Simple and sensitive method for the determination of methylmercury in hair using thin-layer chromatography with thermal decomposition gold amalgamation atomic absorption spectrophotometry.

An improved method is described for the determination of methylmercury (MeHg) in human hair that is based on thin-layer chromatography and thermal decomposition gold amalgamation atomic absorption spectrophotometry. The dithizone extraction and application procedure of this technique were optimized, thus improving its sensitivity and robustness and enabling the use of less toxic solvents than other approaches. The limit of detection was 0.18 ng MeHg (as Hg), corresponding to 0.018 mg MeHg kg-1 of hair relative to a 10 mg sample. This method is appropriate for detecting MeHg in hair at the approximate reference dose level established by the United States Environmental Protection Agency.

Application of the Quartz Crystal Microbalance Method for Measuring Mercury in the Air of Working Environments Involved to Artisanal and Small-scale Gold Mining (ASGM).

Artisanal small gold mining (ASGM) is responsible for approximately 40% of the total Hg emissions into the atmosphere worldwide. In developing countries, many people are engaged in ASGM activities. We developed a small, simple Hg measuring device, which detects Hg in the air based on the change of the oscillation frequency of an Au electrode on a quartz crystal microbalance (QCM). This device is called QCM-Hg. We tested the viability of the QCM-Hg in various work settings including a gold mining area and gold shops. In working environments with an airborne Hg concentration of several µg m-3, the changing rate of the oscillation frequency for either 2 or 3 min corresponded with the Hg concentrations measured using the conventional method of gold amalgamation and cold vapor atomic absorption spectrometer (CVAAS). The results revealed that the QCM-Hg is a useful device for real-time Hg monitoring in actual working environments related to ASGM activities and Hg treatment facilities.

Development of Human Hair Reference Material Supporting the Biomonitoring of Methylmercury.

A certified reference material, NIMD-01, was developed for the analysis of mercury speciation in human hair. We collected the hair of Vietnamese males from a barbershop in Hanoi in 2016 and prepared 1200 bottles containing 3 g of sieved and blended hair powder. The certified value was given on a dry-mass basis, with the moisture content obtained by drying at 85°C for 4 h. Certified values with the expanded uncertainties (coverage factor, k = 2) were as follows: methylmercury, 0.634 ± 0.071 mg kg-1 as mercury; total mercury, 0.794 ± 0.050 mg kg-1; copper, 12.8 ± 1.4 mg kg-1; zinc, 234 ± 29 mg kg-1; selenium, 1.52 ± 0.29 mg kg-1. An indicative arsenic concentration of 0.17 ± 0.03 mg kg-1 was measured. Extended uncertainties were estimated by sample homogeneity, long- and short-term stabilities, and a characterization from measurements made by collaborating laboratories.

The spatial distribution of total mercury in sediments in the Yatsushiro Sea, Japan.

The Yatsushiro Sea in Japan is contaminated with mercury in wastewater discharge from the Chisso Company, which produced acetaldehyde from 1932 onwards. In this study, we investigated the current spatial distribution of total mercury (T-Hg) in sediments, both at the surface and at different depths, using 43 sediment cores and 22 surface sediment samples collected from the Yatsushiro Sea in August 2017. Altogether, 821 surface and core samples were analyzed for their T-Hg concentrations. Most of the mercury that was discharged from Chisso remained either in, or around the exit of, Minamata Bay, but some had been transported from Minamata Bay to the Nagashima Sea. We estimated that almost 51 and 6 tons of mercury had accumulated in the Yatsushiro and the Amakusa Sea areas, respectively. The amount of acetaldehyde produced in Chisso over time was correlated with the T-Hg concentrations in the sediments from the Yatsushiro Sea.

Chemical characteristics of dissolved mercury in the pore water of Minamata Bay sediments.

Methylmercury concentrations in fish from the historically polluted Minamata Bay remain higher than in fish from other coastal seas around Japan. To obtain a better understanding of this phenomenon, the chemical characteristics of pore water from Minamata Bay sediments were investigated. Samples were taken from two stations over a 1-year period. Total average values in the pore water at the two stations for dissolved total mercury and methylmercury concentrations were 6.64±4.93 and 2.69±2.07ng/l, respectively. The pore water was centrifuged at 1000rpm to 3000rpm. The highest ratio of dissolved methylmercury to dissolved total mercury exceeded 60% for pore water centrifuged at 3000rpm. Furthermore, because total average values of Log Kd of total mercury and methylmercury in sediment (St1 and St2) were 5.42 and 2.32 (Lkg-1), methylmercury in Minamata Bay sediment is more eluted than other mercury species.

