabolites, industrial chemicals and other pollutants), was employed. Results show that POCIS can afford a larger number of positive identifications as compared to spot sampling. On the other hand, the best suited retention mechanisms, in terms of identified analytes, are SFC, and followed by RPLC, MMLC and HILIC. However, the best combination (POCIS + SFC) would only allow the identification of 67% of the detected analytes. Thus, the combination of the two sampling strategies, spot and passive sampling, with two orthogonal retention mechanisms, RPLC and SFC, is proposed in order to maximize the number of analytes detected (89%). This strategy was applied to different surface water (river and estuary) samples from Galicia (NW Spain). A total of 155 compounds were detected at a confidence level 2a, from which the major class was pharmaceuticals (61%).

The bioaccumulation and depuration of seven organophosphorus flame retardants (OPFRs) in marine mussel Mytilus galloprovincialis were studied. OPFRs showed to be bioavailable in aquatic environments. When mussels are exposed to environmentally relevant concentrations of OPFRs, uptake kinetics fit well to a first-order model with a single compartment; in contrast depuration rates were generally underestimated by that model, most likely because it does not take into account the biotransformation of OPFRs by the organisms. The highest bioaccumulation rates were observed for tricresyl phosphate (TCrP), triphenyl phosphate (TPhP) and 2-ethylhexyldiphenylphosphate (EHDPP). This could be due to the presence of aryl groups in these compounds, their low solubility in water, and their affinity for fat tissues. According to these findings TCrP, with a BCF value of 4042 L kg−1 wet weight, should be classified in environmental regulations as an accumulative chemical

Plasticizers are chemical compounds used in the production of flexible plastics for a large variety of applications. They are present in most of the environments and, hence, we are highly exposed to them via several routes (ingestion, inhalation, etc). Due to the endocrine disruption potential of some of these chemicals and the unknown toxicological effects of their alternatives, assessing human exposure to these contaminants is an issue of emerging concern. Herein we propose an analytical methodology for the determination of several plasticizer metabolites in wastewater as a non-invasive, cheap, and fast exposure monitoring tool complementary to the analysis of urine. A solid-phase extraction procedure followed by an ultra(high)-performance liquid chromatography-tandem mass spectrometry method was optimized and validated for 21 analytes among phthalate, terephthalate, and di-iso-nonyl cyclohexane-1,2-dicarboxylate metabolites. Method quantification limits ranged from 0.079 to 4.4 ng L−1. The method was applied to the analysis of seven daily composite wastewater samples collected in the NW of Spain. Metabolites of low molecular weight phthalates and of di-2-ethylhexyl phthalate were quantified in all samples, despite the existing regulations limiting the use of phthalates. Metabolites of terephthalates, introduced at the end of the 20th century as phthalate substituents, were also quantified in all samples, being the first time that they were detected in this matrix. Exposure back-calculation highlighted di-2-ethylhexyl terephthalate as the second most common plastic additive after diethyl phthalate in the population considered, reflecting the increasing substitution of di-2-ethylhexyl phthalate by its analogous terephthalate

Several studies show that many water bodies in developing countries are increasingly affected by anthropogenic pressure, such as agricultural activities, domestic and industrial wastewater. However, data is scarce in several of such countries, including Panama. Thus, in this work, the ecotoxicological status of selected rivers in Panama with distinct input sources were evaluated using the zebrafish (Danio rerio) embryo bioassays combined with a liquid chromatography-high resolution mass spectrometry screening of contaminants of emerging concern (CECs), using a library of over 3200 chemicals. A total of 68 CECs, including pharmaceuticals and metabolites, pesticides and several industrial chemicals, could be tentatively identified. Additionally, the zebrafish embryo bioassays showed a significant increase (p < 0.05) in embryo mortality/abnormalities when incubated with water samples from two rivers, Matasnillo and Curundú (47.5% and 32%, respectively). Importantly, a positive correlation between ecotoxicological endpoints and some of the detected CECs was observed. The findings demonstrate that both rivers are under strong anthropogenic pressure, and therefore, management actions are urgently needed to decrease their level of contamination. Overall, this study further supports the use of the zebrafish embryo bioassay as a fast, high throughput approach for screening the toxicity of water samples, and highlights the advantages of combining ecotoxicological assays with high-resolution mass spectrometry to an expedite assessment of the ecotoxicological status of water bodies.

We have completed the characterization of the turbot (Scophthalmus maximus) myeloperoxidase (mpx) gene and protein, which we partially described in a previous study. The turbot mpx gene has 15 exons that encode a protein of 767 aa, with a signal peptide, propeptide and light and heavy chains, and also with haem cavities, a Ca+2-binding motif and several N- and O-glycosylation sites. The mature protein forms homodimers of about 150 kDa and is very abundant in turbot neutrophils. In addition to the mpx (epx2a) gene, another three peroxidase genes, named epx1, epx2b1 and epx2b2, were identified in the turbot genome. Epx1, Epx2b1 and Epx2b2 proteins also have signal peptides and many structural characteristics of mammalian MPO and eosinophil peroxidase (EPX). Mpx was strongly expressed in head kidney, while epx2b1 and epx2b2 were strongly expressed in the gills, and epx1 was not expressed in any of the tissues or organs analysed. In vitro stimulation of head kidney leucocytes with the parasite Philasterides dicentrarchi caused a decrease in mpx expression and an increase in epx2b1 expression over time. In turbot infected experimentally with P. dicentrarchi a significant increase in mpx expression in the head kidney was observed on day 7 postinfection, while the other genes were not regulated. However, mpx, epx2b1 and epx2b2 were downregulated in the gills of infected fish, and epx1 expression was not affected. These results suggest that the four genes responded differently to the same stimuli. Interestingly, BLAST analysis revealed that Epx1 and Mpx showed greater similarity to mammalian EPX than to MPO. Considering the phylogenetic and synteny data obtained, we concluded that the epx/mpx genes of Gnathostomes can be divided into three main clades: EPX1, which contains turbot epx1, EPX2, which contains turbot mpx (epx2a) and epx2b1 and epx2b2 genes, and a clade containing mammalian EPX and MPO (EPX/MPO). EPX/MPO and EPX2 clades share a common ancestor with the chondrich

