Acid extraction is commonly used to analyze arsenic species in rice. During the extraction process, spiked monomethylarsonic acid (MMA) is often transformed into different compounds. A similar phenomenon is observed in the arsenic speciation analysis of seafood. To identify these compounds, we analyzed a previously prepared extract using liquid chromatography-time-of-flight/mass spectrometry in differential analysis and liquid chromatography-inductively coupled plasma-MS. The compound was identified as monomethylmonothioarsonic acid (MMMTA), a thioarsenical, which is estimated to be more cytotoxic than MMA. As MMMTA was readily produced by bubbling hydrogen sulfide through MMA, this suggests that MMA reacts with sulfur in rice during the extraction process. Our data also suggested that dimethylarsinic acid could be transformed into another compound, although the generation rate was low. For reliable arsenic speciation analyses, the transformation of arsenic compounds during extraction must be avoided. This study demonstrates that arsenic compounds can be transformed by dilute acid extraction.

Long term exposure to arsenic through consumption of contaminated groundwater has been a global issue since the last five decades; while from an alternate standpoint, arsenic compounds have emerged as unparallel chemotherapeutic drugs. This review highlights the contribution from arsenic speciation studies that have played a pivotal role in the progression of our understanding of the biological behaviour of arsenic in humans. We also discuss the limitations of the speciation studies and their association with the interpretation of arsenic metabolism. Chromatographic separation followed by spectroscopic detection as well as the utilization of biotinylated pull-down assays, protein microarray and radiolabelled arsenic have been instrumental in identifying hundreds of metabolic arsenic conjugates, while, computational modelling has predicted thousands of them. However, these species exhibit a variegated pattern, which supports more than one hypothesis for the metabolic pathway of arsenic. Thus, the arsenic species are yet to be integrated into a coherent mechanistic pathway depicting its chemicobiological fate. Novel biorelevant arsenic species have been identified due to significant evolution in experimental methodologies. However, these methods are specific for the identification of only a group of arsenicals sharing similar physiochemical properties; and may not be applicable to other constituents of the vast spectrum of arsenic species. Consequently, the identity of arsenic binding partners in vivo and the sequence of events in arsenic metabolism are still elusive. This resonates the need for additional focus on the extraction and characterization of both low and high molecular weight arsenicals in a combinative manner.

The importance of recognizing and quantifying chemical anions/cations found in various types of samples, including environmental and biological samples, has been extensively studied. Recent findings suggest the possibility of health risks caused by organic compound dimethylarsinic acid (DMAs) rather than its inorganic arsenic metabolite.

This article aims to fabricate polymeric-membrane electrochemical sensors with high sensitivity and selectivity for the cacodylic acid sodium salt dimethylarsinate (DMAs) based on silver diethyldithiocarbamate (AgDDTC) and CuIIphthalocyanine (CuPC) as novel neutral carriers and their applications.

DMAs calibration relations and titrations were carried out using a potentiometric workstation equipped with a double-junction reference electrode, in conjunction with the fabricated working electrodes.

Sensors revealed fast and stable anionic response with near-Nernstian slopes (-38.6 ± 0.9 and -31.5 ± 0.6 mV/decade), within concentration ranges (1.7 × 10-5 -1.0 × 10-2 and 3.0 × 10-5 -1.0 × 10-2 M) and detection limits (1.0 × 10-5 and 1.6 × 10-5 M) for AgDDTC- and CuPC-based sensors, respectively. Sensors are characterized by extended life-time, signal stability, high precision and short response times. Selectivity for the cacodylate anion over most common anions was tested for the proposed electrodes. Sensors were satisfactorily applied for DMAs quantification in biological matrices with recoveries ranging between 96.2 and 99.0%. Membrane sensors were interfaced with a flow-through system for continuous monitoring of DMAs. The sensors were tested for the assay of different amino acids based on their reaction with cacodylate, where reaction end points were monitored with the proposed electrodes using direct potentiometric determination and flow injection analysis (FIA).

Potentiometric ion-selective PVC-membrane electrodes based on silver diethyldithiocarbamate (AgDDTC) and CuIIphthalothyanine (CuPC) provide adequate and reliable means for the determination of dimethylarsenate anion (cacodylate anion, DMAs). These membrane electrodes are easy to manufacture, they have the advantages of high selectivity and sensitivity, broad dynamic ranges, low detection limits, quick response times and cost effectiveness. Such properties make these sensors suitable for the assay of DMAs levels in aqueous solutions by direct potentiometry, flow injection and potentiometric titration, as well as in monitoring of the titration end points of the reactions between various amino acids and DMAs anion in aqueous solutions.

