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Dihydrochalcones, phlorizin (PZ) and its aglycone phloretin (PT), have evidenced immunomodulatory effects through several mechanisms. However, the differential metabolic signatures that lead to these properties are largely unknown. Since macrophages play an important role in the immune response, our study aimed to characterise human THP-1 macrophages under PZ and PT exposure. A multiplatform-based untargeted metabolomics approach was used to reveal metabolites associated with the anti-inflammatory mechanisms triggered by the dihydrochalcones in LPS-stimulated macrophages, for the first time. Results showed differential phenotypic response in macrophages for all treatments. Dihydrochalcone treatment in LPS-stimulated macrophages mimics the response under normal conditions, suggesting inhibition of LPS response. Antagonistic effects of dihydrochalcones against LPS was mainly observed in glycerophospholipid and sphingolipid metabolism besides promoting amino acid biosynthesis. Moreover, PT showed greater metabolic activity than PZ. Overall, the findings of this study yielded knowledge about the mechanisms of action PZ and PT at metabolic level in modulating inflammatory response in human cells., This research was funded by EU FEDER funds, Interreg España-Portugal Programme, under the framework of the Project: ref. 0377_IBERPHENOL_6_E; by Xunta de Galicia funds, Spain (CITACA Strategic Partnership, Reference: ED431E 2018/07) and also supported by UIDB/50006/2020 and PTDC/SAU-NUT/30448/2017 with funding from FCT/MCTES through national funds. I. F. acknowledge her Fundação para a Ciência e Tecnologia (FCT) research contract (SFRH/BPD/86173/2012).

In this study, a new selective pressurised liquid extraction (SPLE) methodology was optimised for determining about 70 organic pollutants (OPs) including organochlorine (OCPs), organophosphate (OPPs) and pyrethroid (PYRs) pesticides, polychlorinated biphenyls (PCBs), polybromodiphenyl ethers (PBDEs), as well as, polycyclic aromatic hydrocarbons (PAHs) in wild boar liver samples considering the temperature, pressure and time of contact between the solvent and the matrix as influential variables. Clean-up of extracts was performed by solid-phase extraction (SPE) using EZ-POP cartridges. Detection of OPs was carried out by gas chromatography (GC) coupled to tandem mass spectrometry (QqQ-MS/MS). This new approach offers:•A new non-time consuming SPLE methodology for determining about 70 OPs in wild boar.•Recoveries achieved ranged between 74 to 119 % with RSD less than 20 %.•Detection and quantification limits in the low to mid pg/g range., Graphical abstract Image, graphical abstract

Organic pollutants (OPs) represent a wide range of chemicals that are potentially harmful for human and wildlife health. Many of these pollutants have been identified as endocrine disruptors that can alter hormonal balance producing adverse biological effects such as neurotoxicity, reproductive disorders, carcinogenicity and hepatotoxicity. For years, hair has been selected as a non-invasive source to assess levels of animal contamination. In the present study, a multiclass screening method for determining about 60 organic pollutants in pet hair was designed and validated for qualitative and quantitative purposes. Concentrations from different classes of organochlorine, and organophosphate pesticides (OCPs, and OPPs), polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (NDL-PCBs and DL-PCBs), polybrominated diphenyl ethers (PBDEs) and organophosphate esters (OPEs) were identified in the selected pet hair samples from Ourense (NW, Spain). We detected most of these pollutants in the selected hair pets. The mean concentrations found ranged from 89 to 6556ng/g for OPEs, from 8.6 to 1031ng/g for PAHs, from 8.6 to 256ng/g for PBDEs, from 29 to 184ng/g for OPPs, from 0.29 to 139 for OCPs, from 0.30 to 59ng/g for NDL-PCBs and from 1.2 to 14ng/g for DL-PCBs. To our knowledge, this is the first study to document the presence of OPs in pets from North-West Spain and it could provide baseline information for future monitoring of OPs in the area.