Supercritical fluid chromatography (SFC), combined with mass spectrometry (MS), was employed for the determination of five chiral fungicides, from two different chemical families (acylalanine and triazol) in wine and vineyard soils. The effect of different SFC parameters (stationary phase, chiral selector, mobile phase modifier and additive) in the resolution between enantiomers and in the efficiency of compounds ionization at the electrospray source (ESI) was thorougly described. Under final working conditions, chiral separations of selected fungicides were achieved using two different SFC-MS methods, with an analysis time of 10 min and resolution factors from 1.05 to 2.45 between enantiomers. In combination with solid-phase extraction and pressurized liquid extraction, they permitted the enantiomeric determination of target compounds in wine and vineyard soils with limits of quantification in the low ppb range (between 0.5 and 2.5 ng mL−1, and from 1.3 to 6.5 ng g−1, for wine and soil, respectively), and overall recoveries above 80%, calculated using solvent-based standards. For azolic fungicides (tebuconazole, myclobutanil and penconazole) soil dissipation and transfer from vines to wines were non-enantioselective processes. Data obtained for acylalanine compounds confirmed the application of metalaxyl (MET) to vines as racemate and as the R-enantiomer. The enantiomeric fractions (MET-S/(MET-S+MET-R)) of this fungicide in vineyard soils varied from 0.01 to 0.96; moreover, laboratory degradation experiments showed that the relative dissipation rates of MET enantiomers varied depending on the type of soil

Sludge from sewage treatment plants (STPs) is recognized as a sink of moderate to high lipophilic compounds resistant to biodegradation. Herein, we investigate the presence of emerging pollutants in sewage sludge combining the information provided by mass spectrometry detection, following ultra-performance liquid chromatography (UPLC), with the use of an accurate spectral database of pesticides and pharmaceuticals. In a first step, the performance of matrix solid-phase dispersion, as sample preparation technique, and two non-target data acquisition strategies (data dependent, DDA, and data independent analysis modes, DIA), used in combination with a UPLC quadrupole time-of-flight system, are assessed using a selection of deuterated compounds added either to freeze-dried sludge samples, or to sludge extracts. Possibilities and limitations of both modes are discussed. Following the DDA approach, a group of 68 micropollutants was identified in sludge from different STPs. Some of them are reported in this compartment for the first time. Finally, semi-quantitative concentration data are reported for a group of 37 pollutants in samples obtained from 16 STPs. Out of them, 10 pharmaceuticals, showing detection frequencies and median sludge residues above 50% and 100 ng g−1, respectively; are highlighted as pollutants to be monitored in sludge in order to understand their behaviour during the wastewater treatment

Amphetamine (AMP), methamphetamine (MAMP) and 3,4-methylenedioxymethamphetamine (MDMA) occur in wastewater not only as a result of illicit consumption, but also, in some cases, from prescription drug use or by direct drug disposal into the sewage system. Enantiomeric profiling of these chiral drugs could give more insight into the origin of their occurrence. In this manuscript, a new analytical methodology for the enantiomeric analysis of amphetamine-like substances in wastewater has been developed. The method consists of a solid-phase extraction (SPE) followed by liquid chromatography-triple quadrupole-tandem mass spectrometry (LC-MS/MS), which showed low quantification limits in the 2.4–5.5 ng L−1 range. The LC-MS/MS method was first applied to characterize a total of 38 solid street drug samples anonymously provided by consumers. The results of these analysis showed that AMP and MDMA trafficked into Spain are synthesized as racemate, while MAMP is exclusively produced as the S(+)-enantiomer. Then, the analytical method was employed to analyse urban wastewater samples collected from the wastewater treatment plants (WWTPs) of five different cities in 2018 and 2019. Consumption estimated through normalized population loads in wastewater showed an increased pattern of AMP use in the Basque Country. Furthermore, the enantiomeric profiling of wastewater samples was contrasted to lisdexamfetamine (LIS) and selegiline (SEL) prescription figures, two pharmaceuticals which metabolize to S(+)-AMP, and to R(-)-AMP and R(-)-MAMP, respectively. From this analysis, and considering uncertainties derived from metabolism and adherence to treatment, it was concluded that LIS is a relevant source of AMP in those cases with low wastewater loads, i.e. up to a maximum of 60% of AMP detected in wastewater in some samples could originate from LIS prescription, while SEL does not represent a significant source of AMP nor MAMP. Finally, removal efficiencies could be evaluated for the WWTP

Data obtained from wastewater analysis can provide rapid and complementary insights in illicit drug consumption at community level. Within Europe, Spain is an important country of transit of both cocaine and cannabis. The quantity of seized drugs and prevalence of their use rank Spain at the top of Europe. Hence, the implementation of a wastewater monitoring program at national level would help to get better understanding of spatial differences and trends in use of illicit drugs. In this study, a national wastewater campaign was performed for the first time to get more insight on the consumption of illicit drugs within Spain. The 13 Spanish cities monitored cover approximately 6 million inhabitants (12.8% of the Spanish population). Untreated wastewater samples were analyzed for urinary biomarkers of amphetamine, methamphetamine, MDMA, cocaine, and cannabis. In addition, weekend samples were monitored for 17 new psychoactive substances. Cannabis and cocaine are the most consumed drugs in Spain, but geographical variations showed, for instance, comparatively higher levels of methamphetamine in Barcelona and amphetamine in Bilbao, with about 1-fold higher consumption of these two substances in such metropolitan areas. For amphetamine, an enantiomeric profiling was performed in order to assure the results were due to consumption and not to illegal dumping of production residues. Furthermore, different correction factors for the excretion of cannabis were used to compare consumption estimations. All wastewater results were compared with previously reported data, national seizure data and general population survey data, were a reasonable agreement was found. Daily and yearly drug consumption were extrapolated to the entire Spanish population with due precautions because of the uncertainty associated. These data was further used to estimate the retail drug market, where for instance cocaine illicit consumption alone was calculated to contribute to 0.2–0.5% of the Spanish

This work investigates the reaction of 1-vinyl-2-pyrrolidinone (VP) and 2-piperazin-1-yletanamine (PPE) under UV radiation. Both substances are high-volume production chemicals (production >1000 tons/year) widely used in polymers, coatings and a wide array of applications, which have been classified as mobile chemicals and which can then lead to the formation of persistent and mobile transformation products (TPs). Thus, their reaction with UV light was studied by means of liquid chromatography-quadrupole-time-of-flight-mass spectrometry (LC-QTOF-MS). Both compounds presented a high reactivity, the VP quantum yield was 0.28 mol/E; whereas, PPE had a quantum yield notably higher than 1 (16 mol/E). Five and 7 TPs were identified for VP and PPE, respectively. Some of them had been already reported in literature due to sunlight photodegradation or other oxidation processes, but most of them are reported here for the first time. Finally, the acute and chronical toxicity of precursors and TPs were estimated using two quantitative structure-activity relationship (QSAR) software tools which led to some discrepancies in the estimations, pointing to the need for experimental toxicity assays for these compounds

