RESUMO
BACKGROUND: Diet is a major source of human exposure to hazardous environmental chemicals, including many perfluoroalkyl acids (PFAAs). Several assessment methods of dietary exposure to PFAAs have been used previously, but there is a lack of comparisons between methods. AIM: To assess human exposure to PFAAs through diet by different methods and compare the results. METHODS: We studied the dietary exposure to PFAAs in 61 Norwegian adults (74% women, average age: 42 years) using three methods: i) by measuring daily PFAA intakes through a 1-day duplicate diet study (separately in solid and liquid foods), ii) by estimating intake after combining food contamination with food consumption data, as assessed by 2-day weighted food diaries and iii) by a Food Frequency Questionnaire (FFQ). We used existing food contamination data mainly from samples purchased in Norway and if not available, data from food purchased in other European countries were used. Duplicate diet samples (n=122) were analysed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) to quantify 15 PFAAs (11 perfluoroalkyl carboxylates and 4 perfluoroalkyl sulfonates). Differences and correlations between measured and estimated intakes were assessed. RESULTS: The most abundant PFAAs in the duplicate diet samples were PFOA, PFOS and PFHxS and the median total intakes were 5.6ng/day, 11ng/day and 0.78ng/day, respectively. PFOS and PFOA concentrations were higher in solid than liquid samples. PFOS was the main contributor to the contamination in the solid samples (median concentration 14pg/g food), while it was PFOA in the liquid samples (median concentrations: 0.72pg/g food). High intakes of fats, oils, and eggs were statistically significantly related to high intakes of PFOS and PFOA from solid foods. High intake of milk and consumption of alcoholic beverages, as well as food in paper container were related to high PFOA intakes from liquid foods. PFOA intakes derived from food diary and FFQ were significantly higher than those derived from duplicate diet, but intakes of PFOS derived from food diary and FFQ were significantly lower than those derived from duplicate diet. We found a positive and statistically significant correlation between the PFOS intakes derived from duplicate diet with those using the food diary (rho=0.26, p-value=0.041), but not with the FFQ. Additionally, PFOA intakes derived by duplicate diet were significantly correlated with estimated intakes from liquid food derived from the food diary (rho=0.34, p=0.008) and estimated intakes from the FFQ (rho=0.25, p-value=0.055). CONCLUSIONS: We provide evidence that a food diary or a FFQ-based method can provide comparable intake estimates to PFOS and PFOA intakes derived from a duplicate diet study. These less burdensome methods are valuable and reliable tools to assess dietary exposure to PFASs in human studies.
Assuntos
Bebidas/análise , Exposição Ambiental , Poluentes Ambientais/análise , Fluorocarbonos/análise , Contaminação de Alimentos/análise , Adulto , Cromatografia Líquida , Dieta/estatística & dados numéricos , Monitoramento Ambiental , Feminino , Humanos , Pessoa de Meia-Idade , Noruega , Avaliação Nutricional , Espectrometria de Massas em TandemRESUMO
Meat samples of 84 minke whales (Balaenoptera acutorostrata) mainly from the Barents Sea, collected between 1 May and 16 August 2011, were analyzed for total mercury, methylmercury, cadmium, lead, total arsenic, inorganic arsenic and selenium. The average total mercury concentration found was 0.15 ± 0.09 mg/kg, with a range from 0.05 to 0.49 mg/kg. The molar ratio of selenium to mercury varied between 1.0 and 10.3. Cadmium content ranged from 0.002 to 0.036 mg/kg, while the content of lead in whale meat ranged from <0.01 to 0.09 mg/kg. None of the whale samples exceeded established EU maximum levels for metals in fish muscle, but 4.8% and 6.8% of the samples exceeded Japanese maximum levels for total mercury and methylmercury, respectively, in whale meat. There was only minor variations in element concentrations between whales from different geographical areas, and cadmium was the only element were the concentration increased with increasing length.
