Determination of mercury and methylmercury in hair samples by neutron activation

Mercury and methylmercury in hair samples were determined by neutron activation analysis. Samples were digested in 10M NaOH, and methylmercury was then isolated by solvent extraction with toluene. The isolated methylmercury was then absorbed onto cysteine paper. The dried cysteine paper was activated for six hours in a TRIGA reactor and methylmercury was analysed via 279.2 keV of²⁰³Hg. Methylmercury and total mercury in some standard reference materials were also analysed, and the results were in good agreement with those reported in the literature. Results for hair samples showed that the methylmercury concentration ranged 14–40% of the total mercury. Gas chromatogram showed that methylmercury was only present in the samples analysed. In samples where methylmercury and other organic mercury are presented, the NAA method is good for the determination of the total organic mercury only.     

Hollow fiber liquid phase microextraction combined with graphite furnace atomic absorption spectrometry for the determination of methylmercury in human hair and sludge samples

Two methods, based on hollow fiber liquid–liquid–liquid (three phase) microextraction (HF-LLLME) and hollow fiber liquid phase (two phase) microextraction (HF-LPME), have been developed and critically compared for the determination of methylmercury content in human hair and sludge by graphite furnace atomic absorption spectrometry (GFAAS). In HF-LPME, methylmercury was extracted into the organic phase (toluene) prior to its determination by GFAAS, while inorganic mercury remained as a free species in the sample solution. In HF-LLLME, methylmercury was first extracted into the organic phase (toluene) and then into the acceptor phase (4% thiourea in 1 mol L^− 1 HCl) prior to its determination by GFAAS, while inorganic mercury remained in the sample solution. The total mercury was determined by inductively coupled plasma-mass spectrometry (ICP-MS), and the levels of inorganic mercury in both HF-LLLME and HF-LPME were obtained by subtracting methylmercury from total mercury. The factors affecting the microextraction of methylmercury, including organic solvent, extraction time, stirring rate and ionic strength, were investigated and the optimal extraction conditions were established for both HF-LLLPME and HF-LPME. With a consumption of 3.0 mL of the sample solution, the enrichment factors were 204 and 55 for HF-LLLPME and HF-LPME, respectively. The limits of detection (LODs) for methylmercury were 0.1 μg L^− 1 and 0.4 μg L^− 1 (as Hg) with precisions (RSDs (%), c = 5 μg L^− 1 (as Hg), n = 5) of 13% and 11% for HF-LLLPME–GFAAS and HF-LPME–GFAAS, respectively. For ICP-MS determination of total mercury, a limit of detection of 39 ng L^− 1 was obtained. Finally, HF-LLLME–GFAAS was applied to the determination of methylmercury content in human hair and sludge, and the recoveries for the spiked samples were in the range of 99–113%. In order to validate the method, HF-LLLME–GFAAS was also applied to the analysis of a certified reference material of NRCC DORM-2 dogfish muscle, and the determined values were in good agreement with the certified values.     

气相色谱仪和测汞仪联机测定人发中的甲基汞

本文在前报^[1]工作基础上,将气相色谱仪和测汞仪联机测定有机汞的方法,用于人发中甲基汞的测定。根据Birke等人报导^[2],人发和血液中只查到甲基汞,未查到乙基汞和更高级的烷基汞;同时本工作所用的人发试样,经白求恩医科大学环境医学研究室分析,除甲基汞外未查到乙基汞。因此本法测得的总有机汞能代表甲基汞。目前尚未见到类似方法的报导。     

Determination of total and methyl mercury in human permanent healthy teeth by electrothermal atomic absorption spectrometry after extraction in organic phase