This work studies the chlorination and monochloramination reaction kinetics of two phenazone-type drugs (phenazone – Phe and propyphenazone – PrPhe) and three metabolites of phenazone-type drugs (4-formylaminoantipyrine – FAA, 4-aminoantipyrine - AA and 4-acetoamidoantipyrine - AAA). Kinetics were faster with chlorine (apparent second-order constants between 100 and 66,500 times higher) than with monochloramine. For FAA and AAA, no significant reaction was observed during monochloramination. Further, apparent rate constants decreased as the pH increased from pH 5.7 to 8.3, except during chlorination of AA. The transformation products (TPs) formed were also elucidated by liquid chromatography-high resolution mass spectrometry. The main transformation pathway for Phe and PrPhe consisted of halogenations, hydroxylations and dealkylations, while AAA and FAA were firstly transformed to AA, then followed by pyrazole ring opening and hydroxylations. The extend of the reaction was also tested in real water samples, where, in general, slower reaction kinetics were obtained during monochloramination, while the chlorination reaction showed similar half-lives to ultrapure water. Finally, acute and chronic toxicity of the TPs were estimated using two quantitative structure-activity relationship (QSAR) software (ECOSAR and TEST), showing that some TPs could be more toxic than their precursor compounds

Background In this study, an alternative and complementary method to those approaches currently used to estimate alcohol consumption by the population is described. This method, known as wastewater-based epidemiology (WBE), allows back-calculating the alcohol consumption rate in a given population from the concentrations of a selected biomarker measured in wastewater. Methods Composite (24-h) wastewater samples were collected at the inlet of 17 wastewater treatment plants located in 13 Spanish cities for seven consecutive days in 2018. The sampled area covered 12.8% of the Spanish population. Wastewater samples were analyzed to determine the concentration of ethyl sulfate, the biomarker used to back-calculate alcohol consumption. Results Alcohol consumption ranged from 4.5 to 46 mL/day/inhabitant. Differences in consumption were statistically significant among the investigated cities and between weekdays and weekends. WBE-derived estimates of alcohol consumption were comparable to those reported by its corresponding region in the Spanish National Health Survey in most cases. At the national level, comparable results were obtained between the WBE-derived annual consumption rate (5.7 ± 1.2 L ethanol per capita (aged 15+)) and that reported by the National Health Survey (4.7 L ethanol per capita (aged 15+)). Conclusions This is the largest WBE study carried out to date in Spain to estimate alcohol consumption rates. It confirms that this approach is useful for establishing spatial and temporal patterns of alcohol consumption, which could contribute to the development of health care management plans and policies. Contrary to established methods, it allows obtaining information in a fast and relatively economical way

The hypothesis that exposure to certain environmental chemicals during early life stages may disrupt reproduction across multiple non-exposed generations has significant implications for understanding disease etiology and adverse outcomes. We demonstrate here reproductive multi and transgenerational effects, at environmentally relevant levels, of one of the most prescribed human pharmaceuticals, simvastatin, in a keystone species, the amphipod Gammarus locusta. The transgenerational findings has major implications for hazard and risk assessment of pharmaceuticals and other contaminants of emerging concern given that transgenerational effects of environmental chemicals are not addressed in current hazard and risk assessment schemes. Considering that the mevalonate synthesis, one of the key metabolic pathways targeted by simvastatin, is highly conserved among metazoans, these results may also shed light on the potential transgenerational effects of simvastatin on other animals, including humans.

The presence of fungicide and insecticide residues in wine has been largely investigated. However, few studies have addressed the persistence of these compounds in vineyard soils. In this research, we investigate the residues of a relevant number of fungicides and insecticides in vineyard soils, obtained in the Northwest of Spain, at the beginning of each agriculture campaign. Moreover, the dissipation of species showing high concentrations were monitored during the non-vegetative period of vines, in order to understand their soil evolution between application campaigns. To this end, a multiresidue analytical procedure based on pressurized liquid extraction (PLE) followed by liquid chromatography tandem mass spectrometry (LC-MS/MS) determination was first optimized. Under final working conditions, absolute recoveries in the range from 70 to 130% were achieved for 44 out of 51 selected compounds. The method LOQs remained at the low ng g−1 level (0.2–13 ng g−1) with a linear response range up to 500 ng g−1. Analysis of vineyard soils, collected during a 2-year period, from a geographic area with a high incidence of fungal diseases, demonstrated the presence of relevant concentrations of several fungicides and the insecticide imidacloprid (IMI) in this compartment. Most compounds detected at the end of the application season remained in soil at the beginning of the next year campaign. Among them, six fungicides (dimethomorph, boscalid, myclobutanil, penconazole, pyraclostrobin and pyrimethanil) and IMI showed average dissipation efficiencies below 50%, so they pose a potential to accumulate in this kind of soils

The comprehensive identification of organic species existing in indoor environments is a key issue to understand their impact in human health. This study proposes the analysis of condensed water samples, collected with portable dehumidifiers, to characterize semi-volatile compounds in the gas phase of confined areas. Water samples are concentrated by solid-phase extraction (SPE). The obtained extracts are analysed by gas chromatography (GC) time-of-flight mass spectrometry (TOF-MS), following a non-target screening data mining approach. In first term, spectra of deconvoluted compounds are compared with those in NIST low resolution library; thereafter, tentative identifications are verified using an in-house database of accurate electron ionization (EI) MS spectra. Chromatographic (retention index) and spectral data are combined for unambiguous species identification. The potential of condensed water samples to reflect changes in the composition of indoor atmospheres, the match between data obtained using different dehumidifiers, and the relative concentration efficiency of condensed water compared to that attained by active sampling of moderate air volumes are discussed. A total of 141 semi-volatile compounds were identified (98 confirmed against authentic standards) in a set of 21 samples obtained from different homes and working places. This list contains more than 40 fragrances (including several potential allergens), solvents and intermediates in the production of polymeric materials, plasticizers and flame retardants

