RESUMO
Due to the estrogenic behavior of bisphenol (BP) A, industries have developed many substitutes, such as BPS and BPF. However, due to their structural similarities, adverse effects on reproduction are currently observed in various organisms, including fish. Even if new results have shown impacts of these bisphenols on many other physiological functions, their mode of action remains unclear. In this context, we proposed to better understand the impact of BPA, BPS, and BPF on immune responses (leucocyte sub-populations, cell death, respiratory burst, lysosomal presence, and phagocytic activity) and on biomarkers of metabolic detoxification (ethoxyresorufin-O-deethylase, EROD, and glutathione S-transferase, GST) and oxidative stress (glutathione peroxidase, GPx, and lipid peroxidation with thiobarbituric acid reactive substance method, TBARS) in an adult sentinel fish species, the three-spined stickleback. In order to enhance our understanding of how biomarkers change over time, it is essential to determine the internal concentration responsible for the observed responses. Therefore, it is necessary to explore the toxicokinetics of bisphenols. Thus, sticklebacks were exposed either to 100 µg/L of BPA, BPF or BPS for 21 days, or for seven days to 10 and 100 µg/L of BPA or BPS followed by seven days of depuration. Although BPS has very different TK, due to its lower bioaccumulation compared to BPA and BPF, BPS affect oxidative stress and phagocytic activity in the same way. For those reasons, the replacement of BPA by any substitute should be made carefully in terms of risk assessment on aquatic ecosystems.
RESUMO
Due to the high production volume and persistence in the environment of bisphenol A (BPA) and its substitutes, realistic exposure scenarii were proposed in some species to better understand the relationship between external and internal concentrations. For example, a recent PBTK model has been developed and adapted to BPA ADME (Absorption, Distribution, Metabolization, and Excretion) processes in three-spined stickleback. These substances have an impact on organism physiology including reproductive and immune functions. In this context, physiologically-based toxicokinetic models coupled with toxicodynamics (PBTK-TD) have proven to be valuable tools to fill the knowledge gap between external exposure and effect dynamics. The aim of the current work was to explain the impact of BPA on the immune response by determining its temporality. In addition, the relationship between BPA dose and these responses was investigated using a PBTK-TD model. Two experiments were performed on stickleback to characterize their biomarker responses, (i) a short exposure (14 days) at 0, 10 and 100 µg/L, including a depuration phase (7 days), and (ii) a long exposure (21 days) at 100 µg/L to measure the immunomarker dynamic over a long period. The fish spleens were sampled to analyze immune responses of stickleback at various times of exposure and depuration: leucocyte distribution, phagocytic capacity and efficiency, lysosomal presence and leucocyte respiratory burst index. At the same date, blood, muscle, and liver were sampled to quantify BPA and their metabolites (BPA monoglucuronide and BPA monosulfate). All these data enabled the development of the indirect pharmacodynamic models (PBTK-TD) by implementing the responses of biomarkers in the existing BPA PBTK of stickleback. The results shown a high induction of phagocytosis activity by BPA in the two exposure conditions. Furthermore, the immunomarkers exhibit very different temporal dynamics. This study demonstrates the need of a thorough characterization of biomarker response for a further use in Environmental Biomonitoring.