Assuntos
Arsênio/análise , Monitoramento Ambiental , Carne/análise , Compostos de Metilmercúrio/análise , Baleia Anã , Poluentes Químicos da Água/análise , Animais , Oceano Atlântico , Cádmio , Cetáceos , Peixes , Mercúrio/análise , Músculos/química , SelênioRESUMO
Hg pollution released from the U-864 submarine sunk during WWII and potential introduction of that Hg into the marine food chain have been studied by a combination of quantitative Hg and MeHg determination and Hg isotopic analysis via cold vapor generation multicollector inductively coupled plasma-mass spectrometry (CVG-MC-ICP-MS) in sediment and Cancer pagurus samples. The sediment pollution could be unequivocally linked with the metallic Hg present in the wreck. Crabs were collected at the wreck location and 4 nmi north and south, and their brown and claw meat were analyzed separately. For brown meat, the δ202Hg values of the individuals from the wreck location were shifted toward the isotopic signature of the sediment and, thus, the submarine Hg. Such differences were not found for claw meat. The isotope ratio results suggest direct ingestion of metallic Hg by C. pagurus but do not offer any proof for any other introduction of the submarine Hg into the marine food chain.
Assuntos
Braquiúros , Mercúrio , Animais , Poluição Ambiental , Gases , Isótopos/químicaRESUMO
BACKGROUND: Determining the concentration of nanoparticles (NPs) in marine organisms is important for evaluating their environmental impact and to assess potential food safety risks to human health. OBJECTIVE: The current work aimed at developing an in-house method based on single-particle inductively coupled plasma-mass spectrometry (SP-ICP-MS) suitable for surveillance of NPs in mussels. METHODS: A new low-cost and simple protease mixture was utilized for sample digestion, and novel open-source data processing was used, establishing detection limits on a statistical basis using false-positive and false-negative probabilities. The method was validated for 30 and 60 nm gold NPs spiked to mussels as a proxy for seafood. RESULTS: Recoveries were 76-77% for particle mass concentration and 94-101% for particle number concentration. Intermediate precision was 8-9% for particle mass concentration and 7-8% for particle number concentration. The detection limit for size was 18 nm, for concentration 1.7 ng/g, and 4.2 × 105 particles/g mussel tissue. CONCLUSION: The performance characteristics of the method were satisfactory compared with numeric Codex criteria. Further, the method was applied to titanium-, chromium- and copper-based particles in mussels. HIGHLIGHTS: The method demonstrates a new practical and cost-effective sample treatment, and streamlined, transparent, and reproducible data treatment for the routine surveillance of NPs in mussels.
Assuntos
Bivalves , Espectrometria de Massas , Nanopartículas Metálicas , Animais , Bivalves/química , Espectrometria de Massas/métodos , Nanopartículas Metálicas/química , Limite de Detecção , Ouro/química , Tamanho da Partícula , Contaminação de Alimentos/análise , Alimentos Marinhos/análise , Titânio/análise , Titânio/química , Cobre/análise , Nanopartículas/análise , Nanopartículas/químicaRESUMO
Due to the analytical challenges of detecting and quantifying nanoparticles in seawater, the data on distributions of NPs in the marine environment is limited to qualitative studies or by ensemble measurements subject to various analytical artifacts. Single particle inductively coupled plasma mass spectrometry (SP-ICP-MS) allows determination of individual inorganic NPs at environmentally relevant concentrations, yet only few studies have been conducted on selected elements in surface sea water. Here, a sequential multi-element screening method was developed and implemented to provide a first survey of the horizontal and vertical distributions of inorganic nanoparticles and trace elements in a pristine Norwegian fjord prospect for submarine tailings deposition. Statistical control of false-positive detections while minimizing the size detection limit was ensured using a novel raw signal processing. Scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS) gave confirmative and qualitative information regarding particle morphology and composition. Following SP-ICP-MS screening for particles of 16 elements, particulate Al, Fe, Mn, Pb, Si and Ti were found and determined to mass concentrations in ng/L of 1-399, 1-412, below limit of detection (Assuntos
Nanopartículas Metálicas
, Nanopartículas
, Oligoelementos
, Oligoelementos/análise
, Estuários
, Chumbo/análise
, Microscopia Eletrônica de Varredura
RESUMO
Currently, there is no legal limit for methyl mercury (MeHg) in food; thus, no standardized method for the determination of MeHg in seafood exists within the European jurisdiction. In anticipation of a future legislative limit an inductively coupled plasma isotope dilution mass spectrometry (GC-ICP-ID-MS) method was developed in collaboration with the European Standardization Organization (CEN). The method comprises spiking the tissue sample with Me201Hg, followed by decomposition with tetramethylammonium hydroxide, pH adjustment and derivatization with sodium tetraethylborate, and finally organic extraction of the derivatized MeHg in a hexane phase. Subsequently, the sample is analyzed via GC-ICP-MS and the result calculated using the ID equation. The working range of the method was 0.0005-1.321 mg/kg MeHg in marine tissue, with an internal reproducibility (RSD) of 12-1%. The method was validated based on statistical measures, such as the z-scores, using the commercially available reference materials from National Institute of Standards and Technology Standard Reference Material (NIST SRM) 1566b, NIST SRM 2977 and National Research Council of Canada (NRCC) TORT 2, NRCC, DORM 3, NRCC DOLT 4, and European Reference Material (ERM) CE 464. Z-scores for all standard reference materials, except for NIST SRM 1566b, were better than 11.51. The wide range of marine tissues used during the validation ensures that the method will be applicable for measuring of MeHg in seafood matrixes of all kinds.
