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.     

A new neutron activation technique for simultaneous determination of inorganic and total mercury contents in human hair

A simple method for simultaneous determination of inorganic and total mercury contents in human hair by neutron activation analysis (NAA) has been developed. The method is based on the selective extraction of methylmercury from hair by hydrochloric acid. Thus, the residual phase containing inorganic mercury can be determined by NAA. Further, the methylmercury contents in hair samples are easily calculated by subtracting the inorganic mercury contribution from the total Hg simultaneously given by INAA. Several reference materials of human hair, including IAEA hair RM 085 and 086, Chinese hair RMs GBW 09101 and 07601, were analyzed by this method. Our results show that the method is reliable.     

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.     

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.     

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.     

New human hair certified reference material for methylmercury and trace elements

A new human hair certified reference material (NIES CRM No. 13) for mercury speciation and trace element analysis was prepared at the National Institute for Environmental Studies (NIES), Environmental Agency of Japan. Scalp hair from Japanese males, which is identical with the original material for the previous human hair CRM (NIES CRM No. 5), was used. Special attention was paid to reduce contamination from a grinding vessel during the preparation procedure. A newly-prepared ceramic/Teflon disc mill was used for cryogenic grinding of the hair. 1, 000 bottles (3 g each) were produced after sieving and blending of the hair powder. Certified values for total mercury and methylmercury, as well as other trace elements of toxicological and nutritional significance (antimony, cadmium, copper, lead, selenium, and zinc), were determined based on analyses from extensive collaborations. Reference values for 12 elements (aluminium, arsenic, barium, calcium, cobalt, iron, magnesium, manganese, silver, sodium, sulfur and vanadium) were also given.     

On-line separation for the speciation of mercury in natural waters by flow injection-cold vapour-atomic absorption spectrometry

Inorganic mercury and methylmercury are determined in natural waters by injecting the filtered samples onto a low cost commercial flow injection system in which an anion exchange microcolumn is inserted after the injection loop (FIA-IE). If hydrochloric acid is used as the carrier solution, the HgCl4(2-) species (inorganic mercury) will be retained by the anion exchanger while the CH3HgCI species (methylmercury) will flow through the resin with negligible retention. Four anion exchangers and seven elution agents were checked, in a batch mode, to search for the best conditions for optimal separation and elution of both species. Dowex M-41 and L-cysteine were finally selected. Mercury detection was performed by cold vapour-electrothermal atomic adsorption spectrometry (HG-ETAAS). Both systems were coupled to perform the continuous on-line separation/detection of both inorganic mercury and methylmercury species. Separation and detection conditions were optimized by two chemometric approaches: full factorial design and central composite design. A limit of detection of 0.4 microg L(-1) was obtained for both mercury species (RSD < 3.0% for 20 microg L(-1) inorganic and methylmercury solutions). The method was applied to mercury speciation in natural waters of the Nerbioi-lbaizabal estuary (Bilbao, North of Spain) and recoveries of more than 95% were obtained.     

Mimicking receptor for methylmercury preconcentration based on ion-imprinting

Solid-phase extraction (SPE) based on molecularly imprinted polymers (MIPs) were used to develop selective separation and preconcentration for methylmercury ion from complex matrixes. In this study, an ion-imprinting polymer was prepared to make artificial organomercury lyase preorganizing three methacryloyl-(l)-cysteine methylester (MAC) monomers and one methylmercury ion in a three-coordinate form by template polymerization, with the goal preparing a solid-phase which has the high selectivity for methylmercury ions.Methylmercury-imprinted beads were produced by a dispersion polymerization technique with use methylmercury-methacryloyl-(l)-cysteine (MM-MAC) complex monomer and ethylene glycoldimethacrylate (EDMA). After removal of methylmercury ions, methylmercury-imprinted beads were used for solid-phase extraction and determination of mercury compounds. Methylmercury adsorption and selectivity studies of methylmercury versus other metal ions which Hg(II), Zn(II), Pb(II) and Cd(II) were reported and distribution and selectivity coefficients of these ions with respect to methylmercury were calculated here.ICP-OES and HPLC-DAD determinations of methylmercury and mercury ions in the certified reference, LUTS-1 from the National Research Council of Canada and synthetic sea water showed that the interfering matrix had been almost removed during preconcentration. The methylmercury-imprinted solid-phase as mimic receptor was good enough for methylmercury determination in complex matrixes.     

