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.

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.

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.

Automated dynamic hollow fiber liquid-liquid-liquid microextraction combined with capillary electrophoresis for speciation of mercury in biological and environmental samples.

A simple home-made automatic dynamic hollow fiber based liquid-liquid-liquid microextraction (AD-HF-LLLME) device was designed and constructed for the simultaneous extraction of organomercury and inorganic mercury species with the assistant of a programmable flow injection analyzer. With 18-crown-6 as the complexing reagent, mercury species including methyl-, ethyl-, phenyl- and inorganic mercury were extracted into the organic phase (chlorobenzene), and then back-extracted into the acceptor phase of 0.1% (m/v) 3-mercapto-1-propanesulfonic acid (MPS) aqueous solution. Compared with automatic static (AS)-HF-LLLME system, the extraction equilibrium of target mercury species was obtained in shorter time with higher extraction efficiency in AD-HF-LLLME system. Based on it, a new method of AD-HF-LLLME coupled with large volume sample stacking (LVSS)-capillary electrophoresis (CE)/UV detection was developed for the simultaneous analysis of methyl-, phenyl- and inorganic mercury species in biological samples and environmental water. Under the optimized conditions, AD-HF-LLLME provided high enrichment factors (EFs) of 149-253-fold within relatively short extraction equilibrium time (25min) and good precision with RSD between 3.8 and 8.1%. By combining AD-HF-LLLME with LVSS-CE/UV, EFs were magnified up to 2195-fold and the limits of detection (at S/N=3) for target mercury species were improved to be sub ppb level.

Legacy source of mercury in an urban stream-wetland ecosystem in central North Carolina, USA.

In the United States, aquatic mercury contamination originates from point and non-point sources to watersheds. Here, we studied the contribution of mercury in urban runoff derived from historically contaminated soils and the subsequent production of methylmercury in a stream-wetland complex (Durham, North Carolina), the receiving water of this runoff. Our results demonstrated that the mercury originated from the leachate of grass-covered athletic fields. A fraction of mercury in this soil existed as phenylmercury, suggesting that mercurial anti-fungal compounds were historically applied to this soil. Further downstream in the anaerobic sediments of the stream-wetland complex, a fraction (up to 9%) of mercury was converted to methylmercury, the bioaccumulative form of the metal. Importantly, the concentrations of total mercury and methylmercury were reduced to background levels within the stream-wetland complex. Overall, this work provides an example of a legacy source of mercury that should be considered in urban watershed models and watershed management.

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.

Total and organic mercury concentrations in the muscles of Pacific albacore (Thunnus alalunga) and bigeye tuna (Thunnus obesus).

Muscles of 115 North Pacific albacore (ALB, Thunnus alalunga) and 75 Pacific bigeye tuna (BET, Thunnus obesus), collected from 2001 to 2006, were analyzed. No ALB, but 13 large BET had organic mercury (OH g) concentrations exceeding 1 µg g(-1) wet weight. For both ALB and BET, total mercury (THg) and OH g concentrations were significantly and positively correlated with fork length (FL) and body weight. The muscle Hg bioaccumulation rates of BET were higher than those of ALB, particularly in the adult fish. Moreover, the lines had crossover points among the two species that imply the young BET (FL<110 cm) contains lower muscle Hg concentrations than ALB of the same size. The suggested weekly dietary intake of ALB and small-BET meats is 340 g, and of BET meat it is 150 g for a 60-kg person based on the provisional tolerable weekly intake (PTWI) of methylmercury set by the WHO.

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.

[Evaluation of the mercury accumulating capacity of pepper (Capsicum annuum)]. / Evaluación de la capacidad acumuladora de mercurio del ají (Capsicum annuum).