Assuntos
Técnicas de Química Analítica/métodos , Análise de Alimentos/métodos , Contaminação de Alimentos/análise , Cromatografia Gasosa-Espectrometria de Massas/métodos , Espectrometria de Massas/métodos , Compostos de Metilmercúrio/análise , Alimentos Marinhos/análise , Boratos/química , Técnicas de Química Analítica/normas , Cromatografia/métodos , Europa (Continente) , Hexanos/química , Concentração de Íons de Hidrogênio , Técnicas de Diluição do Indicador , Isótopos/química , Espectrometria de Massas/normas , Padrões de Referência , Reprodutibilidade dos Testes , Estados UnidosRESUMO
Despite the ban of nitrofurans (NFs) for use in food production in many countries in the 1990s, NF metabolites in food are still regularly detected during import control testing. We have developed a confirmatory routine method for the detection and quantification of NF metabolites in seafood using LC-MS/MS and validated the method according to the strict criteria in European legislation and Codex Alimentarius. Method characteristics were found to fulfill the criteria. We report for the first time a new false positive for 1-amino-2,4-imidazolidinedione (AHD), the metabolite of Nitrofurantoin (NFT). By using optimized washing procedures, the non tissue bound false positives can be minimized. The results from the validation on both lean and fatty fish and crustaceans, results from proficiency tests and routine use over many years, demonstrates that the method is fit for purpose to determine NF metabolites in the seafood category.
Assuntos
Cromatografia Líquida/métodos , Contaminação de Alimentos/análise , Nitrofurantoína/análise , Alimentos Marinhos/análise , Espectrometria de Massas em Tandem/métodos , Animais , Reações Falso-Positivas , Análise de Alimentos/métodos , Técnicas de Diluição do Indicador , Nitrofurantoína/metabolismoRESUMO
Organotin compounds are anthropogenic metal species with multiple uses as pesticides, preservatives, antifouling agents, biocides, and catalysts. Butyltins are the main organotin compounds found in biota, and the highest levels are found in marine foodstuffs. In this paper, we present the figures of merit for an in-house validated method for routine analysis of butyltins in seafood using GC inductively coupled plasma isotope dilution MS. The working range of the method spanned several orders of magnitude from 3.3-1013, 2.4-785, and 0.3-900 ng Sn/g dry weight for monobutyltin (MBT), dibutyltin (DBT), and tributyltin (TBT), respectively. The trueness of the method was evaluated by analyzing Certified Reference Materials (CRMs) ERM CRM 477 (Mussel Tissue) and NIES CRM 15 (Scallop). Recoveries, with RSD % in parentheses, were 78 (±14), 80 (±6), and 88% (±8%) for MBT, DBT, and TBT in ERM CRM 477 and 96% (±5%) for TBT in NIES CRM 15. Good agreements were found between experimental uncertainties and uncertainties predicted for single-laboratory validated methods calculated from the maximum standard measurement uncertainty function. The method has proven to be robust, and the wide range of seafood validated ensures that the method is applicable for measuring butyltins in marine tissue.
Assuntos
Contaminação de Alimentos/análise , Cromatografia Gasosa-Espectrometria de Massas/métodos , Compostos Orgânicos de Estanho/análise , Alimentos Marinhos/análise , Compostos de Trialquitina/análise , Animais , Bivalves/química , Braquiúros/química , Decápodes/química , GadiformesRESUMO
Organotin compounds are anthropogenic metal species with multiple uses as pesticides, preservatives, antifouling agents, biocides, and catalysts. Butyltins are the main organotin compounds found in biota, and the highest levels are found in marine foodstuffs. In this paper, we present the figures of merit for an in-house validated method for routine analysis of butyltins in seafood using GC inductively coupled plasma isotope dilution MS. The working range of the method spanned several orders of magnitude from 3.3-1013, 2.4-785, and 0.3-900 ng Sn/g dry weight for monobutyltin (MBT), dibutyltin (DBT), and tributyltin (TBT), respectively. The trueness of the method was evaluated by analyzing Certified Reference Materials (CRMs) ERM CRM 477 (Mussel Tissue) and NIES CRM 15 (Scallop). Recoveries, with RSD % in parentheses, were 78 (±14), 80 (±6), and 88% (±8%) for MBT, DBT, and TBT in ERM CRM 477 and 96% (±5%) for TBT in NIES CRM 15. Good agreements were found between experimental uncertainties and uncertainties predicted for single-laboratory validated methods calculated from the maximum standard measurement uncertainty function. The method has proven to be robust, and the wide range of seafood validated ensures that the method is applicable for measuring butyltins in marine tissue.
