Tetramethylarsonium Ion Research Articles

Sample pre-treatment to eliminate cationic methylated arsenic for determining arsenite on an anion-exchange column by high performance liquid chromatography–inductively coupled plasma mass spectrometry

Co-elution of cationic methylated arsenic, e.g. arsenobetaine may interfere with the determination of arsenite on the Hamilton PRP-X100 anion-exchange column using a phosphate buffer isocratically. Therefore, a sample pre-treatment method with self-packed AG MP-50 cation-exchange cartridges was proposed, which enables the arsenite determination in samples containing arsenobetaine on a PRP-X100 column using a phosphate buffer (pH 5.6) isocratically. Methylated arsenic, including dimethylarsinic acid, trimethylarsine oxide, tetramethylarsonium ion, arsenobetaine and arsenocholine, with concentrations below 1000 μg As L −1, may be completely retained in the AG MP-50 cartridge without any changes of arsenite, arsenate and monomethylarsonic acid speciation. Such retention was independent of the pH and matrix. It is proposed to be based on hydrophobic interaction. With the help of AG MP-50 cartridges, 11 arsenic species were detected in fish (DORM-2), mussels (BCR-477) and red algae ( Porphyra tenera) in 10 min on the PRP-X100 column using a phosphate buffer isocratically. Arsenite was the only minor species (up to 0.9%) among all water extractable arsenic species in fish, mussel and red algae.

Arsenic speciation in plants growing in arsenic-contaminated sites

Concentrations of total arsenic and of arsenic species were determined by ICPMS and HPLC–ICPMS in terrestrial plant samples. The arsenic concentration in plant samples from the contaminated sites ranged from 1.14 to 98.5 mg kg −1 (dry mass). However, a very high value, exceeding largely this range was found in a moss sample growing in the contaminated area (1750 mg kg −1). Plants growing in a non-contaminated area with similar geological characteristics contained 0.06–0.58 mg As kg −1. Plant samples from different species were selected and extracted with water, water/methanol (9 + 1, v/v), and water/methanol (1 + 1, v/v). Water/methanol (9 + 1, v/v) was selected as extractant for the speciation analysis for all the plant samples. The extraction efficiencies ranged from 3.0% to 41.4%, with good agreement between samples from the same plant species. Arsenite and/or arsenate were found in all the plant samples. Additionally, methylarsonate (MA), dimethylarsinate (DMA), trimethylarsine oxide (TMAO) and tetramethylarsonium ion (TETRA) were also identified in several plants, and in some cases MA and DMA were the main species found. TMAO, which is usually found as a trace constituent in organisms, was also a significant arsenical in one of the studied samples, where it constituted 24% of the extracted arsenic. In the present study, the patterns of arsenic species varied with the plant species and much higher proportion of organoarsenicals was found in plants from the more contaminated sites.

Unusual arsenic speciation in sea anemones

Nine species of sea anemones ( Anthopleura asiatica, Actinia equina, Actinodendron arboreum, Phymanthus loligo, Entacmaea actinostoloides, Stichodactyla gigantea, S. haddoni, S. mertensii and Metridium senile) contained arsenic in the range of 1.6–7.0 μg As g −1 (wet mass basis). Irrespective of the species, water-soluble arsenic compounds accounted for more than 80% of the total arsenic. Analysis of water-soluble arsenic compounds by LC/ESI-MS revealed that four arsenicals, arsenobetaine (AB), trimethylarsoniopropionate, arsenocholine (AC) and tetramethylarsonium ion (TEMA), are contained in most species but arsenate, methylarsonic acid, dimethylarsinic acid and trimethylarsine oxide are absent in all species. Interestingly, compositional patterns of the four arsenicals greatly differed from species to species. Only three species ( S. gigantea, S. haddoni and M. senile) contained AB at the highest proportions, similar to the majority of marine animals. However, the remaining six species showed unusual compositional patterns of arsenic compounds; AC was most predominant in A. arboreum and P. loligo and TEMA in A. asiatica, A. equina, E. actinostoloides and S. mertensii. On the whole, high proportions (24.6–87.1% of the water-soluble arsenic) of TEMA appear to be a peculiar characteristic of many species of sea anemones. Thus, sea anemones are an important animal group in the arsenic cycling, especially in that they may be donors of TEMA to predators.

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.

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.

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.

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).