Assuntos
Smegmamorpha , Poluentes Químicos da Água , Animais , Poluentes Químicos da Água/toxicidade , Smegmamorpha/fisiologia , Fagocitose , BiomarcadoresRESUMO
Over the last decade, fluctuations of retinoids (RETs), also known as vitamin A and derivatives, have proved to be useful biomarkers to assess the environmental chemical pressure on a wide variety of non-target vertebrates. This use of RET-based biomarkers is of particular interest in the non-target sentinel species Gammarus fossarum in which RETs were shown to influence crucial physiological functions. To study and probe this metabolism in this crustacean model, a UHPLC-MS/MS method was developed to 1) identify and 2) monitor several endogenous RETs in unexposed females throughout their reproductive cycle. Then, females were exposed in controlled conditions to exogenous all-trans retinoic acid (atRA) and citral (CIT), a RA synthesis inhibitor, to simulate an excess or deficiency in RA. Perturbation of vitamin A metabolism by pesticides was further studied in response to methoprene (MET), a juvenile hormone analog as well as glyphosate (GLY). The developed method allowed, for the first time in this model, the identification of RA metabolites (all-trans 4-oxo and 13-cis 4-oxo RA), RA isomers (all-trans and 13-cis RA) as well as retinaldehyde (RALD) isomers (all-trans, 11-cis, and 13-cis RALD) and showed two distinct phases in the reproductive cycle. Retinoic acid successfully increased the tissular concentration of both RA isomers and CIT proved to be efficient at perturbating the conversion from RALD to RA. Methoprene perturbed the ratios between RA isomers whereas GLY had no observed effects on the RET system of G. fossarum females. We were able to discriminate different dynamics of RET perturbations by morphogens (atRA or CIT) or MET which highlights the plausible mediation of RETs in MET-induced disorders. Ultimately, our study shows that RETs are influenced by exposure to MET and strengthen their potential to assess aquatic ecosystem chemical status.
Assuntos
Metoprene , Vitamina A , Animais , Feminino , Ecossistema , Espectrometria de Massas em Tandem , Tretinoína , Retinoides , Isotretinoína , Retinaldeído/metabolismo , GlifosatoRESUMO
Bisphenol A (BPA) is a chemical of major concern due to its endocrine disrupting function, high production volume, and persistence in the aquatic environment. Consequently, organisms such as fish are subject to chronic exposure to BPA. However, physiologically-based toxicokinetic (PBTK) models, which are valuable tools to improve the understanding of a chemical's fate in an organism, have never been specifically adapted to model BPA toxicokinetics (TK) in fish. In our work, an existing PBTK developed for four different fish species was modified to model BPA ADME processes (absorption, distribution, metabolization and excretion). The metabolization of BPA into BPA-monoglucuronide (BPA gluc) and BPA-monosulfate (BPA sulf) and their TK in various organs was taking into account in the model. Experiments were performed to generate BPA TK data in a model species commonly used in ecotoxicology, the stickleback. The model structure had to include two sites of metabolization to simulate BPA TK accurately in stickleback organs. Thus, the fish liver may not be the only site of the metabolization of BPA: plasma or gills could also play a role in BPA metabolization. The PBTK model predictive performance evaluated on literature data in zebrafish and rainbow trout concurs with this conclusion. Finally, a calibration mixing data from the three species was compared to the calibration on stickleback data only.
Assuntos
Smegmamorpha , Poluentes Químicos da Água , Animais , Compostos Benzidrílicos/toxicidade , Fenóis , Toxicocinética , Poluentes Químicos da Água/toxicidade , Peixe-ZebraRESUMO
During recent decades, ultrasmall inorganic nanoparticles have attracted considerable interest due to their favorable biodistribution, pharmacokinetics and theranostic properties. In particular, AGuIX nanoparticles made of polysiloxane and gadolinium chelates were successfully translated to the clinics. In an aqueous medium, these nanoparticles are in dynamic equilibrium with polysiloxane fragments due to the hydrolysis of Si-O-Si bonds. Thanks to high-performance liquid chromatography coupled with electrospray ionization mass spectrometry, all these fragments were separated and identified.
RESUMO
Despite their very wide use in various fields, knowledge concerning surfactants in environmental solid matrices is generally poor. One of the difficulties encountered in the analysis of surfactants is their very diverse physicochemical properties which require different extraction techniques. The objective of this work was therefore to develop an extraction method in sediments that allows the simultaneous analysis of anionic, cationic and non-ionic surfactants. Different extraction techniques (salting-out, ultrasound), solvents and additives were compared. The optimized method, followed by analysis by coupling liquid chromatography with tandem mass spectrometry, was then validated and applied to real samples in which the analytes were quantified by matrix matched calibration. Optimization of the extraction parameters showed different trends depending on the surfactant family. However, ultrasound assisted extraction with a 90/10 acetonitrile/water mixture at 1% acetic acid and 0.1 M EDTA showed the best results overall. The quantification limits obtained, between 6.4 µg/kg for linear alkylbenzene sulfonate (LAS) C10 and 158 µg/kg for 1-laureth sulfate, allow the analysis of traces in sediments. Eighteen of the 27 targeted surfactants were thus detected. The highest concentrations were found for LAS and quaternary ammoniums. Strong correlations between concentrations of different homologues of the same families of surfactants were observed.
