RESUMO
Untargeted analysis of a composite house dust sample has been performed as part of a collaborative effort to evaluate the progress in the field of suspect and nontarget screening and build an extensive database of organic indoor environment contaminants. Twenty-one participants reported results that were curated by the organizers of the collaborative trial. In total, nearly 2350 compounds were identified (18%) or tentatively identified (25% at confidence level 2 and 58% at confidence level 3), making the collaborative trial a success. However, a relatively small share (37%) of all compounds were reported by more than one participant, which shows that there is plenty of room for improvement in the field of suspect and nontarget screening. An even a smaller share (5%) of the total number of compounds were detected using both liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS). Thus, the two MS techniques are highly complementary. Most of the compounds were detected using LC with electrospray ionization (ESI) MS and comprehensive 2D GC (GC×GC) with atmospheric pressure chemical ionization (APCI) and electron ionization (EI), respectively. Collectively, the three techniques accounted for more than 75% of the reported compounds. Glycols, pharmaceuticals, pesticides, and various biogenic compounds dominated among the compounds reported by LC-MS participants, while hydrocarbons, hydrocarbon derivatives, and chlorinated paraffins and chlorinated biphenyls were primarily reported by GC-MS participants. Plastics additives, flavor and fragrances, and personal care products were reported by both LC-MS and GC-MS participants. It was concluded that the use of multiple analytical techniques was required for a comprehensive characterization of house dust contaminants. Further, several recommendations are given for improved suspect and nontarget screening of house dust and other indoor environment samples, including the use of open-source data processing tools. One of the tools allowed provisional identification of almost 500 compounds that had not been reported by participants.
RESUMO
Because of large-scale production and use of an increasing diversity of chemicals in modern society, estuarine and coastal waters may be contaminated with numerous substances. Some of these compounds have the potential to affect microalgae at the base of the pelagic food chain. Therefore, we identified the main chemical stressors that negatively affect the effective photosystem II efficiency (ÏPSII) in marine microalgae of the Dutch estuarine and coastal waters. An enhanced effect-directed analysis (EDA) was carried out by combining reversed-phase ultra performance liquid chromatography fractionation of extracts from passive samplers, followed by effect assessment using the pulse amplitude modulation fluorometry assay and chemical analysis of biologically active fractions using high-resolution mass spectrometry. This study focuses on a novel microfractionation technique using 96-well plates to enhance throughput in EDA, structure elucidation, and the analytical and effect confirmation of the compounds that are identified. Although there are numerous unknown compounds present in estuarine and coastal waters, our EDA study shows that atrazine, diuron, irgarol, isoproturon, terbutryn, and terbutylazine are the main contributors to the observed effect on the ÏPSII of marine microalgae.
Assuntos
Fracionamento Químico/métodos , Ensaios de Triagem em Larga Escala/métodos , Microalgas/metabolismo , Fotossíntese , Bioensaio , Fluorometria , Herbicidas/toxicidade , Fotossíntese/efeitos dos fármacos , Silicones/química , Triazinas/toxicidade , Ureia/toxicidade , Poluentes Químicos da Água/toxicidadeRESUMO
Over the past decade, Cell and Gene Therapies (C>) have been an emerging therapeutic area with more than twenty C> drug products approved and over 1000 registered trials. The remarkable progress in these modalities brings new challenges for scientists who evaluate manufacturing and storage materials, including risk assessments for extractables and leachables (E&L). Establishing a business process to qualify materials for these applications is an important risk mitigation strategy in support of these assessments. Process validation verifying process performance and product quality requirements using qualified materials also ensures that leachables from the materials do not result in an impact to process and product. The authors provide an overview of available guidelines and publications relevant to E&L risk assessments that can be used to support ex vivo C> products, highlighting gaps and standardization needs in the areas of biocompatibility and extractables conditions. Finally, the authors present leachable testing strategies, relevant to the specific manufacturing and storage conditions of C> products, and safety assessment considerations for organic and inorganic chemical entities.