Transformation of Organoarsenical Species by the Microflora of Freshwater Crayfish

A study of the transformation of arsenic species by the microflora of the freshwater crayfish Procambarus clarkii was carried out. The study of the degradation of AB (arsenobetaine) was performed in aerobic conditions in two culture media (tryptic soy broth and saline medium) at two temperatures (30 and 8 degrees C). The microflora transformed AB into TMAO (trimethylarsine oxide), DMA (dimethylarsinate), MA (methylarsonate), and an unidentified compound (U1). The quickest transformations were carried out by microflora from hepatopancreas incubated in saline medium at 30 degrees C. The individualized study of other arsenic species [AC (arsenocholine), TETRA (tetramethylarsonium ion), TMAO, DMA, and MA] was also performed in saline medium. The only transformation observed was of AC into AB. The bacteria possibly responsible for AB degradation were isolated, identified by phenotypic and genotypic methods, and individually assayed for AB transformation. Only isolates allocated to the species Pseudomonas putida were able to metabolize AB.

Organoarsenical Species Contents in Cooked Seafood

The organoarsenical species arsenobetaine (AB), arsenocholine (AC), tetramethylarsonium ion (TMA+), dimethylarsinic acid (DMA), and monomethylarsonic acid (MMA) were determined in 64 cooked seafood products (fish, bivalves, squid, crustaceans) included in a Total Diet Study carried out in the Basque Country (Spain). For cooking, various treatments were employed (grilling, roasting, baking, stewing, boiling, steaming, microwaving). The results obtained show that in cooked seafood AB is the major species, followed by DMA and TMA+. AC and MMA are minor species. The results in cooked seafood were compared with the arsenic species contents obtained for the same product raw. After cooking there was an increase in DMA for sardines and bivalves and an increase or appearance of TMA+ for meagrim, anchovy, Atlantic horse mackerel, and sardine. The data provided add to the very scant information available about organoarsenical species contents in cooked seafood.

Toxic arsenic compounds in environmental samples: Speciation and validation

Speciation of arsenic (As) compounds in environmental samples requires rigorous analytical procedures at each stage of sample collection, treatment and measurement. In view of the very high number of As species occurring in environmental samples, this article deals with only those that are considered toxic, including As-containing warfare agents and arsenobetaine (AB), because this is the most abundant As species in marine fish. With the exception of the tetramethylarsonium ion, acute toxicity generally decreases with increasing degree of methylation. The warfare agents considered are mostly phenylarsenic compounds. During storage, events, such as changes in oxidation state or induced by microbial activity or losses by volatilization or adsorption, have to be avoided. Extraction of arsenicals from soils and sediments is still a critical step that is not very well controlled. As long as marine animals are considered, extraction efficiencies >90% are commonly obtained [K.A. Francesconi, D. Kuehnelt, Analyst (Cambridge, UK) 129 (2004) 373]. This is logical, since marine animals are rich in AB, and this small molecule is soluble in water and methanol and in mixtures of them. In terrestrial organisms and plants, extraction efficiencies can be much lower and vary according to the extraction conditions. Since most environmental As species are generally present in soluble forms, liquid-separation techniques, first high-performance liquid chromatography (HPLC) but also capillary electrophoresis (CE), are most frequently used for separating As species. Fast separations can be performed with narrow-bore reversed-phase HPLC columns. Pre-column derivatizations with hydride generation [e.g., As(III), As(V), MA, DMA] and mercaptans/dimercaptans (e.g., Clarks I and II and Pfiffikus) allow separation of the arsenicals by gas chromatography (GC). The most popular detectors are inductively coupled mass spectrometry (ICP–MS) and atomic fluorescence spectrometry (AFS), especially after HPLC separation and hydride formation, increasingly replacing atomic absorption spectrometry (AAS), while HPLC–MS or HPLC–MS–MS (HPLC–MS2) is the detector of choice for (less toxic) arsenosugars and arsenolipids. For phenylarsenic warfare species, ultraviolet (UV) detection is also still used frequently. Finally, as other measurement fields, speciation analysis requires the availability of suitable reference materials (RMs) for verification of accuracy and quality assurance. We address these important aspects although the number of certified RMs is rather limited.