New human hair certified reference material for methylmercury and trace elements

 A new human hair certified reference material (NIES CRM No. 13) for mercury speciation and trace element analysis was prepared at the National Institute for Environmental Studies (NIES), Environmental Agency of Japan. Scalp hair from Japanese males, which is identical with the original material for the previous human hair CRM (NIES CRM No. 5), was used. Special attention was paid to reduce contamination from a grinding vessel during the preparation procedure. A newly-prepared ceramic/Teflon disc mill was used for cryogenic grinding of the hair. 1,000 bottles (3 g each) were produced after sieving and blending of the hair powder. Certified values for total mercury and methylmercury, as well as other trace elements of toxicological and nutritional significance (antimony, cadmium, copper, lead, selenium, and zinc), were determined based on analyses from extensive collaborations. Reference values for 12 elements (aluminium, arsenic, barium, calcium, cobalt, iron, magnesium, manganese, silver, sodium, sulfur and vanadium) were also given. Received: 8 February 1996/Accepted: 4 April 1996     

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.     

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 trace amounts of methylmercury in sediment and biological tissue by using water vapor distillation in combination with RP C18 preconcentration and HPLC-HPF/HHPN-ICP-MS

A novel technique has been developed for the determination of trace amounts of methylmercury in sediment and biological tissues. The well known water vapor distillation technique for the isolation of methylmercury from different matrices was coupled with an RP C18 preconcentration using dithiocarbamate complexation. A newly developed HPLC-method allowed the separation of five different mercury species at different mercury masses with HPF/HHPN (High-Performance-Flow/Hydraulic-High-Pressure-Nebulizing) and detection by ICP-MS. The method takes advantage of the ability to measure individual isotopes. Recoveries of the water vapor distillation procedure samples for different mercury compounds from sediment were tested. For methylmercury, the detection limit for a 0.5 g sample was calculated to be 0.025 μg/kg. The new technique was assured using different reference materials.     

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.     

Instrumental neutron activation analysis for quality assurance of a hair reference material for mercury speciation

Instrumental neutron activation analysis (INAA) has been employed in the investigation of mass balance for mercury species analysis in the analytical process. A new human hair reference material (IAEA-085) was analyzed for methylmercury using a solid/liquid extraction procedure, with samples of extracts, residues, and untreated samples being analyzed by INAA. The certified reference material NIES CRM No. 13, human hair, was analyzed in parallel. From the results obtained through the mass balance studies, it was found that the extraction procedure was quantitatively complete, and that there was no difference between the mass balance of Hg and the total Hg in the untreated materials.     

Determination of trace amounts of methylmercury in sediment and biological tissue by using water vapor distillation in combination with RP C18 preconcentration and HPLC-HPF/HHPN-ICP-MS

A novel technique has been developed for the determination of trace amounts of methylmercury in sediment and biological tissues. The well known water vapor distillation technique for the isolation of methylmercury from different matrices was coupled with an RP C18 preconcentration using dithiocarbamate complexation. A newly developed HPLC-method allowed the separation of five different mercury species at different mercury masses with HPF/HHPN (High-Performance-Flow/Hydraulic-High-Pressure-Nebulizing) and detection by ICP-MS. The method takes advantage of the ability to measure individual isotopes. Recoveries of the water vapor distillation procedure samples for different mercury compounds from sediment were tested. For methylmercury, the detection limit for a 0.5 g sample was calculated to be 0.025 μg/kg. The new technique was assured using different reference materials. Received: 23 October 1996 / Revised: 14 February 1997 / Accepted: 16 February 1997     

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.     

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。     

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.     

Determination of Mercury and Selenium in Hair Samples of Brazilian Indian Populations Living in the Amazonic Region by NAA

Biomonitoring of mercury contamination of Brazilian Indian population groups living in the Xingu Park, a reservation situated in the Amazonic region, has revealed very high levels of mercury in hair samples as compared to controls. Total mercury was determined by INAA in most of the tribes living in the Park and methylmercury was determined by CVAAS in samples with total mercury above 10 mg/kg. Due to the fact that selenium seems to protect animals against the toxic effects of methylmercury, it was considered also of interest to determine its concentrations in the hair samples with very high mercury levels. Selenium was determined by INAA via the short-lived radionuclide ⁷⁷mSe (T _1/2 = 17.45 s). The correlations between selenium and mercury concentrations in Brazilian controls and in the Indian population groups are discussed.     

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.