Trimethylarsine Oxide Research Articles

Tissue dosimetry, metabolism and excretion of pentavalent and trivalent dimethylated arsenic in mice after oral administration

Dimethylarsinic acid (DMA(V)) is a rat bladder carcinogen and the major urinary metabolite of administered inorganic arsenic in most mammals. This study examined the disposition of pentavalent and trivalent dimethylated arsenic in mice after acute oral administration. Adult female mice were administered [ 14C]-DMA(V) (0.6 or 60 mg As/kg) and sacrificed serially over 24 h. Tissues and excreta were collected for analysis of radioactivity. Other mice were administered unlabeled DMA(V) (0.6 or 60 mg As/kg) or dimethylarsinous acid (DMA(III)) (0.6 mg As/kg) and sacrificed at 2 or 24 h. Tissues (2 h) and urine (24 h) were collected and analyzed for arsenicals. Absorption, distribution and excretion of [ 14C]-DMA(V) were rapid, as radioactivity was detected in tissues and urine at 0.25 h. For low dose DMA(V) mice, there was a greater fractional absorption of DMA(V) and significantly greater tissue concentrations of radioactivity at several time points. Radioactivity distributed greatest to the liver (1–2% of dose) and declined to less than 0.05% in all tissues examined at 24 h. Urinary excretion of radioactivity was significantly greater in the 0.6 mg As/kg DMA(V) group. Conversely, fecal excretion of radioactivity was significantly greater in the high dose group. Urinary metabolites of DMA(V) included DMA(III), trimethylarsine oxide (TMAO), dimethylthioarsinic acid and trimethylarsine sulfide. Urinary metabolites of DMA(III) included TMAO, dimethylthioarsinic acid and trimethylarsine sulfide. DMA(V) was also excreted by DMA(III)-treated mice, showing its sensitivity to oxidation. TMAO was detected in tissues of the high dose DMA(V) group. The low acute toxicity of DMA(V) in the mouse appears to be due in part to its minimal retention and rapid elimination.

Arsenic Speciation Analysis of Cultivated White Button Mushrooms (Agaricus bisporus) Using High-Performance Liquid Chromatography−Inductively Coupled Plasma Mass Spectrometry, and X-ray Absorption Spectroscopy

Agaricus bisporus mushrooms were grown in compost amended with either arsenic-contaminated mine waste or an arsenate solution, to a final concentration of approximately 200 microg g(-1). Fungi were cultivated at a small-scale mushroom facility in Vineland (ON), where the controlled environment allowed for a large number of fruiting bodies (mushrooms) to be produced. The total arsenic concentrations as well as speciation were examined for each treatment over several harvests (breaks). Total concentrations were determined by acid digestion and inductively coupled plasma mass spectrometry (ICP-MS) detection and ranged from 2.3 to 16 microg g(-1) dry mass in treatment mushrooms. Arsenic compounds were extracted from mushrooms with methanol/water (1:1 v/v), and separated by high-performance liquid chromatography (HPLC, anion/cation exchange) before detection with ICP-MS. Fruiting bodies from all treatments contained arsenite, dimethylarsinic acid (DMA), and arsenobetaine (AB), and to a lesser extent arsenate and trimethylarsine oxide (TMAO). The ratio of arsenic compounds did not vary greatly over the first three harvests. AB was absent in compost not inoculated with A. bisporus supporting the hypothesis that AB is a product of fungal, not microbial, arsenic metabolism. X-ray absorption spectroscopy results lead us to hypothesize that AB plays a role in nutrient translocation within the fruiting body, as well as maintaining turgor pressure to ensure the mushroom cap remains elevated for maximum spore dispersal.

Specific accumulation of arsenic compounds in green turtles ( Chelonia mydas) and hawksbill turtles ( Eretmochelys imbricata) from Ishigaki Island, Japan

Concentrations of total arsenic (As) and individual compounds were determined in green and hawksbill turtles from Ishigaki Island, Japan. In both species, total As concentrations were highest in muscle among the tissues. Arsenobetaine was a major compound in most tissues of both turtles. High concentrations of trimethylarsine oxide were detected in hawksbill turtles. A significant negative correlation between standard carapace length (SCL), an indicator of age, and total As levels in green turtles was found. In contrast, the levels increased with SCL of hawksbill turtles. Shifts in feeding habitats with growth may account for such a growth-dependent accumulation of As. Although concentrations of As in marine sponges, the major food of hawksbill turtles are not high compared to those in algae eaten by green turtles, As concentrations in hawksbill turtles were higher than those in green turtles, indicating that hawksbill turtles may have a specific accumulation mechanism for As.

