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Knowledge about human exposure and health effects associated with non-routinely monitored disinfection by-products (DBPs) in drinking water is sparse., This project has been funded by Ajuntament de Barcelona (Institut de Cultura, Pla Barcelona Ciencia 2019. #19S01446-006), and partly funded by Instituto de Salud Carlos III and co-funded by European Union (ERDF) “A way to make Europe” (PI20/00829). We acknowledge support from the Spanish Ministry of Science and Innovation and State Research Agency through the “Centro de Excelencia Severo Ochoa 2019-2023” Program (CEX2018-000806-S and CEX2018-000794-S, for ISGlobal and IDAEA-CSIC, respectively), and support from the Generalitat de Catalunya through the CERCA Program. MJF acknowledges her Ramón y Cajal fellowship (RyC-2015-17108), from the AEI-MICIU. We thank Alexandra Paraian, Eva Maria Herrera (IDAEA-CSIC), Natalia Lopo, Lourdes Arjona and Antonia Valentín (ISGlobal) for their technical assistance during the sampling, analytical measurements and data analysis. The HAAs analyses in urine were carried out in the framework of the final master’s thesis (Universitat de Barcelona) by Paula Villasante. We also thank Patricia González, Anna Gómez, Sònia Navarro, and Laia Font-Ribera (Public Health Agency of Barcelona) for providing valuable information on Barcelona’s drinking water supplies. We finally would like to acknowledge all the volunteers that participated in the project.

Microbial biofilms communities in mineral waters and hot springs have a particular composition with species belonging to different groups such as epsilonproteobacteria and gammaproteobacteria or different siderobacteria and other chymoautrophic organisms, in addition to certain bacillaryophytes, chlorophytes and especially cyanobacteria. Balneotherapy can cause adverse reactions to the usual doses of application of treatments, that consists of a non-specific clinical picture, the so-called "thermal crisis” or “balneointoxication". Despite its clinical similarity (gastric discomfort, hepatic congestive outbreaks, cutaneous reactions, etc.) with that observed in acute cyanotoxin poisonings, thermal crisis has never been associated with the abundant growth of potentially toxic cyanobacteria in the mineral water sources. The aim of this work was to verify the hypothetical involvement of cyanotoxins in this clinical picture. Samples from mostly sulphurous water sources, with thermal characteristics ranging from cold to hyperthermal waters were analysed. ELISA (both in solution and in cellular matrix samples), LC-ESI-HRMS (in cellular matrix samples), and analysis of potential toxicity by means of a standardized bioassay were carried out. The toxic effect observed in the toxicity bioassays in one third of the sources may be related to the existence of microcystins and nodularins and even with other cyanobacterial peptides detected. In addition, several responses observed in the toxicity analyses reflect a pattern, probably linked to a type of hormetic response (hormesis is an adaptive response to low levels of stress, characterized by a biphasic dose-response curve)., This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

In August 2014, a puffer fish poisoning incidence resulting in one fatality was reported in New Caledonia. Although tetrodotoxin (TTX) intoxication was established from the patients' signs and symptoms, the determination of TTX in the patient's urine, serum or plasma is essential to confirm the clinical diagnosis. To provide a simple cost-effective rapid screening tool for clinical analysis, a maleimide-based enzyme-linked immunosorbent assay (mELISA) adapted for the determination of TTX contents in human body fluids was assessed. The mELISA was applied to the analysis of urine samples from two patients and a response for the presence of TTX and/or structurally similar analogues was detected in all samples. The analysis by LC-MS/MS confirmed the presence of TTX but also TTX analogues (4-epiTTX, 4,9-anhydroTTX and 5,6,11-trideoxyTTX) in the urine. A change in the multi-toxin profile in the urine based on time following consumption was observed. LC-MS/MS analysis of serum and plasma samples also revealed the presence of TTX (32.9 ng/mL) and 5,6,11-trideoxyTTX (374.6 ng/mL) in the post-mortem plasma. The results provide for the first time the TTX multi-toxin profile of human samples from a puffer fish intoxication and clearly demonstrate the implication of TTX as the causative agent of the reported intoxication case. info:eu-repo/semantics/acceptedVersion

Although consumption of Tetraodontidae species is prohibited in the EU, intoxications are still reported. The evaluation of tetrodotoxins (TTXs) by mass spectrometry (LC-MS/MS and LC-HRMS) and a screening immunoassay (mELISA) in tetraodontid fishes caught along the Western Mediterranean Sea revealed high concentrations of TTXs in Lagocephalus sceleratus while no TTXs were identified in L. lagocephalus and Sphoeroides pachygaster individuals. The high TTXs content found in the L. sceleratus analysed herein demonstrate the occurrence of highly toxic puffer fish in the Western Mediterranean Sea. Being L. sceleratus a recent invasive species in the Mediterranean, surveillance, risk assessment and risk management measures are necessary. The strategy used within this research work could be a valuable tool for future food safety monitoring. The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007–2013) through the ECsafeSEAFOOD project (grant agreement no. 311820), the Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA) through the PROMAQUA project (RTA2013-00096-00-00) and CERCA Programme/Generalitat de Catalunya. L.R. acknowledges a scholarship from the ECsafeSEAFOOD project.

This study focuses on the reaction of ozone with constituents of activated sludge liquor with the aim of understanding the behavior of organic micropollutants and to analyze the changes in the mixed liquor sludge matrix. To do so, different sludge sampling campaigns from two different wastewater treatment plants (WWTPs) were treated with different transferred ozone doses (TOD) and analyzed observing the effect on the sludge properties and micropollutants removal. Indeed, the ozone treatment promotes changes in the sludge matrix, increasing the Chemical Oxygen Demand (COD) solubilization until the Immediate Ozone Demand (IOD) completion and decreasing the Sludge Volume Index (SVI). Moreover, significant pharmaceutical removals have been achieved with low transferred ozone doses (TODs), reaching removals up to 99% depending on the compound.