The early exposure to organic pollutants (OPs) related to dietary habit or environmental exposure is a concern of growing interest in environmental health. When OPs enter the body, they can accumulate in fatty tissue and even travel through the bloodstream being able to cross the placenta and reach the fetus through the substance exchange between the mother and the child. Epidemiologic and clinical data affirm that these chemicals increase the risk of adverse effects during childhood. This article reviews and addresses one of the most relevant analytical methods for determining OPs in placenta. We discuss and critically evaluate procedures, such as solid-liquid and selective pressurized liquid extraction (SPLE). Clean-up of extracts was performed by solid-phase extraction (SPE) using EZ-POP cartridges. Detection of OPs was carried out by gas chromatography (GC) coupled to tandem mass spectrometry (MS/MS). Recoveries ranged from 52% to 94% by SPLE with estimated quantification limits between 0.15 and 2.2ng/g for organochlorine pesticides (OCPs), between 0.083 and 0.50 for organophosphate pesticides (OPPs), between 0.025 and 0.15ng/g for polychlorinated biphenyl (PCBs), between 0.015 and 0.10ng/g for polybromodiphenyl ethers (PBDEs), between 0.33 and 0.66ng/g for pyrethroids and between 0.022 and 0.87ng/g for polycyclic aromatic hydrocarbons (PAHs). Most of the target OPs were detected in twenty real placenta samples studied, with pyrethroids, PAHs and OPPs representing most of the 90% of OPs with means of 13-18ng/g versus PCBs, OCPs, and PBDEs with means4.0ng/g. The rsults of this work indicate a prenatal exposure to OPs in Galicia.

Results gained from in vivo metabolism could be more authentic to reflect the biotransformation of drugs than in vitro metabolism. This paper discusses the development and characterization of an analytical method for controlling tiamulin (TIA) and doxycycline (DOX) residues in pig plasma and urine, as well as their in vivo biotransformation during treatment and suppression steps at pig farms. For such a purpose, urine samples were taken daily (24-hr intervals) during treatment and until day 8 after last dose. Plasma samples were collected on the last treatment day and up to days 4 and 7 after the last dose. Only traces of TIA and their metabolites have been still detected 8 days after the last day of therapy, in agreement with the period of suppression of both antibiotics set by the manufacturers in 7 days. It was estimated that TIA and DOX half-lives were of two days and one day, respectively. TIA and DOX metabolites have been identified for the first time in plasma and urine pig samples during treatment and suppression steps.

There are strong evidences that probiotics influence the immune status of the host, in a strain-specific manner, acting in the gastrointestinal tract. On the hypothesis that certain extracellular proteins and peptides from gut bacteria may mediate part of this immunomodulation and assuming they are able to diffuse through the mucus layer and interact with immune cells we have developed this work. Our study attempts to understand the immunomodulatory mechanisms of (i) Pext, the extracellular protein fraction of Lactobacillus acidophilus DSM20079T, (ii) HM14, a peptide encrypted in an extracellular glycoside hydrolase from Bifidobacterium longum NCIMB 8809 and (iii) Escherichia coli O111:B4 lipopolysaccharide (LPS), a well-known pro-inflammatory molecule, over human peripheral blood mononuclear cells (PBMCs). An untargeted LC-ESI-QTOF-MS metabolomics approach was applied to reveal intracellular changes in treated-PBMCs isolated from healthy donors. Differences in NADH arrest, NAD+ concentration reduction, as well as increases in palmitic acid and methanephrin were observed in HM14 and Pext treated-cells compared to those stimulated with LPS. This would support an anti-inflammatory molecular mechanism of action of such proteinaceous molecules. Moreover, this methodology has confirms the importance of metabolomics approaches to better understanding immune cell responses to gut bacterial-derived molecules., This work was funded by the Autonomic Investigadores Emerxentes do Sistema Universitario de Galicia (Grant EM2014/046) and the Spanish Programa Estatal de Investigación, Desarrollo e Innovación Orientada a los Retos de la Sociedad (Grant AGL2013-44039R). This work has received also financial support from the Xunta de Galicia (Plan de mellora do Centro de Investigacións Agroalimentarias CIA3 do Campus de Ourense).

Cortisol and corticosterone in saliva were evaluated as pig stress biomarkers, using pig genotype (Duroc, L62 or Pietrain) and lairage time in the slaughterhouse (0, 2.0, 4.0 or 6.0 h) as controlled variables. Although some pigs were found to be carriers of stress susceptibility, all were healthy heterozygous individuals. Pre-slaughter transport increased cortisol levels in saliva above 3.0 μg/L (medium stress), and 4.0 h of lairage in the slaughterhouse raised them above 6.0 μg/L, whereas corticosterone concentrations exceeded 4 μg/L, which are suggestive of high stress. The highest cortisol levels were detected in the Duroc genotype. Other factors such as food deprivation, background noise, the presence of a large number of animals waiting to be slaughtered, mixing with unfamiliar animals or recent mixing of genders may also influence stress. Corticosterone proved a reliable indicator of high stress only. Meat quality from the pig breeds studied was not affected by lairage in the slaughterhouse for up to 6.0 h.