Simple electrochemical membranes for dimethylarsinate (DMAs) were prepared, based on diethyldithiocarbamate (AgDDTC) and CuIIphthalocyanine (CuPC). - DMAs sensors were fabricated in two different modules: batch (for static) and flow-through (for hydrodynamic) approaches. - Levels of DMAs were determined in spiked biological samples. - AgDDTC-based sensors were successfully applied in the determination of several amino acids via potentiometric titration with DMAs.

Thioarsenicals, such as dimethylmonothioarsinic acid (DMMTA^V) and dimethyldithioarsinic acid (DMDTA^V), have been increasingly discovered as important arsenic metabolites, yet analysis of these unstable arsenic species remains a challenging task. A method based on surface-enhanced Raman spectroscopy (SERS) detection in combination with the coffee ringeffect for separation is expected to be particularly useful for analysis of thioarsenicals, thanks to minimal sample pretreatment and unique fingerprint Raman identification. Such a method would offer an alternative approach that overcomes limitations of conventional arsenic speciation techniques based on high performance liquid chromatography separation and mass spectrometry detection. A novel analytical method based on combination of the coffee ringeffect and SERS was developed for the speciation of thiolated arsenicals. A gold nanofilm (AuNF) was employed not only as a SERS substrate, but also as a platform for the separation of thioarsenicals. Once a drop of the thioarsenicals solution was placed onto the AuNF and evaporation of the solvent and the ring stamp formation onto AuNF began, the SERS signal intensity substantially increased from center to edge regions of the evaporated droplet due to the presence of the coffee ring effect. Through calculating the pKa's of DMMTA^V and DMDTA^V and accordingly manipulating the chemical environment, separation of these thioarsenicals was realized as they travelled different distances during the development of the coffee ring. The migration distances of individual species were influenced by a radial outward flow of a solute, the thioarsenicals-AuNF interactions and a thermally induced Marangoni flow. The separation of DMMTA^V (center) and DMDTA^V (edge) on the coffee ring, in combination with fingerprint SERS spectra, enables the identification of these thioarsenicals by this AuNF-based coffee ring effect-SERS method.

Diet, especially seafood, is the main source of arsenic exposure for humans. The total arsenic content of a diet offers inadequate information for assessment of the toxicological consequences of arsenic intake, which has impeded progress in the establishment of regulatory limits for arsenic in food. Toxicity assessments are mainly based on inorganic arsenic, a well-characterized carcinogen, and arsenobetaine, the main organoarsenical in seafood. Scarcity of toxicity data for organoarsenicals, and the predominance of arsenobetaine as an organic arsenic species in seafood, has led to the assumption of their nontoxicity. Recent toxicokinetic studies show that some organoarsenicals are bioaccessible and cytotoxic with demonstrated toxicities like that of pernicious trivalent inorganic arsenic, underpinning the need for speciation analysis. The need to investigate and compare the bioavailability, metabolic transformation, and elimination from the body of organoarsenicals to the well-established physiological consequences of inorganic arsenic and arsenobetaine exposure is apparent. This review provides an overview of the occurrence and assessment of human exposure to arsenic toxicity associated with the consumption of seafood.

Dimethylated thioarsenicals such as dimethylmonothioarsinic acid (DMMTA^V) and dimethyldithioarsinic acid (DMDTA^V), which are produced by the metabolic pathway of dimethylarsinic acid (DMA^V) thiolation, have been recently found in the environment as well as human organs. DMMTA^V and DMDTA^V can be quantified to determine the ecological effects of dimethylated thioarsenicals and their stability in environmental media. The synthesis method for these compounds is unstandardized, making replicating previous studies challenging. Furthermore, there is a lack of information about storage techniques, including storage of compounds without species transformation. Moreover, because only limited information about synthesis methods is available, there may be experimental difficulties in synthesizing standard chemicals and performing quantitative analysis. The protocol presented herein provides a practically modified synthesis method for the dimethylated thioarsenicals, DMMTA^V and DMDTA^V, and will help in the quantification of species separation analysis using high performance liquid chromatography in conjunction with inductively coupled plasma mass spectrometry (HPLC-ICP-MS). The experimental steps of this procedure were modified by focusing on the preparation of chemical reagents, filtration methods, and storage.