Bioaccessibility and Transport by Caco-2 Cells of Organoarsenical Species Present in Seafood

Organoarsenical standards and raw and cooked seafood (DORM-2, sole, and Greenland halibut) were subjected to in vitro gastrointestinal digestion to estimate arsenic bioaccessibility (maximum soluble concentration in gastrointestinal medium). The in vitro digestion did not modify the chemical form of the organoarsenic species standards. In seafood, bioaccessibility was 67.5-100% for arsenobetaine (AB), 30% for dimethylarsinic acid (DMA), 45% for tetramethylarsonium ion (TETRA), and >50% for trimethylarsine oxide (TMAO). Cooking induced no changes in bioaccessible contents. In addition, transport by Caco-2 cells, an intestinal epithelia model, was evaluated from organoarsenical standards and DORM-2. For standards, transport ranged from 1.7% for AB to 15.5% for TETRA. In DORM-2, transport was observed for only AB (12%), with far higher efficiency than in the case of the standard solution, thus illustrating the interest of using whole foods for studying bioavailability.

HPLC‐ICP‐MS Speciation Analysis of Arsenic in Urine of Japanese Subjects without Occupational Exposure

The toxicity and carcinogenicity of arsenic depend on its species. Individuals living in Japan consume much seafood that contains high levels of organoarsenics. Speciation analysis of urinary arsenic is required to clarify the health risks of arsenic intake. There has been no report of urinary arsenic analysis in Japan using high performance liquid chromatography with inductively coupled plasma mass spectrometry (HPLC-ICP-MS). We performed speciation analysis of urinary arsenic for 210 Japanese male subjects without occupational exposure using HPLC-ICP-MS. The median values of urinary arsenics were as follows: sodium arsenite (AsIII), 3.5; sodium arsenate (AsV), 0.1; monomethylarsonic acid (MMA), 3.1; dimethylarsinic acid (DMA), 42.6; arsenobetaine (AsBe), 61.3; arsenocholine, trimethylarsine oxide, and unidentified arsenics (others), 5.2; and total arsenic (total As), 141.3 microgAs/l. The median creatinine-adjusted values were as follows: AsIII, 3.0; AsV, 0.1; MMA, 2.6; DMA, 35.9; AsBe, 52.1; others 3.5; and total As, 114.9 microgAs/g creatinine. Our findings indicate that DMA and AsBe levels in Japan are much higher than those found in Italian and American studies. It appears that the high levels of DMA and AsBe observed in Japan may be due in part to seafood intake. ACGIH and DFG set the BEI and BAT values for occupational arsenic exposure as 35 microgAs/l and 50 microgAs/l, respectively, using the sum of inorganic arsenic (iAs), MMA, and DMA. In the general Japanese population, the sums of these were above 50 microgAs/l in 115 (55%) samples. We therefore recommend excluding DMA concentration in monitoring of iAs exposure.

Elevation of 8-hydroxydeoxyguanosine and cell proliferation via generation of oxidative stress by organic arsenicals contributes to their carcinogenicity in the rat liver and bladder

Monomethylarsonic acid (MMA V), dimethylarsinic acid (DMA V) and trimethylarsine oxide (TMAO V) are well-documented inorganic arsenic (iAs) methylated metabolites. In our previous studies, DMA V and TMAO V were shown to exert carcinogenicity in the rat bladder and liver, respectively. Furthermore, MMA V, DMA V and TMAO V exhibited promoting activity on rat hepatocarcinogenesis. To clarify mechanisms of arsenical carcinogenicity and compare biological responses in the liver and bladder, male F344 rats were sequentially treated for 5, 10, 15, 20 days with MMA V, DMA V and TMAO V in their drinking water at a dose of 0.02%. Significant increase of P450 total content and generation of hydroxyl radicals in the liver were observed from 10 and 15 days of treatment with arsenicals, respectively, with the highest levels induced by TMAO V. Similarly, elevation of 8-hydroxy-2′-deoxyguanosine (8-OHdG) formation was found in the DNA with significant increase by TMAO V treatment in the liver at days 15 and 20, and DMA V in the bladder after 20 days treatment. In addition, cell proliferation and apoptosis indices were significantly increased by TMAO V in the liver and by DMA V in the bladder of rats. These events were accompanied by differential up-regulation of phase I and II metabolizing enzymes, cyclins D1 and E, PCNA, caspase 3 and FasL. The results indicate that early elevation of 8-OHdG and cell proliferation via generation of oxidative stress by TMAO V and DMA V contributes to their carcinogenicity in the rat liver and bladder.

