Occurrence of trivalent monomethyl arsenic and other urinary arsenic species in a highly exposed juvenile population in Bangladesh.

Following reports of high cytotoxicity and mutagenicity of monomethyl arsonous acid (MMA(III)) and early reports of urinary MMA(III) in arsenic-exposed individuals, MMA(III) has often been included in population studies. Use of urinary MMA(III) as an indicator of exposure and/or health risk is challenged by inconsistent results from field studies and stability studies, which indicate potential artifacts. We measured urinary arsenic species in children chronically exposed to arsenic in drinking water, using collection, storage, and analysis methods shown to conserve MMA(III). MMA(III) was easily oxidized in sample storage and processing, but recoveries of 80% or better in spiked urine samples were achieved. Attempts to preserve the distribution of MMA between trivalent and pentavalent forms using complexing agents were unsuccessful and MMA(III) spiked into treated urine samples actually showed lower stability than in untreated samples. In 643 urine samples from a highly exposed population from the Matlab district in Bangladesh stored for 3-6 months at ≤-70 °C, MMA(III) was detected in 41 samples, with an estimated median value of 0.3 µg/l, and levels of MMA(III) above 1 µg/l in only two samples. The low urinary concentrations in highly exposed individuals and known difficulties in preserving sample oxidation state indicate that urinary MMA(III) is not suitable for use as an epidemiological biomarker.

Involvement of N-6 adenine-specific DNA methyltransferase 1 (N6AMT1) in arsenic biomethylation and its role in arsenic-induced toxicity.

BACKGROUND: In humans, inorganic arsenic (iAs) is metabolized to methylated arsenical species in a multistep process mainly mediated by arsenic (+3 oxidation state) methyltransferase (AS3MT). Among these metabolites is monomethylarsonous acid (MMAIII), the most toxic arsenic species. A recent study in As3mt-knockout mice suggests that unidentified methyltransferases could be involved in alternative iAs methylation pathways. We found that yeast deletion mutants lacking MTQ2 were highly resistant to iAs exposure. The human ortholog of the yeast MTQ2 is N-6 adenine-specific DNA methyltransferase 1 (N6AMT1), encoding a putative methyltransferase. OBJECTIVE: We investigated the potential role of N6AMT1 in arsenic-induced toxicity. METHODS: We measured and compared the cytotoxicity induced by arsenicals and their metabolic profiles using inductively coupled plasma-mass spectrometry in UROtsa human urothelial cells with enhanced N6AMT1 expression and UROtsa vector control cells treated with different concentrations of either iAsIII or MMAIII. RESULTS: N6AMT1 was able to convert MMAIII to the less toxic dimethylarsonic acid (DMA) when overexpressed in UROtsa cells. The enhanced expression of N6AMT1 in UROtsa cells decreased cytotoxicity of both iAsIII and MMAIII. Moreover, N6AMT1 is expressed in many human tissues at variable levels, although at levels lower than those of AS3MT, supporting a potential participation in arsenic metabolism in vivo. CONCLUSIONS: Considering that MMAIII is the most toxic arsenical, our data suggest that N6AMT1 has a significant role in determining susceptibility to arsenic toxicity and carcinogenicity because of its specific activity in methylating MMAIII to DMA and other unknown mechanisms.

Arsenic exposure within the Korean community (United States) based on dietary behavior and arsenic levels in hair, urine, air, and water.

BACKGROUND: Determining arsenic exposure in groups based on geographic location, dietary behaviors, or lifestyles is important, as even moderate exposures may lead to health concerns. OBJECTIVES/METHODS: The Korean community in Washington State, represents a group warranting investigation, as they consume foods (e.g., shellfish, rice, finfish, and seaweed) known to contain arsenic. As part of the Arsenic Mercury Intake Biometric Study, we examined the arsenic levels in hair and urine along with the diets of 108 women of childbearing age from within this community. Arsenic levels in indoor air and drinking water were also investigated, and shellfish commonly consumed were collected and analyzed for total and speciated arsenic. RESULTS: The six shellfish species analyzed (n = 667) contain total arsenic (range, 1-5 microg/g) but are a small source of inorganic arsenic (range, 0.01-0.12 microg/g). Six percent of the individuals may have elevated urinary inorganic arsenic levels (> 10 microg/L) due to diet. Seaweed, rice, shellfish, and finfish are principal sources for total arsenic intake/excretion based on mass balance estimates. Rice consumption (163 g/person/day) may be a significant source of inorganic arsenic. Air and water are not significant sources of exposure. Hair is a poor biometric for examining arsenic levels at low to moderate exposures. CONCLUSIONS: We conclude that a portion of this community may have dietary inorganic arsenic exposure resulting in urine levels exceeding 10 microg/L. Although their exposure is below that associated with populations exposed to high levels of arsenic from drinking water (> 100 microg/L), their exposure may be among the highest in the United States.