OBJECTIVE: To assess the mercury accumulating capacity in contaminated soils from the community of Mina Santa Cruz, in the south of the department of Bolívar, Colombia, of the pepper plant (Capsicum annuum), in order to establish the risk to the health of the consuming population. MATERIALS AND METHODS: Samples were taken from tissues (roots, stems, and leaves) of pepper plants grown in two soils contaminated with mercury and a control soil during the first five months of growth to determine total mercury through cold vapor atomic absorption spectrometry. Total mercury was determined in the samples of pepper plant fruits consumed in Mina Santa Cruz. RESULTS: The mean concentrations of total mercury in the roots were higher than in stems and leaves. Accumulation in tissues was influenced by mercury levels in soil and the growth time of the plants. Mercury concentrations in fruits of pepper plant were lower than tolerable weekly intake provided by WHO. CONCLUSION: Percent of translocation of mercury to aerial parts of the plant were low in both control and contaminated soils. Despite low levels of mercury in this food, it is necessary to minimize the consumption of food contaminated with this metal.

Risks associated with the transfer of toxic organo-metallic mercury from soils into the terrestrial feed chain.

Although the transfer of organo-metallic mercury (OrgHg) in aquatic food webs has long been studied, it has only been recently recognized that there is also accumulation in terrestrial systems. There is still however little information about the exposure of grazing animals to OrgHg from soils and feed as well as on risks of exposure to animal and humans. In this study we collected 78 soil samples and 40 plant samples (Lolium perenne and Brassica juncea) from agricultural fields near a contaminated industrial area and evaluated the soil-to-plant transfer of Hg as well as subsequent trophic transfer. Inorganic Hg (IHg) concentrations ranged from 0.080 to 210mgkg(-1) d.w. in soils, from 0.010 to 84mgkg(-1) d.w. in roots and from 0.020 to 6.9mgkg(-1) d.w. in shoots. OrgHg concentrations in soils varied between 0.20 and 130µgkg(-1) d.w. representing on average 0.13% of the total Hg (THg). In root and shoot samples OrgHg comprised on average 0.58% (roots) and 0.66% (shoots) of THg. Average bioaccumulation factors (BAFs) for OrgHg in relation to soil concentrations were 3.3 (for roots) and 1.5 (for shoots). The daily intake (DI) of THg in 33 sampling sites exceeded the acceptable daily intake (ADI) of THg of both cows (ADI=1.4mgd(-1)) and sheep (ADI=0.28mgd(-1)), in view of food safety associated with THg in animal kidneys. Estimated DI of OrgHg for grazing animals were up to 220µgd(-1) (for cows) and up to 33µgd(-1) (for sheep). This study suggested that solely monitoring the levels of THg in soils and feed may not allow to adequately taking into account accumulation of OrgHg in feed crops and properly address risks associated with OrgHg exposure for animals and humans. Hence, the inclusion of limits for OrgHg in feed quality and food safety legislation is advised.

Sensitive determination method for mercury ion, methyl-, ethyl-, and phenyl-mercury in water and biological samples using high-performance liquid chromatography with chemiluminescence detection.

A sensitive determination method for mercury speciation analysis was developed. Four mercury species, mercury ion, methylmercury, ethylmercury, and phenylmercury, were complexed with emetine-dithiocarbamate (emetine-CS(2)), and then injected onto a HPLC instrument coupled with a tris(2,2'-bipyridine)ruthenium(III) chemiluminescence detection system. The emetine-CS(2) complexing agent was effectively used to measure the concentration in addition to serving as a separation and detection reagent. The calibration curves for these mercury complexes were linear in the range of 0.050 - 10 µg L(-1) (as Hg). The limit of detection for (emetine-CS(2))(2)Hg, emetine-CS(2)-methylmercury, emetine-CS(2)-ethylmercury, and emetine-CS(2)-phenylmercury were 30, 17, 21, and 22 ng L(-1), respectively. The sensitivity of this method enables the determination of mercury species in water samples at sub-ppb levels. Furthermore, the method was applied to biological samples in combination with acid leaching and liquid-liquid extraction using emetine-CS(2) as an extraction reagent. The determination results were in good agreement with the values of the certified reference materials.

Phase transfer membrane supported liquid-liquid-liquid microextraction combined with large volume sample injection capillary electrophoresis-ultraviolet detection for the speciation of inorganic and organic mercury.