Assuntos
Tensoativos , Poluentes Químicos da Água , Cromatografia Líquida/métodos , Extração em Fase Sólida/métodos , Espectrometria de Massas em Tandem/métodos , Água , Poluentes Químicos da Água/análiseRESUMO
Of the large number of emerging pollutants discharged from wastewaters into surface waters, surfactants are among those with the highest concentrations. However, few monitoring in river waters of these substances have already been performed and only on a few families, mostly anionic. This work aimed to develop a multi-family analytical strategy suitable for the quantification of low concentrations of surfactant in surface waters. Twelve families of surfactants, anionic, cationic and non-ionic were selected. Their quantification by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) and their extraction by SPE were optimized by comparing different retention mechanisms. The best performances were obtained with a C18 grafted silica LC column and a hydrophilic-lipophilic balanced (HLB) polymeric SPE cartridge. The final analytical method was validated and applied for the quantification of surfactants in 36 river water samples. Method limits of quantification (LQ), intra and inter days precision and trueness were evaluated. With LQ between 15 and 485 ng/L, and trueness over 80%, this method was suitable for monitoring surfactants in surface water. Application on French river water samples revealed the presence of anionic, cationic and non-ionic surfactants with median concentrations from 24 ng/L for octylphenol ethoxylates (OPEO), up to 4.6 µg/L regarding linear alkylbenzene sulfonates (LAS).
RESUMO
Urban wastewater treatment plants (WWTP) are a major vector of highly ecotoxic contaminants of emerging concern (CECs) for urban and sub-urban streams. Ecotoxicological risk assessments (ERAs) provide essential information to public environmental authorities. Nevertheless, ERAs are mainly performed at very local scale (one or few WWTPs) and on pre-selected list of CECs. To cope with these limits, the present study aims to develop a territorial-scale ERA on CECs previously identified by a "suspect screening" analytical approach (LC-QToF-MS) and quantified in the effluents of 10 WWTPs of a highly urbanized territory during three periods of the year. Among CECs, this work focused on pharmaceutical residue and pesticides. ERA was conducted following two complementary methods: (1) a single substance approach, based on the calculation for each CEC of risk quotients (RQs) by the ratio of Predicted Environmental Concentration (PEC) and Predicted No Effect Concentration (PNEC), and (2) mixture risk assessment ("cocktail effect") based on a concentration addition model (CA), summing individual RQs. Chemical results led to an ERA for 41 CEC (37 pharmaceuticals and 4 pesticides) detected in treated effluents. Single substance ERA identified 19 CECs implicated in at least one significant risk for streams, with significant risks for DEET, diclofenac, lidocaine, atenolol, terbutryn, atorvastatin, methocarbamol, and venlafaxine (RQs reaching 39.84, 62.10, 125.58, 179.11, 348.24, 509.27, 1509.71 and 3097.37, respectively). Mixture ERA allowed the identification of a risk (RQmix > 1) for 9 of the 10 WWTPs studied. It was also remarked that CECs leading individually to a negligible risk could imply a significant risk in a mixture. Finally, the territorial ERA showed a diversity of risk situations, with the highest concerns for 3 WWTPs: the 2 biggest of the territory discharging into a large French river, the Rhône, and for the smallest WWTP that releases into a small intermittent stream.
