Determination,speciation and distribution of mercury in soil in the surroundings of a former chlor-alkali plant: assessment of sequential extraction procedure and analytical technique

Background

The paper presents the evaluation of soil contamination with total, water-available, mobile, semi-mobile and non-mobile Hg fractions in the surroundings of a former chlor-alkali plant in connection with several chemical soil characteristics. Principal Component Analysis and Cluster Analysis were used to evaluate the chemical composition variability of soil and factors influencing the fate of Hg in such areas. The sequential extraction EPA 3200-Method and the determination technique based on capacitively coupled microplasma optical emission spectrometry were checked.

Results

A case study was conducted in the Turda town, Romania. The results revealed a high contamination with Hg in the area of the former chlor-alkali plant and waste landfills, where soils were categorized as hazardous waste. The weight of the Hg fractions decreased in the order semi-mobile?>?non-mobile?>?mobile?>?water leachable. Principal Component Analysis revealed 7 factors describing chemical composition variability of soil, of which 3 attributed to Hg species. Total Hg, semi-mobile, non-mobile and mobile fractions were observed to have a strong influence, while the water leachable fraction a weak influence. The two-dimensional plot of PCs highlighted 3 groups of sites according to the Hg contamination factor. The statistical approach has shown that the Hg fate in soil is dependent on pH, content of organic matter, Ca, Fe, Mn, Cu and SO₄^2- rather than natural components, such as aluminosilicates. Cluster analysis of soil characteristics revealed 3 clusters, one of which including Hg species. Soil contamination with Cu as sulfate and Zn as nitrate was also observed.

Conclusions

The approach based on speciation and statistical interpretation of data developed in this study could be useful in the investigation of other chlor-alkali contaminated areas. According to the Bland and Altman test the 3-step sequential extraction scheme is suitable for Hg speciation in soil, while the used determination method of Hg is appropriate.

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Sequential extraction and thermal desorption of mercury from contaminated soil and tailings from Mongolia

Mercury forms in contaminated environmental samples were studied by means of sequential extraction and thermal desorption from the solid phase. The sequential extraction procedure involved the following fractions: water soluble mercury, mercury extracted in acidic conditions, mercury bound to humic substances, elemental Hg and mercury bound to complexes, HgS, and residual mercury. In addition to sequential extraction, the distribution of mercury species as a function of soil particles size was studied. The thermal desorption method is based on the thermal decomposition or desorption of Hg compounds at different temperatures. The following four species were observed: Hg⁰, HgCl₂, HgS and Hg^(II) bound to humic acids. The Hg release curves from artificial soils and real samples were obtained and their applicability to the speciation analysis was considered.      

Some aspects of speciation and reactivity of mercury in various matrices

Speciation of mercury compounds in environmental and biological samples requires different techniques and different approaches. This speciation is mandatory to explain the toxicity, the reactivity and the bioavailability of mercury. It is dominated by inorganic mercury species Hg(II) and Hg(0), and the organic mercury species CH₃Hg and (CH₃)₂Hg. In this paper, some aspects of mercury speciation are presented in terms of:- mercury reactivity (Hg(II) complexation and reduction),- mercury species distribution in the main compartments of the environment     

Applicability of multisyringe chromatography coupled to cold-vapor atomic fluorescence spectrometry for mercury speciation analysis

In this paper, a novel automatic approach for the speciation of inorganic mercury (Hg(2+)), methylmercury (MeHg(+)) and ethylmercury (EtHg(+)) using multisyringe chromatography (MSC) coupled to cold-vapor atomic fluorescence spectrometry (CV/AFS) was developed. For the first time, the separation of mercury species was accomplished on a RP C18 monolithic column using a multi-isocratic elution program. The elution protocol involved the use of 0.005% 2-mercapthoethanol in 240 mM ammonium acetate (pH 6)-acetonitrile (99:1, v/v), followed by 0.005% 2-mercapthoethanol in 240 mM ammonium acetate (pH 6)-acetonitrile (90:10, v/v). The eluted mercury species were then oxidized under post-column UV radiation and reduced using tin(II) chloride in an acidic medium. Subsequently, the generated mercury metal were separated from the reaction mixture and further atomized in the flame atomizer and detected by AFS. Under the optimized experimental conditions, the limits of detection (3σ) were found to be 0.03, 0.11 and 0.09 μg L(-1) for MeHg(+), Hg(2+) and EtHg(+), respectively. The relative standard deviation (RSD, n=6) of the peak height for 3, 6 and 3 μg L(-1) of MeHg(+), Hg(2+) and EtHg(+) (as Hg) ranged from 2.4 to 4.0%. Compared with the conventional HPLC-CV/AFS hyphenated systems, the proposed MSC-CV/AFS system permitted a higher sampling frequency and low instrumental and operational costs. The developed method was validated by the determination of a certified reference material DORM-2 (dogfish muscle), and was further applied for the determination of mercury species environmental and biological samples.     