Two small Lagocephalus sceleratus juveniles were captured in picarel targeting catches from North Aegean Sea (Greece) in the autumn of 2017. An electrochemical immunosensing tool using magnetic beads as immobilisation support was developed and applied to the rapid screening of tetrodotoxins (TTXs), potent neurotoxins that constitute a food safety hazard when present in seafood. This tool revealed the presence of TTXs in both individuals. Results were compared with those provided by mELISA and LC-HRMS, the latter confirming the presence of TTX. Some of the tissues contained TTX contents close to or above 2 mg/kg. L. sceleratus juveniles had been considered as non-toxic and, to our knowledge, this is the first report of high TTX levels in small L. sceleratus individuals. Such specimens can be mistaken with other edible species, posing a threat to consumers. The availability of low-cost and user-friendly tools for TTXs detection will contribute to guarantee seafood safety. © 2019 Elsevier Ltd, The research leading to these results has received funding from the Ministerio de Ciencia, Innovación y Universidades through the CIGUASENSING (BIO2017-87946-C2-2-R) project. The authors wish to express their gratitude to Dimitris Lachouvaris for collecting the samples and to Dr Nikolaos Lambadariou for providing the map. The authors also acknowledge support from CERCA Programme/Generalitat de Catalunya.

This study aimed to assess the bioaccessibility of different marine biotoxins in naturally contaminated shellfish and fish gonads using an in vitro digestion methodology. In general, hydrophilic toxins (domoic acid, paralytic shellfish poisoning toxins and tetrodotoxins) showed higher bioaccessibility than lipophilic ones (okadaic acid and azaspiracids). The bioaccessibility of toxins from the okadaic acid group ranged from 69% (raw European razor clams) to 74% (raw donax clams). Regarding azaspiracids, 47% of the initial content was bioaccessible in steamed blue mussel. As for hydrophilic toxins, 100% of the initial content was bioaccessible after digestion in raw shellfish and puffer fish gonads. The total tetrodotoxin bioaccessibility in puffer fish gonads decreased significantly after steaming. The profile of tetrodotoxins changed during the digestion process: TTX and 11-norTTX-6S-ol analogues decreased significantly after digestion, but the 5,6,11-trideoxy TTX analogue increased in both raw and steamed puffer fish gonads. These preliminary findings confirm the need to consider bioaccessibility data in future seafood risk assessment, as such information enables a more accurate and realistic estimation of potential seafood hazards, particularly in what concerns lipophilic toxins, therefore, constituting a crucial tool in the refinement of regulatory limits for the presence of biotoxins in seafood. © 2019, The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007–2013) under the ECsafeSEAFOOD project (grant agreement n° 311820) and through the European Regional Development Fund. IRTA members acknowledge support from CERCA Programme/Generalitat de Catalunya. L.R. acknowledges a scholarship from the ECsafeSEAFOOD project. Ana C. Braga has a Doctoral Grant (PD/BD/113484/2015) from the Portuguese Science and Technology Foundation (FCT), whereas Antonio Marques and Pedro R. Costa are supported through the FCT Investigator program (IF). Lagocephalus sceleratus was kindly provided by Andrés Izquierdo-Muñoz from Centro de Investigación Marina de Santa Pola (CIMA) (Spain).

A fast, simple, selective, and sensitive method for the analysis of 11 haloacetic acids (HAAs) in chlorine-treated water has been developed. The method is based on liquid chromatography-electrospray ionization-triple quadrupole tandem mass spectrometry (LC/ESI-QqQ-MS/MS) with direct injection of the aqueous sample. The main novelty of this method over the previously published procedures based on different techniques of mass spectrometry with direct injection is the combination of the simultaneous analysis of three types of HAAs (chlorinated, brominated, and iodinated) with its simplicity and low LODs (0.01–0.6 μg/L), avoiding the use of ion-pairing reagents for LC as well as the complexity and high cost of other techniques such as ion chromatography and capillary electrophoresis coupled to tandem mass spectrometry (IC-MS/MS and CE-MS/MS). The developed method was compared with another procedure carried out in our laboratory based on direct injection-liquid chromatography-electrospray ionization-high-resolution mass spectrometry with an Orbitrap analyzer (LC/ESI-Orbitrap-HRMS). The application of this technique to HAA analysis had not been previously described. LODs achieved by LC-HRMS (0.01–2 μg/L) were higher than the ones obtained by LC-MS/MS. Therefore, the LC/ESI-QqQ-MS/MS method was applied to the analysis of real samples. Quality parameters were calculated with satisfactory results and real samples related to three drinking water treatment plants (DWTPs), tap water, and the drinking water distribution system of Barcelona area (Catalonia, NE Spain) were analyzed. Furthermore, the evolution of HAA concentration along time in a DWTP-treated water sample was studied. © 2019, Springer-Verlag GmbH Germany, part of Springer Nature., This work has been financially supported by AGBAR in the framework of the AGBAR-CSIC R+D agreement on BControl of Organic Micropollutants in Water^ (grant number VATC 20140543).

Foods with high added value, such as Iberian dry-cured products, are susceptible to fraud. Many attempts have been made to differentiate the commercial/quality categories of Iberian dry-cured hams by analytical determinations. However, as discrimination by such means is not fully reliable, legislation to prevent fraudulent practice is based on administrative controls and certification. Here, new analytical approaches based on ultrahigh resolution mass spectrometry (UHRMS) and crystallographic techniques applied to the lipid fraction, in combination with chemometrics, are studied. The results of the triacylglycerol profile determined by UHRMS and the fingerprint provided by the thermograms obtained by differential scanning calorimetry offer the promise of analytic discrimination of Iberian dry-cured ham categories. In addition, these determinations, in combination with chemometrics, may prove extremely useful to authenticate many foods containing high to moderate amounts of lipids.