A simple and sensitive method has been developed for determination of inorganic and methyl mercury in biological samples by ETAAS. For determination of methyl mercury; it was transferred to toluene phase by acid leaching extraction method. For total mercury after digestion of samples; it was extracted to toluene phase by means of the chelating agent diethyldithiocarbamate. Formation of complex between MeHg and diethyldithiocarbamate enhance the MeHg signal and increases the reproducibility. Furthermore, Pd-DDC was used as modifier for both mercury and methyl mercury determinations. The optimization performance was independently carried out by modifying the parameters such as temperature of mineralization, atomization and gas flow rate for methylmercury and inorganic mercury in ETAAS. The limits of detection were 0.15 and 0.12 μg g⁻¹ for methyl mercury and total mercury, respectively. The repeatability of the measurements of whole procedure were 15.8% for methyl mercury and 16.9% for total mercury determination. The accuracy of the method has been investigated by means of spiking different amounts of methylmercury and inorganic mercury to the samples. The recoveries were found within the range of 88-95% for methyl mercury and 85-92% for total mercury. For determination of total mercury, the method was validated by CVAAS. The obtained results by the present procedure were in good agreement with those of the CVAAS. The proposed method was applied for 30 human permanent healthy teeth (without filling) which significant positive correlations were found among number of amalgam filling and total mercury and MeHg.     

Production of hair intercomparison materials for use in population monitoring programmes for mercury and methylmercury exposure

Ten kilograms of hair obtained from India were used as the basis for two intercomparison materials, one with natural low levels of mercury and methylmercury, and one with an elevated level of methylmercury. The latter was produced by labeling the hair with a solution containing methylmercury. To convert the hair into homogeneous powders, cryogenic milling was utilized. 70% of the final material passed through a 0.075 mm sieve. Subsequent studies were carried out to establish the homogeneity of the materials and the stability of the methylmercury label. The materials will be distributed in an international intercomparison, the results from which will be used to obtain recommended values for total mercury and methylmercury.     

Speciation of Mercury in Microalgae by Isotope Dilution-inductively Coupled Plasma Mass Spectrometry

Species-specific stable isotope dilution in combination with gold trap- or gas chromatography (GC)-inductively coupled plasma mass spectrometry (ICP-MS) is reported for the determination of inorganic mercury and methylmercury in diatoms (Chaetoceros curvisetus). The optimum conditions for the separation parameters were established. The isotope dilution analysis was performed using ¹⁹⁹Hg-enriched Hg²⁺ and laboratory-synthesized ²⁰¹Hg-enriched methylmercury. The absolute detection limits obtained with isotope dilution-ICP-MS were 9 pg for total mercury and 0.6 pg for methylmercury. The relative error of 7 Hg isotopic abundances based on the peak area measurements was better than 2.0% for 20 pg of methylmercury (as Hg) and 250 pg of inorganic mercury. The accuracy of the method was validated with a biological certified reference material. The developed method was then applied to investigate the uptake of inorganic mercury and methylmercury by C. curvisetus. Continuous uptake of inorganic mercury and methylmercury was observed during 5 days of incubation.     

Determination of total mercury and methylmercury in hair samples from residents of Kuala Lumpur, Malaysia by neutron activation analysis

to estimate the level of total mercury and methylmercury in Kuala Lumpur residents, 400 hair samples were analysed by neutron activation analysis. Separation of methylmercury from hair samples were carried out prior to neutron activation. The average level of total mercury and methylmercury in hair samples were 3.38 mg^.kg^-1 (in range of 0.59-18.73 mg^.kg^-1) and 1.13 mg^.kg^-1 (in range of 0-4.65 mg^.kg^-1), respectively. The average percentage ratio of methylmercury to total mercury was 31.15% (in range of 0 to 75.81%). This revised version was published online in August 2006 with corrections to the Cover Date.     

Certification of two human hair reference materials issued by the International Atomic Energy Agency

Two new human hair reference materials, with different levels of mercury and methylmercury, have been developed and characterized by the International Atomic Energy Agency, for use in validation of measurements for mercury exposure. The set of materials consists of IAEA-086, with a low level of methylmercury, and IAEA-085, with an elevated methylmercury level. An international intercomparison exercise was carried out, and 68 institutes from 40 countries have contributed data. Based on the evaluation of the results from the intercomparison and analyses by expert laboratories, values of 23.2 and 0.57 mg/kg total mercury are recommended for IAEA-085 and IAEA-086, respectively. Values for methylmercury are recommended at 22.9 mg/kg, MeHg as Hg, for IAEA-085, and at 0.26 mg/kg, MeHg as Hg, for IAEA-086. Recommended and information values are also given for other selected trace elements.     