Determination of mercury species using thermal desorption analysis in AAS

Analytical 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.     

Mercury determination and speciation analysis in surface waters

A sorbent L-cysteine grafted silica gel has been evaluated for separation and enrichment of dissolved inorganic i-Hg(II) and methylmercury CH₃Hg(I) from surface waters at sub-μg L⁻¹ concentrations. Chemical parameters for mercury species enrichment and separation have been optimized. Analytical schemes for the determination of Hg species, using selective column solid phase extraction (SPE) with continuous flow chemical vapor generation atomic absorption spectrometry (CF-CVG-AAS) or inductively coupled plasma-mass spectrometry (ICP-MS) were developed. Possibilities for on-site SPE enrichment were demonstrated as well. The limits of quantification were 1.5 and 5 ng L⁻¹ for dissolved i-Hg(II) and CH₃Hg(I) by CF-CVG-AAS and 1 and 2.5 ng L⁻¹ by ICP-MS with relative standard deviations between 7–12% and 7–14%, respectively. The chemically modified SPE sorbent has demonstrated high regeneration ability, chemical and mechanical stability, acceptable capacity and good enrichment factors. Results for total dissolved mercury were in reasonable agreement with those from independent analyses by direct ICP-MS determinations for river waters and for estuarine water certified reference material.      

Determination of mercury in polluted soils surrounding a chlor-alkali plant: Direct speciation by X-ray absorption spectroscopy techniques and preliminary geochemical characterisation of the area

Soil samples collected in the surroundings of a chlor-alkali plant in the Netherlands were characterised by synchrotron-based techniques and conventional analytical procedures, in order to evaluate the environmental impact of Hg emissions and other heavy metals present in these locations. Analysis of total metal content by inductively coupled plasma-optical spectroscopy (ICP-OES) revealed a heterogeneous contamination of Hg, with concentrations ranging from 4.3 to 1150 μg g⁻¹. In addition, significant concentrations of Cu, Ni, Pb, Zn, Mn and principally Fe were also identified within the studied samples. Direct determination of mercury species by X-ray absorption near edge spectroscopy (XANES) showed inorganic Hg compounds to prevail in all soils, being Cinnabar (HgS_red) and Corderoite (Hg₃S₂Cl₂) the main species. Nevertheless, more soluble mercury compounds, such as HgO and HgSO₄, have been also identified in significant proportion (from 6 to 20% of total mercury content), indicating a potential risk of mercury mobilisation. On the other hand, the application of sequential extraction schemes (SES) revealed large portions of weakly available Hg extracted in the residual fraction, while Hg associated to the exchangeable phase amounts as much as 19% of total Hg, thus, supporting the results obtained by XANES.Finally, synchrotron-based micro X-ray fluorescence (μ-XRF) was applied to identify qualitative trends on elemental associations in sample particles through a systematic mapping of its surface. In this concern, results show a well-defined correlation between Hg and Cu/Ni in the analysed particles. On the other hand, an absence of correlation between Hg and several other elements (Fe, Ti, Ca, Zn, Mn and S) was also observed. These effects have been attributed to chemical and physical interactions of mercury species on both enriched particles and sample matrices.     