Assuntos
Poluentes Químicos da Água , Purificação da Água , Monitoramento Ambiental , Medição de Risco , Eliminação de Resíduos Líquidos , Águas Residuárias/análise , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidadeRESUMO
BACKGROUND: Aryl phosphate esters (APEs) are widely used and commonly present in the environment. Health hazards associated with these compounds remain largely unknown and the effects of diphenyl phosphate (DPhP), one of their most frequent derivatives, are poorly characterized. OBJECTIVE: Our aim was to investigate whether DPhP per se may represent a more relevant marker of exposure to APEs than direct assessment of their concentration and determine its potential deleterious biological effects in chronically exposed mice. METHODS: Conventional animals (FVB mice) were acutely or chronically exposed to relevant doses of DPhP or to triphenyl phosphate (TPhP), one of its main precursors. Both molecules were measured in blood and other tissues by liquid chromatography-mass spectrometry (LC-MS). Effects of chronic DPhP exposure were addressed through liver multi-omics analysis to determine the corresponding metabolic profile. Deep statistical exploration was performed to extract correlated information, guiding further physiological analyses. RESULTS: Multi-omics analysis confirmed the existence of biological effects of DPhP, even at a very low dose of 0.1mg/mL in drinking water. Chemical structural homology and pathway mapping demonstrated a clear reduction of the fatty acid catabolic processes centered on acylcarnitine and mitochondrial ß-oxidation in mice exposed to DPhP in comparison with those treated with vehicle. An interesting finding was that in mice exposed to DPhP, mRNA, expression of genes involved in lipid catabolic processes and regulated by peroxisome proliferator-activated receptor alpha (PPARα) was lower than that in vehicle-treated mice. Immunohistochemistry analysis showed a specific down-regulation of HMGCS2, a kernel target gene of PPARα. Overall, DPhP absorption disrupted body weight-gain processes. CONCLUSIONS: Our results suggest that in mice, the effects of chronic exposure to DPhP, even at a low dose, are not negligible. Fatty acid metabolism in the liver is essential for controlling fast and feast periods, with adverse consequences on the overall physiology. Therefore, the impact of DPhP on circulating fat, cardiovascular pathologies and metabolic disease incidence deserves, in light of our results, further investigations. https://doi.org/10.1289/EHP6826.
Assuntos
Poluentes Ambientais/toxicidade , Fosfatos/toxicidade , Animais , Ésteres/toxicidade , Camundongos , Modelos Químicos , Testes de ToxicidadeRESUMO
Healthcare facility discharges, by their nature, are often considered as non-domestic effluent, which can provide significant pollution comparatively to other domestic sources. In this context, a total of 12 monthly sampling campaigns were collected from a healthcare facility as well as the output of a sewerage system of Site Pilote de Bellecombe (SIPIBEL) observatory. This study focuses more specifically on 12 surfactants and biocides: four anionics, four cationic, two non-ionic, one zwitterionic, and one dispersive agent, among the most commonly used commercial surfactants. Particular attention was also provided to routine wastewater quality parameters. Both effluents were heavily contaminated by most anionic surfactants; they displayed median concentrations up to 1 to 2 mg/L for linear alkylbenzene sulfonates and between 10 and 100 µg/L for other sodium sulfate congeners (lauryl and laureth). Overall, for the majority of surfactants, the healthcare facility contribution to the total flux reaching the wastewater treatment plant ranges between 5 and 9%.
Assuntos
Ácidos Alcanossulfônicos/análise , Tensoativos , Águas Residuárias/análise , Poluentes Químicos da Água/análise , Ácidos Alcanossulfônicos/química , Águas Residuárias/químicaRESUMO
Apiculture and pollination services are seriously threatened by bee weakening and losses phenomena. In this context, a survey was performed on samples from beehives across French areas during the 2012-2016 growing seasons, primarily taken from symptomatic colonies. A total of 488 honeybees, beebread, and wax were analyzed for the presence of pesticide residues. A total of 13 analytes including neonicotinoids and pyrethroids insecticides together with some of their metabolites and the fungicide boscalid were screened within samples. Methodologies based on efficient modified quick, easy, cheap, effective, rugged, and safe extractions followed by an LC-MS/MS quantification were implemented for each matrix. Thirty-eight percent of the 125 bee samples, 61% of the 87 wax samples, and 77% of the 276 beebread samples contained at least one of the targeted pesticides. Beebread was the most contaminated matrix with an average of two pesticide detections by positive sample and a maximum of seven compounds for a sample. Neonicotinoids and boscalid were the most often detected pesticides, whatever the matrix. The comparison of neonicotinoid detections in samples collected before and after the partial neonicotinoid ban in France displays a decrease in the frequency of detections for contamination levels lower than 1 ng/g in beebread. For higher levels and other matrices, no tendency can be drawn.