Hundreds of millions of people around the world are exposed to elevated concentrations of inorganic and organic arsenic compounds, increasing the risk of a wide range of health effects. Studies of the environmental fate and human health effects of arsenic require authentic arsenic compounds. We summarize here the synthesis and characterization of more than a dozen methylated and thiolated arsenic compounds that are not commercially available. We discuss the methods of synthesis for the following 14 trivalent (III) and pentavalent (V) arsenic compounds: monomethylarsonous acid (MMA^III), dicysteinylmethyldithioarsenite (MMA^III(Cys)₂), monomethylarsonic acid (MMA^V), monomethylmonothioarsonic acid (MMMTA^V) or monothio-MMA^V, monomethyldithioarsonic acid (MMDTA^V) or dithio-MMA^V, monomethyltrithioarsonate (MMTTA^V) or trithio-MMA^V, dimethylarsinous acid (DMA^III), dimethylarsino-glutathione (DMA^III(SG)), dimethylarsinic acid (DMA^V), dimethylmonothioarsinic acid (DMMTA^V) or monothio-DMA^V, dimethyldithioarsinic acid (DMDTA^V) or dithio-DMA^V, trimethylarsine oxide (TMAO^V), arsenobetaine (AsB), and an arsenicin-A model compound. We have reviewed and compared the available methods, synthesized the arsenic compounds in our laboratories, and provided characterization information. On the basis of reaction yield, ease of synthesis and purification of product, safety considerations, and our experience, we recommend a method for the synthesis of each of these arsenic compounds.

Dimethylmonothioarsinic acid (DMMTA^V) is a highly toxic, thiolated analogue of dimethylarsinic acid (DMA^V). In comparison, a further thiolated analogue, dimethyldithioarsinic acid (DMDTA^V), and DMA^V both exhibit lower toxicity. To understand the environmental conditions responsible for forming DMMTA^V, the kinetics of DMA^V thiolation are examined. The thiolation of DMA^V is pH-dependent and consists of two consecutive first-order reactions under excess sulfide conditions. The first thiolation of DMA^V to form DMMTA^V is faster than the second one to DMDTA^V. DMMTA^V is therefore an intermediate. The first reaction is first-order in H₂S at pH 6.0 and 20 °C; therefore, the overall reaction is second-order and the rate coefficient in this condition is 0.0780 M^-1 s^-1. The rate coefficient significantly decreases at pH 8.0, indicating that H₂S(aq) triggers the thiolation of DMA^V. The second reaction rate is significantly decreased at pH 2.5; therefore, reaction under strongly acidic conditions leads to accumulation of highly toxic DMMTA^V in the early stages of thiolation. The transformation of DMDTA^V to DMMTA^V is catalyzed in the presence of ferric iron. Formation of DMMTA^V should be considered when assessing risk posed by arsenic under sulfidic or sulfate reducing conditions.

Dimethyldithioarsinic acid (DMDTA(V)), present in such intense sources as municipal landfill leachate, has drawn a great deal of attention due to its abundant occurrence and different aspect of toxicity. The hydrosulfide (HS(-)) concentration in leachate was studied as a major variable affecting the formation of DMDTA(V). To this end, the HPLC-ICPMS system equipped with the reversed-phase C18 column was used to determine DMDTA(V). Simulated landfill leachates (SLLs) were prepared to cover a mature landfill condition with the addition of sodium sulfate and sulfide at varying concentrations in the presence of dimethylarsinic acid (DMA(V)). The concentration of sodium sulfide added in the SLLs generally exhibited a strong positive correlation with the concentration of DMDTA(V). As such, the formation of DMDTA(V) in the SLLs is demonstrated to be controlled by the interactive relationship between DMA(V) and the HS(-).