A methodological study of mercury speciation using dogfish liver CRM (DOLT-2)

The purpose of the study was to optimise analytical methods for determination of the chemical speciation of mercury in studies of protective mechanisms of selenium. Optimisation of the methods was performed using CRM DOLT-2 (Dogfish liver), both in its original form and after separation of various fractions. The sample was homogenised with 10 mM Tris-HCl buffer (pH 7.6) and ultracentrifuged. The soluble phase obtained was applied to a size exclusion chromatography column (Sephadex G-75 column) for separation of various protein fractions. Total mercury (total Hg), monomethyl mercury (MeHg) and selenium (Se) were determined in whole dogfish liver tissue and its soluble and insoluble phases (pellet). Different approaches for determination of total Hg and MeHg were compared. Simultaneous determination of MeHg and inorganic mercury (Hg2+) was based on alkaline dissolution and/or acid leaching, followed by ethylation, room temperature precollection, isothermal gas chromatography (GC), pyrolysis and detection with cold vapour atomic fluorescence spectrometry (CVAFS). The sum of MeHg and Hg2+ was compared to total Hg results obtained by acid digestion and CVAAS detection. The accuracy of MeHg determination was checked by its determination using acid leaching at room temperature, solvent extraction, back extraction into Milli-Q water, ethylation, GC and CVAFS detection. For the insoluble phase it is recommended to use solvent extraction for MeHg and acid digestion CVAAS for total Hg. For determination of MeHg and Hg2+ in the lyophilised sample and water soluble fractions containing low concentrations of mercury species, the simultaneous measurement of MeHg and Hg2+ after alkaline dissolution is the most appropriate method.     

Mercury characterisation in urban particulate matter

A five-step sequential extraction procedure was proposed in order to assess the distribution of mercury (Hg) forms in urban particulate matter (PM): exchangeable, HCl-soluble, organic-bound, elemental and other slightly soluble Hg species, mercury(II) sulphide (HgS), and residual Hg. This process was applied to the analysis of urban dust samples collected at locations in Prague (Czech Republic) with high traffic density. In addition to sequential extractions, thermal desorption analysis was performed. The differences in Hg concentrations between untreated and thermally treated samples were indicated as the thermally releasable amount of Hg. For the study of PM-adsorbing capacity, Hg vapours were passed through the samples as long as the enrichment of materials was observed. The retained elemental Hg was readily released by thermal desorption. All Hg analyses were based on the highly sensitive pyrolysis technique of atomic absorption spectrometry using the mercury analyser AMA-254.     

The behaviour of different organometallic compounds in the presence of inorganic mercury(II): transalkylation of mercury species and their analysis by the GC–MIP–PED system

It is shown that four different mercury species are formed by the abiotic reaction of inorganic mercury with different, ecologically relevant, organolead and organoarsenic compounds. Therefore solutions of tetraethyl-lead, trimethyl-lead chloride and dimethylarsonic acid were prepared and mixed with stock solutions of inorganic mercury at different concentrations. The final solutions were analyzed for their content of newly synthesized mercury species. The analysis was carried out by using a system of solid-phase micro-extraction (SPME): capillary gas chromatography (GC), microwave-induced plasma (MIP) and a plasma-emission detector (PED). We found that transfer of one or two alkyl groups to the inorganic mercury is possible under the conditions mentioned below. The transalkylation rate depends on the kind of organometallic compound and on the pH. The results were confirmed by the reaction with inorganic mercury and analysis of a soil sample, containing tetraethyl-lead and trimethyl-lead, in which not only the monoalkyl compound, but also the dialkylated compound of the relevant inorganic metal, were found. © 1997 John Wiley & Sons, Ltd.     

Mercury speciation in thawed out and refrozen fish samples by gas chromatography coupled to inductively coupled plasma mass spectrometry and atomic fluorescence spectroscopy