Mobile arsenic species in unpolluted and polluted soils

The fate and behaviour of total arsenic (As) and of As species in soils is of concern for the quality of drinking water. To estimate the relevance of organic As species and the mobility of different As species, we evaluated the vertical distribution of organic and inorganic As species in two uncontaminated and two contaminated upland soils. Dimethylarsinic acid (up to 6 ng As g(-1)), trimethylarsine oxide (up to 1.5 ng As g(-1)), 4 unidentified organic As species (up to 3 ng As g(-1)) and arsenobetaine (up to 15 ng As g(-1)), were detected in the forest soils. Arsenobetaine was the dominant organic As species in both unpolluted and polluted forest soils. No organic As species were detected in the contaminated grassland soil. The organic As species may account for up to 30% of the mobile fraction in the unpolluted forest floor, but never exceed 9% in the unpolluted mineral soil. Highest concentrations of organic As species were found in the forest floors. The concentrations of extractable arsenite were highest in the surface horizons of all soils and may represent up to 36% of total extractable As. The concentrations of extractable arsenate were also highest in the Oa layers in the forest soils and decreased steeply in the mineral soil. In conclusion, the investigated forest soils contain a number of organic As species. The organic As species in forest soils seem to result from throughfall and litterfall and are retained mostly in the forest floor. The relative high concentrations of extractable arsenite, one of the most toxic As species, and arsenate in the forest floor point to the risk of their transfer to surface water by superficial flow under heavy rain events.

Interaction of Product Analogues with the Active Site of Rhodobacter Sphaeroides Dimethyl Sulfoxide Reductase

We report a structural characterization using X-ray absorption spectroscopy of Rhodobacter sphaeroides dimethyl sulfoxide (DMSO) reductase reduced with trimethylarsine and show that this is structurally analogous to the physiologically relevant dimethyl sulfide reduced DMSO reductase. Our data unambiguously indicate that these species should be regarded as formal MoIV species and indicate a classical coordination complex of trimethylarsine oxide, with no special structural distortions. The similarity of the trimethylarsine and dimethyl sulfide complexes suggests, in turn, that the dimethyl sulfide reduced enzyme possesses a classical coordination of DMSO with no special elongation of the S-O bond, as previously suggested.

Study on simultaneous speciation of arsenic and antimony by HPLC–ICP-MS

A method was developed for the simultaneous speciation of arsenic and antimony with HPLC–ICP-MS using C30 reversed phase column. Eight kinds of arsenic compounds (As(III), As(V), monomethylarsonic acid (MMAA), dimethylarsinic acid (DMAA), arsenobetaine (AB), arsenocholine (AsC), trimethylarsine oxide (TMAO) and tetramethylarsonium (TeMA)), Sb(III) and Sb(V) were simultaneously separated by the special mobile phase containing ammonium tartrate. Especially for the species of organic As, a C30 column was better than a C18 column in the effect of separation. Limits of detection (LOD) for these elements were 0.2 ng ml −1 for the species of each As, and 0.5 ng ml −1 for the species of each Sb, when a 10 μl of sample was injected, respectively. The proposed method was applied to a hot spring water and a fish sample.