Wastewater-based epidemiology (WBE) has become a very useful tool to monitor a population's drug consumption or exposure to environmental and food contaminants. In this work, WBE has been applied to estimate tobacco consumption in seven Spanish regions. To this end, 24 h composite wastewater samples were taken daily for one week in 17 wastewater treatment plants, covering altogether a population of ca. 6 million inhabitants. The samples were treated by enzymatic deconjugation and the wastewater content of two human-specific nicotine metabolites (namely, cotinine and trans-3′-hydroxycotinine) was measured to estimate the daily consumption of nicotine. The population-weighted average nicotine consumption in the seven analyzed regions was 2.2 g/(day∙1000 inh.), without any daily pattern. This average estimated nicotine consumption value agreed with the value derived from official tobacco sales data. Differences in consumption among the seven studied regions were found, being Galicia, the region with the lowest rate, and the Basque Country and Catalonia those with the highest rates. However, no conclusive correlation was found between those values and the prevalence data taken from two different national surveys, nor sociodemographic and health data. This study demonstrates that this tool can complement other indicators in order to accurately assess tobacco consumption rates at regional and national levels and provides the most extensive application of the approach in the Spanish territory.

A new analytical method for the determination of 22 perfluoroalkylated (carboxylic and sulfonic) acids in water samples is presented. The method's objective was to achieve the simultaneous quantification of compounds with different chain lengths (from C1 to C18). To this end, 500 mL of water were extracted with Oasis WAX solid-phase extraction cartridges and eluted with 3 mL of 5% ammonia in methanol. After evaporation to dryness, extracts were reconstituted in methanol:ultrapure water (1:1) and analyzed by mixed-mode liquid chromatography-tandem mass spectrometry (MMLC-MS/MS) using a weak anion exchange/reversed-phase column. The method provided good results, with limits of quantification lower than 1 ng/L in river water for most of compounds, except the two perfluorocarboxylic acids with the longest alkyl chain (>C14) and trifluoroacetic acid, for which a blank contamination problem was observed. The method proved good trueness and precision in both ultrapure and river water (R ≥ 81%, RSD ≤ 15%). After validation, the method was applied to the analysis of nine water samples where nine perfluoroalkylated acids were quantified. Seven of them were ultrashort- (C1-C4) and short-chain (C4-C8) perfluoroalkylated acids, pointing out the importance of developing methods capable to target such substances for further monitoring

A novel approach based on the direct pulsed irradiation of UV-C light onto ibuprofen (IBP) solutions was evaluated in this work, as proof of concept for the direct removal of micropollutants. The experiments confirmed that laser irradiation is able to completely degrade IBP in 15 min in distilled water, with a DOC depletion of ca. 25% and with transformation products (TPs) remaining in solution and estimated to represent ca. 10% of the initial IBP concentration. In wastewater spiked samples, removal efficiency is slightly lower but still significant (ca. 5% IBP remaining after 15 min). Hence, this work suggests that low power solid state pulsed lasers, emitting at 266 nm wavelength, show promise for the removal of these type of micropollutants from water. These results open new opportunities towards the development of chemical-free water treatment methods based on direct, selective irradiation using state of the art, miniaturized laser devices

An analytical method based on high-resolution quadrupole–time-of-flight (QToF) mass spectrometry has been developed as an alternative to the classical method, using a low-resolution ion trap (IT) analyzer to reduce interferences in N-nitrosamines determination. Extraction of the targeted compounds was performed by solid-phase extraction (SPE) following the United States Environmental Protection Agency (USEPA) -521 method. First, both electron impact (EI) and positive chemical ionization (PCI) using methane as ionization gas were compared, along with IT and QToF detection. Then, parameters such as limits of detection (LOD) and quantification (LOQ), linearity, and repeatability were assessed. The results showed that the QToF mass analyzer combined with PCI was the best system for the determination of the N-nitrosamines, with instrumental LOD and LOQ in the ranges of 0.2–4 and 0.6–11 ng mL−1, respectively, which translated into method LOD and LOQ in the ranges of 0.2–1.3 and 0.6–3.9 ng L−1, respectively. The analysis of real samples showed the presence of 6 of the N-nitrosamines in influent, effluent, and tap water. N-nitrosodimethylamine (NDMA) was quantified in all the analyzed samples at concentrations between 1 and 27 ng L−1. Moreover, four additional nitrosamines were found in tap and wastewater samples

The use of transcriptomics data brings new insights and works as a powerful tool to explore the molecular mode of action (MoA) of transgenerational inheritance effects of contaminants of emerging concern. Therefore, in this dataset, we present the transcriptomic data of the transgenerational effects of environmentally relevant simvastatin levels, one of the most prescribed human pharmaceuticals, in the keystone amphipod species Gammarus locusta. In summary, G. locusta juveniles were maintained under simvastatin exposure up to adulthood (exposed group - F0E) and the offspring of F0E were transferred to control water for the three subsequent generations (transgenerational group - F1T, F2T and F3T). To gain insights into the biological functions and canonical pathways transgenerationally disrupted by simvastatin, a G. locusta de novo transcriptome assembly was produced and the transcriptomic profiles of three individual G. locusta females, per group, over the four generations (F0 to F3) - solvent control groups (F0.C, F1.C, F2.C and F3.C), F0 320 ng/L simvastatin exposed group (F0.320E) and F1 to F3 320 transgenerational group (F1.320T; F2.320T and F3.320T) - were analyzed. Briefly, Illumina HiSeq™ 2500 platform was used to perform RNA sequencing, and due to the unavailability of G. locusta genome, the RNA-seq datasets were assembled de novo using Trinity and annotated with Trinotate software. After assembly and post-processing steps, 106093 transcripts with N50 of 2371 bp and mean sequence length of 1343.98 bp was produced. BUSCO analyses showed a transcriptome with gene completeness of 97.5 % Arthropoda library profile. The Bowtie2, RSEM and edgeR tools were used for the differential gene expression (DEGs) analyses that allowed the identification of a high quantity of genes differentially expressed in all generations. Finally, to identify the main metabolic pathways affected by the transgenerational effects of SIM across all generations, the DGEs genes were blasted o

