Inorganic Mercury Species Research Articles

Effects of ultraviolet radiation on mercury isotope fractionation during photo-reduction for inorganic and organic mercury species

Photo-reduction of mercury (Hg) is an important mechanism for removal of both Hg2+ species and monomethylmercury (MMHg) from surface waters. Large mass independent fractionation (MIF) signatures of Hg isotopes preserved in natural samples are thought to reflect MIF produced during aqueous photo-reduction by the magnetic isotope effect (MIE). Recently, Hg MIF signatures in natural samples are being used to quantify photochemical reduction in aquatic systems. However, the fractionation factors used are from laboratory experiments that did not investigate many of the environmental parameters that may affect MIF during photo-reduction. In this study, the effects of different regions of the solar spectrum on the expression of MIF caused by the MIE during Hg2+ and MMHg photo-reduction were investigated to assess how the type of radiation affects the extent and signature of MIF. Photo-reduction in the presence of DOM was carried out through exposure to the full natural solar spectrum unfiltered, with the ultraviolet B (UVB; 290–320nm) portion removed and with both the UVB and the ultraviolet A (UVA; 320–400nm) removed. There is a clear relationship between the expression and magnitude of MIF and the energy of incident radiation for both Hg2+ and MMHg photo-reduction. The experiments indicate that MIF produced during photo-reduction of Hg2+ is significantly influenced by both UVB and UVA radiation. For MMHg photodemethylation, however, UVB radiation is mostly responsible for the MIF with minor contributions from UVA. Overall, there is a lack of correlation between the observed MIF and total photo-reduction in the experiments conducted in this study, which indicates that indirect and other non-MIF producing pathways of photo-reduction are the dominant pathways by which Hg species are being photo-reduced. Extrapolating these experimental results to natural systems is difficult as the experiments were not performed at realistic Hg/DOM ratios and the distribution of ligands that Hg is bound to in the experiments likely differs from natural systems. However, the strong relationship between MIF and energy of incident radiation in this study supports Hg MIF signatures as promising tools for helping to quantify photochemical cycling of mercury, but this study also highlights the need to understand the link between MIF and total photo-reduction before this tool can be fully utilized.

Mercury speciation in seawater by liquid chromatography-inductively coupled plasma-mass spectrometry following solid phase extraction pre-concentration by using an ionic imprinted polymer based on methyl-mercury–phenobarbital interaction

Trace levels of inorganic mercury, methyl-mercury and ethyl-mercury have been assessed in seawater by high performance liquid chromatography (HPLC) hyphenated with inductively coupled plasma-mass spectrometry (ICP-MS) after solid phase extraction (SPE) pre-concentration with a novel synthesized ionic imprinted polymer. The adsorbent material was prepared by trapping a non-vinylated chelating ligand (phenobarbital) via imprinting of a ternary mixed ligand complex of the non-vinylated chelating agent, the template (methyl-mercury), and the vinyl ligand (metacrylic acid, MAA). Ethylene dimetacrylate (EDMA) and 2,2′-azobisisobutyronitrile (AIBN) were used as cross-linker and initiator reagents, respectively; and the precipitation polymerization technique was used in a porogen of acetonitrile/water (4:1). The best retention properties for methyl-mercury, inorganic mercury and ethyl-mercury species from seawater were obtained when loading 200mL of sample adjusted to pH 8.0 and at a flow rate of 2.0mLmin−1 on a column-packed with 200mg of the material. Quantitative mercury species recoveries were obtained using 4mL of an eluting solution consisting of 0.8% (v/v) 2-mercaptoethanol and 20% (v/v) methanol (pH adjusted to 4.5) pumped at a flow rate of 2.0mLmin−1. Mercury species separation was achieved on a Kinetex C18 column working under isocratic conditions (0.4% (v/v) 2-mercaptoethanol, 10% (v/v) methanol, pH 2.5, flow rate 0.7mLmin−1). ICP-MS detection was performed by monitoring the mercury mass to charge ratio of 202. The limits of quantification of the method were 11, 6.7, and 12ngL−1, for inorganic mercury, methyl-mercury and ethyl-mercury, respectively (pre-concentration factor of 50); whereas, analytical recoveries ranged from 96 to 106%. The developed method was successfully applied to several seawater samples from unpolluted areas.

Neurotoxicity of organic and inorganic mercury species – Effects on and transfer across the blood-cerebrospinalfluid barrier, cytotoxic effects in target cells

Neurotoxicity of organic and inorganic mercury species – Effects on and transfer across the blood-cerebrospinalfluid barrier, cytotoxic effects in target cells

Fumigant methyl iodide can methylate inorganic mercury species in natural waters.