Assuntos
Abelhas/química , Monitoramento Ambiental/métodos , Poluentes Ambientais/análise , Fungicidas Industriais/análise , Resíduos de Praguicidas/análise , Própole/química , Ceras/química , Animais , Cromatografia Líquida/métodos , França , Espectrometria de Massas em Tandem/métodosRESUMO
Boscalid is a new-generation fungicide that has been detected in several bee matrices. The objective of this work was to characterize boscalid metabolites in honeybees based on in vivo experimentation, and next to verify the presence of theses metabolites into honeybees from colonies presenting troubles. A methodology based on complementary mass spectrometric tools, namely ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-QToF) or triple quadrupole mass spectrometry (UHPLC-QqQ) was implemented. Honeybees were sprayed with boscalid, at field rate (to induce the metabolization process) and the parent compound with its generated metabolites were then extracted using modified EU-QuEChERS method. The mass characteristics including exact mass, isotopic profile and mass fragments allowed assuming the structure of several metabolites. Some of them were unambiguously identified by comparison with synthesized analytical standards. The metabolites were resulted from hydroxylation and dechlorination of the parent compound as well as the substitution of a chlorine atom with an hydroxyl group. The metabolites were then quantified in bee samples collected from various beehives located in France. Boscalid and three of its metabolites were present in some samples at a level ranged between 0.2 and 36.3 ng/g.
Assuntos
Abelhas/metabolismo , Compostos de Bifenilo/análise , Compostos de Bifenilo/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Fungicidas Industriais/análise , Espectrometria de Massas/métodos , Niacinamida/análogos & derivados , Animais , França , Niacinamida/análise , Niacinamida/metabolismoRESUMO
Plant protection spray treatments may expose non-target organisms to pesticides. In the pesticide registration procedure, the honey bee represents one of the non-target model species for which the risk posed by pesticides must be assessed on the basis of the hazard quotient (HQ). The HQ is defined as the ratio between environmental exposure and toxicity. For the honey bee, the HQ calculation is not consistent because it corresponds to the ratio between the pesticide field rate (in mass of pesticide/ha) and LD50 (in mass of pesticide/bee). Thus, in contrast to all other species, the HQ can only be interpreted empirically because it corresponds to a number of bees/ha. This type of HQ calculation is due to the difficulty in transforming pesticide field rates into doses to which bees are exposed. In this study, we used a pragmatic approach to determine the apparent exposure surface area of honey bees submitted to pesticide treatments by spraying with a Potter-type tower. The doses received by the bees were quantified by very efficient chemical analyses, which enabled us to determine an apparent surface area of 1.05 cm(2)/bee. The apparent surface area was used to calculate the exposure levels of bees submitted to pesticide sprays and then to revisit the HQ ratios with a calculation mode similar to that used for all other living species. X-tomography was used to assess the physical surface area of a bee, which was 3.27 cm(2)/bee, and showed that the apparent exposure surface was not overestimated. The control experiments showed that the toxicity induced by doses calculated with the exposure surface area was similar to that induced by treatments according to the European testing procedure. This new approach to measure risk is more accurate and could become a tool to aid the decision-making process in the risk assessment of pesticides.