The dynamics of mercury near Idrija mercury mine, Slovenia: Horizontal and vertical distributions of total, methyl, and ethyl mercury concentrations in soils.

The distributions of the total mercury (T-Hg), methylmercury (MeHg), and ethylmercury (EtHg) concentrations in soil and their relationship to chemical composition of the soil and total organic carbon content (TOC, %) were investigated. Core samples were collected from hill slope on the right and left riverbanks of the Idrija River. Former smelting plant is located on the right bank. The T-Hg average in each of the core samples ranged from 0.25 to 1650 mg kg-1. The vertical T-Hg variations in the samples from the left bank showed no significant change with depth. Conversely, the T-Hg varied with depth, with the surface, or layers several centimeters from the surface, tending to show the highest values in the samples from the right bank. Since the right and left bank soils have different chemical compositions, different pathways of mercury delivery into soils were suggested. The MeHg and EtHg concentrations ranged from n.d. (not detected) to 444 µg kg-1 and n.d. to 17.4 µg kg-1, respectively. The vertical variations of MeHg and EtHg were similar to those of TOC, except for the near-surface layers containing TOC greater than 20%. These results suggest that the decomposition of organic matter is closely related to organic mercury formation.

Distribution and characteristics of methylmercury in surface sediment in Minamata Bay.

This study was carried out to evaluate the present-day chemical properties of methylmercury in surface sediment in Minamata Bay where a dredging project was completed 28years ago. Present-day sediment from Minamata Bay consists of sandy silt, and the average loss-on-ignition in surface sediment was 7.0±2.3%. The average methylmercury concentrations in the upper sediment layers were significantly higher than those in the lower sediment layers. Currently, the concentrations in sediments in Minamata Bay do not exceed the Japanese regulatory standard value for mercury. The average concentration of methylmercury in Minamata Bay surface sediment was 1.74±1.0ng/g on a dry weight basis (n=107). The methylmercury concentration in Minamata Bay surface sediment was almost 16 times higher than that in surface sediment from Isahaya Bay surface sediment, which was 0.11±0.045ng/g on a dry weight basis (n=5).

Methylmercury in the breast milk of Japanese mothers and lactational exposure of their infants.

The human fetus is known to be exposed to methylmercury (MeHg), but little is known about the risk of infant exposure via breast milk. To evaluate the lactational exposure to MeHg via breast milk in Japanese infants, the levels of total mercury (THg) and MeHg were determined in breast milk and maternal blood using samples from a birth cohort study at the Tohoku Study of Child Development. Maternal blood and breast milk were collected one day postpartum and one month after delivery, respectively. The median THg (and MeHg) concentrations in maternal RBCs, plasma and breast milk were 17.8 ng g(-1) (17.8 ng g(-1)), 1.51 ng g(-1) (1.33 ng g(-1)) and 0.81 ng g(-1) (0.45 ng g(-1)), respectively (n=27). The median percentage of MeHg in THg was 54% in breast milk. Breast milk contained substantial amounts of MeHg, which was strongly associated with the internal accumulation of MeHg and the lipid content of the milk (r=0.684). The range of lipid contents in milk varied widely from 0.50 to 6.60 g/100 g of milk, with a median of 3.60 g/100 g. The median (range) weekly average intake of MeHg via breast milk was estimated to be 0.63 µg kg(-1) (0.08-1.68 µg kg(-1)) BW/week. Because the MeHg and lipid contents in milk substantially fluctuate, an investigation of the variations of MeHg and lipid content in breast milk may be required for a more precise risk assessment.

Retrospective study on temporal and regional variations of methylmercury concentrations in preserved umbilical cords collected from inhabitants of the Minamata area, Japan.

