Biodegradation of arsenosugars in marine sediment

In the marine environment, arsenic accumulates in seaweed and occurs mostly in the form of arsenoribofuranosides (often called arsenosugars). This study investigated the degradation pathways of arsenosugars from decaying seaweed in a mesocosm experiment. Brown seaweed (Laminaria digitata) was placed on top of a marine sediment soaked with seawater. Seawater and porewater samples from different depths were collected and analysed for arsenic species in order to identify the degradation products using high‐performance liquid chomatography–inductively coupled plasma mass spectrometry. During the first 10 days most of the arsenic found in the seawater and the shallow sediment is in the form of the arsenosugars released from the seaweed. Dimethylarsenoylethanol (DMAE), dimethylarsinic acid (DMA(V)) and, later, monomethylarsonic acid (MMA(V)) and arsenite and arsenate were also formed. In the deeper anaerobic sediment, the arsenosugars disappear more quickly and DMAE is the main metabolite with 60–80% of the total arsenic for the first 60 days besides a constant DMA(V) contribution of 10–20% of total soluble arsenic. With the degradation of the soluble DMAE the solubility of arsenic decreases in the sediment. The final soluble degradation products (after 106 days) were arsenite, arsenate, MMA(V) and DMA(V). No arsenobetaine or arsenocholine were identified in the porewater. Copyright © 2005 John Wiley & Sons, Ltd.     

Liquid chromatography electrospray mass spectrometry with variable fragmentor voltages gives simultaneous elemental and molecular detection of arsenic compounds

A single quadrupole high performance liquid chromatography electrospray mass spectrometry system with a variable fragmentor voltage facility was used in the positive ion mode for simultaneous recording of elemental and molecular mass spectral data for arsenic compounds. The method was applicable to the seven organoarsenic compounds tested: four arsenic-containing carbohydrates (arsenosugars), a quaternary arsonium compound (arsenobetaine), dimethylarsinic acid, and dimethylarsinoylacetic acid. It was not suitable for the two inorganic arsenic species arsenite and arsenate. In the case of arsenosugars, qualifying ion data for a characteristic common fragment (m/z 237) was also simultaneously obtained. The method was used to identify and quantify the major arsenosugars in crude extracts of two brown algae.     

Biotransformation of arsenate to arsenosugars by Chlorella vulgaris

Chlorella vulgaris was cultivated in a growth medium containing arsenate concentration of <0.01, 10, 100 and 1000 mg l^?1. Illumination was carried out in 12 h cycles for 5 days. The health status of the culture was monitored by continuous pH and dissolved oxygen (DO) readings. Destructive sampling was used for the determination of biomass, chlorophyll, total arsenic and arsenic species. The chlorophyll a content, the DO and pH cycles were not significantly different for the different arsenate concentrations in the culture. In contrast, biomass production was significantly (p < 0.05) increased for the arsenic(V) treatment at 1000 mg l^?1 compared with 100 mg l^?1. The arsenic concentration in the algae increased with the arsenate concentration in the culture. However, the bioconcentration factor decreased a hundred‐fold with increase of arsenate from the background level to 1000 mg l^?1. The arsenic species were identified by using strong anion‐exchange high‐performance liquid chromatography–inductively coupled plasma mass spectrometry analysis after methanol/water (1 : 1) extraction. The majority (87–100%) of the extractable arsenic was still arsenate; arsenite was found to be between 1 and 6% of total extractable arsenic in the algae. In addition to dimethylarsinic acid, one unknown arsenical (almost co‐eluting with methylarsonic acid) and three different arsenosugars have been identified for the first time in C. vulgaris growing in a culture containing a mixture of antibiotics and believed to be axenic. The transformation to arsenosugars in the algae is not dependent on the arsenate concentration in the culture and varies between 0.2 and 5% of total accumulated arsenic. Although no microbiological tests for bacterial contamination were made, this study supports the hypothesis that algae, and not associated bacteria, produce the arsenosugars. Copyright © 2003 John Wiley & Sons, Ltd.     

Simultaneous determination of twelve inorganic and organic arsenic compounds by liquid chromatography-ultraviolet irradiation-hydride generation atomic fluorescence spectrometry

A coupling between column liquid chromatography (LC) and atomic fluorescence spectrometry was developed for arsenic speciation. After separation, the compounds are oxidised on-line by UV irradiation, volatilised by hydride-generation and carried to the detector by a stream of argon. A combination of anion-exchange and hydrophobic interactions in a single column (Dionex AS7) was found suitable for the simultaneous separation of organic and inorganic species. Twelve compounds (arsenite, arsenate, monomethylarsonic acid, dimethylarsinic acid, arsenobetaine, arsenocholine, trimethylarsine oxide, tetramethylarsonium ion and four arsenosugars) were separated using an acetate buffer and a nitric acid solution as mobile phases. Limits of detection are 4-22 pg. The technique was applied to three marine samples. Arsenobetaine was detected as the main species in all samples, with concentrations varying from 59 to 1947 ng(As) g(-1) of fresh mass.     