The International Agency for Research on Cancer (IARC) has concluded that dimethylarsinic acid [(CH3)2AsO(OH), DMA(V)], a main metabolite of inorganic arsenic, is responsible for carcinogenesis in urinary bladder and lung in rodents, and various modes of carcinogenic action have been proposed. One theory concerning the mode of action is that the biotransformation of dimethylarsinous acid [(CH3)2AsOH, DMA(III)] from DMA(V) plays an important role in the carcinogenesis by way of reactive oxygen species (ROS) production. Furthermore, dimethylmonothioarsinic acid [(CH3)2AsS(OH), DMMTA(V)], a metabolite of DMA(V), has also been noted because of its higher toxicity. However, the metabolic mechanisms of formation and disappearance of DMA(III) and DMMTA(V), and their toxicity are not fully understood. Thus, the purpose of the present study was to clarify the mechanism of metabolic formation of DMMTA(V) and DMA(V) from DMA(III). The in vitro transformation of arsenicals by treatment with liver homogenate from rodents and sulfur transferase was detected by HPLC-ICP-MS and HPLC-tandem MS. DMMTA(V) is produced from DMA(III) but not DMA(V) by cellular fractions from mouse liver homogenates and by rhodanese from bovine liver in the presence of thiosulfate, a sulfur donor. Not only DMMTA(V) thus produced but also DMA(III) are re-converted into DMA(V) by an in vitro addition of S9 mix. These findings indicate that the metabolic process not only of DMA(III) to DMA(V) or DMMTA(V) but also of DMMTA(V) to DMA(V) consists of a complicated mode of interaction between monooxygenase including cytochrome P450 (CYP) and/or sulfur transferase.

Although inorganic arsenic has long been recognized as a potent toxicant and carcinogen in humans, recent evidence shows that at least some of its effects are mediated by methylated metabolites. Elucidating the conversion of inorganic arsenic to mono-, di-, and trimethylated species has provided insights into the enzymology of this pathway and identified genetic and environmental factors that influence the susceptibility of individuals to this metalloid's adverse health effects. Notably, almost all work on the formation, fate, and effects of methylated arsenicals has focused on oxoarsenicals in which arsenic is bound to one or more oxygen atoms. However, thioarsenicals are a class of arsenicals in which a sulfur atom has replaced one or more oxygens that are bound to arsenic. Thioarsenicals have been identified as urinary metabolites in humans and other animals following exposure to inorganic arsenic. Studies find that methylated thioarsenicals exhibit kinetic behavior and toxicological properties that distinguish them from methylated oxoarsenicals. This perspective considers that formation, fate, and effects of methylated thioarsenicals with an emphasis on examining the linkages between the molecular processes that underlie both methylation and thiolation reactions. Integrating this information will provide a more comprehensive view of the relationship between the metabolism of arsenic and the risk posed by chronic exposure to this environmental contaminant.

Diphenylarsinic acid (DPAA) is a toxic phenylarsenical compound often found around sites contaminated with phenylarsenic chemical warfare agents, diphenylcyanoarsine or diphenylchloroarsine, which were buried in soil after the World Wars. This research concerns the elucidation of the chemical structure of an arsenic metabolite transformed from DPAA under anaerobic sulfate-reducing soil conditions. In LC/ICP-MS analysis, the retention time of the metabolite was identical to that of a major phenylarsenical compound synthesized by chemical reaction of DPAA and hydrogen sulfide. Moreover the mass spectra for the two compounds measured using LC/TOF-MS were similar. Subsequent high resolution mass spectral analysis indicated that two major ions at m/z 261 and 279, observed on both mass spectra, were attributable to C12H10AsS and C12H12AsSO, respectively. These findings strongly suggest that the latter ion is the molecular-related ion ([M+H](+)) of diphenylthioarsinic acid (DPTA; (C6H5)2AsS(OH)) and the former ion is its dehydrated fragment. Thus, our results reveal that DPAA can be transformed to DPTA, as a major metabolite, under sulfate-reducing soil conditions. Moreover, formation of diphenyldithioarsinic acid and subsequent dimerization were predicted by the chemical reaction analysis of DPAA with hydrogen sulfide. This is the first report to elucidate the occurrence of DPAA-thionation in an anaerobic soil.