Analysis of Bioavailable Arsenic in Rice with Whole Cell Living Bioreporter Bacteria

A multiwell plate bioassay was developed using genetically modified bacteria (bioreporter cells) to detect inorganic arsenic extracted from rice. The bacterial cells expressed luciferase upon exposure to arsenite, the activity of which was detected by measurement of cellular bioluminescence. The bioreporter cells detected arsenic in all rice varieties tested, with averages of 0.02-0.15 microg of arsenite equivalent per gram of dry weight and a method detection limit of 6 ng of arsenite per gram of dry rice. This amounted to between approximately 20 and 90% of the total As content reported by chemical methods for the same sample and suggested that a major proportion of arsenic in rice is in the inorganic form. Calibrations of the bioassay with pure inorganic and organic arsenic forms showed that the bacterial cells react to arsenite with highest affinity, followed by arsenate (with 25% response relative to an equivalent arsenite concentration) and trimethylarsine oxide (at 10% relative response). A method for biocompatible arsenic extraction was elaborated, which most optimally consisted of (i) grinding rice to powder, (ii) mixing with an aqueous solution containing pancreatic enzymes, (iii) mechanical shearing, (iv) extraction in mild acid conditions and moderate heat, and (v) centrifugation and pH neutralization. Detection of mainly inorganic arsenic by the bacterial cells may have important advantages for toxicity assessment of rice consumption and would form a good complement to total chemical arsenic determination.

Lithium Ion Attachment Mass Spectrometry for the Direct Detection of Organoarsenic Compounds in the Metallomics Studies

Abstract In this study, three organoarsenic compounds including dimethylarsinic acid, trimethylarsine oxide, and arsenic ethoxide were directly detected by coupling a direct sample insertion probe with a lithium ion attachment mass spectrometer. Previously only gaseous sample could be detected by this method; however, solid and liquid samples can also be detected directly with the use of a direct sample insertion probe. This method will be applicable for the direct analysis of various organometallic compounds in the latest metallomics studies.

Biogeochemistry of Organic and Inorganic Arsenic Species in a Forested Catchment in Germany

Little is known about the fate and behavior of diffuse inputs of arsenic (As) species in forested catchments which often are the sources of drinking water. The objective of this study was to investigate the mobility and transformation of different As species in forest ecosystems to assess the environmental risk related to the diffuse pollution of As. We determined concentrations and fluxes in precipitation, litterfall, soil solutions (Oa horizon and 20- and 90-cm depth), and runoff of organic and inorganic As species and Astotal in a forest ecosystem in NE-Bavaria, Germany. The concentrations of Astotal were mostly <1 microg As L(-1) in aqueous samples and were highest in forestfloor percolates (7.6 microg As L(-1)). In litterfall, the concentrations of As species never exceeded 0.1 microg As g(-1). Arsenate and arsenite were the prevalent As species in all samples. Organic As species, comprising monomethylarsonic acid, dimethylarsinic acid, trimethylarsine oxide, arsenobetaine, and three unidentified organic As species, were mostly found in throughfall reaching up to 45% of Astotal. The total deposition of Astotal (calculated as throughfall + litterfall) was 5.6 g As ha(-1) yr(-1) with 16% contribution of litterfall. The annual Astotal fluxes were 30 g As ha(-1) yr(-1) for forest floor percolates, 8.0 g As ha(-1) yr(-1) at 20-cm soil depth, and 1.4 g As ha(-1) yr(-1) at 90-cm soil depth. The annual runoff of Astotal from the catchment amounted to 3.8 g As ha(-1) yr(-1). The annual fluxes of total organic As species was highest in total deposition (1.1 g As ha(-1) yr(-1)) and decreased largely with depth in the soil profile. The annual runoff of total organic As species was only 0.08 g As ha(-1) yr(-1). Significant correlations in soil solutions and runoff were found between Astotal and dissolved organic C and Fe. Correlations between Astotal concentrations in runoff and water fluxes were seasonally dependent and with a steeper slope in the growing season than in the dormant season. The elevated concentrations of organic As species in throughfall indicate microbial methylation of As in the phyllosphere, but no evidence for methylation in the soil was found. The mass balance of the catchment points out the strong retention and probable degradation or oxidation of organic As species and arsenite but also to mobilizable pools of Astotal and arsenate. The forest floor is presently a source, whereas the mineral soil is a sink for Astotal and arsenate. The As concentrations in runoff seem to be controlled by As mobilization from forest floor and riparian wetland soils during heavy rain events and superficial flow. The risk for excessment of the drinking water threshold concentrations of As in runoff and soil solutions is considered low at our site.