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

Arsenic 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.     

Determination of arsenic species in Solanum Lyratum Thunb using capillary electrophoresis with inductively coupled plasma mass spectrometry

A simple and highly efficient interface to couple capillary electrophoresis with inductively coupled plasma mass spectrometry by a microflow polyfluoroalkoxy nebulizer and a quadruple ion deflector was developed in this study. By using this interface, six arsenic species, including arsenite, arsenate, monomethylarsonic acid, dimethylarsinic acid, arsenobetaine, and arsenocholine, were baseline‐separated and determined in a single run within 11 min under the optimized separation conditions. The instrumental detection limit was in the range of 0.02–0.06 ng/mL for the six arsenic compounds. Repeatability expressed as the relative standard deviation (n = 5) of both migration time and peak area were better than 2.5 and 4.3% for six arsenic compounds. The proposed method, combined with a closed‐vessel microwave‐assisted extraction procedure, was successfully applied for the determination of arsenic species in the Solanum Lyratum Thunb samples from Anhui province in China with the relative standard deviations (n = 5) ≤4%, method detection limits of 0.2–0.6 ng As/g and a recovery of 98–104%. The experimental results showed that arsenobetaine was the main speciation of arsenic in the Solanum Lyratum Thunb samples from different provinces in China, with a concentration of 0.42–1.30 μg/g.     

A gradient anion exchange chromatographic method for the speciation of arsenic in lobster tissue extracts

A gradient anion exchange chromatographic technique was developed for the separation of arsenobetaine (AsB), arsenocholine (AsC), arsenite (As(III)), arsenate (As(V)), monomethylarsonic acid (MMAA) and dimethylarsinic acid (DMAA) in one chromatographic run. This technique used low residue ammonium carbonate buffer and the inductively coupled plasma-mass spectrometry (ICP-MS) chromatograms showed little baseline drift. Gradient elution improved peak shape and peak separation. The separation was completed in approximately 27 min with low detection limits (0.017-0.029 mug As kg(-1)). Baseline resolution of all the arsenic species evaluated was achieved when the concentration of AsC was less than approximately 12.5 mug As kg(-1). This technique was successfully applied to different extracts of a standard reference material, TORT-2, and lobster tissue. AsB was found to be the major arsenic species present. AsC, DMAA, MMAA and As(V) were also found, although MMAA was not detected in all of the TORT-2 extracts. Two unknown peaks found may be due to the presence of arsenosugars or other arsenic species. Discrepancy between extraction recoveries previously determined using flow injection-ICP-MS and the high-performance liquid chromatography-ICP-MS was observed in some cases. The differences may be due to the extraction technique and/or conditions at which the extractions were performed.     

Arsenic speciation in freshwater organisms from the river Danube in Hungary

Total arsenic and arsenic species were determined in a range of freshwater samples (sediment, water, algae, plants, sponge, mussels, frog and fish species), collected in June 2004 from the river Danube in Hungary. Total arsenic concentrations were measured by ICPMS and arsenic species were measured in aqueous extracts of the samples by ion-exchange HPLC-ICPMS. In order to separately determine the efficiency of the extraction method and the column recovery, total arsenic concentrations in the extracts were obtained in three ways: (i) ICPMS determination after acid digestion; (ii) flow injection analysis performed directly on the extract; (iii) the sum of arsenic species eluting from the HPLC column. Extraction efficiencies were low (range 10-64%, mean 36%), but column recovery was acceptable (generally >80%) except for the fish samples, where substantial, currently unexplained, losses were observed. The dominating arsenic species in the extracts of freshwater algae were arsenosugars, whereas arsenate [As(V)] was present only as a minor constituent. On the other hand, plant extracts contained only inorganic arsenic, except for two samples which contained trace amounts of dimethylarsinate (DMA) and the tetramethylarsonium cation (TETRA). The oxo-arsenosugar-phosphate (ca. 35% of extractable arsenic) and the oxo-arsenosugar-glycerol (ca. 20%) as well as their thio-analogues (1-10%) were found in the mussel extracts, while arsenobetaine (AB) was present as a minor species only. In general, fish extracts contained only traces of arsenobetaine, and the oxo-arsenosugar-phosphate was the major arsenic compound. In addition, samples of white bream contained thio-arsenosugar-phosphate; this is the first report of a thio-arsenical in a fish sample. The frog presented an interesting arsenic speciation pattern because in addition to the major species, arsenite [As(III)] (30%) and the tetramethylarsonium cation (35%), all three intermediate methylation products, methylarsonate (MA), dimethylarsinate and trimethylarsine oxide (TMAO), and arsenate were also present. Collectively, the data indicate that arsenobetaine, the major arsenical in marine animals, is virtually absent in the freshwater animals investigated, and this represents the major difference in arsenic speciation between the two groups of organisms.     