Methyl iodide or iodomethane (CH3I) has recently been registered as a fumigant in many countries, although its environmental impacts are not well understood. Here we report the results of a study on the methylation of mercury by CH3I in natural water by incubation experiments using both Hg ((199)HgCl2 and CH3(201)Hg(+))- and hydrogen (CD3I)-stable isotope addition techniques. We find that methylation of Hg(0), Hg2(2+) and Hg(2+) by CH3I can occur in natural water under sunlight, while only Hg(0) and Hg2(2+) can be methylated in deionized water. We propose that the methylation of Hg by CH3I in natural waters is mediated by sunlight and involves two steps, the reduction of Hg(2+) to Hg(0)/Hg2(2+) and the subsequent methylation of Hg(0)/Hg2(2+) by CH3I. Further quantitative assessment suggests that CH3I-involved methylation of inorganic Hg could be an important source of CH3Hg(+) in an environment where CH3I has been used in large amounts as a fumigant.

A global ocean inventory of anthropogenic mercury based on water column measurements.

Mercury is a toxic, bioaccumulating trace metal whose emissions to the environment have increased significantly as a result of anthropogenic activities such as mining and fossil fuel combustion. Several recent models have estimated that these emissions have increased the oceanic mercury inventory by 36-1,313million moles since the 1500s. Such predictions have remained largely untested owing to a lack of appropriate historical data and natural archives. Here we report oceanographic measurements of total dissolved mercury and related parameters from several recent expeditions to the Atlantic, Pacific, Southern and Arctic oceans. We find that deep North Atlantic waters and most intermediate waters are anomalously enriched in mercury relative to the deep waters of the South Atlantic, Southern and Pacific oceans, probably as a result of the incorporation of anthropogenic mercury. We estimate the total amount of anthropogenic mercury present in the global ocean to be 290±80 million moles, with almost two-thirds residing in water shallower than a thousand metres. Our findings suggest that anthropogenic perturbations to the global mercury cycle have led to an approximately 150 per cent increase in the amount of mercury in thermocline waters and have tripled the mercury content of surface waters compared to pre-anthropogenic conditions. This information may aid our understanding of the processes and the depths at which inorganic mercury species are converted into toxic methyl mercury and subsequently bioaccumulated in marine food webs.

Determination of mercury in gasoline by photochemical vapor generation coupled to graphite furnace atomic absorption spectrometry

Quantitation of mercury in gasoline by graphite furnace atomic absorption spectrometry following its photochemical vapor generation is described. Samples were prepared as a mixture of gasoline and propan-2-ol. A high efficiency flow through 19W photochemical vapor generator was used to process prepared samples with in-situ collection of the resultant Hg0 vapor onto reduced palladium in the graphite tube. Response from inorganic and organomercury standards added to the samples did not differ significantly. A limit of detection and characteristic mass of 0.1μgL−1 and 0.6ng were obtained, respectively. Relative standard deviations of replicate measurements of 3.0ngmL−1 added Hg(II) were typically 7–8%. Recoveries of spikes from samples ranged from 90 to 97%. The concentration of mercury in all tested samples of gasoline was below the limit of quantitation (0.3μgL−1). The method was rapid, efficient, green, fit-for-purpose, responsive to both inorganic and organic mercury species and should also respond to any Hg0 endogenously present in the sample.

Method development for the simultaneous determination of methylmercury and inorganic mercury in seafood

This paper reports the method development for the simultaneous determination of methylmercury (MeHg+) and inorganic mercury (iHg) species in seafood samples. The study focused on the extraction and quantification of MeHg+ (the most toxic species) by liquid chromatography coupled to on-line UV irradiation and cold vapour atomic fluorescence spectroscopy (LC-UV-CV-AFS), using HCl 4 mol L−1 as the extractant agent. Accuracy of the method has been verified by analysing three certified reference materials and different spiked samples. The values found for total Hg and MeHg+ for the CRMs did not differ significantly from certified values at a 95% confidence level, and recoveries between 85% and 97% for MeHg+, based on spikes, were achieved. The detection limits (LODs) obtained were 0.001 mg Hg kg−1 for total mercury, 0.0003 mg Hg kg−1 for MeHg+ and 0.0004 mg Hg kg−1 for iHg. The quantification limits (LOQs) established were 0.003 mg Hg kg−1 for total mercury, 0.0010 mg Hg kg−1 for MeHg+ and 0.0012 mg Hg kg−1 for iHg. Precision for each mercury species was established, being ≤ 12% in terms of RSD in all cases. Finally, the developed method was applied to 24 seafood samples from different origins and total mercury contents. The concentrations for Total Hg, MeHg+ and iHg ranged from 0.07 to 2.33, 0.003–2.23 and 0.006–0.085 mg Hg kg−1, respectively. The established analytical method allows to obtain results for mercury speciation in less than 1 one hour including both, sample pretreatment and measuring step.