An integrated high resolution mass spectrometry (HRMS) strategy has been developed for rapid and accurate determination of free and cell-bound microcystins (MCs) and related peptides in water blooms. The natural samples (water and algae) were filtered for independent analysis of aqueous and sestonic fractions. These fractions were analyzed by MALDI-TOF/TOF-MS and ESI-Orbitrap-HCD-MS. MALDI, ESI and the study of fragmentation sequences have been provided crucial structural information. The potential of combined positive and negative ionization modes, full scan and fragmentation acquisition modes (TOF/TOF and HCD) by HRMS and high resolution and accurate mass was investigated in order to allow unequivocal determination of MCs. Besides, a reliable quantitation has been possible by HRMS. This composition helped to decrease the probability of false positives and negatives, as alternative to commonly used LC-ESI-MS/MS methods. The analysis was non-target, therefore covered the possibility to analyze all MC analogs concurrently without any pre-selection of target MC. Furthermore, archived data was subjected to retrospective "post-targeted" analysis and a screening of other potential toxins and related peptides as anabaenopeptins in the samples was done. Finally, the MS protocol and identification tools suggested were applied to the analysis of characteristic water blooms from Spanish reservoirs. © 2015 Elsevier B.V., The authors are grateful to Dr. J. A. Allué (Araclon Biotech, Zaragoza, Spain) and Dra. I. Fernández (University of Barcelona, Spain) for their assistance with MALDI-TOF-MS analysis; to Dra. N. Cortés-Francisco (IDAEA-CSIC, Barcelona, Spain) for their help in the Orbitrap analysis; to Dr. C. O. Miles (Norwegian Veterinary Institute, Norway) for identification of Dhb-containing microcystins in the bloom samples using thiol derivatization and LC–MS method; to Dr. A. Bueres (Miño-Sil, River Basin Authority, Spain) and Dr. A. Quesada (Universidad Autónoma de Madrid, Spain) for kindly providing the natural bloom samples from the Vilasouto and Alcántara reservoir, respectively; and to Dr. F. Ventura (IDAEA-CSIC) for manuscript revision. This work has been partially developed within the framework of the SOE1/P1/E129 ALARMTOX project (Interreg IVB SUDOE, South-West Europe Territorial Cooperation Program) integrated into the first priority of the Program: “Promotion of innovation and constitution of stable cooperation networks in technological matters”.

In the present paper, an electrospray ionization (ESI)-Orbitrap method is proposed for the direct chemical profiling of epicuticular wax (EW) from Olea europaea fruit. It constitutes a rapid and efficient tool suitable for a wide-ranging screening of a large number of samples. In a few minutes, the method provides a comprehensive characterization of total EW extracts, based on the molecular formula of their components. Accurate mass measurements are obtained by ultrahigh resolution mass spectrometry, and compositional restrictions are set on the basis of the information available from previous studies of olive EW. By alternating positive and negative ESI modes within the same analysis, complementary results are obtained and a wide range of chemical species is covered. This provides a detailed compositional overview that otherwise would only be available by applying multiple analytical techniques.

Ultrahigh resolution Fourier transform ion cyclotron mass spectrometry and nuclear magnetic resonance spectroscopy were combined to evaluate the molecular changes of dissolved organic matter (DOM) through an ultrafiltration-nanofiltration (UF-NF) pilot plant, using two dissimilar NF membranes tested in parallel. The sampling was performed on seven key locations within the pilot plant: pretreated water, UF effluent, UF effluent after addition of reagents, permeate NF 1, permeate NF 2, brine NF 1 and brine NF 2, during two sampling campaigns. The study showed that there is no significant change in the nature of DOM at molecular level, when the water was treated with UF and/or with the addition of sodium metabisulfite and antiscaling agents. However, enormous decrease of DOM concentration was observed when the water was treated on the NF membranes. The NF process preferentially removed compounds with higher oxygen and nitrogen content (more hydrophilic compounds), whereas molecules with longer pure aliphatic chains and less content of oxygen were the ones capable of passing through the membranes. Moreover, slight molecular selectivity between the two NF membranes was also observed.

In January 2015, a 20-month-old child and her family took part in recreational activities at Carrasco and Malvín beaches (Montevideo, Uruguay). An intense harmful algae bloom (HAB) was developing along the coast at that time. A few hours after the last recreational exposure episode, the family suffered gastrointestinal symptoms which were self-limited except in the child’s case, who was admitted to hospital in Uruguay with diarrhea, vomiting, fatigue, and jaundice. The patient had increased serum levels of liver enzymes and bilirubin and five days later presented acute liver failure. She was referred to the Italian Hospital in Buenos Aires, being admitted with grade II–III encephalopathy and hepatomegaly and requiring mechanical respiratory assistance. Serology tests for hepatitis A, B, and C, Epstein-Barr virus, and cytomegalovirus were negative. Laboratory features showed anemia, coagulopathy, and increased serum levels of ammonium, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and bilirubin. Autoimmune Hepatitis Type-II (AH-II) was the initial diagnosis based on a liver kidney microsomal type 1 antibodies (LKM-1) positive result, and twenty days later a liver transplant was performed. The liver histopathology had indicated hemorrhagic necrosis in zone 3, and cholestasis and nodular regeneration, which were not characteristic of AH-II. LC/ESI-HRMS (liquid chromatography electrospray ionization high-resolution mass spectrometry) analysis of MCs in the explanted liver revealed the presence of Microsytin-LR (MC-LR) (2.4 ng·gr−1 tissue) and [D-Leu1]MC-LR (75.4 ng·gr−1 tissue), which constitute a toxicological nexus and indicate a preponderant role of microcystins in the development of fulminant hepatitis., This study was supported by the National Council of Scientific and Technical Research (CONICET) PIP No. 112 201101 00959 res 1672, Ministry of Science and Technology PICT No. 2013-0861, and National University of La Plata PPID/X024.