Unexpected Arsenic Compounds in Low-Rank Coals

Nine low-rank coal samples from three different coal basins (Velenje and Trbovlje basins, Slovenia, and Sokolov basin, Czech Republic) were analyzed for their elemental composition and the presence of arsenic compounds. Total arsenic concentrations in the samples were 1.59-5.77 microg g(-1) with one exception, 142 microg g(-1) for a sample from the Sokolov basin. A methanol/water mixture (1:1) extracted 15.1 - 38.7% of the total arsenic from Velenje basin samples but only 2.2-7.1% from Sokolov and Trbovlje basin samples. Extracts from the Velenje basin samples contained mainly the tetramethylarsonium ion (0.14 - 0.92 microg g(-1)) with considerable amounts of arsenate (0.15 - 0.85 microg g(-1)) and monomethyl arsonic acid 0.04 - 0.27 microg g(-1)). In methanol/water extracts from samples from the Sokolov and Trbovlje basins inorganic arsenic (arsenate) prevailed (0.26 - 37.1 microg g(-1)), but at least trace amounts of organic arsenic compounds were found as well. It is likely that biogeochemical degradation of organic material is related to the unexpected organoarsenic compounds found in low-rank coals. However, it should also not be ruled out that abiogenic synthesis could have taken place in the carbon-rich environment under the harsh conditions present (elevated temperature and pressure).

Placental transfer of arsenic to fetus of Dall’s porpoises ( Phocoenoides dalli)

Concentrations of total arsenic and individual arsenic compounds were determined in liver, muscle, kidney and blubber of mother and fetus of Dall’s porpoises collected from off Sanriku, Japan, in the year 2000 to characterize the placental transfer of arsenic to fetus in cetaceans. Arsenic was detected in all the tissues of Dall’s porpoises. Total arsenic concentrations in liver, kidney, muscle and blubber were 0.76, 0.69, 0.35 and 0.55 μg/g wet wt, respectively, for mother and 0.28, 0.23, 0.26 and 0.07 μg/g wet wt, respectively, for fetus. In all the tissues, concentrations of total arsenic in mother Dall’s porpoise were higher than in fetus. Arsenic speciation revealed that arsenobetaine was the major arsenic compound in liver, kidney and muscle of both mother and fetus. The percentage of arsenobetaine to total arsenic ranged from 76.0 to 91.0% in the tissues. Dimethylarsinic acid, arsenocholine, methylarsonic acid and an unidentified arsenic compound were also detected in tissues of both mother and fetus as minor constituents, whereas tetramethylarsonium ion was not detected in tissues of the fetus. These results suggest that arsenobetaine, dimethylarsinic acid, arsenocholine and methylarsonic acid are transferable from mother to fetus in Dall’s porpoises. To our knowledge, this is the first report on placental transfer of arsenic compounds to fetus in marine mammals.

Occurrence and Speciation of Arsenic in Common Australian Coastal Polychaete Species

Environmental Context. In well-oxygenated water and sediments, nearly all arsenic is present as arsenate (AsO43−). As arsenate is a phosphate (PO43−) analogue, organisms living in arsenate-rich environments must acquire the nutrient phosphorus yet avoid arsenic toxicity. Organisms take in and transform arsenic compounds by many means. Three major modes of arsenic biotransformation have been found to occur in the environment—redox transformation between arsenate and arsenite (AsO2−), the reduction and methylation of arsenic, and the biosynthesis of organoarsenic compounds such as arsenobetaine. These biotransformations lead to biogeochemical cycling of arsenic compounds and bioconcentration of arsenic in aquatic organisms and thence into the food web. Abstract. The paper reports the whole-tissue total arsenic concentrations and water-soluble arsenic species in eight common coastal Australian polychaete species. Laboratory experiments showed the period of depuration did not significantly alter the whole-tissue total arsenic concentrations in the two estuarine polychaete species tested. Significant differences were found between the whole-tissue total arsenic concentrations of the eight polychaete species (mean arsenic concentrations ranged from 18 to 101 µg g−1 dry mass). Total arsenic concentrations in polychaete species, grouped on the basis of a combination of their feeding guild and habitat type, were also significantly different with a significant interaction between these factors indicating that both factors simultaneously influence arsenic concentration in polychaetes. A large number of polychaete species contained similar arsenic species with high proportions of arsenobetaine (AB; 57–88%) and relatively low proportions of As3+, As5+, methyarsonic acid, dimethylarsinic acid, arsenocholine, trimethylarsoniopropionate, and tetramethylarsonium ion (not detected to 12%). All polychaete species contained arsenoribosides (5–30%). This study identified two Australian polychaete species with particularly unusual whole-tissue water-soluble arsenic species proportions: Australonuphis parateres contained a very high proportion of trimethylarsoniopropionate (~33%), while Notomastus estuarius had a very low proportion of arsenobetaine (9%) and high proportions of As3+ (~30%), As5+ (~8%), arsenoribosides (30%), and an unknown anionic arsenic species (~4%). Most polychaetes accumulate arsenobetaine, except deposit feeders inhabiting estuarine mud habitats. Thus most polychaetes, which are prey for higher organisms, are a source of arsenobetaine in benthic food webs. Deposit feeders inhabiting estuarine muddy substrates contain appreciable quantities of inorganic arsenic and arsenoribosides that may be metabolized to different end products in higher organisms.