Mechanisms of Hg species induced toxicity in cultured human astrocytes: genotoxicity and DNA-damage response

The toxicologically most relevant mercury (Hg) species for human exposure is methylmercury (MeHg). Thiomersal is a common preservative used in some vaccine formulations. The aim of this study is to get further mechanistic insight into the yet not fully understood neurotoxic modes of action of organic Hg species. Mercury species investigated include MeHgCl and thiomersal. Additionally HgCl2 was studied, since in the brain mercuric Hg can be formed by dealkylation of the organic species. As a cellular system astrocytes were used. In vivo astrocytes provide the environment necessary for neuronal function. In the present study, cytotoxic effects of the respective mercuricals increased with rising alkylation level and correlated with their cellular bioavailability. Further experiments revealed for all species at subcytotoxic concentrations no induction of DNA strand breaks, whereas all species massively increased H2O2-induced DNA strand breaks. This co-genotoxic effect is likely due to a disturbance of the cellular DNA damage response. Thus, at nanomolar, sub-cytotoxic concentrations, all three mercury species strongly disturbed poly(ADP-ribosyl)ation, a signalling reaction induced by DNA strand breaks. Interestingly, the molecular mechanism behind this inhibition seems to be different for the species. Since chronic PARP-1 inhibition is also discussed to sacrifice neurogenesis and learning abilities, further experiments on neurons and in vivo studies could be helpful to clarify whether the inhibition of poly(ADP-ribosyl)ation contributes to organic Hg induced neurotoxicity.

Determination of Trace Inorganic Mercury Species in Water Samples by Cloud Point Extraction and UV-Vis Spectrophotometry

A new micelle-mediated extraction method was developed for preconcentration of ultratrace Hg(II) ions prior to spectrophotometric determination. 2-(2'-Thiazolylazo)-p-cresol (TAC) and Ponpe 7.5 were used as the chelating agent and nonionic surfactant, respectively. Hg(II) ions form a hydrophobic complex with TAC in a micelle medium. The main factors affecting cloud point extraction efficiency, such as pH of the medium, concentrations of TAC and Ponpe 7.5, and equilibration temperature and time, were investigated in detail. An overall preconcentration factor of 33.3 was obtained upon preconcentration of a 50 mL sample. The LOD obtained under the optimal conditions was 0.86 microg/L, and the RSD for five replicate measurements of 100 microg/L Hg(II) was 3.12%. The method was successfully applied to the determination of Hg in environmental water samples.

Determination of mercury species using thermal desorption analysis in AAS

AbstractAnalytical aspects of the determination of inorganic mercury (Hg) species by thermal desorption followed by atomic absorption spectrometry (AAS) detection were investigated in this work. Characteristic Hg release curves of the following species were observed: Hg0, HgCl2, HgO, HgSO4, HgS, and the Hg bound to humic acids. Particular attention was dedicated to the thermal stability and change of bond of Hg0 in the following matrices: sand, kaolinite, granite, peat, power plant ash, and soil. The bond of elemental Hg in environmental materials was described on basis of this experiment. Contaminated soil samples from two locations in the Czech Republic were investigated by thermal desorption analysis. Afterwards, the contents of volatile and plant-available Hg in the studied samples were determined. The determination of Hg0 using the thermal method was related to the results of liquid sequential extraction. The development of Hg speciation and the stability of Hg were assessed on basis of the data obtained. Thus, the analytical procedure used is a suitable tool for the study of inorganic Hg species in contaminated soils.