Optimisation by experimental design of an IEC/ICP-MS speciation method for arsenic in seafood following microwave assisted extraction

An experimental design was used to optimise a speciation method involving ion exchange chromatography coupled to inductively coupled plasma-mass spectrometry (IEC-ICP-MS) for the determination of arsenic species in fish samples following a microwave-assisted extraction (MAE). The effects of several factors (eluent composition and concentration, gradient program, mobile phase flow rate, column temperature and plasma power), and their possible interactions on the arsenic species separation and detection, were evaluated using an experimental design in order to establish the optimal conditions. Two mobile phases (A and B) composed of HNO3 (0.5 and 50 mmol L−1 respectively) and MeOH (1%) were used in a gradient consisting of 100% A for 2.5 min, 10% A for 3 min, and 80% A for 5 min, on a Dionex Ionpac AS 7 high-capacity anion-exchange column maintained at 20 °C, with a mobile phase flow rate of 1.35 mL min−1 and a rf-power to the plasma of 1450 W. Under these optimised conditions, seven different arsenic species (AsIII, AsV, monomethylarsonic acid, dimethylarsinic acid, arsenobetaine, arsenocholine, trimethylarsine oxide) could be separated within 11 min in a single chromatographic run. Limits of quantification ranged from 0.008 to 0.042 mg As kg−1, which were generally similar to those previously reported. Moreover, trueness was estimated on three different certified reference materials of fish after MAE with 10 mL of methanol–water mixture (8 + 2 v/v). In all cases the results were in good agreement with the certified values.

Arsenic accumulation and speciation in freshwater fish living in arsenic-contaminated waters

Environmental context. Inorganic arsenic, a well-known human carcinogen, represents a major worldwide environmental problem because contaminated water supplies have lead to widespread human exposure. This study investigates the arsenic content of freshwater fish from arsenic-contaminated and non-contaminated sites in Thailand, and reports high arsenic concentrations and significant amounts of inorganic arsenic in the edible muscle tissue. The data suggest that freshwater fish may represent a significant source of inorganic arsenic to some human populations. Abstract. Striped snakehead (Channa striata), carnivorous freshwater fish that serve as popular food in Thailand, were collected from a reference site (1.4 µg As L–1) and from two arsenic-contaminated ponds (Pond A, 550 µg As L–1; Pond B, 990 µg As L–1) in southern Thailand and analysed for arsenic by inductively coupled plasma mass spectrometry (ICPMS) and for arsenic species by HPLC-ICPMS performed on aqueous methanol extracts of muscle, liver and gill (n = 3 fish from each site). Mean total arsenic concentration in muscle tissue of C. striata collected from the reference site was 1.9 µg As g–1 (dry mass) while fish from the contaminated sites contained 13.1 µg As g–1 (Pond A) and 22.2 µg As g–1 (Pond B). Liver and gill tissues showed similar increasing arsenic concentrations on going from the reference site to Ponds A and B, with Pond B showing the highest levels. Speciation analysis on the three tissues showed that, although arsenate was the major extractable arsenical in reference fish (e.g. 0.73 µg As g–1 in muscle tissue), dimethylarsinate was by far the dominant arsenic species in fish from the two contaminated sites. Three non-carnivorous fish species (Danio regina, Rasbora heteromorpha and Puntius orphoides), collected from Pond B only, had lower arsenic concentrations (7.9–11.3 µg As g–1 in muscle tissue) than did C. striata, and contained appreciable amounts of trimethylarsine oxide together with two other major arsenicals, arsenate and dimethylarsinate, and smaller quantities of arsenite and methylarsonate. The study shows for the first time a clear effect of water arsenic concentrations on natural fish tissue arsenic concentrations, and is the first report of a freshwater fish species attaining arsenic concentrations comparable with those found in marine fish species. Furthermore, the high concentrations of toxic inorganic arsenic (predominantly arsenate) in the muscle tissue of the edible fish C. striata have human health implications and warrant wider investigations.

Quantification of seven arsenic compounds in seafood products by liquid chromatography/electrospray ionization-single quadrupole mass spectrometry (LC/ESI-MS)

A liquid chromatography/electrospray ionization-single quadrupole mass spectrometry (LC/ESI-MS) method was developed to quantify seven arsenic compounds: arsenate (As(V)), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), arsenobetaine (AB), trimethylarsine oxide (TMAO), arsenocholine (AC) and tetramethylarsonium ion (TEMA), widely found in seafood. The arsenicals separated by anion- or cation-exchange LC were all readily identified under the optimized ESI-MS conditions. Linear calibration curves constructed by plotting the peak area counts of molecular ions against the arsenic concentrations were obtained for all seven arsenic compounds. The limits of quantification (S/N = 10) were 800, 600, 50, 10, 5, 5 and 5 ng ml−1 for As(V), MMA, DMA, AB, TMAO, AC and TEMA, respectively. The LC/ESI-MS method was found to be useful to quantify arsenic compounds in seafood by model experiments using the mid-gut gland and muscle of a shellfish (Buccinid whelks). Spiking experiments verified the accuracy of the method.