Evaluation of two new arsenic field test kits capable of detecting arsenic water concentrations close to 10 microg/L.

Millions of people worldwide are exposed to arsenic-contaminated drinking water. Arsenic field test kits may offer a cost-effective approach for measuring these exposures in the field, although the accuracy of some kits used in the past has been poor. In this study, arsenic concentrations were measured in 136 water sources in western Nevada using two relatively new arsenic test kits and compared to laboratory measurements using atomic fluorescence spectroscopy (AFS). Spearman's rank correlation coefficients comparing the Quick Arsenic and Hach EZ kits to laboratory measurements were 0.96 (p < 0.001) and 0.95 (p < 0.001), respectively. When analyzed in seven exposure categories (0-9, 10-19, 20-49, 50-99, 100-199, 200-499, and > or = 500 microg/L), test kit and AFS measurements were in the same category in 71% (Quick Arsenic) and 62% (Hach EZ) of samples, and within one category of each other in 99% (Quick Arsenic) and 97% (Hach EZ) of samples. Both kits identified all water samples with high arsenic concentrations (> 15 microg/L) as being above the United States Environmental Protection Agency's drinking water standard and the World Health Organization's guideline value for arsenic of 10 microg/L. These results suggestthatthese easily portable kits can be used to identify water sources with high arsenic concentrations and may provide an important tool for arsenic surveillance and remediation programs.

Evaluation of urinary methoxyphenols as biomarkers of woodsmoke exposure.

Urinary methoxyphenols have been proposed as biomarkers for woodsmoke exposure, but the relationship between exposure and urinary methoxyphenol concentrations has not been characterized. We collected personal particulate matter2.5 and urine samples from 9 adults experimentally exposed to smoke from an open wood fire to characterize this relationship. Personal exposures (PM2.5 mean 1500 microg/ m3) varied 3.5-fold. Twenty-two methoxyphenols, levoglucosan, and 17 polynuclear hydrocarbons were quantified by gas chromatography/mass spectrometry assays for personal filter samples and urine samples. Most methoxyphenols had measurable preexposure levels. Propylguaiacol, syringol, methylsyringol, ethylsyringol, and propylsyringol had peak urinary concentrations after the woodsmoke exposure. Eight subjects had peak urinary elimination of methoxyphenols within 6 h (t1/2 3-5 h), whereas one had delayed elimination. Several metrics for urinary excretion were evaluated. Analyte concentration was greatly affected by diuresis. Excretion rate and analyte concentrations normalized by creatinine gave a clearer signal and were equivalent in predictive ability. Twelve-hour average creatinine-normalized concentrations of each of the 5 methoxyphenols gave a Pearson correlation > or = 0.8 with their particle-phase concentration. The sum of urinary concentrations for the 5 methoxyphenols versus levoglucosan on personal filters gave a regression coefficient of 0.75. This sum versus PM2.5 gave a regression coefficient of 0.79. The intercept of this regression suggests that the threshold for detection of an acute exposure event would be approximately 760 microg/m3 particulate matter from woodsmoke. The signal-to-noise (12-h postexposure average/preexposure average) ranged from 1.1 to 8 for the 5 methoxyphenols. Analysis of multiple compounds provided assurance that elevations were not artifactual due to food or other products.

Ethanol-induced increase in the metabolic clearance of 1,1,1-trichloroethane in human volunteers.