Boric acid-assisted anion-exchange chromatography for separating arsenic species

Separation of 11 arsenic species on a single column was achieved by anion-exchange high performance liquid chromatography (HPLC) and selective detection was provided by inductively coupled plasma mass spectrometry (ICPMS). Boric acid was used to selectively modify the retention of arsenosugars. The borate–arsenosugar complexation improved the separation of the following 11 arsenic compounds: tetramethylarsonium ion (TMI), arsenobetaine (AsB), arsenate, arsenite, dimethylarsinic acid (DMA), monomethylarsonic acid (MMA), arsenosugars 1– 4, and an unidentified arsenic compound. The optimised mobile phase composition was 10 mM ammonium phosphate, 30 mM boric acid at pH 8.5. Rice and rice cereal extracts were analyzed using this method revealing the presence of arsenite, arsenate, DMA, and an unidentified arsenic species that has the same retention time as that of arsenosugar 2. Relative standard deviation of retention time from analyses of arsenic in different sample matrices, Fucus serratus, rice, rice cereal, oyster, dogfish, and mushroom extracts, was less than 1.0%, demonstrating the robustness of the separation method.

Identification of dimethylarsinic acid as the major arsenic compound in fish sauce by liquid chromatography/electrospray ionization-mass spectrometry

: Arsenobetaine, the major arsenic compound in marine animals, is substantially non-toxic. It is, however, possible that arsenobetaine undergoes bacterial transformation during manufacturing of fermented fishery products. In the present study, therefore, three types of fish sauce were examined for arsenic concentrations and species compared with those in the raw materials (sardine, Japanese sandfish and Japanese common squid). Arsenic concentrations of the three types of fish sauce were almost equivalent to those in their raw materials, suggesting no accumulation of arsenic during fermentation. Arsenic speciation was performed by a combination of cation-exchange liquid chromatography (LC) and electrospray ionization (ESI)-single quadrupole mass spectrometry (MS). Optimal conditions of LC/ESI-MS were established to analyze seven arsenic compounds (arsenate, monomethylarsonic acid, dimethylarsinic acid, arsenobetaine, trimethylarsine oxide, arsenocholine and tetramethylarsonium ion) found in biological samples. When analyzed by LC/ESI-MS, the major arsenic compound in the raw materials was arsenobetaine as expected, while not arsenobetaine but dimethylarsinic acid was identified as the major arsenic compound in the three types of fish sauce. These results suggest that arsenobetaine in the raw materials is converted to dimethylarsinic acid but not to arsenate with high toxicity by bacterial actions during manufacturing of fish sauce.

Speciation analysis of organoarsenical compounds in biological matrices by coupling ion chromatography to atomic fluorescence spectrometry with on-line photooxidation and hydride generation

The optimisation of an on-line decomposition based on UV photooxidation for the analysis of organoarsenic species by coupling cation-exchange chromatography and atomic fluorescence spectrometry with hydride generation, is described. In this study, special consideration is given to the compatibility of mobile phases with post-column treatments. Results show that the most commonly used mobile phase, aqueous pyridine solutions, decreases species conversion efficiency, leading to a significant loss of sensitivity. New fully-compatible chromatographic conditions are proposed to separate arsenobetaine, arsenocholine, trimethylarsine oxide and tetramethylarsonium ion within 20min. The very low absolute limits of detection, 4–12pg(As), allow speciation at trace levels. Analysis of a certified reference fish tissue (DORM-2) and other seafood samples (French and Chilean oysters and mussel) highlights the robustness and the accuracy of the optimised system.