RESUMO
The aim of this study was to develop and validate a method for the determination of balenine/ophidine (hereafter referred to as "balenine") in whale extracts and muscle samples from Balaenoptera acutorostrata. Further, the goal was to evaluate the method's applicability for the determination of other histidine-containing dipeptides (HCDs): anserine and carnosine and their amino acids π-methylhistidine, τ-methylhistidine, histidine, and ß-alanine. For balenine, the LOD and LOQ were found to be 0.03 and 0.1 mg/g, respectively, and the linear range was validated up to 160 mg/g. Trueness was evaluated by spiking experiments with balenine, and the recovery was found to be 88-90%. A comparison of the results showed that most of the other analytes were within 80-120% of the value found with the previously developed and validated method. Precision and internal reproducibility for balenine was around 0.9 and 2%, respectively, with measurement uncertainties of 2-4%. Therefore, the method was found to be fit for purpose for the determination of balenine and other HCDs and their constituent amino acids in whale meat and extracts.
Assuntos
Anserina/análise , Cromatografia Líquida/métodos , Dipeptídeos/análise , Baleia Anã , Animais , Calibragem , Carnosina/análise , Liofilização , Limite de Detecção , Metilistidinas/análise , Músculos/químicaRESUMO
The level of perfluorinated alkyl substances (PFAS) was determined in North East Arctic cod (Gadus morhua) liver samples from 15 Norwegian fjords and harbors. Five harbors in the eastern part of Norway, six harbors in the western part and four harbours in the northern part. A total of 200 samples were analyzed for 16 PFAS. Determination of PFAS were carried out by LC-MS/MS following sample clean up by solid phase extraction and ultracentrifugation. The predominating PFAS was PFOS, which was found to be higher than the level of quantification (1.5 µg kg-1 wet weight) in 72% of the samples. The highest level of PFOS found was 21.8 µg kg-1 wet weight in a sample from Kragerø in the eastern part of Norway. A significantly higher level of PFOS was found in the eastern fjords and harbors compared to fjords and harbors in the western and northern part of Norway. Within the northern fjords and harbors elevated PFOS levels were found in Narvik, which may indicate a local source there. Variations in PFOS of the cod livers thus reflect differences in levels of pollution between the areas.
Assuntos
Fluorocarbonos/análise , Gadus morhua/metabolismo , Fígado/química , Ácidos Alcanossulfônicos/análise , Animais , Cromatografia Líquida , Monitoramento Ambiental , Feminino , Masculino , Noruega , Espectrometria de Massas em TandemRESUMO
Feed additives are typically used in intensive farming production over long periods, and hence, they can accumulate in farmed animal tissues. Concerns regarding the use of ethoxyquin as an antioxidant feed additive, have recently arisen due to its potential conversion into a series of transformation products (TPs). The aim of this work was to characterize the TPs of ethoxyquin in fish feed by a novel approach based on the use of traveling-wave ion mobility spectrometry (TWIMS) coupled to high-resolution quadrupole time-of-flight mass spectrometry (QTOFMS). First, ethoxyquin was oxidized under controlled conditions and the generated TPs were added to a comprehensive database. Atlantic salmon feeds were then screened for ethoxyquin TPs using both targeted and untargeted approaches. Twenty-seven TPs were tentatively identified during the oxidation experiments, fifteen of them also being present in the feed samples. In addition, ten other potential TPs were detected in fish feed following the untargeted approach. Thirty-one of these TPs have been reported for the first time in this work through the oxidation experiments and the feed samples. Therefore, this study provides valuable information on the oxidative fate of ethoxyquin in feed, which can be used for future evaluations of potential risk related to this additive.