Improved hollow fiber supported liquid–liquid–liquid membrane microextraction for speciation of inorganic and organic mercury by capillary electrophoresis

An improved hollow fiber supported liquid–liquid–liquid membrane microextraction (HF-LLLMME) method is described for the simultaneous extraction of inorganic and organic mercury species with high efficiency prior to speciation analysis by capillary electrophoresis. The HF-LLLMME was developed by filling cyclohexane and L-cysteine aqueous solution in the pores of the hollow fiber wall and its lumen as organic liquid membrane and acceptor phase, respectively. Inorganic mercury (Hg2+), methylmercury (MeHg), ethylmercury (EtHg) and phenylmercury (PhHg) firstly formed hydrophobic complexes with 1-(2-pyridylazo)-2-naphthol in donor solutions, wherein the hollow fiber was immersed. The complexes were then extracted into the organic liquid membrane and further transferred into the lumen of the hollow fiber to form hydrophilic complexes with L-cysteine. The factors affecting the extraction efficiency were carefully investigated. With the consumption of 50 mL of sample solution, the enrichment factor was 103, 265, 511 and 683 for Hg2+, MeHg, EtHg and PhHg, respectively. The limits of detection were in the range of 0.07–1.0 ng mL−1 (as Hg) with precisions (RSDs, n = 6) ranging from 1.7 to 4.4%. The proposed method was validated by analyzing a certified reference material (DORM-2, dogfish-muscle) and an environmental water sample with satisfactory results.

ChemInform Abstract: A Sensitive Colorimetric and Ratiometric Fluorescent Probe for Mercury Species in Aqueous Solution and Living Cells.

AbstractA new fluorescent probe for inorganic and organic mercury species based on mercury triggered cleavage reaction is developed.

A sensitive colorimetric and ratiometric fluorescent probe for mercury species in aqueous solution and living cells

A highly sensitive and selective fluorescent probe for inorganic and organic mercury species displays colorimetric and ratiometric response in a buffer solution via mercury promoted cleavage reaction. The probe is demonstrated to detect CH(3)HgCl in living cells.

Determination of Organic and Inorganic Mercury Species as Emetine Dithiocarbamate Complexes by High-Performance Liquid Chromatography with Electrogenerated Tris(2,2′-bipyridine)ruthenium(III) Chemiluminescence Detection

A sensitive method for simultaneous determination of organic and inorganic mercury species has been developed and is presented in this study. The method is based on complex formation of mercury species with the emetine dithiocarbamate (emetine-CS2) ligand, HPLC separation, and tris(2,2′-bipyridine)ruthenium(III) chemiluminescence detection. The complexation reactions of the mercury species and emetine-CS2 ligand occurred instantaneously upon the addition of emetine-CS2 solution to the solution containing the mercury species. The complete separation of these complexes was achieved using an ODS column with 20 mM NaH2PO4-acetonitrile (52:48, v/v) containing 30 mM NaClO4 as an ion-pair reagent. The calibration graphs of these complexes were linear in the range from 1–100 µg/L. The detection limits were 0.27 µg/L, 0.33 µg/L, 0.39 µg/L, and 0.17 µg/L for methylmercury, ethylmercury, phenylmercury, and the mercury ion, respectively, at a signal-to-noise ratio of 3. The developed technique was validated by analyzing certified reference materials, CRM7402-a (cod fish, NMIJ) and CE464 (tuna fish, ERM), in combination with sonication-assisted acid leaching and liquid-liquid extraction. The emetine-CS2 ligand has been used for extraction, separation, and detection of mercury species. The results determined using the proposed method were in good agreement with the values of the certified reference materials. The MeHg+ and EtHg+ recoveries for the spiked samples were found to be almost 100%.

Chemical Form Matters: Differential Accumulation of Mercury Following Inorganic and Organic Mercury Exposures in Zebrafish Larvae

Mercury, one of the most toxic elements, exists in various chemical forms each with different toxicities and health implications. Some methylated mercury forms, one of which exists in fish and other seafood products, pose a potential threat, especially during embryonic and early postnatal development. Despite global concerns, little is known about the mechanisms underlying transport and toxicity of different mercury species. To investigate the impact of different mercury chemical forms on vertebrate development, we have successfully combined the zebrafish, a well-established developmental biology model system, with synchrotron-based X-ray fluorescence imaging. Our work revealed substantial differences in tissue-specific accumulation patterns of mercury in zebrafish larvae exposed to four different mercury formulations in water. Methylmercury species not only resulted in overall higher mercury burdens but also targeted different cells and tissues than their inorganic counterparts, thus revealing a significant role of speciation in cellular and molecular targeting and mercury sequestration. For methylmercury species, the highest mercury concentrations were in the eye lens epithelial cells, independent of the formulation ligand (chloride versusl-cysteine). For inorganic mercury species, in absence of l-cysteine, the olfactory epithelium and kidney accumulated the greatest amounts of mercury. However, with l-cysteine present in the treatment solution, mercuric bis-l-cysteineate species dominated the treatment, significantly decreasing uptake. Our results clearly demonstrate that the common differentiation between organic and inorganic mercury is not sufficient to determine the toxicity of various mercury species.