Severe food poisoning events after the consumption of sharks have been reported since the 1940s; however, there has been no clear understanding of their cause. Herein, we report for the first time the presence of ciguatoxins (CTXs) in sharks. The identification by mass spectrometry of CTXs, including two new analogues, in a bull shark (Carcharhinus leucas) that was consumed by humans, causing the poisoning and death of 11 people in Madagascar in 2013 is described. Typical neurotoxic ciguatera symptoms were recorded in patients, and toxicological assays on extracts of the shark demonstrated CTX-like activity. These results confirm this episode as a ciguatera poisoning event and expand the range of pelagic fish species that are involved in ciguatera in the Indian Ocean. Additionally, gambieric acid D, a molecule originally described in CTX-producing microalgae, was identified for the first time in fish. This finding can contribute to a better understanding of trophic relations within food webs. The present work confirms that consumption of sharks from the Indian Ocean should be considered a ciguatera risk, and actions should be taken to evaluate its magnitude and risk in order to manage shark fisheries. © 2017 The Author(s)., We kindly acknowledge Dr. Bernard Reche, the veterinarian in charge of the shark autopsies in La Réunion and who confirmed the viscera sample was stomach, and also Erwan Lagadec (CRVOI) for performing the DNA extraction. We also thank the World Health Organization (WHO) for facilitating the transfer of samples from Madagascar to Reunion Island. The authors acknowledge Dr. Chris Rodgers for his critical review of the manuscript and Dr. Melissa Dunkle for assistance with the English. The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007–2013) under the ECsafeSEAFOOD project (grant agreement no. 311820) and from CERCA/Generalitat de Catalunya program.

Objective: To show the toxicological and phylogenetic characterization of a native Microcystis aeruginosa (M. aeruginosa) strain (named CAAT 2005-3) isolated from a water body of Buenos Aires province, Argentine. Methods: A M. aeruginosa strain was isolated from the drainage canal of the sewage treatment in the town of Pila, Buenos Aires province, Argentina and acclimated to laboratory conditions. The amplification of cpcBA-IGS Phcocyanin (PC, intergenic spacer and flanking regions) was carried out in order to build a phylogenetic tree. An exactive/orbitrap mass spectrometer equipped with an electrospray ionization source (Thermo Fisher Scientific, Bremen, Germany) was used for the LC/ESI-HRMS microcystins analysis. The number of cell/mL and [D-Leu1 ] Mcyst-LR production obtained as a function of time was modelled using the Gompertz equation. Results: The phylogenetic analysis showed that the sequence clustered with others M. aeruginosa sequences obtained from NCBI. The first Argentinian strain of M. aeruginosa (CAAT 2005-3) growing under culture conditions maintains the typical colonial architecture of M. aeruginosa with profuse mucilage. M. aeruginosa CAAT 2005-3 expresses a toxin variant, that was identified by LC-HRMS/Orbitrapas as [D-Leu1 ] microcystin-LR ([M+H]+=1037.8 m/z). Conclusions: [D-Leu1 ] microcystin-LR has been also detected in M. aeruginosa samples from Canada, Brazil and Argentina. This work provides the basis for technological development and production of analytical standards of toxins present in our region., Facultad de Ciencias Exactas, Facultad de Ciencias Naturales y Museo

A simple, reliable, selective and sensitive method for the determination of volatile thiols at trace levels in lipid matrices was developed. The method consisted of a single-step derivatization/extraction procedure followed by electrospray ionization-high performance liquid chromatography-high resolution mass spectrometry (ESI-HPLC-HRMS) analysis using ebselen as a derivatization agent. The analytical conditions were optimized in a lipid model system (triolein) and virgin olive oil (VOO), both spiked with seven representative volatile thiols. The method was validated and the sensitivity, precision, accuracy and selectivity were evaluated. The experimental limit of quantification (LOQ) of the tested thiols in the VOO matrix ranged from 0.05 to 0.5ng/kg. Moreover, the intra-day relative standard deviation (RSD) was in general lower than 10%, except for esters, which reached RSD values of around 13%. The inter-day RDS ranged between 9.6% and 36.6%. The recovery in VOO ranged from 79% to 20% for derivatives with the highest and lowest polarity, respectively. The methodology was applied to the analysis of some VOO samples. Within this first probe sample, some of the VOO analyzed presented concentrations of 4-methoxy-2-methyl-2-butanethiol above the LOQ, ranging between 0.2 and 1.9ng/kg. The advantage of using full-scan acquisition mode when working with HRMS is that it allows the use of a non-target approach based on the fragmentation of thiol derivatives yielding a main product ion at m/z 275.9922 [C13H10ONSe](+) and that corresponded to the ebselen moiety of the derivatives. Any positive finding satisfied the identification and confirmation criteria established during the validation: retention time drift

Perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) are two emerging contaminants that have been detected in all environmental compartments. However, while most of the studies in the literature deal with their presence or removal in wastewater treatment, few of them are devoted to their detection in treated drinking water and fate during drinking water treatment. In this study, analyses of PFOS and PFOA have been carried out in river water samples and in the different stages of a drinking water treatment plant (DWTP) which has recently improved its conventional treatment process by adding ultrafiltration and reverse osmosis in a parallel treatment line. Conventional and advanced treatments have been studied in several pilot plants and in the DWTP, which offers the opportunity to compare both treatments operating simultaneously. From the results obtained, neither preoxidation, sand filtration, nor ozonation, removed both perfluorinated compounds. As advanced treatments, reverse osmosis has proved more effective than reverse electrodialysis to remove PFOA and PFOS in the different configurations of pilot plants assayed. Granular activated carbon with an average elimination efficiency of 64. ±. 11% and 45. ±. 19% for PFOS and PFOA, respectively and especially reverse osmosis, which was able to remove ≥. 99% of both compounds, were the sole effective treatment steps. Trace levels of PFOS (3.0-21. ng/L) and PFOA (, This work has been partially developed within the framework of the SOSTAQUA Project (led by AGBAR-Aigües de Barcelona and financed by CDTI, Ingenio 2010 Programme under the CENIT call ). Appendix A