Fish oil has been identified as one of the most important contributors to the levels of polychlorinated biphenyls (PCBs) in food and feed products. In this study, PCB adsorption from fish oil onto activated carbon (AC), other sustainable adsorbents (mussel shell and wood waste ashes) and organic solvent such as ethanol were compared and optimized. Regarding to adsorbents, PCBs were extracted from fish oil by a 2.0% adsorbent material dose, during 6.0 h at 25 °C. Solvent extraction was carried out using 2 × 5.0 mL ethanol by manually stirring for 3.0 min, and then by Ultrasound-Assisted Solvent Extraction (UASE) for 5.0 min. The results showed that removal rates obtained by using adsorbent materials ranged from 0.0 to 10% for marker PCBs, from 0.0 to 37% for mono-ortho-PCBs, from 0.0 to 74% for PCB11 and from 0.0 to 95% for non-ortho-PCBs. Regarding to solvent extraction, ethanol was used by manually stirring and then by Ultrasound-Assisted Solvent Extraction (UASE). The samples were then centrifuged (2000 rpm/10 min) and the alcoholic phase was removed. With this method, removal efficiencies were much better (85-116%); nevertheless, high eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) removal rates (70-78 and 71-79%, respectively) were detected. We can conclude that adsorption with adsorbents depends on the geometry of PCB congeners, as well as both type of adsorption material and their origin, and that several sorption cycles are needed. Adsorption with ethanol could be the most effective methodology but nutritional quality was impaired, what makes necessary to look for other not so polar removal solvents.

Summary An improved extraction (2.5% HPO3, 5 mm dithiothreitol) and HPLC quantification methodology using a C–18 column at 35 °C and 0.1 m acetic acid (98%) and acetonitrile (2%) mobile phase was developed to quantify total ascorbic acid (AA) in commercial whole/semi-skim/skim raw/pasteurised/UHT milk packaged in opaque bags, transparent plastic, cardboard and Tetra Brik™. AA content ranged from 0.21 to 10 and from 3.4 to 16 mg L−1 in milk from retail outlets and processing plants, respectively, and was higher in organic milk. For same processor/lot samples, pasteurised milk showed higher AA content than UHT milk. This was not true for retail outlets samples. AA content was similar for whole/semi-skim and semi-skim/skim milk, but not for whole/skim comparisons. Among UHT samples, the AA content trend was whole

Animal nutrition constitutes an important issue for the animal production industry. Products intended for animal feed may contain undesirable substances which could endanger animal health or, because of their presence in livestock products, human health or the environment. In this sense, several incidents related with the presence of persistent organic pollutants, particularly with polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs), have happen in food and feed additives. Simple, fast, quantitative, and economic methods for determining PAHs and PCBs in potentially polluting feed ingredients were developed and evaluated. The methods involved liquid-liquid extraction (LLE), clean-up, and detection by liquid chromatography with fluorescence detection (LC-FD) for PAHs and gas chromatography (GC) coupled to electron capture detection (ECD) for PCBs, using tandem mass spectrometry (MS/MS) as confirmatory technique. The overall method quantification limits range from 0.020– 4.0 μg/Kg fo...

Vinasses coming from the five CBOs of Galicia, north-western Spain, were characterized, and successfully employed as economic nutritional supplements for xylitol production by Debaryomyces hansenii . All fermentations can be modelled showing kinetic patterns fairly described by the mathematical models. No negative effect of the phenolic compounds in the liquid phase on the initial volumetric rate of product formation ( r P 0 ) was observed. Multiple linear regression analysis was used to describe the effect of metals and initial xylose acting on P max and Y P / S . Zn was the most influential variable. Besides, partial least-squares regression models show a clear separation, based on the first two principal components, between the whole vinasses and the liquid fractions, which provided the higher P max , with the exception of CBO 4, where P max = 40.4 g/L, was achieved using the solid and liquid fraction.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous contaminants in the marine environment derived from uncontrolled petroleum spills, marine transports, discharges from ships, and urban runoff. The NW coast of Spain (Galicia) has a characteristic hydrography, defined by a continuous suite of estuarine systems called “Rias Gallegas”. Some of these estuaries support important industrial and urban centres while others are preserved from human influence. The PAH contents in invertebrate organisms, such as bivalves, have been widely investigated because they are good bioindicators of pollution due to their ability to filter the water. 12 PAHs were determined in canned mussels (Mytilus galloprovincialis), clams (Venerupis pullastra), cockles (Cerastoderma edule) and knives (Ensis ensis) from the Galician “Rias Gallegas” to perform a possible risk assessment on these contaminants. Pyrene, benzo[b]fluoranthene, benzo[k]fluoranthene and benzo[a]pyrene were detected in all the selected samples by liquid chromatography coupled with fluorescence spectroscopy. With regard to benzo[a]pyrene, which represents the most potentially carcinogenic PAHs, European Regulation 1881/2006/EC fixed at 10 μg/kg of wet weight the MRL (maximum residual level) in bivalve molluscs. None of these marine invertebrates showed values higher than the MRL and therefore, they do not suppose risk for human health. Mussels hydrocarbon body burden levels were highest than exhibited by others bivalves. Some factors may determine this feature, as lipid content or habitat. Lower PAH levels of five and six benzene rings were detected in canned mussels in pickle sauce (vegetable oil, vinegar, paprika, salt and other spices) than in mussels in natural sauce (water, salt and other spices). PAHs migration from mussels to the vegetable oil in pickle sauce was verified to be the reason.

