Ion-induced PCR strategy for mercury detection.

Mercury contamination is one of the most serious environmental problems. It can cause serious effects on the human health, such as case damage in the brain, nervous system, immune system, and kidney failure. Therefore, development of an accurate, sensitive, and simple operational detection method for mercury is very necessary. Herein, we report a new strategy for mercury ion detection based on commonly used PCR technique. High selectivity and sensitivity were achieved by the formation of the thymine-Hg-thymine (T-Hg-T) unnatural base pair at the 3'-end of PCR primers. The detection results of PCR amplification in presence of mercury ion could be reported either by using agarose gel analysis or through real-time fluorometric dye tracing for different detection purposes. To our knowledge, this study represents the first application of PCR based technique to the detection of metal ions.

Assessment of mobility and bioavailability of mercury compounds in sewage sludge and composts.

Content of heavy metals, including mercury, determines the method of management and disposal of sewage sludge. Excessive concentration of mercury in composts used as organic fertilizer may lead to accumulation of this element in soil and plant material. Fractionation of mercury in sewage sludge and composts provides a better understanding of the extent of mobility and bioavailability of the different mercury species and helps in more informed decision making on the application of sludge for agricultural purposes. The experimental setup comprises the composing process of the sewage sludge containing 13.1mgkg-1 of the total mercury, performed in static reactors with forced aeration. In order to evaluate the bioavailability of mercury, its fractionation was performed in sewage sludge and composts during the process. An analytical procedure based on four-stage sequential extraction was applied to determine the mercury content in the ion exchange (water soluble and exchangeable Hg), base soluble (Hg bound to humic and fulvic acid), acid soluble (Hg bound to Fe/Mn oxides and carbonates) and oxidizable (Hg bound to organic matter and sulphide) fractions. The results showed that from 50.09% to 64.55% of the total mercury was strongly bound to organo-sulphur and inorganic sulphide; that during composting, increase of concentrations of mercury compounds strongly bound with organic matter and sulphides; and that mercury content in the base soluble and oxidizable fractions was strongly correlated with concentration of dissolved organic carbon in those fractions.

Accumulation properties of inorganic mercury and organic mercury in the red-crowned crane Grus japonensis in east Hokkaido, Japan.

The red-crowned (Japanese) crane Grus japonensis is native to east Hokkaido, Japan, in contrast to the East Asia mainland. Previously, we reported that red-crowned cranes in Hokkaido were highly contaminated with mercury in the 1990s and that the contamination rapidly decreased to a moderate level in the 2000s. In the present study, we determined levels of organic mercury (O-Hg) in the liver and kidney of cranes in east Hokkaido in comparison with levels of total mercury (T-Hg). T-Hg levels in the kidneys were higher than those in the livers in adults but not in subadults and juveniles; however, the reverse was the case for O-Hg even for adults. The ratio of O-Hg to T-Hg in both the liver and kidney decreased as T-Hg increased in the three developmental stages. While the ratios of O-Hg to T-Hg in the liver and kidney of adults were significantly lower than those of juveniles, the ratios were similar for adults and juveniles in a lower range of T-Hg. The ratio of selenium (Se) to T-Hg decreased as T-Hg increased in both the liver and kidney, irrespective of stages. Mercury burdens in feathers were about 59% and 67% of the total body burdens for juveniles and adults, respectively. Furthermore, ratios of carbon and nitrogen stable isotopes to T-Hg varied greatly, with no relation to mercury level in the liver. The results suggest slow accumulation of inorganic mercury in the kidney of red-crowned cranes in east Hokkaido, Japan.

Speciation analysis of mercury in water samples by dispersive liquid-liquid microextraction coupled to capillary electrophoresis.