In this paper, a novel sample pretreatment technique termed phase transfer based liquid-liquid-liquid microextraction (PT-LLLME) was proposed for the simultaneous extraction of inorganic and organic mercury species. In PT-LLLME, an intermediate solvent (acetonitrile) was added into the donor phase to improve the contacting between target mercury species and complexing reagent. Meanwhile, a membrane supported (MS)-LLLME unit was designed to realize the PT-LLLME procedure. By using nylon membrane as supporting carrier, larger than 50 µL of acceptor solution could be hung up. Following PT/MS-LLLME, the acceptor solutions were directly analyzed by large volume sample stacking capillary electrophoresis/ultraviolet detection (LVSS-CE/UV). Accordingly, a new method of PT/MS-LLLME combined with LVSS-CE/UV was developed for the simultaneous speciation of inorganic and organic mercury species. Parameters affecting the extraction efficiency of PT/MS-LLLME were investigated in details. Under the optimized conditions, enrichment factors (EFs) ranging from 160- to 478-fold were obtained for the extraction of target mercury species by PT/MS-LLLME. By combining PT/MS-LLLME with LVSS-CE/UV, EFs were magnified up to 12,138-fold and the limits of detection (at a signal-to-noise ratio of 3) were at sub ppb level. The established approach of PT/MS-LLLME-LVSS-CE/UV was successfully applied to simultaneous determination of inorganic and organic mercury species in biological samples and environmental water samples.

Fish consumption and risk of contamination by mercury---considerations on the definition of edible parts based on the case study of European sea bass.

In the present study, the risk to humans by consuming European sea bass (Dicentrarchus labrax), captured at three sites along a Hg contamination gradient, was evaluated by comparing muscle and kidney total Hg (T-Hg) levels with the European regulations for marketed fish. Moreover, T-Hg and organic Hg (O-Hg) levels in muscle were compared with the Provisional Tolerable Weekly Intake (PTWI) and the Reference Dose (RfD). Although T-Hg levels in muscle were below the European value allowable for marketed fish, kidney's levels were higher than the set value, stressing the importance of redefining the concept of edible tissue and which tissues should be considered. Mercury weekly ingestion in the contaminated areas was higher than the PTWI, and O-Hg daily ingestion rates were higher than the RfD in all sampling sites. Thus, populations consuming sea bass from the contaminated sites may be at risk, with particular relevance for children and pregnant women.

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.

Amphiphilic porphyrin assembly as a highly selective chemosensor for organic mercury in water.

Homogeneously sized nanoparticles were successfully constructed based on amphiphilic porphyrin-cholesterol arrays, showing unique spectral and colourimetric response to organic mercury in water, even in the presence of Hg(2+).

Organic and total mercury levels in bigeye tuna, Thunnus obesus, harvested by Taiwanese fishing vessels in the Atlantic and Indian Oceans.

Muscle samples of 121 and 110 bigeye tuna (Thunnus obesus) caught by Taiwanese long-line fishing vessels in the Atlantic and Indian Oceans, respectively, were used to analyze total mercury (THg) and organic mercury (OHg) content. The overall THg and OHg concentrations were 0.786 ± 0.386 (0.214-3.133) and 0.595 ± 0.238 (0.143-2.222) mg kg⁻¹ wet weight, respectively, similar to the results of previous studies. Our findings, however, reflected the highest THg and OHg concentrations for the species in each ocean among the published data. Mean THg and OHg concentrations in Atlantic tuna were significantly (p < 0.05) higher than those in Indian tuna. Two of 121 samples of tuna from the Atlantic Ocean, but no samples from the Indian Ocean, had levels of OHg above 2 mg kg⁻¹ wet weight set by the Department of Health Taiwan, and 13 of 121 samples of tuna from the Atlantic Ocean and three of 110 samples from the Indian Ocean had levels of OHg above 1 mg kg⁻¹ wet weight set by US FDA and WHO. Accordingly, for adult Taiwanese men and women with average body weight of 65 and 55 kg, respectively, the maximum allowable weekly intake of bigeye tuna is suggested to be 170 and 145 g, respectively.

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.