The present study was conducted to investigate the historical time-course changes and regional distribution of methylmercury concentrations in preserved umbilical cords collected from Minamata-area inhabitants born between 1947 and 1989. The data from Miyazaki, Tottori, Akita, Tsushima (Nagasaki), Fukuoka and Tokyo were used as controls. A total of 325 data were analyzed to estimate the temporal and spatial distribution of methylmercury among inhabitants born in the Minamata area. Elevated methylmercury concentrations (>or=1 microg/g) were mainly observed in inhabitants born between 1947 and 1968. That peak coincided with the peak of acetaldehyde production in Minamata. The methylmercury concentrations started to decrease in keeping with the decline of acetaldehyde production, which ceased in 1968, and thereafter the methylmercury levels gradually decreased to the control levels. Elevated methylmercury concentrations were first observed in the districts of Minamata, followed by Izumi, Tsunagi and Ashikita, indicating the time-course-dependent regional distributions of methylmercury pollution.

Head hair total mercury and methylmercury levels in some Ghanaian individuals for the estimation of their exposure to mercury: preliminary studies.

The extent of human exposure to mercury in some individuals in Ghana was evaluated by analysing samples of human head hair for total mercury and methylmercury. The average level of total mercury was 0.843 microg g(-1) (in range of 0.119-4.140, n = 123) and that of methylmercury was 0.787 microg g(-1) (in range of 0.208-1.847, n = 42). Mercury was present in the hair samples almost completely in the methylated form. The average percentage ratio of methylmercury to total mercury was 97.2% (in range of 88.5%-107.6%). The results of this study indicate low levels of exposure to methylmercury and does not pose a significant risk to the individuals and to a greater extent the general population.

Correlations between mercury concentrations in umbilical cord tissue and other biomarkers of fetal exposure to methylmercury in the Japanese population.

Methylmercury (MeHg) is one of the most risky substances to affect humans through fish consumption, and the fetus is known to be in the most susceptible group. Our objective in this study is to examine the relationships of total mercury (THg) and MeHg concentrations between umbilical cord tissue and other tissues as biomarkers of fetal exposure to MeHg in the Japanese population. In total, 116 paired samples were collected in three Japanese districts, the Tsushima Islands, Fukuoka City, and Katsushika ward of metropolitan Tokyo. THg was measured for hair and THg and MeHg were measured in cord tissues, maternal blood, and cord blood. The relationships among tissues in Hg concentrations were similar among districts. Therefore, we analyzed the relationships using all the samples. More than 90% of Hg in cord tissue, cord blood, and maternal blood was MeHg. THg and MeHg in cord blood was about two times higher than in maternal blood. A strong correlation was found between THg and MeHg in cord tissue. The cord tissue THg and MeHg showed a strong correlation with cord blood Hg, which is recognized as the best biomarker for fetal exposure to MeHg. The findings of this study indicate the significance of cord tissue THg and MeHg as biomarkers for fetal exposure to MeHg at parturition.

Environmental and human exposure assessment monitoring of communities near an abandoned mercury mine in the Philippines: a toxic legacy.