The coexistence of the anilinopyrimidine fungicides pyrimethanil (PYR) and cyprodinil (CYP), and suspected metabolites in wine samples was investigated by liquid chromatography (LC) with tandem mass spectrometry (MS/MS), based on triple quadrupole (QqQ) and quadrupole time-of-flight (QTOF) MS instruments. For the first time, quantitative data obtained after solid-phase extraction (SPE) of wine samples have demonstrated the systematic presence of 4-hydroxyanilino derivatives of PYR and CYP in wines containing residues of parent fungicides, at concentrations from 0.2 to 58 ng mL−1. Higher concentration ratios (hydroxylated derivative/active fungicide) were measured in red than in white wines, particularly in case of PYR. On average, the concentrations of PYR-4OH were twice those measured for PYR in red wines. A targeted search of hydroxyl derivatives in wine extracts by LC-QTOF-MS showed the existence of additional hydroxylation positions in the pyrimidine ring and/or in the alkyl substituents bond to this cycle in the structure of both anti-botrytis fungicides. Moreover, free and glycosylated forms of the hydroxylated metabolites for both fungicides coexist in wine samples. In case of CYP, it is proved that hydroxylated and glycosylated metabolites are already present in grapes before vinification

El análisis de aguas residuales con fines epidemiológicos es actualmente una herramienta fiable y complementaria a las metodologías basadas en indicadores tradicionales para el control de diferentes sustancias entre las que cabe destacar las drogas. Si bien varios países europeos la utilizan como herramienta de trabajo para la monitorización de drogas de abuso, en España su uso se limita principalmente a estudios realizados por diferentes grupos de investigación, tal y como se describe en el caso práctico puesto de ejemplo en el artículo. Sin embargo, el potencial de la metodología ha quedado evidenciado en los estudios científicos llevados a cabo tanto a nivel español como internacional y, aunque son necesarios más estudios para llegar a conocer todo su potencial, se prevé pueda ser incorporada como herramienta de trabajo complementaria a las que habitualmente se utilizan. En este sentido, la Red Española de Análisis de Aguas Residuales (ESAR-Net), creada en 2017 y formada por diferentes grupos de investigación españoles, pretende contribuir al conocimiento y aplicación de esta metodología en España a través de actividades científicas y de divulgación.

Herein, a method for the simultaneous determination of the currently prescribed sartan drugs (eprosartan, EPR; olmesartan, OLM; losartan, LOS; candesartan, CAN; telmisartan, TEL; irbesartan, IRB; and valsartan, VAL), and the biodegradation product valsartan acid (VALA), in water samples (raw and treated wastewater, river and tap water) was developed. Solid-phase extraction (SPE) and ultra-performance liquid chromatography (UPLC) tandem mass spectrometry (MS/MS) were employed as concentration and determination techniques, respectively. Different sorbents and elution solvents were tested for sample preparation. Under optimized conditions, samples at neutral pH (6–8 units) were concentrated using mixed-mode (reversed-phase and anionic exchange) cartridges. Thereafter, the sorbent was washed with 5 mL of a methanol: water (1:1) solution, dried under a nitrogen stream and compounds were eluted with 2 mL of methanol: NH3 (98:2). The accuracy of the method (accounting for SPE efficiency and matrix effects during electrospray ionization) was investigated using solvent-based calibration standards. Global recoveries, obtained for different water matrices (tap, river, treated and raw wastewater), ranged from 82% to 134%, with standard deviations between 2 and 18%. LOQs varied from 2 to 50 ng L−1. Analysis of un-spiked samples confirmed: (1) the incomplete removal of sartans at sewage treatment plants (STPs), (2) the formation of VALA during municipal water treatment, and (3) the presence of VALA in the processed tap water samples. Additional findings of the current study are the detection of hydroxylated derivatives of the sartan drugs IRB and LOS in wastewater, and the E-Z isomerization of EPR in environmental water samples

The presence of persistent and mobile organic contaminants (PMOC) in aquatic environments has become a matter of concern due to their ability of breaking through natural and anthropogenic barriers, even reaching drinking water. The presence of many of these compounds in surface and drinking water has been reported in screening studies, but there is still a lack of analytical methods capable of quantifying them. Herein, we propose a method combining mixed-mode-solid-phase extraction (MM-SPE) as preconcentration technique and mixed-mode liquid chromatography (MMLC) coupled to tandem mass spectrometry as a determination technique for the quantitative determination of 23 target PMOCs in surface and drinking water samples. When compared to reversed-phase liquid chromatography, the MMLC protocol has proven to be superior in both retentive capabilities and peak shape for ionic compounds, while performing also well for neutrals. The overall method performance was satisfactory with limits of quantification under 50 ng L–1 for most of analytes in both surface and drinking water. The relative standard deviation was lower than 20%, and the average recovery was 78 and 80% in surface and drinking water, respectively. The method was applied to 15 water samples collected in Spain, where 17 out of the 23 target PMOCs were quantified in at least one sample. Among them, 6 chemicals (e.g., benzyltrimethylammonium) are reported and/or quantified here for the first time

Cow routines and behavioral responses are altered substantially following the installation of robot milking. The present study was designed to analyze the effect that switching from milking parlor to automatic milking system (AMS) had on the culling rate (due to various causes) of dairy cattle. For this purpose, culling records and causes for culling were tracked in 23 dairy farms in the Galicia region (NW Spain). The animals in these farms were monitored for 5 years. For the present study, that length of time was divided into three different stages, as follows: 2 years before switching from a milking parlor to AMS (stage 1), the 1st year following the implementation of AMS (stage 2) and the 2nd and 3rd years succeeding the implementation of AMS (stage 3). Cox models for survival analysis were used to estimate the time to culling due to different reasons during stage 1 in relation to stages 2 and 3. The data indicated that the risk of loss due to death or emergency slaughter decreased significantly following the installation of AMS. In contrast, the risk of culling due to low production, udder problems, infertility or lameness increased significantly. Low-production cows (such as cows in advanced lactation due to infertility) or sick cows (such as mastitic or lame cows) allegedly have a noticeable effect both on the performance and the amortization of the cost of AMS, which in turn would lead to a higher probability of elimination than in conventional systems