Assuntos
Contaminação de Medicamentos , Embalagem de Medicamentos , Contaminação de Medicamentos/prevenção & controle , Medição de Risco , Proteínas , Preparações FarmacêuticasRESUMO
BACKGROUND: The potential impact of natural and synthetic estrogens on aquatic ecosystems has attracted considerable attention because it is currently accepted that their joint effects are more severe when they are present in mixtures. Although it is well-known that they occur as mixtures in the marine environment, there is little information about the combined effects of estrogenic chemicals on marine biota. OBJECTIVE: In 14-day tests with juvenile sea bass, we analyzed singly and in combination the estrogenic activity of estradiol (E(2)), ethynylestradiol (EE(2)), and bisphenol A (BPA) using vitellogenin induction as an end point. METHODS: Fish were exposed to each compound, and on the basis of these concentration-response data, we predicted mixture effects by applying the model of concentration addition. The mixtures were tested using a fixed-ratio design, and the resulting mixture effects were compared to the predictions. RESULTS: EE(2) was the most potent steroid, with an EC(50) (median effective concentration) of 0.029 microg/L, 3.6 times more potent than E(2) (EC(50) = 0.104 microg/L); BPA was the least potent chemical, with an EC(50) of 77.94 microg/L. The comparative assessment yielded a good agreement between observed and predicted mixture effects. CONCLUSIONS: This study demonstrates the potential hazard of these compounds to seawater life by their ability to act together in an additive manner. It provides evidence that concentration addition can be used as a predictive tool for assessing the combined effects of estrogenic chemicals in marine ecosystems.
Assuntos
Estrogênios/toxicidade , Vitelogeninas/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Animais , Bass , Compostos Benzidrílicos , Relação Dose-Resposta a Droga , Interações Medicamentosas , Monitoramento Ambiental/métodos , Estradiol/administração & dosagem , Estradiol/toxicidade , Estrogênios/administração & dosagem , Etinilestradiol/toxicidade , Fenóis/administração & dosagem , Fenóis/toxicidade , Medição de Risco , Vitelogeninas/biossíntese , Poluentes Químicos da Água/administração & dosagemRESUMO
The exposure to and effects of estrogenic compounds in male breams from Dutch freshwater locations were investigated. Ovotestis was observed infrequently (maximum frequency 16%). However, plasma vitellogenin (VTG) concentration was elevated highly at some locations. Estrogenic activities in male bream plasma, liver, and in gastrointestinal content were measured in the estrogen-responsive chemical-activated luciferase gene expression (ER-CALUX) assay. Plasma concentrations of vitellogenin correlated very well with the estrogenic activities in gastrointestinal content. The ER-CALUX activity in gastrointestinal content thus could provide a biomarker for recent exposure to estrogenic compounds, and the gastrointestinal content was chosen as investigative matrix for the toxicity identification and evaluation ([TIE]; bioassay-directed fractionation) of estrogenic compounds in bream. The approach consisted of a reversed-phase high-performance liquid chromatography fractionation of gastrointestinal content extract, directed by ER-CALUX and followed by gas chromatography analysis. The estrogenic hormones 17beta-estradiol and its metabolite estrone were identified as major contributors to the activity at all locations (except the reference location), independent of the presence or absence of a known source of estrogenic activity, such as a sewage treatment plant. Chemical screening showed the presence of other pollutants, such as a lower chlorinated dioxin and the disinfectants clorophene and triclosan. However, these compounds did not have high estrogenic potencies and their concentrations were not high enough to contribute significantly to the observed estrogenic activity.
Assuntos
Bioensaio , Monitoramento Ambiental , Estrogênios/metabolismo , Dourada/metabolismo , Poluentes Químicos da Água/metabolismo , Animais , Cromatografia Líquida de Alta Pressão , Diclorofeno/análogos & derivados , Diclorofeno/análise , Diclorofeno/metabolismo , Diclorofeno/toxicidade , Dioxinas/análise , Dioxinas/metabolismo , Dioxinas/toxicidade , Desinfetantes/análise , Desinfetantes/metabolismo , Desinfetantes/toxicidade , Estradiol/análise , Estradiol/metabolismo , Estradiol/toxicidade , Estrogênios/análise , Estrogênios/toxicidade , Estrona/análise , Estrona/metabolismo , Estrona/toxicidade , Trato Gastrointestinal/química , Trato Gastrointestinal/metabolismo , Luciferases/genética , Luciferases/metabolismo , Triclosan/análise , Triclosan/metabolismo , Triclosan/toxicidade , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidadeRESUMO
Organochlorine pesticides (OCPs) have been used to preserve the integrity of historical buildings or to protect collections of artefacts at potentially large volumes and often without detailed application records. Previous research has focused on the efficiency of remediation at contaminated sites (where identified), as well as improvement of preservation techniques and workplace health and safety. Few studies have assessed the human health risks from occupational exposure to OCPs in buildings of cultural and historical importance. Thus, potential risks may remain unidentified. In the present study, OCPs in indoor air were measured in a baroque theatre and a natural history museum in the Czech Republic, both of which had suspected past indoor application. In the theatre attic p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) levels in air were up to 190ngm-3, confirming past indoor use of p,p'-dichlorodiphenyltrichloroethane (p,p'-DDT). There was also evidence of γ-hexachlorocyclohexane (γ-HCH) use in the theatre (max γ-HCH in air of 56ngm-3). Yet, the cancer risk (CR) from occupational exposure via inhalation (Expi) to OCPs in the theatre was low (CR<4.0×10-6). γ-HCH was found at elevated levels in air of the museum (max γ-HCH in air of 15,000ngm-3). CR from Expi in the museum was moderate to high (>1×10-4). Our results show the CR through Expi to OCPs in buildings, such as museums can still be significant enough to warrant mitigation measures, e.g., remediation.