Different sub-sampling procedures were applied for the determination of mercury species (as total mercury Hg, methylmercury MeHg⁺ and inorganic mercury Hg²⁺) in frozen fish meat. Analyses were carried out by two different techniques. After the sample material was pre-treated by microwave digestion, atomic fluorescence spectroscopy (AFS) was used for the determination of total Hg. Speciation analysis was performed according to the following procedure: dissolution of sample material in tetramethylammonium hydroxide (TMAH), derivatisation with sodium tetraethylborate (NaBEt₄), extraction into isooctane and measurement with gas chromatography inductively coupled plasma mass spectrometry (GC-ICPMS) for the identification and quantification of methylmercury (MeHg⁺) and inorganic mercury (Hg²⁺). The concentration range of total Hg measured in the shark fillets is between 0.9 and 3.6 g g^–1 thawed out shark fillet. Speciation analysis leads to 94% Hg present as MeHg⁺. Homogeneity, storage conditions and stability of analytical species and sample materials have great influence on analytical results. Sub-sampling of half-frozen/partly thawed out fish and analysis lead to significantly different concentrations, which are on average a factor of two lower.     

Characterization of mercury species in soils by HPLC-ICP-MS and measurement of fraction removed by diffusive gradient in thin films

The properties and behaviour of Hg depend on both the oxidation state and the chemical form: the bioavailability, toxicity, persistence and accumulation of mercury in the food web are strongly influenced by chemical speciation. The present work aims to determine the chemical forms of mercury present in soil and to evaluate the fraction of mercury in soil solution available to plants. In order to do this, we analyzed eight samples of contaminated soils with Hg concentrations ranging from 1.31 to 21.7 mg kg⁻¹, collected from different depths (0–10 and 40–50 cm) close to an abandoned industrial site in Val Basento (southern Italy). Two innovative analytical techniques were used: HPLC–ICP-MS and diffusive gradient in thin films (DGT). The analytical procedure was validated using ERM 580-certified sediment and spiked samples in the case of HPLC–ICP-MS, and by a performance test in the case of DGT. In all samples, the only species found in soil and soil solution was MeHg⁺ and Hg²⁺. In soil, the MeHg⁺/Hg_tot ratio ranged from 0.05% to 0.82%; total mercury in soil solution was less than 0.01% of total mercury in soil. The percentage of MeHg⁺ in soil solution varied considerably (from 0% to 50%), with a maximum concentration of 0.02 mg L⁻¹. The root available concentration evaluated by DGT is comparable to the total mercury content of the soil solution measured by HPLC–ICP-MS. The DGT results suggest that all mercury in solution is available for uptake in DGT, and that mercury is supplied from soil to solution. However, for all samples the soluble and root available (DGT-labile) fractions of mercury are generally very low with respect to the total mercury concentration. This study confirmed that both HPLC–ICP-MS and DGT techniques are suitable tools for the estimation of Hg root availability and in assessing the risk from contaminated soils.     

Determination of the different states of mercury in seawater near the Vlora and Durres Bays

Because of the toxicity and mobility of organic mercury, there is a need for determination of organic and inorganic mercury at very low concentrations in the environment, especially in seawater. A procedure for extraction and determination of low concentrations of the different states of mercury - total, inorganic, and organic - in seawater, by cold vapour atomic absorption spectroscopy (CVAAS) is described.Inorganic mercury only was directly reduced to the Hg(0) state by tin(II) chloride under strongly acid conditions (H(2)SO(4)) and mercury metal (Hg(0)) was determined by cold vapour atomic absorption spectroscopy. Organic and inorganic mercury were extracted, with toluene, as the bromide derivatives and re-extracted, together, into ammonium chloride solution. Organic mercury was converted into inorganic mercury by thermal digestion at 80-90 degrees C in the presence of strong oxidants. These two states of mercury were determined together as total mercury. Inorganic mercury was measured directly after pre-concentration of the sample by toluene extraction. Toluene dissolved in aqueous phase after re-extraction of the sample was removed by heating for 30 min at 80-90 degrees C. Organic mercury was calculated as the difference between total and inorganic mercury.The sensitivity of the method is 0.0001 ng mL(-1) Hg, depending on sample volume.     