Dimethylarsinic acid (DMA(V)) was pre-concentrated from water samples using a strong cation exchange (SCX) disk functionalized with sulfonic groups, before being analyzed by wavelength dispersive X-ray fluorescence spectrometry (WDXRF). The adsorption of DMA(V) occurred preferentially on the surface of the SCX disk, regardless of pH levels, probably due to interactions with the sulfonic functional groups. However, no other arsenic species, such as arsenate (iAs(V)), arsenite (iAs(III)), and monomethylarsonic acid (MMA(V)), were retained. The SCX-WDXRF method produced a strongly linear calibration curve (R(2)=0.9996) with its limit of detection at 0.218 μgL(-1) when a one-liter water sample was used for pre-concentration. The As intensity of the system was sensitive to the Pb content retained on the SCX disk owing to the proximity of the As-Kα and Pb-Lα lines. To compensate for this interference, a correction factor was developed by considering the calibration slope ratio between the X-ray intensity measured at a Bragg angle of 48.781° and the Pb content of the SCX disks. The results of spike tests for iAs(V), iAs(III), MMA(V), and DMA(V) with and without the addition of Pb in synthetic landfill leachate exhibited reasonable recoveries (i.e., 98-105%) after the spectral adjustment for the Pb interference.

Arsenic is a worldwide environmental pollutant and a human carcinogen. It is well recognized that the toxicity of arsenicals largely depends on the oxidoreduction states (trivalent or pentavalent) and methylation levels (monomethyl, dimethyl, and trimethyl) that are present during the process of metabolism in mammals. However, presently, the specifics of the metabolic pathway of inorganic arsenicals have yet to be confirmed. In mammals, there are two possible mechanisms that have been proposed for the metabolic pathway of inorganic arsenicals, oxidative methylation, and glutathione conjugation. Oxidative methylation, which was originally proposed in fungi, is based on findings that arsenite (iAs(III)) is sequentially converted to monomethylarsonic acid (MMA(V)) and dimethylarsinic acid (DMA(V)) in both humans and in laboratory animals such as mice and rats. However, recent in vitro observations have demonstrated that arsenic is only methylated in the presence of glutathione (GSH) or other thiol compounds, which strongly suggests that arsenic is methylated in trivalent forms. The glutathione conjugation mechanism is supported by findings that have shown that most intracellular arsenicals are trivalent and excreted from cells as GSH conjugates. Since non-conjugated trivalent arsenicals are highly reactive with thiol compounds and are easily converted to less toxic corresponding pentavalent arsenicals, the arsenic-glutathione conjugate stability may be the most important factor for determining the toxicity of arsenicals. In addition, "being a non-anionic form" also appears to be a determinant of the toxicity of oxo-arsenicals or thioarsenicals. The present review discusses both the metabolism of arsenic and the toxicity of arsenic metabolites.

The occurrence, distribution, speciation, and biotransformation of arsenic in aquatic environment (marine and freshwater) have been studied extensively by several research groups during last couple of decades. However, most of those studies have been conducted in marine waters, and the results are available in a number of reviews. Speciation, bioaccumulation, and biotransformation of arsenic in freshwaters have been studied in recent years. Although inorganic arsenic (iAs) species dominates in both marine and freshwaters, it is biotransformed to methyl and organoarsenic species by aquatic organisms. Phytoplankton is considered as a major food source for the organisms of higher trophic levels in the aquatic food chain, and this autotrophic organism plays important role in biotransformation and distribution of arsenic species in the aquatic environment. Bioaccumulation and biotransformation of arsenic by phytoplankton, and trophic transfer of arsenic in marine and freshwater food chains have been important concerns because of possible human health effects of the toxic metalloid from dietary intake. To-date, most of the studies on arsenic biotransformation, speciation, and trophic transfer have focused on marine environments; little is known about these processes in freshwater systems. This article has been reviewed the bioaccumulation, biotransformation, and trophic transfer of arsenic in marine and freshwater food chain.

The microbial transformation of arsenic species in municipal landfill leachate (MLL) was investigated with the objective to highlight arsenic transformation in the landfill system. Across the 43 day incubation in MLL, more than 90% arsenate (iAs(V)) was found to reduce to arsenite (iAs(III)) within 20 days, while iAs(III) was comparably stable although a fraction of iAs(III) was temporarily oxidated to iAs(V) in the first 3 days. Transformation of monomethylarsonic acid (MMA(V)) to dimethylarsinic acid (DMA(V)) in MLL was slow with only 5% MMA(V) methylated to DMA(V) after 43 days incubation. A portion of DMA(V) and MMA(V) in MLL was demonstrated to transform into thiol-organoarsenic and monomethylarsonous acid (MMA(III)), which were identified to include dimethyldithioarsinic acid (DMDTA(V)), dimethylmonothioarsinic acid (DMMTA(V)) and monomethyldithioarsonic acid (MMDTA(V)) by HPLC-ICPMS and LC-ESI-MS/MS. The microbial formation of DMDTA(V), DMMTA(V) and MMDTA(V) is postulated to relate to hydrogen sulfide generated by bacteria in MLL. Differences in arsenic transformation in sterilised and non-sterilised MLLs demonstrate bacteria play a crucial role in arsenic transformation in the landfill body. This study reveals the complexity of arsenic speciation and highlights the potential risk of forming highly toxic thiol-organoarsenic and MMA(III) in the landfill environment.