Determination of methylmercury in human hair by ethylation followed by headspace solid-phase microextraction-gas chromatography-cold-vapour atomic fluorescence spectrometry

An analytical procedure for the determination of methylmercury in human hair after acid digestion using aqueous ethylation, headspace solid-phase microextraction sampling and final gas chromatography-cold-vapour atomic fluorescence spectrometry detection is described. Acid digestion, extraction procedure and chromatographic conditions were optimised. An optimal linear range using standard mercury solutions was found and concentration detection limits for the mercury species, MeHg and Hg2+, were about 50 and 80 ng/g, respectively, for 100 mg of human hair. The reproducibility of the developed analytical procedure assessed for hair samples with incurred MeHg was better than 18% (n=5). A certified reference material from the National Institute of Environmental Studies (Japan) was used for validation. Analysis of human hair collected from urban inhabitants was performed and the mean value of methylmercury content in hair samples was 0.764 +/- 0.732 microg/g for the population tested. The developed analytical method is simple, fast and a suitable procedure for the monitoring and screening of human exposure to methylmercury.     

Evaluation of various sample pre-treatment methods for total and inorganic mercury determination in biological certified reference materials by CVAAS technique

Sample preparation methods for non-separation cold vapor atomic absorption spectrometry (CVAAS) sequential inorganic mercury speciation in biological certified reference materials (CRMs) were investigated. The methylmercury concentration was calculated as the difference between total and inorganic mercury. Microwave-assisted decomposition method, and three ultrasonic extraction procedures based on acid leaching with HCl and HCOOH and solubilization with TMAH were employed as sample preparation methods. The replacement of a sample decomposition procedure by extraction prior to analysis by CVAAS, as well as the aspect of speciation analysis is discussed. The limits of detection in the sample were determined as 50 and 10 ng L⁻¹ for inorganic and total mercury, which corresponds to absolute detection limits of 40 and 8 ng g⁻¹ for inorganic and total mercury, respectively. The results were in good agreement with the 95% confidence level t-test of the certified values for total and inorganic mercury in the reference materials investigated. From the analysis of the CRMs, it was evident that the difference between the total and inorganic mercury concentrations agrees with the methylmercury concentration. The relative standard deviation was better than 11% for most of the samples.      

Simple mercury fractionation in biological samples by CV AAS following microwave-assisted acid digestion or TMAH pre-treatment

A simple and reliable method to determine total and inorganic mercury in biological certified reference material (CRM) by cold vapor atomic absorption spectrometry (CV AAS) is proposed. After the CRM treatment at room temperature with tetramethylammonium hydroxide (TMAH), inorganic mercury is determined by CV AAS. Total mercury is measured by the same technique, after sample acid digestion in a microwave oven. Organic mercury, basically methylmercury, is obtained by difference. In both procedures, the quartz tube is kept at room temperature. By means of analysis of the following reference materials: pig kidney, lobster hepatopancreas, dogfish liver and mussel tissue, it was clear that the difference between the total and inorganic mercury concentrations agrees with the methylmercury concentration. Only one calibration curve against aqueous standards in acidic medium was carried out for both procedures. The concentrations obtained by both procedures are in agreement with the certified values according to the t-test at a 95% confidence level. The relative standard deviations were lower than 3.0% for digested CRM and 6.0% for CRM treated with TMAH for most of the samples. The limits of detection in the samples were 0.02 µg g^− 1 and 0.04 µg g^− 1 for inorganic and total Hg, respectively, since the sample mass for total mercury was half of that for inorganic mercury determination. Simplicity and high efficiency without using chromatographic techniques are some of the qualities of the proposed method, being adequate for fractionation analysis of mercury in biological samples.     