This study evaluated the effect of moderate doses of ethanol over a short period of time on the toxicokinetics of an organic solvent, 1,1,1-trichloroethane. A group of 10 moderate drinkers were recruited and exposed via inhalation for 2 h to a low concentration of 1,1,1-trichloroethane (175 ppm) on two separate occasions. Subjects were administered ethanol (0.35 g/kg body weight) on each of the 7 days preceding one of the exposures. Blood and urine samples were collected during and following each exposure, with blood analyzed for 1,1,1-trichloroethane and urine analyzed for the metabolites of 1,1,1-trichloroethane: trichloroethanol and trichloroacetic acid. Prior ethanol consumption resulted in a significant increase in apparent metabolic clearance of 1,1,1-trichloroethane (mean increase = 25.4%). The results of this study demonstrate that ethanol consumption over time can affect the rate at which an organic solvent is cleared through metabolism in humans. For chemicals with toxic metabolic products, this inductive effect of ethanol consumption on the rate of biotransformation could be potentially harmful to exposed individuals. Metabolic clearance of compounds with high hepatic extraction may not be affected by enzyme induction as it is likely that these compounds are essentially completely metabolized while passing through the liver.

Reverse osmosis filter use and high arsenic levels in private well water.

Inorganic arsenic causes cancer, and millions of people worldwide are exposed to arsenic-contaminated water. Regulatory standards for arsenic levels in drinking water generally do not apply to private domestic wells. Reverse osmosis (RO) units commonly are used by well owners to reduce arsenic concentrations, but may not always be effective. In a survey of 102 homes in Nevada, 19 used RO devices. Pre- and post-RO filtration arsenic concentrations averaged 443 microg/l and 87 microg/l, respectively. The average absolute and percent reductions in arsenic concentrations after filtration were 356 microg/l and 79%, respectively. Postfiltration concentrations were higher than 10 microg/l in 10 homes and higher than 100 microg/l in 4 homes. These findings provide evidence that RO filters do not guarantee safe drinking water and, despite regulatory standards, some people continue to be exposed to very high arsenic concentrations.

Determination of methoxyphenols in ambient atmospheric particulate matter: tracers for wood combustion.

Combustion of wood and other biomass fuels produces source-specific organic compounds arising from pyrolysis of lignin, including substantial amounts of 4-substituted methoxylated phenolic compounds (methoxyphenols). These compounds have been used as atmospheric markers to determine the contribution of wood smoke to ambient atmospheric fine particulate matter (PM). However, reliable quantification of methoxyphenols represents an analytical challenge because these compounds are polar, semi-volatile, and somewhat reactive. We reportherein an improved gas chromatographic-mass spectrometric (GC/MS) method for the sensitive and reliable determination of methoxyphenols in low-volume ambient PM samples. Deuterated standard compounds are added to the environmental samples prior to extraction to determine analyte recoveries in each sample. Analytical figures of merit for the assay, as applied to ambient PM2.5 and PM10 samples are as follows: recovery = 63-100%; precision = 2-6%; analytical limit of detection (S/N 2) = 0.002 microg/mL; limit of quantitation = 0.07-0.45 ng/m3 (assuming a 14 m3 sample). The improved method was applied to ambient PM samples collected between 1999 and 2000 in Seattle, WA. Particle-bound methoxyphenol concentrations in the range <0.1 to 22 ng/m3 were observed and the methoxyphenols were present almost exclusively in the fine (PM2.5) size fraction. We also demonstrated that XRF analysis of samples of atmospheric PM collected on Teflon filters significantly reduced the levels of methoxyphenols measured in the PM samples in subsequent assay of the same filters. Therefore, XRF analysis of filters, commonly undertaken to obtain trace element concentrations for use in source apportionment analyses, would preclude the subsequent analysis of those filters for methoxyphenols and other similarly semivolatile or reactive organic chemicals.

Determination of levoglucosan in atmospheric fine particulate matter.