With the aim to enhance characterization of virgin olive oil (VOO), high resolution mass spectrometry (HRMS) and high resolution tandem mass spectrometry (HRMS/MS), in positive and negative electrospray ionization (ESI) modes, coupled to fused-core reverse phase chromatography, were applied to distinct VOO phenolic extracts after the optimization of chromatographic conditions, ESI and fragmentation parameters. HRMS, but also HRMS/MS resulted fundamental to progress in secoiridoids structural elucidation. The former revealed that the secoiridoid composition of VOO was far more complicated than previously reported, while the latter helped clarify product ion elemental composition allowing new fragmentations, in addition to those reported in the literature, to be put forward. In particular, for the first time, different product ions with the same nominal mass were unequivocally identified in the spectra of secoiridoid compounds, confirming the greater capacity of HRMS/MS to clarify structure than low-resolution MS. Furthermore, and differing from previous studies, the multiple isomers of the main VOO secoiridoids could be differentiated on the basis of their HR product ion spectra in positive mode.

Sensitive and unequivocal determination of analytes/contaminants in complex matrices is a challenge in the field of food safety control. In this study, various acquisition modes (Full MS/AIF, Full MS+tMS/MS, Full MS/dd MS/MS and tSIM/ddMS/MS) and parameters of a quadrupole-orbitrap hybrid mass spectrometer (Q Exactive) were studied in detail. One of the main conclusions has been that, reducing the scan range for Full MS (using the quadrupole) and targeted modes give higher signal-to-noise (S/N) ratios and thereby better detection limits for analytes in matrix. The use of Q Exactive in a complex case, for the confirmatory analysis of hormones in animal urine is presented. A targeted SIM data dependent MS/MS (tSIM/ddMS/MS) acquisition method for determination of eight synthetic hormones (trenbolone, 17α ethinylestradiol, zeranol, stanozolol, dienestrol, diethylstilbestrol, hexestrol, taleranol) and a naturally occurring hormone (zearalenone) in animal urine were optimized to have sensitive precursors from targeted SIM mode and trigger MS/MS scans over the entire chromatograph peak. The method was validated according to EC/657/2002. CCα (decision limit) for the analytes ranged between 0.11 μg L(-1) and 0.69 μg L(-1) and CCβ (detection capability) ranged between 0.29 μg L(-1) and 0.90 μg L(-1).

International audience; There has been increasing interest in the use of selected non-Saccharomyces yeasts in co-culture with Saccharomyces cerevisiae. The main reason is that the multistarter fermentation process is thought to simulate indigenous fermentation, thus increasing wine aroma complexity while avoiding the risks linked to natural fermentation. However, multistarter fermentation is characterised by complex and largely unknown interactions between yeasts. Consequently the resulting wine quality is rather unpredictable. In order to better understand the interactions that take place between non-Saccharomyces and Saccharomyces yeasts during alcoholic fermentation, we analysed the volatile profiles of several mono-culture and co-cultures. Candida zemplinina, Torulaspora delbrueckii and Metschnikowia pulcherrima were used to conduct fermentations either in mono-culture or in co-culture with S. cerevisiae. Up to 48 volatile compounds belonging to different chemical families were quantified. For the first time, we show that C. zemplinina is a strong producer of terpenes and lactones. We demonstrate by means of multivariate analysis that different interactions exist between the co-cultures studied. We observed a synergistic effect on aromatic compound production when M. pulcherrima was in co-culture with S. cerevisiae. However a negative interaction was observed between C. zemplinina and S. cerevisiae, which resulted in a decrease in terpene and lactone content. These interactions are independent of biomass production. The aromatic profiles of T. delbrueckii and S. cerevisiae in mono-culture and in co-culture are very close, and are biomass-dependent, reflecting a neutral interaction. This study reveals that a whole family of compounds could be altered by such interactions. These results suggest that the entire metabolic pathway is affected by these interactions.

The number of cytostatic drugs used in cancer treatments is wide and increases every year; therefore, tools have been developed to predict their concentration in the environment to prioritize those for monitoring studies. In the present study, the predicted environmental concentrations (PECs) were calculated according to consumption data in Catalonia (NE Spain) for 2014. According to PECs and to the most widely reported compounds, 19 cytostatics were monitored in two sampling campaigns performed along the Besos River. A total of seven drugs were detected at levels between 0.5 and 656 ng L−1. PEC and measured environmental concentrations (MECs) were compared in order to validate PECs. The PEC/MEC ratio presented a good agreement between predicted and measured concentrations confirming the PEC estimations. Mycophenolic acid, prioritized as the compound with the highest PEC, was detected at the highest concentrations (8.5–656 ng L−1) but showed no risk for aquatic organisms (risk quotient

The epicuticular wax (EW) layer is located on the surface of most of plants organs. It provides the cuticle with most of its properties and is the primary barrier against biotic and abiotic stress. Despite the importance of Olea europaea cultivation, few studies have characterized the EW covering leaves and olives, which could be involved in resistance to both infection and environmental conditions. In the present study, wide-ranging screening was carried out using direct-injection electrospray ionisation coupled to high-resolution mass spectrometry to analyse EW in developing olives of nine varieties. The proportions of EW fractions (wax esters (WEs), diacylglycerols, triacylglycerols (TAGs), triterpenic acids and aldehydes) strongly depended on the olive cultivar, and in only a few cases were they influenced by the sampling date. The specific compositions of the major fractions, WEs and TAGs, were strictly related to the cultivar, while the degree of unsaturation and the chain length of the WEs evolved throughout the four weeks prior to the olive colour turning.