The presence of pollutants in estuary and oceanic systems is a global problem and a serious concern to human and environmental health. Usually, environmental monitoring studies consider classical persistent organic pollutants (POPs). However, the lists of POPs keep continuously growing and new POPs and other emerging pollutants should be considered in new monitoring programs. So, this study aimed to investigate the distribution and profile of classical POPs (polychlorinated biphenyl (PCBs), organochlorine pesticides (OCPs), and polycyclic aromatic hydrocarbons (PAHs)), new POPs and emerging pollutants (polybrominated diphenyl ethers (PBDEs), perfluorinated compounds (PFCs), novel halogenated flame retardants (NFRs) and UV filters) in bivalve mollusc samples (both raft-cultivated and wild mussel, Mytilus galloprovincialis; cockle, Cerestoderma edule; and clam, Ruditapes descussatus) collected in nine Galician Rias during the period February 2012 to February 2013. A predominance of PAHs (6.8–317 ng/g dry weight (dw)) followed by PCBs (0.47–261 ng/g dw), UV filters (1.4–157 ng/g dw), PFCs (0.53–62 ng/g dw), OCPs (0.07–29 ng/g dw), PBDEs (0.31–6.6 ng/g dw) and NFRs (0.07–3.2 ng/g dw) was found in the studied bivalves, being the UV filter octocrylene the compound found at the highest concentration (141 ng/g dw in a cockle sample), while the PAHs chrysene and benzo(b)fluoranthene were the compounds with the highest average concentration (20 and 14 ng/g dw, respectively). Inter-species, temporal and geographical variations on pollutants concentration were assessed by multifactorial analysis of variance. Statistically significant differences among the type of mollusc were observed for levels of organochlorinated and organobrominated pollutants considered (PCBs, OCPs and PBDEs), which were detected at higher concentrations in wild mussel. On the other hand, the main PFCs and UV filters showed a higher detection frequency in cockle samples. Location played significant role fo

Global production of synthetic polymers, led by polyethylene (PE), rose steadily in the last decades, and marine ecosystems are considered as a global sink. Although PE is not biodegradable, in coastal areas it fragments into microplastics (MP) readily taken up by biota, and have been postulated as vectors of hydrophobic chemicals to marine organisms. We have tested this hypothesis using two organisms representative of the marine plankton, the holoplanktonic copepod Acartia clausi, and the meroplanktonic larva of the Paracentrotus lividus sea-urchin, and two model chemicals with similar hydrophobic properties, the 4-n-Nonylphenol and the 4-Methylbenzylidene-camphor used as plastic additive and UV filter in cosmetics. Both test species actively ingested the MP particles. However, the presence of MP never increased the bioaccumulation of neither model chemicals, nor their toxicity to the exposed organisms. Bioaccumulation was a linear function of waterborne chemical disregarding the level of MP. Toxicity, assessed by the threshold (EC10) and median (EC50) effect levels, was either independent of the level of MP or even in some instances significantly decreased in the presence of MPs. These consistent results challenge the assumption that MP act as vectors of hydrophobic chemicals to planktonic marine organisms

Lagrangian ocean analysis, where virtual parcels of water are tracked through hydrodynamic fields, provides an increasingly popular framework to predict the dispersal of water parcels carrying particles and chemicals. We conduct the first direct test of Lagrangian predictions for emerging contaminants using (1) the latitude, longitude, depth, sampling date, and concentrations of UV filters in raft cultured mussel (Mytilus galloprovincialis) of the estuary Ria de Arousa, Spain (42.5°N, 8.9°W); (2) a hydrodynamic numerical model at 300 m spatial resolution; and (3) a Lagrangian dispersion scheme to trace polluted water parcels back to pollution sources. The expected dispersal distances (mean ± SD) are 2 ± 1 km and the expected dispersal times (mean ± SD) are 6 ± 2 h. Remarkably, the probability of dispersal of UV filters from potential sources to rafts decreases 5-fold over 5 km. In addition to predicting dispersal pathways and times, this study also provides a framework for quantitative investigations of concentrations of emerging contaminants and source apportionment using turbulent diffusion. In the coastline, the ranges of predicted concentrations of the UV-filters 4-methylbenzylidene-camphor, octocrylene, and benzophenone-4 are 3.2 × 10–4 to 0.023 ng/mL, 2.3 × 10–5 to 0.009 ng/mL, and 5.6 × 10–4 to 0.013 ng/mL, respectively. At the outfalls of urban wastewater treatment plants these respective ranges increase to 8.9 × 10–4 to 0.07 ng/mL, 6.2 × 10–5 to 0.027 ng/mL, and 1.6 × 10–3 to 0.040 ng/mL.

This works investigates the chlorination and bromination of two rubber and polymer related chemicals, which have emerged as relevant water contaminants, i.e. 1,3-di-o-tolylguanidine (DTG) and 1,3-diphenylguanidine (DPG). Kinetic constants at different pH values were obtained and modelled, taking into account the pKa values of DTG/DPG and HClO, showing that the maximum reaction rate (kapp > 104 M−1 s−1) is obtained at pH values 8.8 for DPG and 9.1 for DTG. Bromination is also very fast, although unlike chlorination, deviation from the model was observed at neutral pH, which was attributed to formation of metastable transformation product (TP). A total of 35 TPs, corresponding to halogenation, hydroxylation, formation of monophenylguanidine derivatives and cyclization reactions, were tentatively identified. Furthermore it was found that chloroform can be formed up to a 25% molar yield, while dichloroacetonitrile was formed into less than a 3% yield. Several ecotoxicological endpoints were predicted by quantitative structure–activity relationship models (QSAR) for the TPs, some of which were predicted to be more toxic than DPG/DTG. Also a chlorinated solution investigated by a Vibrio Fisheri acute toxicity test, confirmed that toxicity increases with chlorination.

This work provides a new analytical method for the determination of cocaine, its metabolites benzoylecgonine and cocaethylene, the pyrolytic products anhydroecgonine and anhydroecgonine methyl ester, and the pharmaceutical levamisole in wastewater. Samples were solid‐phase extracted and extracts analyzed by liquid chromatography–tandem mass spectrometry using, for the first time in the illicit drug field, a stationary phase that combines reversed‐phase and weak cation‐exchange functionalities. The overall method performance was satisfactory, with limits of detection below 1 ng/L, relative standard deviations below 21%, and percentages of recovery between 93% and 121%. Analysis of 24‐hour composite raw wastewater samples collected in Santiago de Compostela (Spain) and Brasilia (Brazil) highlighted benzoylecgonine as the compound showing the highest population‐normalized mass loads (300–1000 mg/day/1000 inhabitants). In Brasilia, cocaine and levamisole loads underwent an upsurge on Sunday, indicating a high consumption, and likely a direct disposal, of cocaine powder on this day. Conversely, the pyrolytic product resulting from the smoke of crack, anhydroecgonine methyl ester, and its metabolite anhydroecgonine were relatively stable over the four days, agreeing with a non‐recreational‐associated use of crack