Simple mercury fractionation in biological samples by CV AAS following microwave-assisted acid digestion or TMAH pre-treatment

A simple and reliable method to determine total and inorganic mercury in biological certified reference material (CRM) by cold vapor atomic absorption spectrometry (CV AAS) is proposed. After the CRM treatment at room temperature with tetramethylammonium hydroxide (TMAH), inorganic mercury is determined by CV AAS. Total mercury is measured by the same technique, after sample acid digestion in a microwave oven. Organic mercury, basically methylmercury, is obtained by difference. In both procedures, the quartz tube is kept at room temperature. By means of analysis of the following reference materials: pig kidney, lobster hepatopancreas, dogfish liver and mussel tissue, it was clear that the difference between the total and inorganic mercury concentrations agrees with the methylmercury concentration. Only one calibration curve against aqueous standards in acidic medium was carried out for both procedures. The concentrations obtained by both procedures are in agreement with the certified values according to the t-test at a 95% confidence level. The relative standard deviations were lower than 3.0% for digested CRM and 6.0% for CRM treated with TMAH for most of the samples. The limits of detection in the samples were 0.02 µg g^{− 1} and 0.04 µg g^{− 1} for inorganic and total Hg, respectively, since the sample mass for total mercury was half of that for inorganic mercury determination. Simplicity and high efficiency without using chromatographic techniques are some of the qualities of the proposed method, being adequate for fractionation analysis of mercury in biological 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 (100 nM). The assay was optimized to be performed in 3 h but results can already be read after 1 h 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 75 ng/l (300 pM) allows measurement of methylmercury even from natural samples.     

Determination of total mercury and methylmercury in human hair by graphite-furnace atomic absorption spectrophotometry using 2,3-dimercaptopropane-1-sulfonate as a complexing agent.

For the determination of total mercury in hair, an amount (25.0 mg) of hair sample was digested with conc. HNO3 (400 microl) at 90 degrees C for 10 min in a 7-ml teflon microreaction vessel. After digestion, the pH of the acidic hair mixture was adjusted to 5.0-6.0 by NaOH and was then passed through a clean-up Sep-Pak C18 cartridge. To the eluate, 2,3-dimercaptopropane-1-sulfonate (DMPS) and sodium acetate buffer (pH = 6.0) were added to form a mercury-DMPS complex. This complex was preconcentrated on two Sep-Pak C18 cartridges in series, and each cartridge was eluted with methanol and adjusted to 2.00 ml. A portion (50 microl) was introduced into a graphite cuvette and then atomized according to a temperature program. The method detection limit (MDL, 3sigma) was 0.064 (microg g(-1)); the calibration graph was linear up to 7.52 microg g(-1). Good accuracies were obtained when testing two human hair certified reference materials (GBW 09101 and BCR-397). Six real samples were analyzed, and the recoveries were 95.8 - 98.2% with a relative standard deviation (RSD, n = 3) < 2.1%. For the determination of methylmercury (CH3Hg+), 25.0 mg of hair sample was extracted with 2.0 mol dm(-3) HCl (1.0 ml) by ultrasonicating for 1 h. The supernatant solution was used for CH3Hg+ analysis and the hair residue was used for the analysis of inorganic mercury (Hg2+). The MDL of CH3Hg+ was 0.068 microg g(-1); the calibration graph was linear up to 6.00 microg g(-1). Six real samples were analyzed, and the recoveries were 96.0-99.2% with RSD (n = 3) < 2.3%. The sum of the concentrations of CH3Hg+ and Hg2+ was very close to that of the total mercury measured with a relative error within 3.6%. The proposed method can be accurately applied to the measurement of CH3Hg+, Hg2+, and total mercury in hair samples.     

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).     

Determination of total and inorganic mercury in biological and environmental samples with on-line oxidation coupled to flow injection-cold vapor atomic absorption spectrometry