The conventional scheme for arsenic methylation accounts for methylated oxyarsenical production but not for thioarsenical formation. Here, we report that in vitro anaerobic microbiota of mouse cecum converts arsenate into oxy- and thio- arsenicals. Besides methylarsonic acid (MMA(V)), arsenate was transformed into six unique metabolites: mono-, di-, and trithio-arsenic acid, monomethyldithio- and monomethyltrithio-arsonic acid, and dimethyldithioarsonic acid. Thioarsenicals were found in soluble and particulate fractions of reaction mixtures, suggesting interactions with anaerobic microbiota. Metabolism of ingested arsenate to oxy- and thio-arsenicals before absorption across the gastrointestinal barrier could affect bioavailability, systemic distribution, and resulting toxicity.

Arsenic species in municipal landfill leachates (MLL) were investigated by HPLC-DRC-ICPMS and LC-ESI-MS/MS. Various arsenic species including arsenate (iAs(V)), arsenite (iAs(III)), monomethylarsonic acid (MMA(V)), dimethylarsinic acid (DMA(V)), as well as sulfur-containing organoarsenic species were detected. Two sulfur-containing arsenic species in a MLL were positively identified as dimethyldithioarsinic acid (DMDTA(V)) and dimethylmonothioarsinic acid (DMMTA(V)) by comparing their molecular ions, fragment patterns and sulfur/arsenic ratios with in-house synthesised thiol-organoarsenic compounds. The findings demonstrated the potential for transformation of DMA(V) to DMDTA(V) and DMMTA(V) in a DMA(V)-spiked MLL in a landfill leachate environment.

The arsenic (+3 oxidation state) methyltransferase (As3mt) gene encodes a 43 kDa protein that catalyzes methylation of inorganic arsenic. Altered expression of AS3MT in cultured human cells controls arsenic methylation phenotypes, suggesting a critical role in arsenic metabolism. Because methylated arsenicals mediate some toxic or carcinogenic effects linked to inorganic arsenic exposure, studies of the fate and effects of arsenicals in mice which cannot methylate arsenic could be instructive. This study compared retention and distribution of arsenic in As3mt knockout mice and in wild-type C57BL/6 mice in which expression of the As3mt gene is normal. Male and female mice of either genotype received an oral dose of 0.5 mg of arsenic as arsenate per kg containing [(73)As]-arsenate. Mice were radioassayed for up to 96 h after dosing; tissues were collected at 2 and 24 h after dosing. At 2 and 24 h after dosing, livers of As3mt knockouts contained a greater proportion of inorganic and monomethylated arsenic than did livers of C57BL/6 mice. A similar predominance of inorganic and monomethylated arsenic was found in the urine of As3mt knockouts. At 24 h after dosing, As3mt knockouts retained significantly higher percentages of arsenic dose in liver, kidneys, urinary bladder, lungs, heart, and carcass than did C57BL/6 mice. Whole body clearance of [(73)As] in As3mt knockouts was substantially slower than in C57BL/6 mice. At 24 h after dosing, As3mt knockouts retained about 50% and C57BL/6 mice about 6% of the dose. After 96 h, As3mt knockouts retained about 20% and C57BL/6 mice retained less than 2% of the dose. These data confirm a central role for As3mt in the metabolism of inorganic arsenic and indicate that phenotypes for arsenic retention and distribution are markedly affected by the null genotype for arsenic methylation, indicating a close linkage between the metabolism and retention of arsenicals.