Sales of bottled drinking water have shown a large growth during the last two decades due to the general belief that this kind of water is healthier, its flavour is better and its consumption risk is lower than that of tap water. Due to the previous points, consumers are more demanding with bottled mineral water, especially when dealing with its organoleptic properties, like taste and odour. This work studies the compounds that can generate obnoxious smells, and that consumers have described like swampy, rotten eggs, sulphurous, cooked vegetable or cabbage. Closed loop stripping analysis (CLSA) has been used as a pre-concentration method for the analysis of off-flavour compounds in water followed by identification and quantification by means of GC-MS. Several bottled water with the aforementioned smells showed the presence of volatile dimethyl selenides and dimethyl sulphides, whose concentrations ranged, respectively, from 4 to 20 ng/L and from 1 to 63 ng/L. The low odour threshold concentrations (OTCs) of both organic selenide and sulphide derivatives prove that several objectionable odours in bottled waters arise from them. Microbial loads inherent to water sources, along with some critical conditions in water processing, could contribute to the formation of these compounds. There are few studies about volatile organic compounds in bottled drinking water and, at the best of our knowledge, this is the first study reporting the presence of dimethyl selenides and dimethyl sulphides causing odour problems in bottled waters.

Several factors affect virgin olive oil (VOO) phenolic profile. The aim of this study was to monitor olive hydrolytic (β-glucosidase) and oxidative (peroxydase, POX, and polyphenoloxydase, PPO) enzymes during olive ripening and storage and to determine their capacity to shape VOO phenolic profile. To this end, olives from the cultivars Chetoui and Arbequina were stored at 4°C or 25°C for 4weeks and their enzymatic activities and oil phenolic profiles were compared to those of ripening olives. We observed different trends in enzymes activities according to cultivar and storage temperature. Secoiridoid compounds, determined by high resolution mass spectrometry (HRMS), and their deacetoxylated, oxygenated, and deacetoxy-oxygenated derivatives were identified and their contents differed between the cultivars according to olive ripening degree and storage conditions. These differences could be due to β-glucosidase, POX and PPO activities changes during olive ripening and storage. Results also show that oxidised phenolic compounds could be a marker of VOO ''freshness".

In the present study, accurate mass measurements by ultrahigh resolution mass spectrometry with Orbitrap-Exactive working at resolving power R: 100,000 (m/z 200, full width at half maximum) with an accuracy better than 2 ppm in all the mass range (m/z 200 to 2000) were used to show a detailed molecular composition of diverse edible oils and fats. Flow injection was used to introduce samples into the mass spectrometer, obtaining a complete analysis of each sample in less than 10 min, including blanks. Meticulous choice of organic solvents and optimization of the ion source and Orbitrap mass analyzer parameters were carried out, in order to achieve reproducible mass spectra giving reliable elemental compositions of the lipid samples and to prevent carry over. More than 200 elemental compositions attributable to diacylglycerols, triacylglycerols (TAGs), and their oxidation products have been found in the spectra of food lipids from different origin. Several compounds with very close molecular mass could only be resolved through ultrahigh resolution, allowing detailed and robust TAG profiling with a high characterization potential. Copyright © 2012 John Wiley & Sons, Ltd.

In view of the significance of glycerolipid fingerprint for virgin olive oil (VOO) characterization, and for the importance of triacylglycerol (TAG) and diacylglycerol (DAG) composition for oil physico-chemical and physiological properties, the modifications induced in the oil glycerolipid profile by olive ripening were monitored by MALDI-TOF MS. Differences in oils from two olive varieties grown in the same area and at different olive ripening stages were clearly evidenced, in particular in TAG rather than in DAG profile. Although the olive variety was the main determinant in VOO glycerolipid profile, the olive maturity stage showed a significant effect that should be taken into account when considering VOO fingerprinting for authentication and characterization purposes.

According to the high incidence of cancer worldwide, the amount of cytostatic drugs administered to patients has increased. These compounds are excreted to wastewaters, and therefore become potential water contaminants. At this stage, very little is known on the presence and elimination of cytostatic compounds in wastewater treatment plants (WWTP). The aim of this study was to develop a liquid chromatography-high-resolution mass spectrometry (LC-Orbitrap-MS) method for the determination of cyclophosphamide and epirubicin in wastewaters. These compounds represent two outmost used cytostatic agents.Extraction and analytical conditions were optimized for cyclophosphamide and epirubicin in wastewater. Both solid-phase extraction using Oasis 200 mg hydrophilic-lipophilic balanced (HLB) cartridges and direct injection analysis were evaluated. Mass spectral characterization and fragmentation conditions were optimized at 50,000 resolving power (full width at half maximum, m/z 200) to obtain maximum sensitivity and identification performance. Quality parameters (recoveries, limits of detection, and repetitivity) of the methods developed were determined, and best performance was obtained with direct water analysis of the centrifuged wastewater. Finally, this method was applied to determine the presence of cyclophosphamide and epirubicin in wastewaters from a hospital effluent, an urban effluent, and influents and effluents from three WWTP.Cyclophosphamide and epirubicin were recovered after 50 mL preconcentration on solid-phase extraction 200 mg Oasis HLB cartridges (87% and 37%, respectively), and no breakthrough was observed by extracting 500 mL of water. Limits of detection were of 0.35 and 2.77 ng/L for cyclophosphamide and epirubicin, respectively. On the other hand, direct injection of water spiked at 1 μg/L provided recoveries of 107% for cyclophosphamide and 44% for epirubicin and limits of detection from 3.1 to 85 ng L(-1), respectively. The analysis of wastewaters using direct injection analysis revealed the presence of cyclophosphamide and epirubicin in WWTP influents and hospital and urban effluents at levels ranging from 5.73 to 24.8 μg L(-1).The results obtained in this study demonstrate the capability of LC-Orbitrap-MS for accurate trace analysis of these very polar contaminants. This method permitted to identify cyclophosphamide and epirubicin in wastewaters and influents of WWTP, but no traces were detected in WWTP effluents. The methodology herein developed is sensitive and robust and applicable for screening of a large number of samples since no preconcentration is needed.