Eleven polycyclic aromatic hydrocarbons (PAHs) have been analysed in commercial milk formulae and infant cereals. Two hydroxylated PAHs metabolites (1-OH-Pyr and 3-OH-B[a]P) and their conjugates were also analysed in milk samples. To determine the selected PAH metabolites, a simple, fast quantitative and economic method was developed. This method comprising ultrasound-assisted solvent extraction, enzymatic hydrolysis, solid-phase clean-up and detection by liquid chromatography with fluorescence detection (LC–FD) and liquid chromatography tandem mass spectrometry (LC–MS/MS) as confirmatory technique. The method was evaluated by constructing calibration curves, measurement of recovery, precision and the limits of detection. The purpose of this survey was to determine the selected analytes, to assess the exposure of babies and infants and to produce data for comparison with proposed limits that were being considered at the time of the survey. The results showed that not only no samples would have exceeded the limit for benzo[ a ]pyrene which is used as an indicator for the presence of PAHs, but also no hydroxy PAH metabolites have been detected.

Summary Thermal waters are certainly a substantial asset of the Galicia region of Spain. They can be regarded as worth developing because of their human health implications and, if thermal tourism is promoted, their importance to the local economy. In this paper the chemistry of major and trace inorganic elements in about 45 thermal springs and wells discharging in the same hydrographical system are presented and discussed. For handling the results of all measurements, graphical representations of B/Li vs. SO 4 2 - /Cl− ratios, Hill–Piper diagram, discriminant analysis (DA) and principal component analysis (PCA) were employed. All this with the intention to classify, based on their inorganic pattern, both thermal springs and wells waters, but also waters circulating through adamellite and granodiorite rocks. The results of the hydrogeochemistry analysis showed three main water families: sulphated, chlorinated and bicarbonated waters. The results show also the presence of saline materials with chloride influence in the deeper aquifer, allowing its classification in deeper and younger/shallow waters.

The purpose of this work was to examine the relationship of the total polyphenol index (TPI) as measured spectrophotometrically and individual phenols as determined by HPLC to antioxidant activity in red wines made from two Vitis vinifera grape varieties grown in NW Spain (viz. Mencia and Brancellao) during bottled storage in the dark for 12 months. Antioxidant activity was determined by using various methods based on inhibition of the coupled oxidation of the β-carotene/linoleic acid mixture. Also, free radical scavenging activity was determined with 2,2-diphenyl-1-picrylhydrazyl (DPPH). Antioxidant activity as measured by DPPH after storage for 3 months was higher in Mencia wines than in Brancellao wines (4.5 ± 0.60 vs. 3.7 ± 0.30 mmol/L trolox equivalents). Beyond 3 months of storage, radical scavenging activity increased in both types of wine (from 4.5 ± 0.60 to 6.0 ± 0.80 mmol/L in Mencia wines and from 3.7 ± 0.30 to 4.7 ± 0.71 mmol/L trolox equivalents in Brancellao wines). The significant correlation found between antioxidant activity and TPI in all wines (r > 0.88) is indicative of the significance of condensation and polymerisation products to the total antioxidant activity of the wines. The relatively high correlation of total flavonols (r = 0.89) and acylated anthocyanins (r = 0.70) as measured by HPLC with to the overall antioxidant capacity suggests that these two polyphenol classes can substantially influence the antioxidant properties of these wines. The linoleic acid/β-carotene assay exposed a higher antioxidant capacity in Brancellao wines than in Mencia wines after 3 months of storage (6.0 ± 0.80 vs. 4.7 ± 0.70). Most of the studied wines lost an average 45% antioxidant activity during bottled storage. Antioxidant activity as measured with the linoleic acid/β-carotene assay was closely correlated with both hydroxycinnamic acids (r = 0.90) and flavanols (r = 0.71 and 0.61 for monomeric and polymeric forms, respectively).