Abandoned mines are an important global concern and continue to pose real or potential threats to human safety and health including environmental damage/s. Very few countries had government mine regulation and reclamation policies until the latter part of the century where legal, financial and technical procedures were required for existing mining operations. Major reasons for mine closure may be mainly due to poor economies of the commodity making mining unprofitable, technical difficulties and national security. If the mine is abandoned, more often than not it is the government that shoulders the burden of clean-up, monitoring and remediation. The topic of abandoned mines is complex because of the associated financial and legal liability implications. Abandoned mercury mines have been identified as one of the major concerns because of their significant long-term environmental problems. Primary mercury production is still ongoing in Spain, Kyrgzystan, China, Algeria, Russia and Slovakia while world production declined substantially in the late 1980s. In the Philippines, the mercury mine located southeast of Manila was in operation from 1955 to 1976, before ceasing operation because of the decline in world market price for the commodity. During this time, annual production of mercury was estimated to be about 140,000 kg of mercury yearly. Approximately 2,000,000 t of mine-waste calcines (retorted ore) were produced during mining and roughly 1,000,000 t of these calcines were dumped into nearby Honda Bay to construct a jetty to facilitate mine operations where about 2000 people reside in the nearby three barangays. In October, 1994 the Department of Health received a request from the Provincial Health Office for technical assistance relative to the investigation of increasing complaints of unusual symptoms (e.g. miscarriages, tooth loss, muscle weakness, paralysis, anemia, tremors, etc.) among residents of three barangays. Initial health reports revealed significant elevation of blood mercury levels exceeding the then recommended exposure level of 20ppb in 12 out of the 43 (27.9%) residents examined. The majority of the volunteers were former mine workers. In this study the abnormal findings included gingivitis, mercury lines, gum bleeding and pterydium. The most common neurologic complaints were numbness, weakness, tremors and incoordination. Anemia and elevated liver function tests were also seen in a majority of those examined. The assessment also revealed a probable association between blood mercury level and eosinophilia. The same association was also seen between high mercury levels and the presence of tremors and working in the mercury mine. To date, there are very limited environmental and health studies on the impact of both total and methylmercury that have been undertaken in the Philippines. Thus, this area of study was selected primarily because of its importance as an emerging issue in the country, especially regarding the combined effects of total and methylmercury low-dose and continuous uptake from environmental sources. At present the effects of total mercury exposure combined with MeHg consumption remain an important issue, especially those of low-dose and continuous uptake. Results of the study showed that four (4) species of fish, namely ibis, tabas, lapu-lapu and torsillo, had exceeded the recommended total mercury and methylmercury levels in fish (NV>0.5 microg/gf.w., NV>0.3 microg/gf.w., respectively). Saging and kanuping also exceeded the permissible levels for methylmercury. Total and methylmercury in canned fish, and total mercury in rice, ambient air and drinking water were within the recommended levels, however, additional mercury load from these sources may contribute to the over-all body burden of mercury among residents in the area. Surface water quality at the mining area, Honda Bay and during some monitoring periods at Palawan Bay exceeded total mercury standards (NV>0.002 ng/mL). Soil samples in two sites, namely Tagburos and Honda Bay, exceeded the EPA Region 9 Primary Remediation Goal recommended values for total mercury for residential purposes (NV>23 mg/kg). The hand to mouth activity among infants and children is another significant route for mercury exposure. Statistically significant results were obtained for infants when comparing the results after one year of monitoring for methylmercury levels in hair for both exposed and control sub-groups. Likewise, comparing the initial and final hair methylmercury levels among pregnant women/mothers in the exposed group showed statistically significant (p<0.05) results. Comparing the exposed and control sub-groups' mercury hair levels per sub-group showed statistically significant results among the following: (a) initial and final total mercury hair levels among children, (b) initial and final methylmercury hair levels among children, (c) final total mercury hair levels among pregnant women, (d) initial and final total mercury hair levels among mothers, and (e) initial and final methyl hair levels among mothers.

Health and environmental assessment of mercury exposure in a gold mining community in Western Mindanao, Philippines.

The small-scale gold mining activities using mercury began in the late 1980s in Sibutad, Western Mindanao. It is located very near the Murcielagos Bay with tailing ponds directly discharging into bodies of water. This cross-sectional study aimed to determine the health and environmental effects of gold mining activities on the community. Residents were randomly selected and classified into two groups, namely, the directly exposed and indirectly exposed populations using a set inclusion criteria. Complete medical and laboratory examinations were performed. Environmental and biologic samples were collected for total mercury and methylmercury determinations. The results showed that the directly exposed group had significantly higher mean blood total mercury and methylmercury levels in comparison with the indirectly exposed population. Although there were no significant differences between hair total mercury and methylmercury levels, there was a trend for higher levels of these biomarkers among the directly exposed residents as compared with the unexposed group. The absence of statistically significant differences may be attributable to the small sample size. Ambient air quality monitoring for mercury exceeded the allowable levels. However, levels of mercury in drinking water and sediments were within allowable limits. Frequency of gastrointestinal complaints was significantly associated with elevated hair methylmercury levels (p=0.02). Also, there appears to be a trend towards higher blood total mercury levels and frequency of gastrointestinal complaints (p=0.09). An interesting finding in this study was the increasing incidence of elevated diastolic blood pressure with elevated hair total mercury levels (p=0.07). Mercury storage at home is a risk factor.

Spatial variations of mercury in sediment of Minamata Bay, Japan.

