6. WELFARE QUALITY ® APLICADO A CERDOS DE ENGORDE
6.3 R ECOPILACIÓN DE DATOS PARA CERDOS DE FINALIZACIÓN EN EL MATADERO
448
Quantification of Carbon Nanotubes in Complex Matrices: Possibilities of Electron Microscopy
R. Kaegi, A. Gogos, Eawag Swiss Federal Institute of Aquatic Science and Technology
Fascinating properties of Carbon nanotubes (CNTs) allow the development of novel materials with increased functionalities (e.g. reduced weight, increased strength). Nevertheless, CNTs do pose potential environmental and human health risks and reliable methods to quantify CNTs at low concentration in complex matrices are still lacking. We therefore developed a method, based on the unique shape of the CNTs to quantify these materials in complex matrices. Multiwalled CNTs (IRMM 382) suspended in either ultrahigh quality (UHQ) water or in soil
98 SETAC Europe 28th Annual Meeting Abstract Book
extracts were directly centrifuged on transmission electron microscopy (TEM) grids, resulting in an even distribution of the CNTs on the grids. Samples were investigated with a scanning electron microscope (SEM, Magellan XHR 400, FEI) operated at an acceleration voltage of 20 kV in emersion mode and using a bright field transmission electron detector. A ridge detection algorithm implemented in the image processing software Fiji was used to detect and characterize individual CNTs on recorded images. The concentration of the CNTs in the suspension was calculated based on the total length of all CNTs (provided by the ridge detection algorithm) detected on the images in combination with their thickness (20 nm), their density (1.4 gcm-3) and the well-defined volume of suspension that was centrifuged on the TEM grids. CNTs were well separated on the TEM grids and an increasing number of CNTs was observed on images with increasing concentrations of CNTs in suspension. Plotting the calculated concentration of CNTs in suspension against the nominal CNT concentrations (10 µgL-1 – 100 µgL-1) resulted in a linear relationship. The calculated and the nominal CNT concentrations were in good agreement at low CNT concentrations, but at high concentrations, the calculated concentrations underestimated the nominal values by a factor of ~2. Almost identical results were obtained from CNTs in UHQ water and in soil extracts (5 mgL-1) indicating that the detection of the CNTs was not compromised by the presence of soil particles. Future experiments will focus on a selective removal of the soil particles by an additional treatment with diluted hydrofluoric acid. Initial experiments are promising and suggest that the detection limit of the methods can be lowered to 1 mg(CNT) / kg(soil), which would represent huge step forward in detecting of CNTs in complex matrices.
449
Monitoring for perfluorinated compounds, insecticides, and brominated flame retardants in the water of Daechung lake and Geum river basin
H. LEE, National Institute of Environmental Research (NIER / Geum River Water Environment Research Center; Y. Cho, J. Khan, National Institute of
Environmental Research NIER / Geum river environment research center; B. Lee, National Institute of Environmental Research NIER / Han river water environment research center; B. Seol, M. Chae, S. Cheon, National Institute of Environmental Research NIER / Geum River Water Environment Research Center
A multiresidue analytical method using LC-MS/MS was developed for perfluorinated compounds (PFCs), insecticides, and brominated flame retardants (BFRs) in water samples with the simultaneous SPE method. The ranges of recoveries were 19.7 ~ 135.0 % (PFCs), 95.0 ~ 117.2 % (Insecticides), and 72.5 ~ 86.4% (BFRs), with coefficients of variation of less than 15%. Method detection limit (MDLs) of PFCs, insecticides, and BFRs were 0.3 ~ 7.1 ng/L, 3.0 ~ 3.7 ng/L, and 5.1 ~ 11.7 ng/L, respectively while limit of quantifications (LOQs) were 0.9 ~ 21.4 ng/L (PFCs), 9.0 ~ 11.0 ng/L (Insecticides), and 15.4 ~ 35.0 ng/L (BFRs). For understanding the background levels of PFCs, insecticides, and BFRs in the river water, those compounds were monitored in Geum river main stream, So-ok stream, Juwon stream, and Daechung Lake (Dam) every month (March to December) utilizing the developed method. The compounds of the highest detection frequency were PFOA, PFHxA, and dinotefuran (Insecticide), whereas BFRs were detected only in March and December, except for main stream. In conclusion, the trends were not observed on periodical and spatial characteristics and the background levels were secured for PFCs, insecticides, and BFRs in Geum river basin.