Like other ciliates, Philasterides dicentrarchi, the scuticociliate parasite of turbot, produces a feeding-only or growing stage called a trophont during its life cycle. Exposure of the trophonts to heat-inactivated serum extracted from the turbot host and containing specific antibodies that induce agglutination/immobilization leads to the production of a mucoid capsule from which the trophonts later emerge. We investigated how these capsules are generated, observing that the mechanism was associated with the process of exocytosis involved in the release of a matrix material from the extrusomes. The extruded material contains mucin-like glycoproteins that were deposited on the surface of the cell and whose expression increased with time of exposure to the heat-inactivated immune serum, at both protein expression and gene expression levels. Stimulation of the trophonts with the immune serum also caused an increase in discharge of the intracellular storage compartments of calcium necessary for the exocytosis processes in the extrusomes. The results obtained suggest that P. dicentrarchi uses the extrusion mechanism to generate a physical barrier protecting the ciliate from attack by soluble factors of the host immune system. Data on the proteins involved and the potential development of molecules that interfere with this exocytic process could contribute to improving the prevention and control of scuticociliatosis in turbot

In this work, fabric phase sorptive extraction (FPSE) is investigated for the extraction and preconcentration of ultra-trace level residues of fungicides (19 compounds) and insecticides (3 species) in wine samples. Subsequently, the preconcentrated analytes are selectively determined using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Parameters affecting the efficiency and repeatability of the extraction are evaluated in depth; moreover, the proposed method is characterized in terms of linear response range, trueness, precision and limits of quantification (LOQs). The set-up of the extraction process and the type of coating were the variables exerting the most prominent effects in the repeatability and the yield of the extraction, respectively. Under optimized conditions, samples (10 mL of wine diluted with the same volume of ultrapure water) were extracted with a small amount of cellulose fabric (3 discs with 4 mm of diameter: total surface 0.38 cm2) coated with a sol-gel polyethylene glycol sorbent (sorbent amount 3.3 mg), immersed in the diluted sample, without being in direct contact with the PTFE covered magnetic stir bar. Following the overnight extraction step, analytes were quantitatively recovered using only 0.3 mL of an ACN-MeOH (80:20) mixture. Under equilibrium sampling conditions, the linear response range of the method varied from 0.2 to 200 ng mL−1, with limits of quantification (LOQs) between 0.03 and 0.3 ng mL−1. Relative recoveries ranged from 77 ± 6% to 118 ± 4%, and from 87 ± 4% to 121 ± 6% for red and white wines, respectively. Application of the optimized method to commercial wines demonstrated the existence of up to 9 out of 22 investigated compounds in the same wine sample. The compound identified at the highest concentration was iprovalicarb (IPR), with a value of 130 ± 9 ng mL−1 in a commercial white wine

Here we propose a single acid digestion (SAD) sample preparation method for ICP-MS analysis of animal serum samples to determine trace element contents. The method was evaluated in comparison with a commonly used procedure involving dilution of samples in an alkaline solution (AKD). In the SAD procedure, aliquots (1 mL) of bovine serum samples were treated at low temperature with a mixture of concentrated nitric acid and hydrogen peroxide. Trace elements (As, B, Ba, Cd, Co, Cr, Cu, Fe, Hg, Li, Mn, Mo, Ni, Pb, Sb, Se, Sr, U, and Zn) were directly determined by ICP-MS analysis of diluted solutions of samples. Both methods were sufficiently sensitive to enable quantification of most trace elements, with the exception of the AKD method for Cd, Hg and Pb. The quality of the data was verified by using certified reference material. Good results were obtained for the SAD procedure and all elements, but recoveries were unacceptable with the AKD procedure for Se (recovery: 57%), Cd (154%) and Fe (139%). Strong associations (R2 >0.90, P = 0.000) between the data obtained by both methods were demonstrated for the elements considered. The proposed SAD sample preparation method produced satisfactory results for determining most toxic and essential trace elements targeted in monitoring studies.

The analysis of wastewater for the determination of human biomarkers of exposure (human metabolites) is a non-intrusive, economic and complementary alternative to the analysis of urine in the monitoring of human exposure to chemicals of concern. This study provides the first gas chromatography-based method for the determination of three metabolites of chlorinated organophosphorous flame retardants (OPFRs: bis(2-chloroethyl) phosphate, bis(chloropropyl) phosphate and bis(1,3-dichloro-2-propyl) phosphate) in wastewater. A solid-phase extraction procedure based on the use of mixed-mode reversed-phase weak anion exchange sorbents was optimized including a fractionated elution of OPFRs and their metabolites. Analytes derivatization was investigated by comparing two silylating reagents, N-tert-butyldimethylsilyl-N-methyltrifluoroacetamide and N-methyl-N-(trimethylsilyl)trifluoroacetamide, the first one providing better results. Determination was performed by gas chromatography-high resolution mass spectrometry with a quadrupole-time-of-flight system (GC-QTOF) in order to improve selectivity. Furthermore, the use of GC-QTOF combined with the specific ion obtained from silylated metabolites (m/z 154.9924) can be exploited to screen for other phosphate ester metabolites. Under final conditions, the overall method performance was satisfactory, affording method detection limits ranging from 1.1 to 4.6 ng/L, percentages of recovery from 90% to 110%, and relative standard deviations below 13%. The analysis of composite raw wastewater samples collected over 24 h in the NW of Spain allowed to quantify, for the first time in this matrix, the metabolite bis(chloropropyl) phosphate at levels over 60 ng/L

Background: Solar water disinfection (SODIS) is an appropriate technology for household treatment of drinking water in low-to-middle-income communities, as it is effective, low cost and easy to use. Nevertheless, uptake is low due partially to the burden of using small volume polyethylene terephthalate bottles (1.5–2 L). A major challenge is to develop a low-cost transparent container for disinfecting larger volumes of water. (2) Methods: This study examines the capability of transparent polypropylene (PP) buckets of 5 L- and 20 L- volume as SODIS containers using three waterborne pathogen indicators: Escherichia coli, MS2-phage and Cryptosporidium parvum. (3) Results: Similar inactivation kinetics were observed under natural sunlight for the inactivation of all three organisms in well water using 5 L- and 20 L-buckets compared to 1.5 L-polyethylene-terephthalate (PET) bottles. The PP materials were exposed to natural and accelerated solar ageing (ISO-16474). UV transmission of the 20 L-buckets remained stable and with physical integrity even after the longest ageing periods (9 months or 900 h of natural or artificial solar UV exposure, respectively). The 5 L-buckets were physically degraded and lost significant UV-transmission, due to the thinner wall compared to the 20 L-bucket. (4) Conclusion: This work demonstrates that the 20 L SODIS bucket technology produces excellent bacterial, viral and protozoan inactivation and is obtained using a simple transparent polypropylene bucket fabricated locally at very low cost ($2.90 USD per unit). The increased bucket volume of 20 L allows for a ten-fold increase in treatment batch volume and can thus more easily provide for the drinking water requirements of most households. The use of buckets in households across low to middle income countries is an already accepted practice.