Mercury-contaminated effluent was discharged into Minamata Bay from a chemical plant over a period of approximately 40 years until 1968. In October 1977, the Minamata Bay Pollution Prevention Project was initiated to dispose of sedimentary sludge containing mercury concentrations higher than 25 mg kg(-1). In March 1990, the project was completed. In an effort to estimate current contamination in the bay, the vertical and horizontal distributions of mercury in sediment were investigated. Sediment core samples were collected on June 26, 2002 at 16 locations in Minamata Bay and Fukuro Bay located in the southern part of Minamata Bay. The sediment in Fukuro Bay had not been dredged. The total mercury concentration in surface sediment was 1.4-4.3 mg kg(-1) (2.9+/-0.9 mg kg(-1), n=9) for the dredged area of Minamata Bay and 0.3-4.8 mg kg(-1) (3.6+/-1.6 mg kg(-1), n=4) for Fukuro Bay. In the lower layers of long cores taken from both areas, the total mercury concentration decreased with depth and finally showed relatively uniform low values. These values can be considered to represent the background concentration absent of anthropogenic influence, which was estimated for the study area to be 0.068+/-0.012 mg kg(-1) (n=10). From the surface, the total mercury concentration in Fukuro Bay increased with depth and reached a maximum at 8-14 cm. In Minamata Bay, several centimeters from the surface the total mercury concentration did not change significantly having considerably higher values than the background level. At six stations, the methylmercury concentration was determined. Although the vertical variations were similar to those for total mercury, Fukuro Bay sediment showed a higher concentration of methylmercury than Minamata Bay sediment.

Tagum study II: follow-up study at two years of age after prenatal exposure to mercury.

OBJECTIVES: To correlate the presence and levels of total mercury (THg) in cord blood and meconium indicating prenatal exposure with developmental milestones at 2 years and to compare these subjects with controls of comparable age using cognitive adaptive test and clinical linguistic auditory milestone scale (CAT/CLAMS). METHODS: In 48 of the original Tagum (T) subjects, cord blood and meconium Hg levels, head circumference (HC) at birth, and duration of breastfeeding were correlated with CAT/CLAMS at 2 years. At 2 years, THg levels using cold atomic vapor absorption spectrometry were determined in the hair of 46 T subjects and 88 Saranggani (S) controls; THg levels in blood were tested in 48 T subjects and 45 S controls. These levels were correlated with CAT/CLAMS. Both groups had standard physical and neurologic examinations, hearing screen using transitory evoked otoacoustic emissions, serum glutamate pyruvate transaminase, and routine urinalysis. A prevalidated Socioeconomic Means Test was given to both groups. RESULTS: The Hg level in cord blood was negatively correlated with CAT/CLAMS at 2 years. The HC at birth was negatively correlated with levels of Hg in hair of T subjects 2 years later. HC at birth and 2 years hence were positively correlated with CAT/CLAMS. The following were significantly higher in S controls than in T subjects: expressive language quotient 82.569 +/- 2.21 versus 71.57 +/- 2.61; CLAMS 87.96 +/- 2.43 versus 77.67 +/- 2.51; CAT 90.57 +/- 2.22 versus 83.15 +/- 1.43; and full-scale developmental quotient 89.31 +/- 2.14 versus 80.56 +/- 1.86. Fifteen percent of T subjects had global delay (full-scale developmental quotient

Declining risk of methylmercury exposure to infants during lactation.