In this study, a method of pretreatment and speciation analysis of mercury by dispersive liquid-liquid microextraction along with CE was developed. The method was based on the fact that mercury species including methylmercury (MeHg), ethylmercury (EtHg), phenylmercury (PhHg), and Hg(II) were complexed with 1-(2-pyridylazo)-2-naphthol to form hydrophobic chelates and l-cysteine could displace 1-(2-pyridylazo)-2-naphthol to form hydrophilic chelates with the four mercury species. Factors affecting complex formation and extraction efficiency, such as pH value, type, and volume of extractive solvent and disperser solvent, concentration of the chelating agent, ultrasonic time, and buffer solution were investigated. Under the optimal conditions, the enrichment factors were 102, 118, 547, and 46, and the LODs were 1.79, 1.62, 0.23, and 1.50 µg/L for MeHg, EtHg, PhHg, and Hg(II), respectively. Method precisions (RSD, n = 5) were in the range of 0.29-0.54% for migration time, and 3.08-7.80% for peak area. Satisfactory recoveries ranging from 82.38 to 98.76% were obtained with seawater, lake, and tap water samples spiked at three concentration levels, respectively, with RSD (n = 5) of 1.98-7.18%. This method was demonstrated to be simple, convenient, rapid, cost-effective, and environmentally benign, and could be used as an ideal alternative to existing methods for analyzing trace residues of mercury species in water samples.

Microextraction techniques used in the procedures for determining organomercury and organotin compounds in environmental samples.

Due to human activities, the concentrations of organometallic compounds in all parts of the environment have increased in recent decades. The toxicity and some biochemical properties of mercury and tin present in the environment depend on the concentration and chemical form of these two elements. The ever-increasing demand for determining compounds at very low concentration levels in samples with complex matrices requires the elimination of interfering substances, the reduction of the final extract volume, and analyte enrichment in order to employ a detection technique, which is characterised by high sensitivity at low limits of quantification. On the other hand, in accordance with current trends, the analytical procedures should aim at the miniaturisation and simplification of the sample preparation step. In the near future, more importance will be given to the fulfilment of the requirements of Green Chemistry and Green Analytical Chemistry in order to reduce the intensity of anthropogenic activities related to analytical laboratories. In this case, one can consider the use of solvent-free/solvent-less techniques for sample preparation and microextraction techniques, because the use of the latter leads to lowering the quantity of reagents used (including solvents) due to the reduction of the scale of analysis. This paper presents an overview of microextraction techniques (SPME and LPME) used in the procedures for determining different chemical forms of mercury and tin.

Localizing organomercury uptake and accumulation in zebrafish larvae at the tissue and cellular level.

Using synchrotron x-ray fluorescence mapping, we have examined the uptake and localization of organic mercury in zebrafish larvae. Strikingly, the greatest accumulation of methyl and ethyl mercury compounds was highly localized in the rapidly dividing lens epithelium, with lower levels going to brain, optic nerve, and various other organs. The data suggest that the reported impairment of visual processes by mercury may arise not only from previously reported neurological effects, but also from direct effects on the ocular tissue. This novel approach is a powerful tool for directly investigating the molecular toxicology of heavy metals, and should be equally applicable to the study of a wide range of elements in developing embryos.

Mercury speciation in fish tissues from a Mediterranean River basin: the Tagus River (central Spain) as a case study.

An assessment of mercury (Hg) accumulation in fish from the Tagus River aquatic system (central Spain), which has been influenced by pollution from industrial and urban development, was performed. Total Hg (THg), inorganic Hg (IHg), and monomethylmercury (MMHg) were determined in muscle and liver of different fish species, including Cyprinus carpio, Ameiurus melas, and Chondrostoma miegii, sampled from three locations. Although concentrations of THg and Hg species showed wide variability among the fish species, they were also found to be considerably dependent on location and fish tissue. Relative contents of MMHg to THg in muscle varied from 60 to 88%, whereas those found in liver ranged from 7 to 59%. Mean THg concentrations ranged from 126 to 810 ng/g (dry weight [dw]) in liver and from 159 to 1057 ng/g dw in muscle. Therefore, the mean THg concentration in all fish muscle samples was far lower than the maximum residue level recommended by the European Union for fishery products. Nevertheless, the concentrations of Hg in fish muscle reported in this study were somewhat increased compared with other areas geographically distant from most major anthropogenic Hg sources and, in some cases, even greater than those previously reported elsewhere in more polluted areas. In contrast, Hg contents in liver were lower than those found in Hg-contaminated areas, but they were within the range found in other areas exposed to diffuse sources of pollution by Hg. Thus, this article provides an overview of the concentration and distribution of Hg species in fish muscle and liver tissues samples taken from a freshwater system in the Mediterranean River basin.

Influence of mercury bioaccessibility on exposure assessment associated with consumption of cooked predatory fish in Spain.