Analysis of arsenic compounds by capillary electrophoresis using indirect UV and mass spectrometric detections

CE with indirect UV and mass-spectrometric detection was used for the simultaneous determination of arsenic acid (As(V)), arsenous acid (As(III)), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), trimethylarsine oxide (TMAO), tetramethylarsonium ion (TMA(+)), arsenobetaine (AB), and arsenocholine (AC). In the CE-indirect UV analysis, a baseline separation of arsenic species was successfully achieved by using a basic background solution (BGS) for anions and an acidic BGS for cations, respectively. The LOD values in CE-indirect UV for the individual analytes were 7.8, 12.5, 7.8, 12.5, 62.5, 125, 250, and 62.5 ppm, respectively. To achieve sensitive and selective analysis, CE coupled with ESI-MS was applied to the determination of arsenic compounds. The organic arsenic species were successfully separated with a higher sensitivity by CE-MS using the acidic BGS. The LODs in CE-MS for MMA, DMA, TMAO, TMA(+), AB, and AC were 1.0, 0.1, 0.01, 0.1, 0.01, and 0.01 ppm, respectively. In contrast, the analysis of inorganic arsenic species (As(V) and As(III)) resulted in a lower detectability in CE-MS compared to that obtained with the CE-indirect UV analysis. However, the speciation of eight arsenics by CE-MS was successfully achieved in a single run by switching the ESI polarity during MS detection.

Arsenic speciation in moso bamboo shoot — A terrestrial plant that contains organoarsenic species

Arsenic is predominantly found as an inorganic species in most terrestrial plants. However, we found that a significant proportion of organic arsenic was present in moso bamboo (Phyllostachys pubescens Mazel) shoot in a market survey of arsenic species in edible terrestrial plants. Moso bamboo shoots from different producing areas in China were collected for analysis to confirm the ubiquity of methylated arsenic species. The total arsenic concentrations of bamboo shoots were determined by hydride generation coupled atomic fluorescence spectrometry (HG-AFS), ranging from 27.7 to 94.0 microg/kg. Information about arsenic species was acquired from cold trap-hydride generation-atomic absorption spectrometry (CT-HG-AAS). Dimethylarsinic acid (DMA) was present in the amount of 13.9% to 44.9% of sum of the arsenic species in all these samples. Monomethylarsonic acid (MMA) and trimethylarsine oxide (TMAO) were also detected in certain samples in the range of 4.2-16.5% and 11.8-18.4%, respectively. In addition, bamboo shoots collected in winter were found to have more total arsenic and organic arsenic than those collected in spring. To investigate the source of the organic arsenic in moso bamboo shoots, arsenic species in the rhizosphere soils of the plants were examined. The absence of organic arsenic in soils would suggest the possibility of formation of methylated arsenic in the plants. In addition, studies of arsenic speciation in the peel and core of winter bamboo shoots showed that all the cores contained organic arsenic while no organic arsenic was detected in the peels. The study provides useful information for better understanding of the distribution of arsenic species in terrestrial plants.

Dynamics of organic and inorganic arsenic in the solution phase of an acidic fen in Germany