Ion chromatography–inductively coupled plasma mass spectrometry determination of arsenic species in marine samples

AbstractArsenic speciation analysis in marine samples was performed using ion chromatography (IC) with inductively coupled plasma mass spectrometry (ICP‐MS) detection. The separation of eight arsenic species, viz. arsenite, monomethyl arsonic acid, dimethylarsinic acid, arsenate, arsenobetaine, tetramethylarsine oxide, arsenocholine and tetramethylarsonium ion was achieved on a Dionex AS4A (weaker anion exchange column) by using a nitric acid pH gradient eluent (pH 3.3 to 1.3). The entire separation was accomplished in 12 min. The detection limits for the eight arsenic species by IC–ICP‐MS were in the range 0.03–1.6 µ g l−1, based on 3σ of the blank response (n = 6). The repeatability and day‐to‐day reproducibility were calculated to be less than 10% (residual standard deviation) for all eight species. The method was validated by analyzing a certified reference material (DORM‐2, dogfish muscle) and then successfully applied to several marine samples, e.g. oyster, fish muscle, shrimp and marine algae. The low power microwave digestion was employed for the extraction of arsenic from seafood products. Copyright © 2004 John Wiley & Sons, Ltd.

Arsenic speciation in marine certified reference materials

This study describes the analysis of arsenicals in three marine based certified reference materials (CRMs), DORM-2, DOLT-2 and TORT-2, all produced by the National Research Council of Canada. Extraction protocols involving accelerated solvent extraction (ASE) and sonication were investigated and arsenicals were determined on-line by anion-exchange and cation-exchange high-performance liquid chromatography (HPLC). The major species in all three tissues was found to be arsenobetaine (AsB), corresponding to 90%, 67% and 73% of the water-soluble arsenic in DORM-2, DOLT-2 and TORT-2, respectively. Other species quantified were arsenocholine (AsC+), tetramethylarsonium ion (TMAs+), trimethylarsonioproprionate (TMAP), dimethylarsenic acid (DMA), monomethylarsenic acid (MMA), arsenosugar D (As-sug D), arsenate (AsV) and arsenosugar A (As-sug A). The major species detected in all three materials were AsB and DMA, which together accounted for 97.4% (DORM-2), 95.4% (DOLT-2) and 91.7% (TORT-2) of the sum of the water soluble As species. The agreement for the determination of AsB by the different methods was good for all three materials, whereas some discrepancy was evident in the results for DMA. When an ASE approach with 50% acetic acid in methanol was used for the extraction, between 22% (DOLT-2) and 58% (TORT-2) more DMA was determined compared with extraction by sonication with water. Some discrepancy was evident between the sum of the species extracted and both the total As determined in the extracts and the certified As content based on complete digestion of the materials. The sum of the species recovered by sonication corresponded to 84% (DOLT-2) to 94% (DORM-2) of the total As determined in the aqueous extract. In comparison with the certified As content of the three materials, the sums of the extracted species range from 46% (DOLT-2) to 76% (TORT-2) and 102% (DORM-2).

Hyphenation of high performance liquid chromatography with sector field inductively coupled plasma mass spectrometry for the determination of ultra-trace level anionic and cationic arsenic compounds in freshwater fishElectronic Supplementary Information (ESI) available: analytical figures of merit obtained with a standard set-up, the LOD for the high resolution mode, LODs reported by other authors and tissue concentrations for the Northern pike.

We describe a hyphenation technique between HPLC and ICP-SFMS for ultra-trace arsenic speciation analysis. Exceptional analytical performance was achieved using a MicroMist nebulizer preceded by a high-pressure splitter. Despite a 1 ∶ 7.5 flow splitting, the detection limits in the range of 1.2 to 2.4 pg mL−1 were about two times lower than those obtained with a concentric nebulizer without any flow splitting, demonstrating the applicability of coupling conventional HPLC system (1.5 mL min−1 eluent flow) with microflow nebulizer for ultra-trace arsenic speciation analysis. This set-up offers an advantage for on-line fraction collection for either multidimensional chromatographic separation of co-eluting As compounds or for structural identification of unknown compounds without sacrificing analytical sensitivity. In addition, this system showed good accuracy and repeatability. The method was applied to the determination of arsenic compounds in freshwater fish samples from an arsenic-rich lake, Moira Lake, Canada. Using cation-exchange chromatography, tetramethylarsonium ion (Tetra) was detected in freshwater fish samples for the first time. Moreover, in pumpkinseed, Tetra was found to be the major arsenic species, indicating that the biomethylation pathway in freshwater ecosystems may include the tetramethyl stage.