BACKGROUND: Predatory fish tend to accumulate high levels of mercury (Hg). Food safety assessment of these fish has been carried out on the raw product. However, the evaluation of the risk from Hg concentrations in raw fish might be modified if cooking and bioaccessibility (the contaminant fraction that solubilises from its matrix during gastrointestinal digestion and becomes available for intestinal absorption) were taken into account. Data on Hg bioaccessibility in raw predatory fish sold in Spain are scarce and no research on Hg bioaccessibility in cooked fish is available. The aim of the present study was to evaluate Hg bioaccessibility in various kinds of cooked predatory fish sold in Spain to estimate their health risk. RESULTS: Both Hg and bioaccessible Hg concentrations were analysed in raw and cooked fish (swordfish, tope shark, bonito and tuna). There were no changes in Hg concentrations during cooking. However, Hg bioaccessibility decreased significantly after cooking (42 ± 26% in raw fish and 26 ± 16% in cooked fish), thus reducing in swordfish and tope shark the Hg concentration to which the human organism would be exposed. CONCLUSION: In future, cooking and bioaccessibility should be considered in risk assessment of Hg concentrations in predatory fish.

Levels of organic and inorganic mercury in human blood predicted from measurements of total mercury.

The toxicologically relevant mercury species inorganic and organic Hg in blood are frequently determined by separate measurements of total Hg and of inorganic Hg, with their difference indicating organic Hg. It is shown that the different partition of inorganic and organic Hg between erythrocytes and plasma (e/p ratio) can be used to calculate the concentrations of either Hg species in either blood constituent from measurement of total Hg only. This was tested on the blood of different groups of volunteers. The calculated concentrations of inorganic and organic Hg in cells and plasma were then compared by linear regression with their previously measured counterparts. An accurate prediction has been found for individual levels of inorganic Hg in plasma and organic Hg in cells. These calculated levels were little affected by variations of the e/p ratios. The coincidence between calculated and measured levels of inorganic Hg in cells and organic Hg in plasma was more sensitive to alterations of the e/p ratios. In conclusion, the relevant concentrations of inorganic Hg in plasma and organic Hg in cells can reliably be calculated from measurements of total Hg and from assumed e/p ratios. This means a sizeable reduction of analytical work, and also provides specific information in cases of low-level co-exposure to both Hg species. Besides the possibility to introduce automated analyses of total Hg in mercury speciation in blood, the proposed calculation scheme has the potential to easily enlarge the data base in epidemiological and toxicological surveys of mercury exposure.

Mercury speciation by CE: an update.

This review provides an update on mercury speciation by CE. It includes a brief discussion on physicochemical properties, toxicity and transformation pathways of mercury species (i.e. methyl-, ethyl-, phenyl- and inorganic mercury) and outlines recent trends in Hg speciation by CE. CE is presented as a complementary technique to chromatographic separation techniques, especially in cases when speed, high efficiency and low sample volumes are required. The development of suitable sample preconcentration/isolation (sample stacking, ion exchange, liquid-liquid-liquid extraction, dual-cloud point extraction) to achieve low LODs for analysis of trace concentrations of mercury species in real samples is emphasized. Hyphenation of CE to element specific detectors (i.e. electrothermal atomic absorption spectrometry, atomic fluorescence spectrometry, inductively coupled plasma-optical emission spectrometry, inductively coupled plasma-mass spectrometry) is discussed as well as a potential of CE in interaction studies that may provide useful information on interaction of various Hg species with selected bio-macromolecules.

Mercury emissions from geothermal power plants.

Geothermal steam used for power production contains significant quantities of volatile mercury. Much of this mercury escapes to the atmosphere as elemental mercury vapor in cooling tower exhausts. Mercury emissions from geothermal power plants, on a per megawatt (electric) basis, are comparable to releases from coal-fired power plants.

DNA engineered copper oxide-based nanocomposites with multiple enzyme-like activities for specific detection of mercury species in environmental and biological samples.