Wetland soils play a key role for the transformation of heavy metals in forested watersheds, influencing their mobility, and ecotoxicity. Our goal was to investigate the mechanisms of release from solid to solution phase, the mobility, and the transformation of arsenic species in a fen soil. In methanol–water extracts, monomethylarsonic acid, dimethylarsinic acid, trimethylarsine oxide, arsenobetaine, and two unknown organic arsenic species were found with concentrations up to 14 ng As g −1 at the surface horizon. Arsenate is the dominant species at the 0–30 cm depth, whereas arsenite predominated at the 30–70 cm depth. Only up to 2.2% of total arsenic in fen was extractable with methanol–water. In porewaters, depth gradient spatial variation of arsenic species, pH, redox potentials, and the other chemical parameters along the profile was observed in June together with high proportion of organic arsenic species (up to 1.2 μg As L −1, 70% of total arsenic). Tetramethylarsonium ion and an unknown organic arsenic species were additionally detected in porewaters at deeper horizons. In comparison, the arsenic speciation in porewaters in April was homogeneous with depth and no organic arsenic species were found. Thus, the occurrence of microbial methylation of arsenic in fen was demonstrated for the first time. The 10 times elevated total arsenic concentrations in porewaters in June compared to April were accompanied by elevated concentrations of total iron, lower concentrations of sulfate and the presence of ammonium and phosphate. The low proportion of methanol–water extractable total arsenic suggests a generally low mobility of arsenic in fen soils. The release of arsenic from solid to solution phases in fen is dominantly controlled by dissolution of iron oxides, redox transformation, and methylation of arsenic, driven by microbial activity in the growing season. As a result, increased concentrations of total arsenic and potentially toxic arsenic species in fen porewaters were found in the growing season, suggesting an enhancing risk of arsenic transport of ground- and surface-waters under these conditions.

Chronic Exposure to Methylated Arsenicals Stimulates Arsenic Excretion Pathways and Induces Arsenic Tolerance in Rat Liver Cells

Although inorganic arsenicals are toxic and carcinogenic in humans, inorganic arsenite has recently emerged as a highly effective chemotherapeutic agent for acute promyelocytic leukemia (APL). Inorganic arsenicals are enzymatically methylated to monomethylarsonic acid (MMAs(V)), dimethylarsinic acid (DMAs(V)), and trimethylarsine oxide (TMAs(V)O) in mammals. We examined the effects of chronic exposure to methylated arsenicals on arsenic tolerance by using rat normal liver TRL 1215 cells. TRL 1215 cells were exposed for 20 weeks to MMAs(V), DMAs(V), or TMAs(V)O at levels that produced submicromolar cellular concentrations of arsenic. On chronic exposure to these methylated arsenicals, the cells acquired tolerance to acute arsenic cytolethality. Cellular arsenic uptake was reduced in these cells compared to passage-matched control cells. The long-term arsenic exposure increased glutathione S-transferase (GST) activity and cellular glutathione (GSH) levels. Glutathione S-transferase, multidrug resistance-associated proteins (Mrps; efflux transporters encoded by Mrp genes), and P-glycoprotein [P-gp; efflux transporter encoded by multidrug resistance gene (MDR)] had also increased in these cells at the transcript and protein levels. The depletion of cellular GSH and the inhibition of Mrps and P-gp functions increased cellular arsenic uptake and reduced arsenic tolerance in these cells. These results indicate that chronic exposure to methylated arsenicals induces a generalized arsenic tolerance that is caused by increased arsenic excretion. Because accumulation of methylated arsenicals may occur in patients with chronic arsenic poisoning and arsenic-treated APL patients, this study may provide important information regarding chronic arsenic poisoning and the latent risk of developing multidrug resistance in APL therapy using inorganic arsenite.

Determination of Arsenic Species in Marine Samples by HPLC-ICP-MS

Arsenic speciation analysis in marine samples was performed using high performance liquid chromatography (HPLC) with ICP-MS detection. The separation of eight arsenic species viz. arsenite (As(III)), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), arsenate (As(V)), arsenobetaine, trimethylarsine oxide (TMAO), arsenocholine and tetramethylarsonium ion (TeMAs) was achieved on a Shiseido Capcell Pak C18 column by using an isocratic eluent (pH 3.0), in which condition As(III) and MMA were co-eluted. The entire separation was accomplished in 15 min. The detection limits for 8 arsenic species by HPLC/ICP-MS were in the range of 0.02 - 0.10 microg L(-1) based on 3sigma of blank response (n=9). The precision was calculated to be 3.1-7.3% (RSD) for all eight species. The method then successfully applied to several marine samples e.g., oyster, scallop, fish, and shrimps. For the extraction of arsenic species from seafood products, the low power microwave digestion was employed. The extraction efficiency was in the range of 52.9 - 112.3%. Total arsenic concentrations were analyzed by using the microwave acid digestion. The total arsenics in the certified reference materials (DORM-2 and TORT-2) were analyzed and agreed with the certified values. The concentrations of arsenics in marine samples were in the range 6.6 - 35.1 microg g(-1).