PAHs from a polluted atmosphere are generally transferred to plants by particle-phase deposition on the waxy leaf cuticle or by uptake in the gas phase through stomata. Thus, they are also present in ingredients for animal feed. Generally, toxic substances are metabolized before or after absorption through the intestinal tract. This is the case of 3-hydroxybenzo[a]pyrene, which can be found free or in its glucuronide and/or sulphate conjugate forms. This article develops a procedure to monitor the carry-over of PAHs from feed to food of animal origin based on the analysis of animal manure. Eleven PAHs and 3-hydroxybenzo[a]pyrene were measured in four animal manures (cow, horse, rabbit and pig) by liquid chromatography with fluorescence detection. benzo[a]pyrene, benzo[b]fluoranthene and benzo[k]fluoranthene were found in all the selected samples, but their total levels were not alarming (benzo[a]pyrene equivalents (microg/kg)

Very challenging analytical problems arise from the continuous introduction in agriculture of chemical pesticides. Particularly, diquat (DQ), paraquat (PQ) and difenzoquat (DF) are a difficult group of quaternary ammonium herbicides to analyze. This article reviews and addresses the most relevant analytical methods for determining the selected herbicides in soil. We discuss and critically evaluate procedures, such as digestion-based methods, shaking extraction and microwave-assisted extraction (MAE). Clean-up of extracts was performed by solid-phase extraction (SPE) using silica cartridges. Detection of these herbicides was carried out by liquid chromatography (LC) coupled to UV detection and mass spectrometry (MS) as confirmatory technique. Recoveries ranged from 98% to 100% by digestion, from no recovered to 61% by shaking, and from 102% to 109% by MAE with estimated quantification limits between 1.0 microg/kg and 2.0 microg/kg by digestion and 5.0 mug/kg and 7.5 microg/kg by MAE using LC/MS-MS as detection technique. The recoveries obtained under the optimum conditions are compared and discussed with those obtained from digestion extraction and MAE.

The purpose of this work is to study the evolution, over three selected harvests, of phenolic compounds and colour stability of red wines produced in the north of Portugal by means of two wine-making processes (conventional maceration/fermentation and fermentation after initial carbonic maceration), with and without the use of four different fining agents (polyvinylpolypyrrolidine, gelatine, egg albumin, and casein). In general, it was observed that wines obtained by conventional maceration/fermentation (PO) present the highest colour intensity and polyphenolic content (total and monomeric anthocyanins, flavan-3-ol monomers and polymers), and the lowest orange–red hue, immediately following vinification. Nevertheless, carbonic maceration (CM) afforded wines with most stability in colour density, for 26 months’ storage. Different evolutions of anthocyanins and of flavan-3-ol monomers (catechins) and polymers (procyanidins) in PO and CM protocols during storage were observed. Decrease of anthocyanins and flavan-3-ol dimmers were, in general, more remarkable in PO wines. Monomers and trimers underwent a rise, especially in CM wines. Wines treated with fining agents tended to have somewhat lower anthocyanins levels, and especially in the case of PVPP, less intense colouration than untreated wines. Principal component analysis (PCA) revealed that catechins and some procyanidins are mainly responsible for the separation and evolution of wines during storage according polyphenolic composition. Partial least squares (PLS) regression models confirm that colour density in wines from PO and CM protocols explains a high proportion of the variance in hue, chemical age, IFC and anthocyanins and a prediction model has been built.