Assuntos
Ração Animal/análise , Etoxiquina/química , Aditivos Alimentares/análise , Espectrometria de Mobilidade Iônica , Espectrometria de Massas em Tandem , Animais , Antioxidantes/química , PeixesRESUMO
Eight laboratories participated in an inter-laboratory method-performance (collaborative) study of a method for the determination of mono methylmercury (MMHg) in foodstuffs of marine origin by gas chromatography inductively coupled plasma isotope dilution mass spectrometry (GC-ICP-IDMS) after dissolution, derivatisation and extraction of the species. The method was tested on seven seafood products covering both a wide concentration range and variations in the MMHg concentrations as well as matrix compositions. The samples were mussel tissue, squid muscle, crab claw meat, whale meat, cod muscle, Greenland halibut muscle and dogfish liver (NRCC DOLT-4), with MMHg concentrations ranging from 0.035 to 3.58mg/kg (as Hg) dry weight. Repeatability relative standard deviations (RSDr) for MMHg ranged from 2.1% to 8.7%. Reproducibility relative standard deviations (RSDR) ranged from 5.8% to 42%. All samples showed HorRat value below 1.0, except for the sample with the lowest MMHg content, mussel tissue, with a HorRat value of 1.6.
Assuntos
Análise de Alimentos/métodos , Contaminação de Alimentos/análise , Mercúrio/análise , Compostos de Metilmercúrio/análise , Alimentos Marinhos/análise , Animais , Bivalves/química , Cromatografia Gasosa , Cação (Peixe) , Linguado , Concentração de Íons de Hidrogênio , Técnicas de Diluição do Indicador , Isótopos , Fígado/química , Espectrometria de Massas , Músculos/química , Reprodutibilidade dos TestesRESUMO
The aim of this paper is to evaluate the food safety of the red king crab from Norwegian waters and obtain information on possible geographical and gender differences. Samples of claw and leg meat of 185 red king crabs (Paralithodes camtschaticus), collected from 23 positions in the Barents Sea, were analysed for dioxins, furans, non-ortho and mono-ortho PCBs, non dioxin-like PCBs, polybrominated diphenyl ethers, and perfluorinated alkyl substances and elements, such as arsenic, cadmium, mercury and lead. The concentrations of persistent organic pollutants and metals were low compared to maximum levels laid down in European regulations. Hence, red king crab is a safe food. Significant differences in the concentrations of metals among different areas, and between male and female crabs, were found. Positive correlations were found between carapace length and mercury, methylmercury and cadmium concentrations, and between fat and arsenic and inorganic arsenic concentrations.
Assuntos
Braquiúros/química , Metais Pesados/análise , Bifenilos Policlorados/análise , Frutos do Mar/análise , Animais , Federação RussaRESUMO
Arsenolipids are the major arsenic species present in marine oils. Several structures of arsenolipids have been elucidated the last 5 years, demonstrating the chemical complexity of this trace element in the marine environment. Several commercial fish oils and marine oils, ranging in total arsenic concentrations from 1.6 to 12.5 mg kg(-1) oil, were analyzed for arsenolipids using reversed-phase high performance liquid chromatography coupled with inductively coupled plasma mass spectrometry (HPLC-ICP-MS). The arsenolipids were quantified using three different arsenic-containing calibration standards; dimethylarsinate (DMA), triphenylarsinoxide (Ph3AsO) and a synthesized arsenic-containing hydrocarbon (AsHC) (dimethylarsinoyl nonadecane; C21H43AsO). The observed variation in signal intensity for arsenic during the gradient elution profile in reversed-phase HPLC was compensated for by determining the time-resolved response factors for the arsenolipids. Isotopes of germanium ((74)Ge) and indium ((115)In) were suited as internal standards for arsenic, and were used for verification of the arsenic signal response factors during the gradient elution. Dimethylarsinate was the most suitable calibration standard for the quantification of arsenolipids, with recoveries between 91% and 104% compared to total arsenic measurements in the same extracts. A range of marine oils was investigated, including oils of several fish species, cod liver and seal, as well as three commercial fish oils. The AsHCs - C17H38AsO, C19H42AsO and C23H38AsO - were identified as the major arsenolipids in the extracts of all oils by HPLC coupled with quadrupole time-of-flight mass spectrometry (qTOF-MS). Minor amounts of two arsenic-containing fatty acids (AsFAs) (C23H38AsO3 and C24H38AsO3) were also detected in the oils. The sum of the AsHCs and the AsFAs determined in the present study accounted for 17-42% of the total arsenic in the oils.