The activity of olive microbiota during the oil extraction process could be a critical point for virgin olive oil quality. With the aim to evaluate the role of microbiological activity during the virgin olive oil extraction process, just before oil extraction freshly collected healthy olive fruits were immersed in contaminated water from an olive mill washing tank. The oils extracted were then compared with control samples from the same batch of hand-picked olives. The presence of lactic and enteric bacteria, fungi and Pseudomonas on the surface of olives was proved to be much higher in washed than in control olives, with increments in cfu/g between 2 and 3 orders of magnitude. The biogenesis of volatile compounds and the extraction of olive polyphenols and pigments were significantly influenced by the microbiological profile of olives even without any previous storage. In most cases the effect of olive microbiota on oil characteristics was greater than the effect exerted by malaxation time and temperature. Oils from microbiologically contaminated olives showed lower amounts of C5 volatiles and higher levels of C6 volatiles from the lipoxygenase pathway and some fermentation products. On the other hand, a decrease of chlorophylls, pheophytins, xanthophylls and the ratio chlorophyll/pheophytin was observed in these oils. Likewise, the microbiological activity during oil extraction led to significantly lower amounts of polyphenols, in particular of oleuropein derivatives. These differences in olive oil chemical composition were reflected in oil sensory characteristics by the decrease of the green and bitter attributes and by the modification of the oil color chromatic ordinates.

10 páginas, 5 tablas, 4 figuras, In the present work, the potential use of several antioxidant and detoxification biomarkers in the digestive gland of wild mussels (Mytilus galloprovincialis) for biomonitoring the marine pollution induced by the Don Pedro oil spill has been investigated. Two locations from the East to South-East of Eivissa (Ibiza) and Formentera islands were selected, one extensively affected by the oil spill and the other one not affected and considered as the control area. Mussels were sampled one, two and six months after the Don Pedro accident. Polycyclic aromatic hydrocarbon (PAH) levels were significantly increased in the soft tissues of mussels in the affected area one month after the disaster, returning to normal values after six months. Markers of oxidative damage in lipids – malondialdehyde, and in proteins – carbonyl derivates, and antioxidant enzyme – catalase, superoxide dismutase and glutathione peroxidase, activities significantly increased as result of the spill oil after one month, returning to basal values at two month sampling time. Glutathione/glutathione disulfide ratio (GSH/GSSG), as a marker of the redox status, was reduced after one and two months indicating a more oxidized situation. Markers of detoxification – glutathione-Stransferase and cytochrome P4501A activities and metallothionein gene expression – were significantly increased by the oil spill onemonthafter the accident, returning to the basal values attwomonthsampling time. In conclusion, the Don Pedro accident induced a transient situation of PAHs pollution resulting in enhanced antioxidant and detoxification defense systems in the wild mussel M. galloprovincialis returning to normal levels six months from the spill. The selected biomarkers are a useful tool for biomonitoring the response to acute exposure to pollutants in marine mussels., This work supported by grants by DG Pesca, Conselleria d’Agricultura i Pesca, Govern de les Illes Balears.

The Mytilos project was carried during the last 3 years (2004-2006) in order to evaluate the level of chemical contamination along the coasts of the western basin of the Mediterranean Sea. Active mussel watch consisted of caged (12 weeks) Mytilus galloprovincialis at 123 stations along the Mediterranean coasts of Spain, France, Italy, North Tunisia, Algeria and Morocco. Results indicated total PAHs levels in the range 22-106 μg/kg dry weight. All large towns or industrial areas from all countries of the basin were affected. Analysis of the origin of contamination demonstrated mainly a pyrolytic origin except in some areas such as Maddalena, Napoli, Alger, and Cornigliano (Genova). Statistical analysis of the PAHs accumulation data enabled the characterisation and ranking of each coastal area. The overall results give scientific and technical basis for large-scale monitoring of the PAH contamination in the entire western Mediterranean basin. © 2010 Springer Science+Business Media B.V., This research was supported by EEC (Interreg/medoccIIIC/MYTILOS). INRH (Morocco), ISMAL (Algeria) and INSTM (Tunisia) were supported by UNEP MEDPOL.

A method based on automated solid-phase extraction (SPE) and isotope dilution gas chromatography/high resolution mass spectrometry (GC/HRMS) has been developed for the analysis of nine nitrosamines in water samples. The combination of automated SPE and GC/HRMS for the analysis of nitrosamines has not been reported previously. The method shows as advantages the selectivity and sensitivity of GC/HRMS analysis and the high efficiency of automated SPE with coconut charcoal EPA 521 cartridges. Low method detection limits (MDLs) were achieved, along with a greater facility of the procedure and less dependence on the operator with regard to the methods based on manual SPE. Quality requirements for isotope dilution-based methods were accomplished for most analysed nitrosamines, regarding to trueness (80-120%), method precision (

8 pages, 3 figures., Headspace solid-phase microextraction (HS-SPME), simultaneous distillation/extraction (SDE) and closed-loop stripping analysis (CLSA) coupled to gas chromatography/mass spectrometry (GC/MS) were used to study virgin olive oil, with the goal of detecting large numbers of characteristic volatile and semi-volatile compounds. More than one hundred compounds were detected in the olive oil extracts, and their percent amounts obtained by each technique were calculated. Qualitative and quantitative differences of virgin olive oil volatile profiles were observed applying the three extraction techniques. SPME showed a higher affinity for alcohols and ketones, while CLSA achieved the highest percentages of esters and hydrocarbons. Finally, the highest extraction of total terpenoid compounds occurred with SDE and CLSA, where CLSA allowed extracting the highest percentages of the most of them. SDE extraction caused the thermal degradation of the oil sample, which resulted in a high percentage of aldehydic compounds., This study was supported by the Generalitat de Catalunya, project 2005SGR00156; by the Ministerio de Ciencia y Tecnología (MCYT), project AGL2005-03451/ALI; and by the Secretaría de Estado de Educación y Universidades (Ministerio de Educación y Ciencia), Ref. SB2003-0118.