A method for determination of inorganic and total mercury by flow injection-cold vapor atomic absorption spectrometry (FI-CVAAS) with on-line oxidation was developed. Potassium peroxodisulphate and sulphuric acid were used as oxidizing agents so that decomposition of organomercury compounds could be achieved. Depending on the temperature selected, inorganic or total mercury could be determined with the same FI manifold. In order to assess the method performance, synthetic wastewater, wastewater, urine and saline water samples were spiked with inorganic mercury, methylmercury and phenylmercury. Quantitative recoveries were obtained for the three mercury species, except with the synthetic wastewater when the chemical oxygen demand value was higher than 1000 mg l⁻¹. In most cases, the standard addition method was usually needed for calibration. LODs calculated as 3 σ/m were 0.47 μg l⁻¹ for inorganic mercury and 0.45 μg l⁻¹ for total mercury. R.S.D. values corresponding to peak height measurements were 1.5 and 2.2% for inorganic mercury and total mercury, respectively. The accuracy of the method was tested by analyzing 5 mol l⁻¹ hydrochloric acid extracts of seven biological and environmental CRMs. LODs in the solid CRMs ranged from 0.032 to 0.074 μg g⁻¹.     

Preliminary study of the effects of some physical parameters on the stability of methylmercury in biological samples.

The effects of storage conditions (long-term storage of wet samples in a deep-freeze or thermal cycling), freeze-drying and gamma-irradiation at 1 and 5 Mrad on the stability of methylmercury in some biological samples were investigated. Methylmercury was determined by volatilization separation followed by gas chromatography and by ion-exchange separation of inorganic and organic species followed by measurement by cold vapour atomic absorption spectrometry (CVAAS). Total mercury was determined by CVAAS. Biological samples studied included fish and shellfish tissues, human hair and blood samples and appropriate reference materials. From the preliminary results obtained it can be concluded that fresh and dried fish muscle and fish certified reference materials show good stability with time and against temperature cycling. Shellfish and blood should not be repeatedly frozen and unfrozen otherwise possible losses of methylmercury can occur. Losses of methylmercury of up to 30% from wet mussels occurred on prolonged storage in a deep-freeze. Gamma-irradiation reduced the methylmercury content of the fish and shellfish only for hake (Merluccius merluccius). Further experiments should be carried out to confirm this and to investigate if this effect is species dependent. Apparent losses of methylmercury on freeze-drying of blood need to be reconfirmed on further samples.     

A new pyrrolidinedithiocarbamate screening method for the determination of methylmercury and inorganic mercury relation in hair samples by HPLC-UV-PCO-CVAAS

A new analytical screening technique for the determination of methylmercury and inorganic mercury in hair samples by HPLC-PCO-CVAAS has been developed. It is based on the extraction of mercury compounds by a buffered sodium pyrrolidinedithiocarbamate solution, separation by reversed-phase HPLC, post column oxidation by UV-irradiation, reduction with alkaline sodium borohydride, and determination by cold vapour atomic absorption detection. The standard deviation was 7% and recoveries were 90% for both compounds. The limit of detection (S/N = 3) for both compounds was calculated to be about 4 ppb.     

A new pyrrolidinedithiocarbamate screening method for the determination of methylmercury and inorganic mercury relation in hair samples by HPLC-UV-PCO-CVAAS

A new analytical screening technique for the determination of methylmercury and inorganic mercury in hair samples by HPLC-PCO-CVAAS has been developed. It is based on the extraction of mercury compounds by a buffered sodium pyrrolidinedithiocarbamate solution, separation by reversed-phase HPLC, post column oxidation by UV-irradiation, reduction with alkaline sodium borohydride, and determination by cold vapour atomic absorption detection. The standard deviation was 7% and recoveries were 90% for both compounds. The limit of detection (S/N = 3) for both compounds was calculated to be about 4 ppb.     

Inter‐laboratory validation of EPA method 3200 for mercury speciation analysis using prepared soil reference materials

Determinations of the concentration of individual mercury species from environmental samples have increased significantly over the past decade. The techniques used for the determination of mercury species in soils or sediments generally involve a series of analytical steps (extraction, separation, detection) that may all be prone to systematic errors. An inter‐laboratory validation study of the EPA draft method 3200 was conducted under the auspices of the United States Environmental Protection Agency on two specifically prepared soil matrices. The study was performed successfully by a limited number of participating laboratories. Evaluation of the data demonstrates that the method is more highly efficient for extracting the highly toxic methylmercury than inorganic mercury. The proposed method does not induce transformation of methylmercury to inorganic mercury. Copyright © 2004 John Wiley & Sons, Ltd.