A microanalytical method suitable for the quantitative determination of the sugar anhydride levoglucosan in low-volume samples of atmospheric fine particulate matter (PM) has been developed and validated. The method incorporates two sugar anhydrides as quality control standards. The recovery standard sedoheptulosan (2,7-anhydro-beta-D-altro-heptulopyranose) in 20 microL solvent is added onto samples of the atmospheric fine PM and aged for 1 hr before ultrasonic extraction with ethylacetate/ triethylamine. The extract is reduced in volume, an internal standard is added (1,5-anhydro-D-mannitol), and a portion of the extract is derivatized with 10% by volume N-trimethylsilylimidazole. The derivatized extract is analyzed by gas chromatography/mass spectrometry (GC/MS). The recovery of levoglucosan using this procedure was 69 +/- 6% from five filters amended with 2 microg levoglucosan, and the reproducibility of the assay is 9%. The limit of detection is approximately 0.1 microg/mL, which is equivalent to approximately 3.5 ng/m3 for a 10 L/min sampler or approximately 8.7 ng/m3 for a 4 L/min personal sampler (assuming 24-hr integrated samples). We demonstrated that levoglucosan concentrations in collocated samples (expressed as ng/m3) were identical irrespective of whether samples were collected by PM with aerodynamic diameter < or = 2.5 microm or PM with aerodynamic diameter < or = 10 microm impactors. It was also demonstrated that X-ray fluorescence analysis of samples of atmospheric PM, before levoglucosan determinations, did not alter the levels of levoglucosan.

Family correlations of arsenic methylation patterns in children and parents exposed to high concentrations of arsenic in drinking water.

We investigated the evidence of a familial contribution to urinary methylation patterns in families ingesting arsenic in drinking water. Arsenic methylation can be assessed by measuring urinary levels of inorganic arsenic (InAs) and its methylated metabolites, monomethylarsonate (MMA), and dimethylarsinate (DMA). Methylation activity is reflected in the ratios: InAs/methylated arsenic (InAs/metAs) and MMA/DMA. Eleven families from Chile were selected because of their long-term exposure to very high levels of arsenic in drinking water (735-762 microg/L). Each family consisted of a father, a mother, and two children. We measured urinary arsenic and its methylated metabolites for each participant (n = 44). The intraclass correlation coefficients showed that 13-52% of the variations in the methylation patterns were from being a member of a specific family. Family correlations were calculated for father-mother, parent-child, and sibling-sibling pairs. Methylation patterns correlated strongly between siblings [r = 0.78 for InAs/metAs, 95% confidence interval (CI), 0.34-0.94; r = 0.82 for MMA/DMA, 95%CI, 0.43-0.95] compared to lower correlations in father-mother pairs (r = 0.18, r = -0.01, respectively), after adjustment for total urinary arsenic, age, and sex. Family correlations were not notably altered when adjustments were made for specific blood micronutrients (methionine, homocysteine, folate, vitamin B6, selenium, and vitamin B12 potentially related to methylation. We also report on a family pedigree with high prevalence of arsenic-induced effects. Participants from this family had low InAs/metAs values, which is consistent with increased toxicity of trivalent methylated arsenic species. Despite our small sample size, we observed that methylation patterns aggregate in families and are correlated in siblings, providing evidence of a genetic basis for the variation in arsenic methylation. Larger studies with more extensive pedigrees will need to be conducted to confirm these findings.

Toluene metabolites as biological indicators of exposure.

The measurement of exhaled and excreted xenobiotics and their metabolites can provide accurate, non-invasive, and time-flexible measurements of internal dose. We analyzed rates of exhaled (2)H(8)-toluene and excreted urinary metabolites from 33 exposures of men to 50 ppm of (2)H(8)-toluene for 2 h at rest. The total dose was distributed as follows: exhaled (2)H(8)-toluene, 13 +/- 6.2%; (2)H(5)-hippuric acid, 75 +/- 6.4%; (2)H(7)-o-cresol, 0.31 +/- 0.22%; (2)H(7)-m-cresol, 0.53 +/- 0.44%; and (2)H(7)-p-cresol, 11 +/- 3.8%. Interindividual variability was assessed using the coefficients of variation for peak exhalation or excretion rates, and fractions of dose excreted: (2)H(8)-toluene, c.v.=60, 47%; (2)H(5)-hippuric acid, 29, 8.6%; (2)H(7)-o-cresol, 80, 73%; (2)H(7)-m-cresol, 37, 83%; and (2)H(7)-p-cresol, 38, 34%. Excretion rates of the cresols were stable over the first 5 h post-exposure, and o-cresol was determined to be the best urinary indicator of exposure, given the lower background levels of this isomer. The hippuric acid/cresol rate ratios for the first 5 h post-exposure could be described by single exponential terms, and thus provided a means for estimating time since exposure for any finite toluene duration/exposure combination.