Mountain lakes are excellent indicators for airborne organic pollutant transport and accumulation studies. However, very few studies have been focused on lake water mass. Also, there is a lack of studies performed after 2004–2009 period when the enforcement of most international organic pollutant banning policies (e.g., The Stockholm convention) occurred. We report concentration of 66 different semi-volatile organic compounds (SOCs) in the Sabocos tarn water mass (a Spanish Pyrenees cirque glacial lake) from 2011 to 2014. Analyzed SOCs include compounds of current and past use with different sources (i.e., current and historic-use pesticides, industrial and urban-use compounds, and combustion by-products) and chemical properties (i.e., volatility, water solubility, and persistence in the environment). Most banned chemicals such as dichlorodiphenyl trichloroetanes (DDTs), aldrins, most polybromodiphenyl ether (PBDE) congeners, endosulfans, triazine, and simazine were detected in consistently low concentrations. In contrast, chemicals still in use such as BDE-209, terbutilazine, diuron, metolachlor, and chlorpyrifos were found in higher and more variable amounts. Polychlorobiphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) were high as compared to previous studies in other Pyrenean and European lakes suggesting airborne pollution from nearby anthropogenic sources. Overall, levels of hexacyclohexanes (HCHs) during most of the sampling period were low according to other studies in similar environments. However, the Sabocos water mass recorded a concentration peak of some HCH isomers in summer 2014 related with the HCH residues transfer operation at the nearby (20 km) Bailin landfill site (Sabinanigo, Spain).

A simple, selective and sensitive method for the analysis of the strong mutagen 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX) and its brominated analogues (BMXs) in chlorine-treated water has been developed. The method is based on gas chromatography coupled to triple quadrupole tandem mass spectrometry (GC-QqQ-MS/MS), previous liquid-liquid extraction (LLE) of a smaller sample volume compared to other methods and on-line derivatization with a silylation reactive. GC-QqQ-MS/MS has been raised as an alternative easier to perform than gas chromatography coupled to high resolution mass spectrometry (GC-HRMS) for the analysis of MX and BMXs, and it allows to achieve low LODs (0.3 ng/L for MX and 0.4-0.9 ng/L for BMXs). This technique had not been previously described for the analysis of MX and BMXs. Quality parameters were calculated and real samples related to 3 drinking water treatment plants (DWTPs), tap water and both untreated and chlorinated groundwater were analyzed. Concentrations of 0.3-6.6 ng/L for MX and 1.0-7.3 ng/L for BMXs were detected. Results were discussed according to five of the main factors affecting MX and BMXs formation in chlorine-treated water (organic precursors, influence of bromide ions, evolution of MX and BMXs in the drinking water distribution system, groundwater chlorination and infiltration of water coming from chlorination processes in groundwater).

In the present paper, we present a simple, reliable, selective and sensitive method for the identification and quantification of volatile thiols at trace levels in coffee brews. A simultaneous derivatization/ extraction procedure followed by liquid chromatography electrospray high-resolution mass spectrometry is proposed and adapted to coffee brew matrix, and the performance of the method is evaluated. The linearity, sensitivity, recovery and both the intra-day and inter-day accuracy were all satisfactory. According to established identification criteria, seven target and nine non-target thiols were identified and quantified in coffee brew samples. Several of them are reported here for the first time in coffee brews, and our results are in agreement with previously reported results for coffee powder analyzed using similar analytical approach.

To identify critical points during olive mill pre-processing operations, the effect of the closed circuit washing stage on olive microbiological contamination, and the influence of the successive short-term storage on olives and virgin olive oil (VOO) quality were evaluated. Microbiological, physical, and chemical parameters were assessed in olives and oils at three mill pre-processing stages: reception, washing, and short-term storage. Olive washing in closed loop systems was shown to be a critical control point at the olive mill due to microbiological cross-contamination and fruit physical damage. Moreover, when the olives were short-term stored before oil extraction, positive VOO sensory attributes decreased by as much as one point of intensity, as justified by the changes observed in phenolic and lipoxygenase derived volatile compounds. These results confirm the high risk of fruit cross-contamination due to the poor hygiene of the water used in olives mills to wash olives, and point out the effect on VOO quality of a common practice such as short term silo storage of olives. Practical applications: The identification of new critical control points within pre-processing operations in ordinary usage, and commonly accepted as good production practices, will contribute to enhance virgin olive oil quality. Preventive actions can be undertaken on the basis of the reported results, such as the control of water hygiene and short term storage conditions. The figure shows the microbiological contamination of olives during washing with recycled water and during short-term silo storage, and the loss of sensory quality in the correspondent virgin olive oils.

A new method for the analysis of lipophilic marine biotoxins (okadaic acid, dinophysistoxins, azaspiracids, pectenotoxins, yessotoxins, spirolids) in fresh and canned bivalves has been developed. A QuEChERS methodology is applied; i.e. the analytes are extracted with acetonitrile and clean-up of the extracts is performed by dispersive solid phase extraction with C-18. The extracts are analyzed by ultra-high performance liquid chromatography coupled to a hybrid quadrupole-Orbitrap mass spectrometer, operating in tandem mass spectrometry mode, with resolution set at 70,000 (m/z 200, FWHM). Separation of the analytes, which takes about 10min, is carried out in gradient elution mode with a BEH C-18 column and mobile phases based on 6.7 mM ammonia aqueous solution and acetonitrile mixtures. For each analyte the molecular ion and 1 or 2 product ions are acquired, with a mass accuracy better than 5 ppm. The quantification is performed using surrogate matrix matched standards, with eprinomectin as internal standard. The high-throughput method, which has been successfully validated, fulfills the requirements of European Union legislation, and has been implemented as a routine method in a public health laboratory. (c) 2015 Elsevier B.V. All rights reserved.