In this paper, we report the synthesis and application of enzyme-like DNA-copper oxide/platinum nanoparticles for the separate quantification of inorganic and organomercury species in various real samples. We synthesized a series of poly(thymine) (T60)-copper oxide/metal nanocomposites (T60-CuxO/M NCs; M = Au, Ag or Pt) that exhibited enzyme-like activities [oxidase (OX), peroxidase (POX), and catalase (CAT)]. The enzyme-like activities are tunable due to the incorporation of various metals into the NCs. Among a series of synthesized CuxO/M NCs, T60-copper oxide-platinum nanocomposites (T60-CuxO/Pt NCs) exhibited the highest OX-like activity via the O2-mediated oxidation of substrates, such as Amplex Red (AR), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), o-phenylenediamine (OPD), and 3,3',5,5'-tetramethylbenzidine (TMB), to form fluorescent or colored products. Interestingly, inorganic mercury ions (Hg2+) and organomercury species, such as methylmercury (MeHg+), ethylmercury (EtHg+), and phenylmercury (PhHg+), significantly inhibited the OX-like activity of T60-CuxO/Pt NCs. For the selective detection of mercury species, we used ABTS in the T60-CuxO/Pt NCs system, and the ABTS/T60-CuxO/Pt NCs-based assay allowed for the detection of mercury ions at nanomolar concentrations based on the decrease in the catalytic activity caused by the mercury ions. To separately quantify the inorganic and organomercury species in a sample, we employed selenium nanoparticles (Se NPs) as a masking agent, as they preferentially bind with inorganic mercury species. The ABTS/T60-CuxO/Pt NCs-based assay with the masking agent of Se NPs further provided specificity for the detection of organomercury species in environmental water samples (tap water, river water, and seawater) and fish muscle samples (dogfish muscle DORM-II).

Synthesis and application of ion-imprinted polymer for the determination of mercury II in water samples.

In this study, an innovative analytical methodology capable of selectively identifying and quantifying mercury contamination by the association of solid-phase extraction using ion-imprinted polymers as a sorbent phase and differential pulse anodic stripping voltammetry is proposed. To this end, the ion-imprinted polymers were synthesized and characterized by infrared spectroscopy and atomic force microscopy. The sorption capacities and the selectivity of the ion-imprinted polymers were compared to the ones related to the non-imprinted ones. Next, the experimental parameters of this solid-phase extraction method (IIP-SPE) were evaluated univariately. The selectivity of this polymeric matrix against other cations (Cd II, Pb II, and Cu II) was also evaluated. Limits of detection (LOD) and quantification (LOQ) obtained for the here proposed methodology were 0.322 µg L-1 and 1.08 µg L-1, respectively. Also, the precision of 4.0% was achieved. The method was finally applied to three water samples from different sources: for the Piratininga and Itaipu Lagoon waters, Hg II concentrations were below the LOQ and for Vargem River waters a concentration equal to 1.35 ± 0.07 mg L-1 was determined. These results were confirmed by recovery tests, resulting in a recovery of 96.2 ± 4.0%, and by comparison with flame atomic absorption spectrometry, resulting in statistical conformity between the two methods at 95% confidence level.

Simple interface of high-performance liquid chromatography-atomic fluorescence spectrometry hyphenated system for speciation of mercury based on photo-induced chemical vapour generation with formic acid in mobile phase as reaction reagent.

Photo-induced chemical vapour generation (CVG) with formic acid in mobile phase as reaction reagent was developed as interface to on-line couple HPLC with atomic fluorescence spectrometry for the separation and determination of inorganic mercury, methylmercury (MeHg), ethylmercury (EtHg) and phenylmercury (PhHg). In the developed procedure, formic acid in mobile phase was used to decompose organomercuries and reduce Hg(2+) to mercury cold vapour under UV irradiation. Therefore, no post-column reagent was used and the flow injection system in traditional procedure is omitted. A number of operating parameters including pH of mobile phase, concentration of formate, flow rate of mobile phase, length of PTFE reaction coil, flow rate of carrier gas and Na(2)S(2)O(3) in sample matrix were optimized. The limits of detection at the optimized conditions were 0.085, 0.033, 0.029 and 0.038 microg L(-1) for inorganic mercury, MeHg, EtHg and PhHg, respectively. The developed method was validated by determination of certified reference material DORM-2 and was further applied in analyses of seafood samples from Yantai port, China. The UV-CVG with formic acid simplifies the instrumentation and reduces the analytical cost significantly.

Investigation into mercury bound to biothiols: structural identification using ESI-ion-trap MS and introduction of a method for their HPLC separation with simultaneous detection by ICP-MS and ESI-MS.