Assuntos
Poluição do Ar em Ambientes Fechados/análise , Hidrocarbonetos Clorados/análise , Exposição por Inalação/análise , Neoplasias/epidemiologia , Praguicidas/análise , República Tcheca , Monitoramento Ambiental , Humanos , Museus , Medição de RiscoRESUMO
The identity of compounds responsible for estrogenic and dioxin-like activities in sediment from the harbour of the small town Zierikzee in Zeeland, The Netherlands, was investigated using a bioassay directed fractionation approach with the in vitro estrogen and dioxin responsive reporter gene assays ER- and DR-CALUX. For identification of compounds exhibiting activity in the bioassays, either one or two-dimensional GC in combination with quadrupole (MSD), ion trap (ITD) or time-of-flight mass spectrometric detection (ToF-MS) was used, depending on the biological and chemical characteristics and the complexity of the fractions. The natural estrogenic hormone 17-beta-estradiol and its metabolite estrone were identified with GC-ITD as the main contributors to the estrogenic activity. After successive rounds of fractionation, the dioxin-like activity could be explained by the presence of various polycyclic aromatic hydrocarbons identified with GC-MSD and two-dimensional comprehensive GC x GC-ToF-MS. Some estrogenic activity of a relatively non-polar nature remained unidentified.
Assuntos
Cromatografia Gasosa/métodos , Dioxinas/análise , Estrogênios/análise , Sedimentos Geológicos/química , Poluentes Químicos da Água/análise , Cromatografia Líquida de Alta PressãoRESUMO
In Oman, DDT was sprayed indoors during an intensive malaria eradication program between 1976 and 1992. DDT can remain for years after spraying and is associated with potential health risk. This raises the concern for human exposure in areas where DDT was used for indoor spraying. Twelve houses in three regions with a different history of DDT indoor spraying were chosen for a sampling campaign in 2005 to determine p,p'-dichlorodiphenyltrichloroethane (p,p'-DDT), p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) and p,p'-dichlorodiphenyldichloroethane (p,p'-DDD) levels in indoor air, dust, and outdoor soil. Although DDT was only sprayed indoor, p,p'-DDT, p,p'-DDE and p,p'-DDD were also found in outdoor soil. The results indicate that release and exposure continue for years after cessation of spraying. The predicted cancer risk based on concentrations determined in 2005, indicate that there was still a significant cancer risk up to 13 to 16years after indoor DDT spraying. A novel approach, based on region-specific half-lives, was used to predict concentrations in 2015 and showed that more than 21years after spraying, cancer risk for exposure to indoor air, dust, and outdoor soil are acceptable in Oman for adults and young children. The model can be used for other locations and countries to predict prospective exposure of contaminants based on indoor experimental measurements and knowledge about the spraying time-schedule to extrapolate region-specific half-lives and predict effects on the human population years after spraying.
Assuntos
Poluição do Ar em Ambientes Fechados/estatística & dados numéricos , DDT/análise , Exposição Ambiental/estatística & dados numéricos , Inseticidas/análise , Adulto , Poluição do Ar em Ambientes Fechados/análise , Criança , Humanos , Controle de Mosquitos/métodos , Controle de Mosquitos/estatística & dados numéricos , OmãRESUMO
Due to their specific effect on photosynthesis, herbicides pose a potential threat to coastal and estuarine microalgae. However, comprehensive understanding of the hazard and risk of these contaminants is currently lacking. Therefore the aim of the present study was to investigate the toxic effects of four ubiquitous herbicides (atrazine, diuron, Irgarol(®)1051 and isoproturon) and herbicide mixtures on marine microalgae. Using a Pulse Amplitude Modulation (PAM) fluorometry based bioassay we demonstrated a clear species and herbicide specific toxicity and showed that the current environmental legislation does not protect algae sufficiently against diuron and isoproturon. Although a low actual risk of herbicides in the field was demonstrated, monitoring data revealed that concentrations occasionally reach potential effect levels. Hence it cannot be excluded that herbicides contribute to observed changes in phytoplankton species composition in coastal waters, but this is likely to occur only occasionally.