Anion-exchange chromatography was utilized for speciation of arsenite (As(III)), arsenate (As(V)), dimethylarsinic acid (DMA(V)), monomethylarsonic acid (MMA(V)), monomethylarsonous acid (MMA(III)), and the new As species monomethylthioarsonic acid (MMTA), using inductively coupled plasma mass spectrometric (ICPMS) detection. MMA(III) and MMTA were identified for the first time in freeze-dried carrot samples that were collected over 25 years ago as part of a joint U.S. EPA, U.S. FDA, and USDA study on trace elements in agricultural crops. The discovery of MMA(III) and MMTA in terrestrial foods necessitated the analytical characterization of synthetic standards of both species, which were used for standard addition in carrot extracts. The negative ion mode, high-resolution electrospray mass spectrometry (HR-ESI-MS) data produced molecular ions of m/z 122.9418 and 154.9152 for MMA(III) and MMTA, respectively. However, ESI-MS was not sensitive enough to directly identify MMA(III) and MMTA in the carrot extracts. Therefore, to further substantiate the identification of MMA(III) and MMTA, two additional separations using an Ion-120 column were developed using the more sensitive ICPMS detection. The first separation used 20 mM tetramethylammonium hydroxide at pH 12.2 with MMA(III) eluting in less than 7 min. In the second separation, MMTA eluted at 11.2 min by utilizing 40 mM ammonium carbonate at pH 9.0. Oxidation of MMA(III) and MMTA to MMA(V) with hydrogen peroxide was observed for standards and carrot extracts alike. Several samples of carrots collected from local markets in 2006 were also analyzed and found to contain low levels of inorganic arsenic species.

Analyses of arsenic (As) species in tissues and body fluids of individuals chronically exposed to inorganic arsenic (iAs) provide essential information about the exposure level and pattern of iAs metabolism. We have previously described an oxidation state-specific analysis of As species in biological matrices by hydride-generation atomic absorption spectrometry (HG-AAS), using cryotrapping (CT) for preconcentration and separation of arsines. To improve performance and detection limits of the method, HG and CT steps are automated and a conventional flame-in-tube atomizer replaced with a recently developed multiple microflame quartz tube atomizer (multiatomizer). In this system, arsines from As(III)-species are generated in a mixture of Tris-HCl (pH 6) and sodium borohydride. For generation of arsines from both As(III)- and As(V)-species, samples are pretreated with L-cysteine. Under these conditions, dimethylthioarsinic acid, a newly described metabolite of iAs, does not interfere significantly with detection and quantification of methylated trivalent arsenicals. Analytical performance of the automated HG-CT-AAS was characterized by analyses of cultured cells and mouse tissues that contained mono- and dimethylated metabolites of iAs. The capacity to detect methylated As(III)- and As(V)-species was verified, using an in vitro methylation system containing recombinant rat arsenic (+3 oxidation state) methyltransferase and cultured rat hepatocytes treated with iAs. Compared with the previous HG-CT-AAS design, detection limits for iAs and its metabolites have improved significantly with the current system, ranging from 8 to 20 pg. Recoveries of As were between 78 and 117%. The precision of the method was better than 5% for all biological matrices examined. Thus, the automated HG-CT-AAS system provides an effective and sensitive tool for analysis of all major human metabolites of iAs in complex biological matrices.

Arsenic speciation was determined by anion-exchange chromatography-inductively coupled plasma-mass spectrometry (AEC-ICP-MS) in groundwater samples collected from an aquifer impacted by methylated As pesticides. Besides the four expected arsenic species AsO3(3-), As4(3-), (CH3)AsO3(2-) and (CH3)2AsO2-, up to nine other arsenic species were encountered, which constituted a major fraction of the total arsenic concentration in most samples. We then synthesized the thio-derivatives of (CH3)AsO3(2-) and (CH3)2AsO2-, and characterized the formed products by electrospray-tandem mass spectrometry (ES-MS-MS). The presence of (CH3)AsO2S2-, (CH3)AsOS2(2-), (CH3)2AsOS- and (CH3)2AsS2-, was confirmed in the groundwater by retention time matching plus ES-MS-MS in collected AEC fraction, and the presence of the trivalent methylated arsenic species (CH3)AsO2(2-) was suggested based on retention time matching only. These arsenic species have not been observed in ambient waters before, and are likely to occur in many environments containing methylated arsenic species and reduced sulfur compounds. They can persist in some of these particular samples for periods of up to six months without preservation, but tend to convert into the corresponding pentavalent oxy-species. Acidification with HCI shifted speciation equilibria rapidly, and is thus unsuitable for stabilizing samples containing these novel arsenic species; cryofreezing or no sample preservation avoided this artifact.