This study presents the development and validation of a new analytical method based on matrix solid-phase dispersion (MSPD), integrating sample extraction and clean-up in one single step, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the simultaneous determination of 18 organophosphorus flame retardants and/or plasticizers (OPEs) in marine mussel (Mytilus edulis and Mytilus galloprovincialis) samples. Among these OPEs, 5 (tetraethyl 1,2-ethanediylbis(phosphonate), 6H-dibenzo[c,e] [1,2]oxaphosphinine 6-oxide, tris(2,3-dibromopropyl) phosphate, 2,2-propanediyldi-4,1-phenylene bis(phosphate) and resorcinol bis(diphenyl phosphate)) are considered here for the first time in marine samples. Different parameters affecting the MSPD (clean-up sorbent and elution solvent) were optimized to obtain a good compromise between analyte recoveries and extract clean-up. Also, particular attention was paid to tackle blank issues. The overall method was validated in terms of trueness, precision and detection and quantification limits. Percentages of recovery varied from 69% to 122% with relative standard deviations below 24%. Detection limits ranged from 0.06 to 5 ng g−1 and quantification limits from 0.19 to 17 ng g−1 dry weight. Finally, the method was applied to the analysis of 7 mussel samples collected in the coast of Galicia (Spain). 8 OPEs were detected in these samples at concentrations ranging from the LOQ to 291 ng g−1 dry weight.

En este artículo se presenta la metodología de análisis de aguas residuales con fines epidemiológicos (wastewater-based epidemiology, WBE) y su potencial para abordar diversos aspectos relacionados con la salud pública. Esta metodología permite obtener datos a una escala temporal y espacial relativamente pequeña (típicamente datos diarios-semanales sobre un municipio) de hábitos de consumo de sustancias de abuso, ilegales (como la cocaína o el cannabis) o legales (como el alcohol) a través de la determinación de biomarcadores de consumo (el compuesto original no metabolizado o alguno de sus metabolitos) en el agua residual. Aparte de discutir los fundamentos, ventajas y limitaciones de WBE, se comentan los precedentes más relevantes a nivel internacional, y las actividades más destacables en España en este ámbito. Finalmente, se exponen, los objetivos de la Red Española de Análisis de Aguas Residuales con Fines Epidemiológicos (ESAR-Net), una “Red de Excelencia” que agrupa a investigadores españoles con amplia experiencia en el área de WBE, así como las perspectivas de futuro de esta metodología puede tener para mejorar las competencias de la Salud Pública en España, This manuscript introduces Wastewater-Based Epidemiology (WBE) and its potential in the assessment of diverse aspects related to public health. This methodology can provide data in a relatively short temporal and local scale (typically dialy-weekly at the municipal level) on consumption patterns of illicit drugs (e.g. cocaine or cannabis), licit substances of abuse (e.g. alcohol) by measuring their consumption biomarkers (i.e. the original unmetabolized substance or some of its metabolite) in wastewater. Besides discussing the fundaments, advantages and shortcomings of WBE, it reviews some of the main precedents at international level and most remarkable activities that have been taken place in this field in Spain. Finally, the Spanish Network of Wastewater-Based Epidemiology (ESAR-Net) as is presented. ESAR-Net is an Excellence Network that sums up the efforts of the most relevant Spanish researchers in the field of WBE, aiming to investigate future perspectives of this methodology and its impact on Public Health competences in Spain

Simvastatin (SIM), a hypocholesterolaemic compound, is among the most prescribed pharmaceuticals for cardiovascular disease prevention worldwide. Several studies have shown that acute exposure to SIM causes multiple adverse effects in aquatic organisms. However, uncertainties still remain regarding the chronic effects of SIM in aquatic ecosystems. Therefore, the present study aimed to investigate the effects of SIM in the model freshwater teleost zebrafish (Danio rerio) following a chronic exposure (90 days) to environmentally relevant concentrations ranging from 8 ng/L to 1000 ng/L. This study used a multi-parameter approach integrating distinct ecologically-relevant endpoints, i.e. survival, growth, reproduction and embryonic development, with biochemical markers (cholesterol and triglycerides). Real Time PCR was used to analyse the transcription levels of key genes involved in the mevalonate pathway (hmgcra, cyp51, and dhcr7). Globally, SIM induced several effects that did not follow a dose-response relationship; embryonic development, biochemical and molecular markers, were significantly impacted in the lower concentrations, 8 ng/L, 40 ng/L and/or 200 ng/L, whereas no effects were recorded for the highest tested SIM levels (1000 ng/L). Taken together, these findings expand our understanding of statin effects in teleosts, demonstrating significant impacts at environmentally relevant concentrations and highlight the importance of addressing the effects of chemicals under chronic low-level concentrations

This study presents a new multi-residue analytical method for the simultaneous determination of 38 psychoactive drugs (including benzodiazepines, antidepressants and drugs of abuse) and related metabolites in raw wastewater. Potential analyte losses during sample filtration and stability in wastewater were evaluated. Analyte losses, especially for 12 compounds, were observed during filtration, indicating a strong sorption onto the filter material. In order to overcome this effect, filtered water samples were combined with methanolic washes of the corresponding filters and the resulting solutions were solid-phase extracted on mixed-mode (reverse-phase plus cation-exchange) sorbents. Extracts were analyzed by ultra-high performance liquid chromatography-tandem mass spectrometry. Quantification was performed by the internal standard method with isotopic labeled analogs. Recovery percentages varied between 65% and 137%; method quantification limits ranged between 0.2 and 22 ng/L in ultrapure water and between 0.3 and 30 ng/L in wastewater for all the analytes but three (for which they were ∼60–80 ng/L). The analysis of 24 h-composite samples collected during one week in the city of Santiago de Compostela demonstrated the ubiquity of 31 analytes, which were positively quantified in all samples. The highest concentrations were found for some of the antidepressants, with mean and maximum levels exceeding, in some cases, the levels previously reported in literature. This fact could be related to the additional washing step of the filters using methanol, which allowed to desorb retained analytes highlighting the importance of this step during the sample preparation protocol