450
Impacts of Contaminants of Emerging Concern on Terrestrial Organisms
S. Dudley, UC Riverside / Environmental Toxicology; M. Pennington, UCR / Environmental Toxicology; C. Sun, UC Riverside / Environmental Science; J. Trumble, University of California, Riverside / Entomology; J. Gan, University of California, Riverside / Department of Environmental Sciences
Impacts of Contaminants of Emerging Concern on Terrestrial Organisms
Stacia Dudley1, Marcus Pennington1, Chenliang Sun2, John Trumble3,Jay Gan2 1Environmental Toxicology Graduate Program, University of California, Riverside, CA 2Department of Environmental Sciences, University of California, Riverside, CA 3Department of Entomology, University of California, Riverside, CA Reclaimed water is a historically underutilized resource. However, with increased population growth and global climate change placing increased pressure on fresh water resources, reclaimed water is evolving into an economical and sustainable means to meet the needs of citizens, industries, and agriculture. The use of recycled water for agriculture comes with the potential risk of environmental and food contamination by contaminants of emerging concern (CECs). These compounds pose a potential threat to the health of ecosystems because they are designed to be biologically active at low concentrations and are considered “pseudo-persistent” due to their continuous release into the environment. Using high resolution mass spectrometry, 14C tracing, enzyme extraction and Illumnia sequencing techniques we evaluated a wide range of biological effects in terrestrial organism caused by exposure to CECs. Organisms in these studies included, the cabbage looper (Trichoplusia ni), an earthworm (Eisiena fetida), a model plant (Arabidopsis
thaliana) and cucumbers (Cucumis sativus), radishes (Raphanus raphanistrum sativus) and tomatoes (Solanum lycopersicum). These studies have revealed a
multitude of effects including increased mortality and development time, reduction in weight, changes to the microbiome and up-regulation of enzymes associated with oxidative stress. Further, the study has highlighted the potential for higher plants to
take up, translocate and detoxify CECs.
451
Occurrence of pharmaceuticals and their metabolites in Euthynnus alletteratus bile
J.M. Peña Herrera, Institute of Environmental Assessment and Water Research (IDAEA-CSIC) / Environmental Chemistry; N. Montemurro, IDAEA CSIC Barcelona / Dipartimento di scienze agro-ambientali e territoriali; E. Navarro, CSIC - Spanish National Research Council / Dept. Recursos marins renovables; J. Navarro, ICM-CSIC / Dept. Recursos marins renovables; M. Solé, ICM-CSIC; S. Perez, IDAEA CSIC / Environmental Chemistry; D. Barcelo, IIQAB-CSIC / Department of Environmental Chemistry
The primary source of pharmaceuticals in surface waters has been attributed to the effluents of WWTP among others. The widespread occurrence of pharmaceuticals in the aquatic environment has raised concerns about their potential adverse effects on exposed wildlife. Little is currently known on exposure levels of drugs in fish, but some studies reported the detection of pharmaceuticals and endocrine disrupting compounds in this type of biota. Due to possible accumulation processes, pharmaceuticals and metabolites could be thousand times more concentrated in fish than in polluted living waters. By other hand, fish are known to possess a hepatic detoxification system which are likely capable of metabolizing pharmaceuticals taken up from polluted waters. Some studies proposed the analysis of bile from fish to evaluate pharmaceuticals exposure including the identification of metabolites by UPLC-HRMS. In this context, we propose the evaluation of the metabolism of frequently detected drugs in fish, performing a rapid screening of bile by HR-MS for the presence of stable intermediates. Fish were collected from different regions in the Mediterranean coast of Spain. Afterwards, their bile was isolated from the fish and analyzed for the detection of parent drugs and some metabolites listed in an in-house suspected list. A sampling campaign was planned to collect tuna fish, Euthynnus alletteratus, from the Mediterranean coast of Spain: Tarragona, Cartagena and Ceuta catched by spearfishing between 2015-2017. Their bile samples were analyzed directly by UPLC-HRMS after a protein precipitation. The HRMS data allowed screening for suspected pharmaceuticals and their metabolites and provided plausible chemical formulae. The comparison of MS/MS spectra of the parent compounds and their metabolites allowed to propose chemical structures for possible metabolites in fish bile. With this analytical methodology some metabolites, corresponding to different reactions that includes products of hidroxylation, glucuronide conjugates were identified. The suspect analysis of bile samples allowed the detection of several pharmaceuticals. Psycho-active drugs were one of the most commonly detected drugs. Their identities were proposed by matching their accurate MS and MS/MS data against different libraries. Finally, authentic standards were employed to confirm the proposed drug identities and to determine analyte concentrations in the fish samples.