Assuntos
Abelhas/efeitos dos fármacos , Modelos Teóricos , Praguicidas/toxicidade , Animais , Abelhas/fisiologia , Superfície Corporal/veterinária , Cromatografia Gasosa , Exposição Ambiental , Dose Letal Mediana , Praguicidas/análiseRESUMO
Beebread is among the matrices suspected of contaminating honeybee. To better understand this contamination, the study aimed to develop an efficient, sensitive and reliable analytical method for the trace analysis of pesticides in beebread. This study focuses specifically on the insecticides pyrethroids and neonicotinoids and some of their metabolites. It describes the development and validation of an original analytical approach that consists of one simple extraction method based on modified QuEChERS followed by a selective and sensitive analysis by UHPLC-MS/MS to determine the target compounds in beebread. The method was validated using a quadratic fit. RSD values below 20% were obtained, except for 5-hydroxy-imidacloprid and imidacloprid at 0.5 ng/g, which exhibited RSDs of 25% and 21%, respectively. The intra-day precision was less than 10% for many of the investigated compounds. The inter-day precision varied between 2% and 36%, depending on the compound and the concentration. The recoveries varied from 53% to 119%, with averages of 83, 81 and 77% for the extraction of beebread samples spiked at 0.5, 5 and 10 ng/g, respectively. The LOD values for all the substances were below ng/g, with the exception of 6-chloronicotinic acid (LOD=1.7 ng/g). The method was then applied to the analysis of 32 beebread samples and revealed the presence of 7 of the target substances. The most frequently detected pesticides belonged to the neonicotinoid family and were generally present at low concentrations, but in some cases exceeded 170 ng/g (acetamiprid and thiacloprid). Some pyrethroids were also detected (lambda-cyhalothrine and bifenthrine), but at very low levels.
Assuntos
Anabasina/análise , Cromatografia Líquida de Alta Pressão/métodos , Inseticidas/análise , Extração Líquido-Líquido/métodos , Própole/química , Piretrinas/análise , Acetonitrilas , Anabasina/isolamento & purificação , Heptanos , Inseticidas/isolamento & purificação , Limite de Detecção , Piretrinas/isolamento & purificação , Reprodutibilidade dos Testes , Espectrometria de Massas em Tandem/métodosRESUMO
The formation of several by-products from the photodegradation of testosterone was previously observed under laboratory conditions. The objectives of the manuscript were to complete the identification of testosterone's photoproducts and to develop an analytical method for the detection of testosterone as well as its three main photoproducts in natural sunlit surface waters. To accomplish these tasks, an efficient extraction method was developed based on solid-phase extraction, followed by the use of liquid chromatography coupled to tandem mass spectrometry, a selective and sensitive detection method. This analytical procedure has the capability of detecting target analytes in the ng/L range, with recoveries above 80 %. The methodology was successfully applied to the analysis of testosterone and its photoproducts in several surface waters.
Assuntos
Extração em Fase Sólida/métodos , Testosterona/análise , Testosterona/química , Poluentes Químicos da Água/análise , Cromatografia Líquida/métodos , Fotólise , Sensibilidade e Especificidade , Espectrometria de Massas em Tandem/métodos , Poluentes Químicos da Água/químicaRESUMO
This study used batch-equilibrium techniques to evaluate the soil sorption and desorption of target steroids, representative of the estrogen, androgen, progestogen and corticosteroid families. The sorption kinetics exhibited an immediate rapid sorption followed by a slow process with a duration dependent on soil particle size. Adsorption and desorption isotherms could be described by the Freundlich equation, in a non-linear form (1/n < 1). High organic carbon-water partitioning coefficient (K(OC)) values indicate that a large mass fraction of the hormones will be associated with the sorbed phase. As a result of the poor linear correlation between log K(OC) and log K(OW) (octanol-water partition coefficient), it could be assumed that both hydrophobic and hydrogen bonding interactions contribute to the sorption mechanisms of the selected steroids. A hysteresis phenomenon was observed, varying with particle size, indicating strong irreversible interactions between some steroids and soil.