Mercury in plants or animal tissue is supposed to occur in the form of complexes formed with biologically relevant thiols (biothiols), rather than as free cation. We describe a technique for the separation and molecular identification of mercury and methylmercury complexes derived from their reactions with cysteine (Cys) and glutathione (GS): Hg(Cys)(2), Hg(GS)(2), MeHgCys, MeHgGS. Complexes were characterised by electrospray mass spectrometry (MS) equipped with an ion trap and the fragmentation pattern of MeHgCys was explained by using MP2 and B3LYP calculations, showing the importance of mercury-amine interactions in the gas phase. Chromatographic baseline separation was performed within 10 min with formic acid as the mobile phase on a reversed-phase column. Detection was done by online simultaneous coupling of ES-MS and inductively coupled plasma MS. When the mercury complexes were spiked in real samples (plant extracts), no perturbation of the separation and detection conditions was observed, suggesting that this method is capable of detecting mercury biothiol complexes in plants.

Optimisation of the headspace-solid phase microextraction for organomercury and organotin compound determination in sediment and biota.

Headspace solid-phase microextraction was optimised for the simultaneous preconcentration of methylmercury (MeHg+), monobutyltin, dibutyltin, tributyltin, monophenyltin (MPhT), diphenyltin (DPhT), and triphenyltin (TPhT) from sediments and biota. Extraction time (3-24 min), extraction temperature (20-90 degrees C), desorption time (1-10.4 min), desorption temperature (152-260 degrees C), and sample volume (5-22 mL) were simultaneously optimised, while variables such as fibre type (30 microm polydimethylsiloxane, PDMS), pH (acetic acid/sodium acetate, HOAc/NaOAc, 2 mol/L, pH approximately 4.8), the concentration of the derivatisation agent (sodium tetraethylborate, NaBEt4, 0.1% m/v), and the ionic strength (fixed by the buffer solution) were kept constant. The variables were optimised according to the experiments proposed by the MultiSimplex program and the responses were considered in order to establish the optimum conditions. The repeatability (relative standard deviation, RSD, 5-20.6%) and limits of detection (LODs, 0.05-0.97 ng/g) of the overall method were also estimated. The lowest precisions were obtained for DPhT and TPhT. The optimised preconcentration method was applied to the determination of MeHg+), butyl- and phenyltins in certified reference materials (IAEA-405 MeHg+) in estuarine sediment, BCR-646 butyl- and phenyltins in marine sediment, BCR-463 MeHg+ in tuna fish, DOLT-2 MeHg+ in dogfish liver, and BCR-477 butyltins in mussel tissue) by GC with microwave-induced plasma/atomic-emission detection.

Total mercury, organic mercury and selenium in liver and kidney of a South American coastal dolphin.

Selenium and total and organic mercury were determined in the liver and kidney of franciscana dolphin (Pontoporia blainvillei) incidentally caught in fishing nets along two Brazilian coastal areas (southeast and south). Regional differences in the concentrations of these contaminants were observed in P. blainvillei. Liver showed the highest organic and total mercury. In general, samples of individuals collected at the southern of Brazil had the highest concentrations of selenium and total and organic mercury. No significant gender differences were observed. Growth stage influenced the accumulation of these contaminants in both organs, and hepatic concentrations increased with the body length, according to the sampling area. Molar mercury and selenium concentrations in liver were significantly correlated, with a Se:Hg ratio close to 4. The among-site differences we found may be related to differences in preferred prey, bioavailability in the marine environment, environmental conditions, or these individuals may belong to distinct populations.

Dual-cloud point extraction as a preconcentration and clean-up technique for capillary electrophoresis speciation analysis of mercury.

A novel dual-cloud point extraction (dCPE) technique is proposed in this paper for the sample pretreatment of capillary electrophoresis (CE) speciation analysis of mercury. In dCPE, cloud point was carried out twice in a sample pretreatment. First, four mercury species, methylmercury (MeHg), ethylmercury (EtHg), phenylmercury (PhHg), and inorganic mercury (Hg(II)) formed hydrophobic complexes with 1-(2-pyridylazo)-2-naphthol (PAN). After heating and centrifuging, the complexes were extracted into the formed Triton X-114 surfactant-rich phase. Instead of the direct injection or analysis, the surfactant-rich phase containing the four Hg species was treated with 150 microL 0.1% (m/v) l-cysteine aqueous solution. The four Hg species were then transferred back into aqueous phase by forming hydrophilic Hg-l-cysteine complexes. After dCPE, the aqueous phase containing the Hg-l-cysteine complexes was subjected into electrophoretic capillary for mercury speciation analysis. Because the concentration of Triton X-114 in the extract after dCPE was only around critical micelle concentration, the adsorption of surfactant on the capillary wall and its possible influence on the sample injection and separation in traditional CPE were eliminated. Plus, the hydrophobic interfering species were removed thoroughly by using dCPE resulted in significant improvement in analysis selectivity. Using 10 mL sample, 17, 15, 45, and 52 of preconcentration factors for EtHg, MeHg, PhHg, and Hg(II) were obtained. With CE separation and on-line UV detection, the detection limits were 45.2, 47.5, 4.1, and 10.0 microg L(-1) (as Hg) for EtHg, MeHg, PhHg, and Hg(II), respectively. As an analysis method, the present dCPE-CE with UV detection obtained similar detection limits as of some CE-inductively coupled plasma mass spectrometry (ICPMS) hyphenation technique, but with simple instrumental setup and obviously low costs. Its utilization for Hg speciation was validated by the analysis of the spiked natural water and tilapia muscle samples.