Preconcentration speciation method for mercury compounds in water samples using solid phase extraction followed by reversed phase high performance liquid chromatography

A simple and rapid method for in situ preconcentration of inorganic and organic mercury compounds in water samples, based on solid phase extraction using dithizone immobilised on a reversed-phase C18 cartridge, has been developed. The adsorbed complexes were stable on the cartridge for at least 2 weeks. The speciation analysis of methylmercury (MeHg), phenylmercury (PhHg) and inorganic mercury (Hg (II)) were done by reversed-phase high performance liquid chromatography. The calibration graphs of MeHg, PhHg and Hg (II) were linear (r>0.999) from the detection limits (0.58, 0.66 and 0.54 ng) to 38, 25 and 26 ng of Hg, respectively. The average recoveries of MeHg, PhHg and Hg (II) from spiked samples (0.3-48.0 mug l(-1) Hg) were 98+/-3, 99+/-1 and 100+/-7%, respectively. By applying SPE procedure a 200-fold concentration of the sample was obtained.     

A methodological study of mercury speciation using Dogfish liver CRM (DOLT-2)

The purpose of the study was to optimise analytical methods for determination of the chemical speciation of mercury in studies of protective mechanisms of selenium. Optimisation of the methods was performed using CRM DOLT-2 (Dogfish liver), both in its original form and after separation of various fractions. The sample was homogenised with 10 mM Tris-HCl buffer (pH 7.6) and ultracentrifuged. The soluble phase obtained was applied to a size exclusion chromatography column (Sephadex ¶G-75 column) for separation of various protein fractions. Total mercury (total Hg), monomethyl mercury (MeHg) and selenium (Se) were determined in whole dogfish liver tissue and its soluble and insoluble phases (pellet). Different approaches for determination of total Hg and MeHg were compared. Simultaneous determination of MeHg and inorganic mercury (Hg²⁺) was based on alkaline dissolution and/or acid leaching, followed by ethylation, room temperature precollection, isothermal gas chromatography (GC), pyrolysis and detection with cold vapour atomic fluorescence spectrometry (CVAFS). The sum of MeHg and Hg²⁺ was compared to total Hg results obtained by acid digestion and CVAAS detection. The accuracy of MeHg determination was checked by its determination using acid leaching at room temperature, solvent extraction, back extraction into Milli-Q water, ethylation, GC and CVAFS detection. For the insoluble phase it is recommended to use solvent extraction for MeHg and acid digestion CVAAS for total Hg. For determination of MeHg and Hg²⁺ in the lyophilised sample and water soluble fractions containing low concentrations of mercury species, the simultaneous measurement of MeHg and Hg²⁺ after alkaline dissolution is the most appropriate method.     

Determination of very low levels of dissolved mercury(II) and methylmercury in river waters by continuous flow with on-line UV decomposition and cold-vapor atomic fluorescence spectrometry after pre-concentration on a silica gel-2-mercaptobenzimidazol sorbent

A new, simple and sensitive method for the simultaneous determination of mercury(II) and methylmercury chloride at sub-ng l⁻¹ levels in river waters is described. Inorganic and organic mercury were preconcentrated from fresh water samples simultaneously on a laboratory-made column containing 2-mercaptobenzimidazol loaded on silica gel and then quantitatively eluted with 0.05 M KCN solution and 2.0 M HCl to desorp inorganic and methylmercury species, respectively. After irradiation with an intensive UV source, MeHg⁺ was decomposed and mercury vapours were generated from inorganic and organic mercury using an acidic SnCl₂ solution in a continuous flow system and were subsequently determined with a cold vapour atomic fluorescence (CV-AFS) spectrometer. Detection limits (3σ) were 0.07 and 0.05 ng l⁻¹ (as Hg) for mercury(II) chloride and methylmercury chloride, respectively. Relative standard deviations of method (%R.S.D.) were 8.8 and 10 for inorganic and organomercuric species in the river water, respectively. The analysis of real samples, taken from different rivers, showed that inorganic mercury levels ranged from 4.0±0.6 to 12±1 ng l⁻¹ (as Hg and 95% confidence limit) and methylmercury levels at 0.2±0.02 ng l⁻¹(as Hg).     