Assuntos
Poluentes do Solo/química , Esteroides/química , AdsorçãoRESUMO
This study describes the development of acetylcholinesterase (AChE), carboxylesterases (CaE1, CaE2, CaE3), glutathion-S-transferase (GST), alkaline phosphatase (ALP) and catalase (CAT) as enzyme biomarkers of exposure to xenobiotics such as thiamethoxam in the honey bee Apis mellifera. Extraction efficiency, stability under freezing and biological variability were studied. The extraction procedure achieved good recovery rates in one extraction step and ranged from 65 percent (AChE) to 97.3 percent (GST). Most of the enzymes were stable at -20°C, except ALP that displayed a slight but progressive decrease in its activity. Modifications of enzyme activities were considered after exposure to thiamethoxam at the lethal dose 50 percent (LD(50), 51.16 ng bee(-1)) and two sublethal doses, LD(50)/10 (5.12 ng bee(-1)) and LD(50)/20 (2.56 ng bee(-1)). The biomarker responses revealed that, even at the lowest dose used, exposure to thiamethoxam elicited sublethal effects and modified the activity of CaEs, GST, CAT and ALP. Different patterns of biomarker responses were observed: no response for AChE, an increase for GST and CAT, and differential effects for CaEs isoforms with a decrease in CaE1 and CaE3 and an increase in CaE2. ALP and CaE3 displayed contrasting variations but only at 2.56 ng bee(-1). We consider that this profile of biomarker variation could represent a useful fingerprint to characterise exposure to thiamethoxam in the honey bee A. mellifera. This battery of honey bee biomarkers might be a promising option to biomonitor the health of aerial and terrestrial ecosystems and to generate valuable information on the modes of action of pesticides.
Assuntos
Abelhas/efeitos dos fármacos , Biomarcadores/análise , Inseticidas/toxicidade , Nitrocompostos/toxicidade , Oxazinas/toxicidade , Tiazóis/toxicidade , Xenobióticos/toxicidade , Acetilcolinesterase/metabolismo , Animais , Abelhas/enzimologia , Catalase/metabolismo , Ativação Enzimática/efeitos dos fármacos , Congelamento , Glutationa Transferase/metabolismo , Dose Letal Mediana , Neonicotinoides , TiametoxamRESUMO
One of the factors that may explain nowadays honeybees' colonies losses is the increasing presence of chemicals in the environment. The aim of this study is to obtain a global view of the presence of environmental contaminants in beehives and, develop a fast, cheap and sensitive tool to analyze environmental contaminants in apiarian matrices. A multi residue analysis was developed to quantify 80 environmental contaminants, pesticides and veterinary drugs, belonging to different chemical classes, in honeys, honeybees and pollens. It consists in a single extraction, based on a modified "QuEChERS method", followed by gas chromatography coupled with Time of Flight mass spectrometry (GC-ToF) and liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The "QuEChERS method" combines salting-out liquid-liquid extraction with acetonitrile and a dispersive-SPE clean up. It was adjusted to honey and especially to honeybee and pollen, by adding a small fraction of hexane in acetonitrile to eliminate lipids that interfere with mass spectrometry analysis. This method, combined with accurate and sensitive detection, allowed quantification and confirmation at levels as low as 10 ng/g, with recoveries between 60 and 120%. Application to more than 100 samples of each matrix was achieved for a global view of pesticide presence in the honeybee environment. Relatively high percentages of honeys, honeybees and pollens were found to be contaminated by pesticides used to combat varroa but also by fungicides like carbendazim and ubiquitous contaminants.
Assuntos
Abelhas/química , Cromatografia Gasosa/métodos , Cromatografia Líquida/métodos , Poluentes Ambientais/análise , Mel/análise , Pólen/química , Extração em Fase Sólida/métodos , Espectrometria de Massas em Tandem/métodos , Animais , Resíduos de Drogas/análise , Sensibilidade e EspecificidadeRESUMO
The study proposes an analytical method to quantify 10 alkylphenols and 12 pesticides at ultra-trace levels by liquid chromatography in reverse mode coupled to positive electrospray ionisation-tandem mass spectrometry. The extraction procedure from environmental solids was optimised by pressurised liquid extraction using acetonitrile:isopropanol (1:1, v/v). The influence of several extraction experimental factors, temperature, pressure, duration and number of cycles, related to the PLE was investigated by an original and efficient chemometric approach. The optimised extraction method (80 degrees C, 40 bar, 10 min, 1 cycle) exhibited recoveries between 67 and 127% with RSD mostly under 13%. The whole method was applied to real samples: sludge, suspended materials, atmospheric fallouts and roof deposit. Pollutant levels were between 1 microg kg(-1) and 5.9 mg kg(-1).