Hollow fiber-based liquid-liquid-liquid microextraction combined with high-performance liquid chromatography for the speciation of organomercury.

A novel and extremely highly sensitive method for the speciation of organomercury in sea food and environmental water samples was developed by hollow fiber liquid-liquid-liquid microextraction (LLLME) coupled to high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection at 254 nm. By the use of toluene filled the pores of the hollow fiber wall as the organic phase and Na(2)S(2)O(3) aqueous solution in the lumen of hollow fiber as acceptor phase, the simultaneous preconcentration of methylmercury (MeHg), ethylmercury (EtHg), and phenylmercury (PhHg) could be realized, and the target mercury species in the post-extraction acceptor phase were directly injected into HPLC for their separation. The factors affecting on the liquid-liquid-liquid microextraction of three organomercury species, including organic solvent, the pH of donor phase, the concentration of complexing agent, extraction time and stirring rate were investigated and the optimal extraction conditions were established. With the consumption of 3.8 mL of sample solution, the enrichment factors were about 120, 215 and 350 for MeHg, EtHg, and PhHg, respectively. The limits of detection were in the range of 0.3-3.8 ng mL(-1) (as Hg) with precisions (RSDs (%), c=30 ng mL(-1) (as Hg), n=7) ranging from 6.4 to 8.9%. The developed technique was validated by analyzing a certified reference material (DORM-2, dogfish-muscle, NRCC) and an environmental water sample, and the analytical results were satisfactory.

Mercury in human brain, blood, muscle and toenails in relation to exposure: an autopsy study.

BACKGROUND: The main forms of mercury (Hg) exposure in the general population are methylmercury (MeHg) from seafood, inorganic mercury (I-Hg) from food, and mercury vapor (Hg0) from dental amalgam restorations. While the distribution of MeHg in the body is described by a one compartment model, the distribution of I-Hg after exposure to elemental mercury is more complex, and there is no biomarker for I-Hg in the brain. The aim of this study was to elucidate the relationships between on the one hand MeHg and I-Hg in human brain and other tissues, including blood, and on the other Hg exposure via dental amalgam in a fish-eating population. In addition, the use of blood and toenails as biological indicator media for inorganic and organic mercury (MeHg) in the tissues was evaluated. METHODS: Samples of blood, brain (occipital lobe cortex), pituitary, thyroid, abdominal muscle and toenails were collected at autopsy of 30 deceased individuals, age from 47 to 91 years of age. Concentrations of total-Hg and I-Hg in blood and brain cortex were determined by cold vapor atomic fluorescence spectrometry and total-Hg in other tissues by sector field inductively coupled plasma-mass spectrometry (ICP-SFMS). RESULTS: The median concentrations of MeHg (total-Hg minus I-Hg) and I-Hg in blood were 2.2 and 1.0 microg/L, and in occipital lobe cortex 4 and 5 microg/kg, respectively. There was a significant correlation between MeHg in blood and occipital cortex. Also, total-Hg in toenails correlated with MeHg in both blood and occipital lobe. I-Hg in both blood and occipital cortex, as well as total-Hg in pituitary and thyroid were strongly associated with the number of dental amalgam surfaces at the time of death. CONCLUSION: In a fish-eating population, intake of MeHg via the diet has a marked impact on the MeHg concentration in the brain, while exposure to dental amalgam restorations increases the I-Hg concentrations in the brain. Discrimination between mercury species is necessary to evaluate the impact on Hg in the brain of various sources of exposure, in particular, dental amalgam exposure.