Various analytical methods based on the HPLC-DAD technique were used to determine 38 phenolic compounds in red wines. While anthocyanins and hydroxycinnamic acids were determined by direct injection of wine samples, hydroxybenzoic acids, catechins, procyanidins and flavonols required an analytical pretreatment involving liquid–liquid partitioning with ethyl ether followed by solid-phase extraction on C18 mini-columns. The proposed analytical methods were used to establish the phenol composition of Mencia and Brancellao, two varietal young red wines, and its influence on colour stability during storage in bottles for one year. At the end of malolactic fermentation, Mencia wine was found to contain much greater amounts of anthocyan pigments than was Brancellao wine. This resulted in a higher colour density but a weaker hue in Mencia wine than in Brancellao wine. Phenolic compounds evolved similarly in both wines during storage; changes in such compounds involved a decrease in the levels of monomeric anthocyanins, phenolic acids, epicatechin and flavonols, and an increase in those of procyanidins. The absence of a relationship between the changes in colour density and monomeric anthocyanins in both wines suggests that copigmentation and polymerization with other phenolic compounds (viz. phenolic acids, catechins and/or flavonols) prevail over degradation of the pigments.

The effect of marinating meat with Pilsner beer, nonalcoholic Pilsner beer, and Black beer (coded respectively PB, P0B, and BB) on the formation of polycyclic aromatic hydrocarbons (PAHs) in charcoal-grilled pork was evaluated and compared with the formation of these compounds in unmarinated meat. Antiradical activity of marinades (DPPH assay) was assayed. BB exhibited the strongest scavenging activity (68.0%), followed by P0B (36.5%) and PB (29.5%). Control and marinated meat samples contained the eight PAHs named PAH8 by the EFSA and classified as suitable indicators for carcinogenic potency of PAHs in food. BB showed the highest inhibitory effect in the formation of PAH8 (53%), followed by P0B (25%) and PB (13%). The inhibitory effect of beer marinades on PAH8 increased with the increase of their radical-scavenging activity. BB marinade was the most efficient on reduction of PAH formation, providing a proper mitigation strategy.

Dioxins include polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and part of polychlorinated biphenyls (PCBs). Only the compounds that are chlorinated at the 2,3,7, and 8 positions have characteristic dioxin toxicity. PCDDs, PCDFs and PCBs accumulate in the food chain due to their high lipophilicity, high stability, and low vapor pressure. They are not metabolized easily; however their hydroxylated metabolites are detected in feces. They cause a wide range of endocrine disrupting effects in experimental animals, wildlife, and humans. Endocrine related effects of PCDDs, PCDFs and PCBs on thyroid hormones, neurodevelopment and reproductive development were referenced. In addition, some studies of contamination of foods, bioaccumulation, dietary exposure assessment, as well as challenges of scientific research in these compounds were reviewed.

The kinetic bioconcentration of N-heterocyclic aromatic hydrocarbons and polycyclic aromatic hydrocarbons in mussels (Mytilus galloprovincialis) after short waterborne exposure was studied. Benzo[a]pyrene (BaP), its analogue azaarene 10-azabenzo[a]pyrene (AzaBaP), and their mixture (Mix), were selected to monitor the changes in water concentrations over three days. Decay of both PAHs concentrations in water after 24 h of waterborne exposure to mussels at levels of 10 and 100 μg/L follows a first order kinetic with half-lives of 4–5 h, with residual levels of PAHs below 7%. While steady-state scenarios are well studied, there is a lack of information of what happens under non-steady-state conditions, the main purpose of our paper. A synergistic bioconcentration of the mixture was found (around 800 in the mix vs. around 400 for individual PAHs at 100 μg/L of waterborne exposure). It could be explained by the following reasons. The most polar AzaBaP does not compete with the most non-polar BaP for the same tissue compartments. Whereas BaP aggregate in hydrophobic areas, AzaBaP can also do in hydrophilic areas. Moreover, a chance for complex formation between them by charge-transfer stabilization mechanisms could make possible a higher bioaccumulation as a mixture. Instead, toxicological results suggest an additive behaviour in the mixture performance, dominated by BaP, which is the key PAH controlling phase I metabolization in mussels, since is approx. three times more toxic. These experiments provide useful indications for a rapid assessment of PAHs kinetic bioconcentration in mussels.