Assuntos
Herbicidas/toxicidade , Microalgas/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Atrazina/toxicidade , Diurona/toxicidade , Fluorometria , Compostos de Fenilureia/toxicidade , Fotossíntese/efeitos dos fármacos , Fitoplâncton/efeitos dos fármacos , Medição de RiscoRESUMO
Pulse Amplitude Modulation (PAM) fluorometry, based on chlorophyll a fluorescence, is a frequently used technique in algal bioassays to assess toxicity of single compounds or complex field samples. Several test conditions can influence the test results, and because a standardized test protocol is currently lacking, linking the results of different studies is difficult. Therefore, the aim of the present study was to gain insight into the effects of test conditions of laboratory algal bioassays using PAM fluorometry on the outcome of toxicity tests. To this purpose, we described the results from several pilot studies on test development in which information is provided on the effects of the main test factors during the pretest phase, the test preparation, the exposure period, and the actual measurement. The experiments were focused on individual herbicides and complex field samples and included the effects of culturing conditions, cell density, solvent concentration, exposure time, and the presence of actinic light. Several of these test conditions were found to influence the outcome of the toxicity test, and the presented information provides important background information for the interpretation of toxicity results and describes which test conditions should be taken into account when using an algal bioassay with PAM fluorometry. Finally, the application of PAM fluorometry in algal toxicity testing is discussed.
Assuntos
Clorófitas/efeitos dos fármacos , Fluorometria/métodos , Herbicidas/toxicidade , Testes de Toxicidade/métodos , Bioensaio/métodos , Clorofila/química , Clorofila A , Fluorescência , LuzRESUMO
The extent to which chemical stressors affect primary producers in estuarine and coastal waters is largely unknown. However, given the large number of legacy pollutants and chemicals of emerging concern present in the environment, this is an important and relevant issue that requires further study. The purpose of our study was to extract and identify compounds which are inhibitors of photosystem II activity in microalgae from estuarine and coastal waters. Field sampling was conducted in the Western Scheldt estuary (Hansweert, The Netherlands). We compared four different commonly used extraction methods: passive sampling with silicone rubber sheets, polar organic integrative samplers (POCIS) and spot water sampling using two different solid phase extraction (SPE) cartridges. Toxic effects of extracts prepared from spot water samples and passive samplers were determined in the Pulse Amplitude Modulation (PAM) fluorometry bioassay. With target chemical analysis using LC-MS and GC-MS, a set of PAHs, PCBs and pesticides was determined in field samples. These compound classes are listed as priority substances for the marine environment by the OSPAR convention. In addition, recovery experiments with both SPE cartridges were performed to evaluate the extraction suitability of these methods. Passive sampling using silicone rubber sheets and POCIS can be applied to determine compounds with different structures and polarities for further identification and determination of toxic pressure on primary producers. The added value of SPE lies in its suitability for quantitative analysis; calibration of passive samplers still needs further investigation for quantification of field concentrations of contaminants.
Assuntos
Fracionamento Químico/métodos , Estuários , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/isolamento & purificação , Poluentes Químicos da Água/toxicidade , Água/química , Complexo de Proteína do Fotossistema II/antagonistas & inibidores , Extração em Fase SólidaRESUMO
In bioassays, exposure concentrations of test compounds are usually expressed as nominal concentrations. As a result of various processes, such as adsorption, degradation, or uptake, the actual freely dissolved concentration of the test compound may differ from the nominal concentration. The goal of the present study was to develop a method to dose passively the freely dissolved fraction of organic chemicals in an in vitro bioassay with adherent cells. To this end, a polydimethylsiloxane (PDMS) film-based method was developed for a reporter gene assay for dioxin-like compounds in a rat liver cell line. Polydimethylsiloxane films loaded with test compounds ensure that the concentration during exposure is in equilibrium and that the ratio between the concentration on the film and the concentration in medium is constant. Benzo[k]fluoranthene (BkF) was used as a model compound to develop the passive dosing method in transwell plates, which was further tested with a complex mixture, i.e., an extract prepared from a contaminated sediment. A higher dioxin-like activity was found when extracts were dosed by passive dosing with PDMS than when directly added to medium. Comparison with analysis of the concentration of BkF in medium shows that passive dosing of individual chemicals may not be necessary if freely dissolved concentrations are known. Use of PDMS for passive dosing of complex samples may represent a more realistic method for exposure in in vitro bioassays.