452
Accumulation and fate of 12 human drugs through the soil-root-leaf system
N. Montemurro, IDAEA CSIC Barcelona / Dipartimento di scienze agro-ambientali e territoriali; C. Postigo, IDAEA, CID-CSIC / Environmental Chemistry; S. Perez, IDAEA CSIC / Environmental Chemistry; D. Barcelo, IIQAB-CSIC / Department of Environmental Chemistry
Crop irrigation with reclaimed water has become an extended practice in many countries worldwide where the water scarcity and excessive exploitation of agriculture are forcing local authorities to look for alternative resources. Despite this practice increases local water resources and contributes to nutrient recycling, using reclaimed water for irrigation, however, represents a primary source of emerging organic contaminants resilient to wastewater treatment processes, such as some pharmaceuticals and personal care products [1]. These pollutants can be retained in the soil, directly uptaken by crops or translocated from soil to plant tissues above the ground [2,3]. The present work aimed to evaluate the transfer and the bioaccumulation of organic contaminants of emerging concern (mainly pharmaceuticals) in lettuce tissues and soil. The distribution of twelve relevant wastewater-derived pollutants was evaluated in lettuce tissues (leaves and root system) and soil. This list included nine prescription drugs (diclofenac,
trimethoprim, carbamazepine, oxcarbazepine, lamotrigine, cis-diltiazem, valsartan, midazolam, and methadone), an illegal drug (cocaine) and two transformation products (acridone and valsartan acid). Lettuce plants were grown in pots in a controlled environment and irrigated with artificial spiked water containing the 12 compounds during the entire growing period (60 days). Control was irrigated with tap water. Afterwards, a set of new lettuce plants were grown in the same soil pots and irrigated with rainwater or with tap water, if necessary. At the end of each experiment, leaves, roots and soil samples have been collected for each plot. All pharmaceutically active compounds were extracted by ultrasonic liquid extraction (USE) [4]. The detection of the target analytes was performed using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The results from the first growing season evidenced the presence of all analytes in all investigated matrices. Carbamazepine was the analyte that accumulated the most in lettuce plants (leaves and root system), whereas cis-diltiazem, methadone, and midazolam were preferentially accumulated in the plant root system and the soil. Concentrations of the target analytes in the plant-root-soil system after the second growing season were significantly lower than those measured after the first growing
season, but still detectable for most of the compounds.
453
Root-uptake and dissipation of atenolol, sulfamethoxazole and carbamazepine applied as a single compound solution or in mixture of all compounds in three soils and five plants
R. Kodesova, A. Klement, Czech University of Life Sciences Prague / Dept of Soil Science and Soil Protection; O. Golovko, University of South Bohemia in Ceske Budejovice / South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses; M. Fer, Czech University of Life Sciences Prague / Dept of Soil Science and Soil Protection; O. Koba, University of South Bohemia in Ceske Budejovice / South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses; M. Kocarek, A. Nikodem, Czech University of Life Sciences Prague / Dept of Soil Science and Soil Protection; R. Grabic, University of South Bohemia in Ceske Budejovice / South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses
This study was focused on a root uptake of carbamazepine, atenolol and sulfamethoxazole from 3 soils: Haplic Chernozem, Haplic Cambisol and Arenosol Epieutric. Five plants (radish, arugula, lettuce, spinach and green peas) planted in those soil were initially irrigated by fresh water and next with water contaminated by a single compound or their mixture. After 3 or 4 weeks, each plant was divided into separate parts: roots (including bulbs of radish), leaves, stems (green peas) and pods (green peas). Plant parts and soils were freeze-dried and dry-masses and concentrations of pharmaceuticals and their metabolites were measured. Despite that atenolol and sulfamethoxazole relatively rapidly dissipate from soils, they and metabolites of atenolol were detected in all plants. Carbamazepine is very stable in soils and fractions of its metabolites are usually low. However, very high concentrations in all plants were measured not only for carbamazepine but also for its metabolites. The degree of compounds’ transformation depended on a plant family. Considerably higher concentrations of atenolol, sulfamethoxazole and metabolites of atenolol were measured in roots in comparison to those in leaves and soils. In the case of carbamazepine, the highest concentrations were measured in leaves followed by roots and soils. Both indicate a high potential of plants to accumulate studied pharmaceuticals in their bodies and a high ability to transform studied compounds. Particularly in the case of carbamazepine, the considerably higher concentrations of metabolites were measured in leaves in comparison to concentrations in roots and very low or negligible concentrations in soils. Transformation of compounds in plant bodies is attributed to enzymes CYP450. Larger concentrations of carbamazepine metabolites were measured in leaves of lettuce, spinach and green peas than in leaves of radish and arugula (Order – Brassicales, Family – Brassicaceae). Oxcarbazepine was detected only in plants (not in soils). The impact of soil type on compound’s uptake was not proven for all tested plants. The impact of application (single compound versus compounds’ mixture) differed for different plants. Antibiotic sulfamethoxazole likely reduced dissipation of other two compounds in soils, which increased relative
concentrations of compounds in plants (i.e., concentrations of compound in plant divided by compound loads in soils that is a total amount of applied solute divided by a dry mass of soil).