Methylmercury (MeHg) can be transferred to infants through milk, in addition to passage through the placenta during intrauterine life. The higher MeHg accumulation and susceptibility to toxicity in the fetus than in the mother during the gestation period is well known. However, the contribution of Hg exposure through breast milk to the MeHg concentration in infants is not clear. Our objective in this study was to examine the changes in MeHg levels in infants who were reared on breast milk to evaluate the risks between fetal and breast-feeding periods based on Hg concentrations in red blood cells (RBCs-Hg). RBCs-Hg and plasma-Hg concentrations (Plasma-Hg) in seven pairs of maternal and infant blood samples were compared at birth and 3 months after parturition. RBCs to Plasma-Hg ratio was approximately 8:1 for mothers at parturition and after 3 months, suggesting that their Hg source is MeHg from consumed fish as is general in Japanese. In all seven cases, RBCs-Hg in the umbilical cords were higher than those in the mothers at parturition. The geometric mean of RBCs-Hg in umbilical cords (10.6ng/g) was about 1.4 times higher than that in the mothers (7.1ng/g). There was a strong correlation in RBCs-Hg in mothers and umbilical cords. However, all the infants showed declines in Hg concentrations throughout the breast-feeding period. The geometric mean RBCs-Hg at 3 months of age was 5.8ng/g, accounting for 54% of that in the umbilical cords. Consequently, maternal RBCs-Hg surpassed that of infants at 3 months, opposite to the situation at parturition. The decline in infant RBCs-Hg during the breast-feeding period can be explained by the low Hg transfer through breast milk and the rapid growth of infants after birth. The geometric mean Milk-Hg was low (0.21ng/g), around 20% of that in maternal Plasma-Hg. The average body weight of infants at 3 months increased to about 1.9 times of that at birth. Thus, offspring are subjected to MeHg exposure through both the gestation and the breast-feeding periods; the risk is especially high during gestation but may decrease during breast-feeding.

Simultaneous determination of inorganic mercury and methylmercury compounds in natural waters.

The purpose of the present work was to develop a simple, rapid, sensitive and accurate method for the simultaneous determination of inorganic mercury (Hg(2+)) and monomethylmercury compounds (MeHg) in natural water samples at the pg L(-1) level. The method is based on the simultaneous extraction of MeHg and Hg(2+)dithizonates into an organic solvent (toluene) after acidification of about 300 mL of a water sample, followed by back extraction into an aqueous solution of Na(2)S, removal of H(2)S by purging with N(2), subsequent ethylation with sodium tetraethylborate, room temperature precollection on Tenax, isothermal gas chromatographic separation (GC), pyrolysis and cold vapour atomic fluorescence spectrometric detection (CV AFS) of mercury. The limit of detection calculated on the basis of three times the standard deviation of the blank was about 0.006 ng L(-1) for MeHg and 0.06 ng L(-1) for Hg(2+)when 300 mL of water was analysed. The repeatability of the results was about 5% for MeHg and 10% for Hg(2+). Recoveries were 90-110% for both species.

Evaluation of changes in methylmercury accumulation in the developing rat brain and its effects: a study with consecutive and moderate dose exposure throughout gestation and lactation periods.

Methylmercury (MeHg) can be transferred to the fetus through the placenta and to newborn offspring through breast milk. The higher mercury (Hg) accumulation and susceptibility to toxicity in the fetus than in the mother during the gestation period is well known. However, the contribution of MeHg exposure through breast milk to the brain Hg concentration in offspring is not clear. The purposes of this study were to evaluate the changes in Hg concentration in the brain of offspring and its effects on the developing rat brain, based on consecutive and moderate doses of MeHg throughout gestation and lactation. Adult female rats were given a diet containing 5 ppm Hg (as MeHg) for 8 weeks. The administration level was thought not to cause adverse effects in adult rats. The rats were then mated and subsequently given the same diet throughout gestation and after parturition. The newborn offspring were placed with the mothers until postnatal day 30. The offspring were exposed to MeHg throughout their intrauterine life through the placenta, and during the postnatal developing phase via contaminated milk. Furthermore, they were given the same diet containing MeHg for 2 months following weaning. On the day of parturition, the concentration of Hg in the brains of newborns was 1.4 times higher than that in the mothers. During the suckling period the concentration in the brain of the offspring rapidly declined to 1/5 of that at birth, suggesting that MeHg transport by milk was limited while the brain and body volumes increased rapidly. The concentration increased gradually again after the offspring started the contaminated diet. In behavioral tests performed at 5 and 6 weeks of age, MeHg-exposed rats showed a significant deficit in motor coordination in the rotarod test and a learning disability in the passive avoidance response test, compared with controls. Histopathologically, focal cerebellar dysplasia, including the heterotopic location of Purkinje cells and granule cells, was observed. These abnormalities may be induced by the effect of highly accumulated MeHg in the brain during the gestation period. Thus, although offspring are subjected to consecutive and moderate dose MeHg exposure throughout both the gestation and suckling periods, the risk is especially high during gestation but may decrease during lactation.