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.

Luminescent bacteria-based sensing method for methylmercury specific determination

A bacterial biosensor method for the selective determination of a bioavailable organomercurial compound, methylmercury, is presented. A recombinant luminescent whole-cell bacterial strain responding to total mercury content in samples was used. The bacterial cells were freeze-dried and used as robust, reagent-like compounds, without batch-to-batch variations. In this bacteria-based sensing method, luciferase is used as a reporter, which requires no substrate additions, therefore allowing homogenous, real-time monitoring of the reporter gene expression. A noninducible, constitutively light-producing control bacterial strain was included in parallel for determining the overall cytotoxicity of the samples. The specificity of the total mercury sensor Escherichia coli MC1061 (pmerRBlux) bacterial resistance system toward methylmercury is due to a coexpressed specific enzyme, organomercurial lyase. This enzyme mediates the cleavage of the carbon-mercury bond of methylmercury to yield mercury ions, which induce the reporter genes and produce a self-luminescent cell. The selective analysis of methylmercury with the total mercury strain is achieved by specifically chelating the inorganic mercury species from the sample using an optimized concentration of EDTA as a chelating agent. After the treatment with the chelating agent, a cross-reactivity of 0.2% with ionic mercury was observed at nonphysiological ionic mercury concentrations (100nM). The assay was optimized to be performed in 3h but results can already be read after 1h incubation. Total mercury strain E. coli MC1061 (pmerRBlux) has been shown to be highly sensitive and capable of determining methylmercury at a subnanomolar level in optimized assay conditions with a very high dynamic range of two decades. The limit of detection of 75ng/l (300 pM) allows measurement of methylmercury even from natural samples.

Measurements of methylmercury, ethylmercury, and inorganic mercury species in a whole blood standard reference material: SRM 955c—Toxic Elements in Caprine Blood

Trace elemental speciation analysis is assuming more critical importance in nutritional and toxicological assessments, and despite the necessity to quantitatively differentiate elemental species in environmental and clinical samples, there are a limited number of reference materials suitable for this purpose. A whole blood certified reference material, SRM 955c Toxic Elements in Caprine Blood was developed for the determination of toxic metals in whole blood including mercury. The certification of methylmercury, ethylmercury, and inorganic mercury was carried out by triple spike speciated isotope dilution inductively coupled plasma mass spectrometry (TS-SID-GC/ICP-MS) and represents the first whole blood material certified for three mercury species. The certified values (±expanded uncertainty) of methylmercury, ethylmercury, inorganic mercury and total mercury in SRM 955c are 4.5 ± 1.0 µg L−1, 5.1 ± 0.5 µg L−1, 9.0 ± 1.3 µg L−1, and 17.8 ± 1.6 µg L−1 respectively.

Differential immunotoxic effects of inorganic and organic mercury species in vitro

Despite the fact that humans are exposed to multiple forms of mercury (elemental, inorganic, and organic), most research on mercury toxicity has focused on methylmercury (MeHg) and on neurotoxic outcomes and mechanisms. Recent work has indicated that the immunotoxic effects of mercury compounds may be significant contributors to human disease as well as mechanistically relevant to other target organ toxicities. In this study, we compared the effects of inorganic Hg (iHg) to organic Hg species (MeHg and ethylmercury, EtHg) in human peripheral blood mononuclear cells (PBMCs) in vitro at sub-cytotoxic concentrations, using methods developed to characterize response of human PBMCs to iHg in vitro. PBMCs were isolated from six volunteer blood donors (three males and three females) and cultured in the presence and absence of lipopolysaccharide (LPS) and low levels (up to 200 nM of each Hg species, separately) for 24 h in culture. Cell culture supernatants were analyzed for cytokine concentrations with a bead-based multiplex assay. We report that iHg and MeHg both increase pro-inflammatory cytokine release in LPS-stimulated PBMCs, while EtHg decreases IFN-γ release as well pro-inflammatory cytokine release. IL-17 release is significantly increased only in response to iHg treatment. Levels of anti-inflammatory cytokines (IL-1Ra and IL-10) were not significantly altered by any Hg treatment. These results indicate that both organic and inorganic species of Hg can affect the human immune system, but that they may exert different effects on immune function.

A ‘turn-on’ fluorescent probe that selectively responds to inorganic mercury species

A fluorescent probe selectively senses inorganic mercury in the turn-on mode through a mercury ion-promoted hydrolysis reaction that leads to a coumarin, among various other metal species except Au(III).