Mercury speciation in sea food by flow injection cold vapor atomic absorption spectrometry using selective solid phase extraction

An on-line inorganic and organomercury species separation, preconcentration and determination system consisting of cold vapor atomic absorption spectrometry (CV-AAS or CV-ETAAS) coupled to a flow injection (FI) method was studied. The inorganic mercury species was retained on a column (i.d., 3 mm; length 3 cm) packed to a height of 0.7 cm with a chelating resin aminopropyl-controlled pore glass (550 A) functionalized with [1,5-bis (2 pyridyl)-3-sulphophenyl methylene thiocarbonohydrazyde] placed in the injection valve of a simple flow manifold. Methylmercury is not directly determined. Previous oxidation of the organomercurial species permitted the determination of total mercury. The separation of mercury species was obtained by the selective retention of inorganic mercury on the chelating resin. The difference between total and inorganic mercury determined the organomercury content in the sample. The inorganic mercury was removed on-line from the microcolumn with 6% (m/v) thiourea. The mercury cold vapor generation was performed on-line with 0.2% (m/v) sodium tethrahydroborate and 0.05% (m/v) sodium hydroxide as reducing solution. The determination was performed using CV-AAS and CV-ETAAS, both approaches have been used and compared for the speciation of mercury in sea food. A detection limit of 10 and 6 ng l(-1) was achieved for CV-AAS and CV-ETAAS, respectively. The precision for 10 replicate determinations at the 1 microg l(-1) Hg level was 3.5% relative standard deviation (R.S.D.), calculated from the peak heights obtained. Both approaches were validated with the use of two certified reference materials and by spiking experiments. By analyzing the two biological certified materials, it was evident that the difference between the total mercury and inorganic mercury corresponds to methylmercury. The concentrations obtained by both techniques were in agreement with the certified values or with differences of the certified values for total Hg(2+) and CH(3)Hg(+), according to the t-test for a 95% confidence level. It is amazing how this very simple method is able to provide very important information on mercury speciation.     

Computational studies of the coordination stereochemistry, bonding, and metal selectivity of mercury

Knowledge of the bonding and selectivity of organic mercury, [H3C-Hg]+ (MeHg+), and inorganic Hg2+ for protein and DNA functional groups is important for understanding the mechanism of heavy metal poisoning. Herein, we elucidate (1) the differences between inorganic Hg2+ and organic MeHg+ in their interactions with different ligands of biological interest, (2) the protein and DNA functional groups that Hg2+ and MeHg+ target in aqueous solution, and (3) the likelihood of "soft" Hg2+ displacing the "borderline" Zn2+ bound to "harder" nitrogen/oxygen-containing side chains such as His and Asp/Glu. The results reveal that, relative to Hg2+, the lower positive charge on MeHg+ results in a longer and weaker bond with a given ligand, in accord with the observed kinetic lability of MeHg+ complexes. They also indicate that negatively charged or polar amino acid side chains containing S-/O-/S/N donors could coordinate to both organic MeHg+ and inorganic Hg2+. In addition, Gua and Cyt could also coordinate to MeHg+ and disrupt Gua...Cyt base pairing. A key novel finding is that Hg2+ is a far better electron acceptor than Zn2+, and can thus accept more negative charge from the Zn ligands than the native Zn2+, thus enhancing Hg-ligand interactions and enabling Hg2+ to displace the native cofactor from zinc essential enzymes and "structural" Zn proteins. The results herein support several possible mechanisms for Hg poisoning. Ways that mercury poisoning could be prevented in cells are discussed.     

UV irradiation controlled cold vapor generation using SnCl2 as reductant for mercury speciation.

A simple and ultrasensitive method, which was based on cold vapor generation (CVG) coupled to atomic fluorescence spectrometry (AFS), was proposed for speciation analysis of inorganic mercury (Hg2+) and methylmercury (MeHg) in water samples. In the presence of UV irradiation, all the mercury (MeHg+Hg2+) in a sample solution can be reduced to Hg0 by SnCl2; without UV irradiation, only Hg2+ species can be determined. So the concentration of MeHg can be obtained from the difference of the total mercury and Hg2+ concentration; thus, speciation analysis of Hg2+ and MeHg was simply achieved without chromatographic separation. Under the optimized experimental conditions, the limits of detection were 0.01 ng mL-1 for both Hg2+ and MeHg. The sensitivity and limit of detection were not dependent on the mercury species, and a simple Hg2+ aqueous standard series can be used for the determination of both Hg2+ and MeHg.