Fish oil has been identified as one of the most important contributors to the level of Persistent Organic Pollutants (POPs) in feed products. The determination of polycyclic aromatic hydrocarbons (PAHs) in fish oils is complicated due to the fat matrix, which affects both extraction efficiency and analytical quality. This article reviews and addresses two of the most relevant analytical methods for determining 11 mutagenic and carcinogenic PAHs, as well as two EPA indicator PAHs in fish oils. We discuss and critically evaluate two different extraction procedures, such as ultrasound-assisted solvent extraction (USAE) and ultrasound-assisted emulsification-microextraction (USAEME). Clean-up of extracts was performed by solid-phase extraction using C18 and glass columns containing silica gel and florisil for USAE or only C18 for USAEME. Detection of the selected PAHs was carried out by high-performance liquid chromatography coupled with fluorescence detection for determination. Optimization of the variables affecting extraction by the selected extraction techniques was conducted and recoveries ranged from 70% to 100% by USAE and from 70% to 108% by USAEME with estimated quantification limits between 0.020 and 2.6 μg/kg were achieved. Moreover, the applicability of the selected methods was evaluated by the analysis of real samples. To our knowledge, this is the first time that USAEME has been applied to the determination of PAHs in food matrices, such as oil fish samples. The methods proposed were applied to the determination of the target PAHs in fish samples from different countries, and it was found that the low PAH contamination of the selected fish oils could mainly occur by atmospheric sources.

Animal nutrition constitutes an important issue for the animal production industry. Products intended for animal feed may contain undesirable substances which could endanger animal health or, because of their presence in livestock products, human health or the environment. In this sense, several incidents related with the presence of persistent organic pollutants, particularly with polychlorinated biphenyls (PCBs), have happen in food and feed additives. Animal feed and feed components are challenging matrices for the determination of residues and contaminants. The variability of these matrices is enormous. It ranges from relatively simple ones like those based on wheat to all kinds of by-products from agro and food industry, such as cereal oils. Firstly, this article reviews and addresses the extraction efficiency of ultrasonic assisted solvent extraction (UASE) and focused ultrasonic solvent extraction (FUSE) for determining selected PCBs in animal feed and ingredients. Detection of these pollutants was carried out by gas chromatography (GC) coupled to electron capture detection (ECD); tandem mass spectrometry (MS/MS) was used as confirmatory technique. Recoveries ranged from 70% to 98% by UASE and from 75% to 106% by FUSE with estimated quantification limits between 0.11 and 0.3 μg/kg in feeds and ingredients and between 0.2 and 0.75 μg/kg in fats. Once the method was optimised, it was applied to 18 feed samples as well as 16 ingredients. PCBs were detected in almost all the selected samples. As expected, the samples of animal origin as shell powder and fish oil showed the highest concentrations of 56 and 29 ng/g, which are equivalent to toxicological concentrations of 123 and 18 ng WHO-TEQDL-PCBs/kg, respectively. Feeds and ingredients from vegetable origin ranged from non-detected to 7.1 μg/kg. PCB 77 and 169 were the discriminant congeners in the selected samples of feed and ingredients. Samples showed that the pattern of PCBs depends on the sources of contamination.

The presence of anthocyanins and flavonols in three selected red grape varieties was investigated, in order to use their polyphenolic characterisation as a fingerprint. Berry skins of Gran Negro grapes were characterised by the presence of high content of malvidin- and peonidin-3-O-glucoside; Mouratón grapes, by the presence of high content of petunidin- and delphinidin-3-O-glucoside; and Brancellao grapes, by the presence of high content of cyanidin-3-O-glucoside. The main flavonols found included the 3-O-glucosides of quercetin, myricetin, kaempferol, laricitrin, isorhamnetin and syringetin. Using cluster analysis and principal components analysis, Gran Negro could be characterised by their content of isorhamnetin-3-O-glucoside and syringetin-3-O-glucoside and, along with Mouratón, by their myricetin conjugates. Flavonol profile could not provide a fingerprint of Brancellao variety. Stepwise discriminant analysis was performed in order to find the polyphenolic compounds, which characterised the selected grape varieties. Finally, anthocyanin and flavonol profiles in red grapes were compared and results confirmed that biosynthesis of flavonols is closely related to that of anthocyanins. © 2011 Elsevier Ltd. All rights reserved., This work was granted by EU FEDER funds, but also by the INCITE09383322PR contract from the Autonomous Community Government in N.W. Spain.

Studies on the temporal deposition of polycyclic aromatic hydrocarbons (PAHs) in peatlands are scarce, and none have been carried out in the Iberian Peninsula. To address this gap, ten PAHs were determined in a short peat core (spanning the last 1000 years) sampled in NW Iberian Peninsula, by HPLC-fluorescence. Fluoranthene, pyrene, benzo[b]fluoranthene and indeno[1,2,3-cd]pyrene predominated in the upper layers (10 cm), whereas fluoranthene and pyrene were the most abundant in the lower layers (40 cm), which showed an absence of high molecular weight PAHs (benzo[ghi]perylene and Indeno[1,2,3-cd]pyrene). Although increased PAH contents have been detected since 1700 A.D., coinciding with the beginning of the Metallurgical and Industrial Revolution, high levels of fluoranthene and pyrene were present in peat samples dating back to the 12th century A.D. The results suggest that changes in sources, type of emission (global or local) and transport could be responsible for the different PAH content and composition of the peat core. These changes are consistent with the history of the use of natural resources in the NW of the Iberian Peninsula.