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 [(14)C]-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 [(14)C]-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.

Traditional Chinese medicines (TCMs) often contain significant levels of potentially toxic elements, including arsenic. Niu Huang Jie Du Pian pills were analyzed to determine the concentration, bioaccessibility (arsenic fraction soluble in the human gastrointestinal system) and chemical form (speciation) of arsenic. Arsenic excretion in urine (including speciation) and facial hair were studied after a one-time ingestion. The pills contained arsenic in the form of realgar, and although the total arsenic that was present in a single pill was high (28 mg), the low bioaccessibility of this form of arsenic predicted that only 4% of it was available for absorption into the bloodstream (1 mg of arsenic per pill). The species of arsenic that were solubilized were inorganic arsenate (As(V)) and arsenite (As(III)) but DMAA and MMAA were detected in urine. Two urinary arsenic excretion peaks were observed: an initial peak several (4-8) hours after ingestion corresponding to the excretion of predominantly As(III), and a larger peak at 14 h corresponding predominantly to DMAA and MMAA. No methylated As(III) species were observed. Facial hair analysis revealed that arsenic concentrations did not increase significantly as a result of the ingestion. Arsenic is incompletely soluble under human gastrointestinal conditions, and is metabolized from the inorganic to organic forms found in urine. Bioaccessible arsenic is comparable to the quantity excreted. Facial hair as a bio-indicator should be further tested.

Thioarsenicals are newly found arsenic metabolites in man and animals, and also in marine organisms. Dimethylmonothioarsinic acid (DMMTA(V)) and dimethyldithioarsinic acid (DMDTA(V)) are the only two thioarsenic metabolites detected in man and/or animals. However, their toxicological and biological significance is not known yet. The present study was performed to gain an insight into the significance of DMMTA(V) and DMDTA(V) in the metabolism of arsenic. The two thioarsenicals were synthesized chemically and injected intravenously into rats at the dose of 0.5 mg As/kg body weight. The distributions of arsenic in organs/tissues and body fluids were determined at 10 min and 12 h after the injection, and arsenic in liver and kidney supernatants, urine, plasma and red blood cell (RBC) lysates was subjected to speciation analysis by HPLC-ICP MS on a gel filtration GS 220 HQ column. Although both thioarsenicals are pentavalent arsenicals, they were distributed in organs/tissues and body fluids differently from the corresponding non-thiolated pentavalent arsenicals, and also from each other. Namely, DMMTA(V) was first found in organs/tissues at 10 min, and then redistributed and retained mostly in RBCs at 12 h, as in the case of trivalent dimethylarsinous acid (DMA(III)). On the other hand, although DMDTA(V) was also found in organs/tissues at 10 min, it had been efficiently excreted in urine in its intact form at 12 h. Thus, DMMTA(V) was unexpectedly distributed in and taken up by organs/tissues in a manner similar to DMA(III) rather than DMA(V), whereas DMDTA(V) was distributed similarly to DMA(V) as expected, but was much more efficiently excreted in urine.

Dimethylthioarsinic acid (DMTA(V)) has recently been identified in biological, dietary and environmental matrices. The relevance of this compound to the toxicity of arsenic in humans is unknown and further exposure assessment and metabolic studies are difficult to conduct because of the unavailability of a well characterized standard. The synthesis of DMTA(V) was accomplished by the reaction of dimethylarsinic acid (DMA(V)) with hydrogen sulfide. The initial reaction product produced is DMTA(V) but multiple products over the course of the reaction are also observed. Therefore, a chromatographic separation was developed to monitor the reaction progress via LC-ICP-MS. In this synthesis, conversion of DMA(V) to DMTA(V) was not taken to completion to avoid the production of side products. The product was isolated from the starting material by standard organic techniques. Single crystal diffraction demonstrated that solid DMTA(V) is present in the form of the oxygen-bridged dimethylthioarsinic anhydride. Dissolution of the anhydride in water produces the acid form of DMTA(V) and the aqueous phase DMTA(V) provided a characteristic molecular ion of m/z 155 by LC-ESI-MS. The synthesis and isolation of dimethylthioarsinic anhydride provides a stable crystalline standard suitable for identification, toxicological study and exposure assessment of dimethylthioarsinic acid.