Assuntos
Arsênio/análise , Cromatografia Líquida de Alta Pressão/métodos , Ácidos Graxos/química , Óleos de Peixe/química , Hidrocarbonetos/química , Espectrometria de Massas/métodosRESUMO
This study is one of several baseline studies that will provide basic and reliable information about the content of undesirable substances in important species of fish caught in Norwegian waters. Concentrations of metals in the muscle and liver of more than 800 Northeast Arctic cod caught at 32 sites in the Barents Sea are reported. The highest concentration of both mercury in the muscle and cadmium in the liver was found in cod caught in the western part of the Barents Sea, while the highest concentration of total arsenic was found in cod from the eastern part. The arsenic concentrations varied greatly among individual fish, ranging from 0.3 to 170 mg kg(-1) wet weight in the muscle. Such high levels of total arsenic have never previously been reported in any fish, and the primary factor for these high concentrations is likely to be the shrimp in the cod diet.
Assuntos
Monitoramento Ambiental , Gadus morhua/metabolismo , Metais/metabolismo , Poluentes Químicos da Água/metabolismo , Animais , Arsênio/metabolismo , Arsênio/normas , Cádmio/metabolismo , Cádmio/normas , Contaminação de Alimentos/estatística & dados numéricos , Chumbo/metabolismo , Chumbo/normas , Mercúrio/metabolismo , Mercúrio/normas , Metais/normas , Alimentos Marinhos/análise , Alimentos Marinhos/estatística & dados numéricos , Poluentes Químicos da Água/normasRESUMO
The contents of total arsenic and inorganic arsenic were determined in fillet samples of Northeast Artic cod, herring, mackerel, Greenland halibut, tusk, saithe and Atlantic halibut. In total, 923 individual fish samples were analysed. The fish were mostly caught in the open sea off the coast of Norway, from 40 positions. The determination of total arsenic was carried out by inductively coupled plasma mass spectrometry following microwave-assisted wet digestion. The determination of inorganic arsenic was carried out by high-performance liquid chromatography-ICP-MS following microwave-assisted dissolution of the samples. The concentrations found for total arsenic varied greatly between fish species, and ranged from 0.3 to 110 mg kg(-1) wet weight. For inorganic arsenic, the concentrations found were very low (<0.006 mg kg(-1)) in all cases. The obtained results question the assumptions made by the European Food Safety Authority (EFSA) on the inorganic arsenic level in fish used in the recent EFSA opinion on arsenic in food.
Assuntos
Arsênio/análise , Dieta , Exposição Ambiental/análise , Peixes , Contaminação de Alimentos/análise , Alimentos Marinhos/análise , Animais , Humanos , NoruegaRESUMO
The molecular mechanisms underlying the neurotoxicity of methylmercury (MeHg), a ubiquitous environmental contaminant, are not yet fully understood. Furthermore, there is a lack of biomarkers of MeHg neurotoxicity for use in environmental monitoring. We have undertaken a proteomic analysis of brains from Atlantic cod (Gadus morhua) exposed to 0, 0.5 and 2 mg/kg MeHg administered by intraperitoneal injection. The doses were given in two injections, half of the dose on the first day and the second half after 1 week, and the total exposure period lasted 2 weeks. Using 2-DE coupled with MALDI-TOF MS and MS/MS, we observed the level of 71 protein spots to be 20% or more significantly altered following MeHg exposure, and successfully identified 40 of these protein spots. Many of these proteins are associated with main known molecular targets and mechanisms of MeHg-induced neurotoxicity in mammals, such as mitochondrial dysfunction, oxidative stress, altered calcium homeostasis and tubulin/disruption of microtubules. More interestingly, several of the affected proteins, with well-established or recently demonstrated critical functions in nervous system-specific processes, have not previously been associated with MeHg exposure in any species. These proteins include the strongest up-regulated protein, pyridoxal kinase (essential for synthesis of several neurotransmitters), G protein (coupled to neurotransmitter receptors), nicotinamide phosphoribosyl-transferase (protection against axonal degeneration), dihydropyrimidinase-like 5 (or collapsin response mediator protein 5, CRMP-5) (axon guidance and regeneration), septin (dendrite development), phosphatidylethanolamine binding protein (precursor for hippocampal cholinergic neurostimulating peptide) and protein phosphatase 1 (control of brain recovery by synaptic plasticity). The results of the present study aid our understanding of molecular mechanisms underlying MeHg neurotoxicity and defense responses, and provide a large panel of protein biomarker candidates for aquatic environmental monitoring.