Foliar runoff is one of the most important processes affecting off-target movement of fungicides. In this way, Ridomil® Gold Plus and Ridomil® Gold MZ are two types of wettable powder technical formulations which contain metalaxyl and they are used for such a purpose. A method for quantitative determination of metalaxyl in pesticide formulas has been developed, validated, and subsequently applied to Ridomil® Gold Plus and Ridomil® Gold MZ. The method employs liquid–liquid extraction followed by liquid chromatography coupled with UV detection (LC–UV), using gas chromatography coupled with mass spectrometry as confirmation technique and to carry out a screening of organic adjuvants of these two selected pesticide formulas. Metalaxyl of 26.5 and 41 g/kg was detected in Ridomil® Gold Plus and Ridomil® Gold MZ, close to the manufacture specified level of 25 and 40 g/kg, respectively. Activator and utility adjuvants were detected in these two wettable powder technical formulations. Only methyl-ester-based surfactants were found within the group of nonionic surfactants, but the long-term fates of most adjuvants in soils and elsewhere in the environment are largely unknown, partially because of the lack of long-term monitoring data.

Some polycyclic aromatic hydrocarbons (PAHs), particularly those with a high molecular weight, have been classified as probably carcinogens to humans by the International Agency for Research on Cancer (IARC). The significance of the determination of PAHs is reflected by the special attention of the European Union, which is paying to regulate the maximum allowed levels of PAHs in foodstuffs such as smoked foods. Like other thermally processed foodstuffs, toasted bread can contain these carcinogenic chemicals, not only due to a contamination at source but also during toasting. In order to check PAHs generated from toasting in sandwich bread, several treatment conditions were evaluated: direct toasting (flame-toasting, coal-grilling or gas oven-toasting) or indirect toasting (electric oven-toasting). PAHs were extracted by solid-liquid extraction (SLE) and determined by liquid chromatography with fluorescence detection (LC-FD). Based on the results, the used toasted technique would strongly affect in PAH levels in the final product. No samples obtained by electric oven and toaster were polluted; otherwise the samples toasted by charcoal and flame grilling presented very important levels. Up to 350μg/kg of total PAHs were detected in toasted samples by wood flame. Differences between different ways of toasting could be ascribed to deposition of PAHs from smoke. Finally, several commercial toasted samples of bread were tested to determine PAHs. Overall, the PAH levels were very low. Benzo[a]pyrene ranged from no detectable to 0.23μg/kg.

Retention of the cationic herbicides paraquat (PQ), diquat (DQ), and difenzoquat (DFQ) in two vineyard soils with a different management history and retention capacity was examined. The influence of copper on the ability of the soils to retain the herbicides was determined by comparing the results of adsorption and desorption tests on untreated and Cu-enriched soil samples, and also on soils that were previously treated with EDTA to extract native copper. The three herbicides were strongly adsorbed by both soils. Soil 1 exhibited linear adsorption isotherms for PQ and DFQ with partition coefficients, KD, of 1.28 x 103 and 1.37 x 103 L kg-1, respectively, and a Freundlich-type isotherm for DQ with a linearized partition coefficient, KD*, of 1.01 x 103 L kg-1. On the other hand, soil 2 exhibited curved isotherms and smaller KD* values (viz. 106, 418, and 28 L kg-1 for PQ, DQ, and DFQ, respectively). Using EDTA to extract copper from the soils released new sites for the herbicides to bind. The three herbicides exhibited strong hysteresis in the adsorption-desorption process. Extracting copper decreased the percent desorption of PQ and DQ; on the other hand, it decreased the affinity of DFQ for the resulting vacant adsorption sites. Similarly, competitive adsorption tests with copper and the herbicides revealed that the metal was only capable of displacing DFQ from adsorption sites. The behavior of this herbicide in the soils was consistent with a specific adsorption model. The disparate behavior of the two soils toward the herbicides was a result of the adsorption sites in soil 1 being less extensively occupied than those of soil 2 in the adsorption tests. The effect of copper on the adsorption of DFQ in the two soils was acceptably reproduced by an adsorption model involving Coulombic and specific sorption with competition from the metal.