High-performance liquid chromatography with hydride generation/atomic absorption spectrometry for the determination of arsenic species with application to some water samples

High-performance liquid chromatography (h.p.l.c.) is used for separation of arsenite, arsenate, monomethylarsinate (MMA) and dimethylarsonate (DMA) followed by continuous sodium tetrahydroborate reduction and atomic absorption spectrometric detection. Sample preconcentration, offering improved detection limits for the individual species and the removal of matrix interferences, is achieved with a pellicular anion-exchange column. The arsenic species are then separated on a strong anion-exchange column placed in series with the preconcentration column. Detection limits of 2 ng (as arsenic) for arsenite, arsenate and MMA, and 1 ng for DMA. Results for arsenic species in soil waters and commercial bottle waters are given.     

Benefits of high resolution IC-ICP-MS for the routine analysis of inorganic and organic arsenic species in food products of marine and terrestrial origin

A recently developed and validated method for simultaneous determination of 17 inorganic and organic arsenic compounds in marine biota has been successfully applied to routine analysis of different food products, including fish, shellfish, edible algae, rice, and other types of grain. During one year, approximately 250 food samples were analyzed, mostly fish and rice. Long-term stability and robustness of the system was observed and reproducible results for certified reference materials were ensured by means of control charts. The separation was performed by ion-pair chromatography on an anion-exchange column to separate anionic, neutral, and cationic arsenic species in one chromatographic run. Hyphenation to ICP–MS allowed element-specific and sensitive detection of the different arsenic species with a detection limit as low as 8 ng As L^–1 in the sample extract, which is equivalent to 2 ng As g^–1 in the original sample. Special emphasis was laid on the analysis of marine algae and rice samples. These food types can contain elevated levels of the very toxic inorganic arsenic species (up to 90% in rice) and therefore are the focus of interest in the food industry. In marine algae, inorganic arsenic was mainly present as arsenate whereas in rice arsenite predominated.     

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).     

Speciation analysis of arsenic compounds in edible oil by ion chromatography-inductively coupled plasma mass spectrometry

An inductively coupled plasma mass spectrometer (ICP-MS) was used as an ion chromatographic (IC) detector for the speciation analysis of arsenic in edible oil. The arsenic species studied include arsenite, arsenate, monomethylarsonic acid, dimethylarsinic acid, arsenobetaine and arsenocholine. Gradient elution using (NH(4))(2)CO(3) and methanol at pH 8.5 allowed the chromatographic separation of all species in less than 8 min. Effluents from the IC column were delivered to the nebulizer of ICP-MS for the determination of arsenic. The concentrations of arsenic species have been determined in several used and fresh vegetable oil samples. In this study, a microwave-assisted extraction method was used for the extraction of arsenic species from oil samples. The extraction efficiency was better than 92% and the recoveries from spiked samples were in the range of 90-105%. The precision between sample replicates was better than 8% for all determinations. The limits of detection were in the range of 0.008-0.024 ng mL(-1) for various arsenic species based on peak height, which corresponded to 0.08-0.24 ng g(-1) in the original oil sample. The major arsenic species in the used oil samples varied based on the food items cooked.     

Evaluation of atomic fluorescence spectrometry as a sensitive detection technique for arsenic speciation

The potential of coupling anion-exchange high-performance liquid chromatography, hydride generation and atomic fluorescence spectrometry (HPLC–HG–AFS) for arsenic speciation is considered. The effects of hydrochloric acid and sodium tetrahydroborate concentrations on signal-to-background ratio, as well as argon and hydrogen flow rates, were investigated. Detection limits for arsenite, dimethylarsinic acid (DMA), monomethylarsonic acid (MMA) and arsenate were 0.17, 0.45, 0.30 and 0.38 μg l⁻¹, respectively, using a 20-μl loop. Linearity ranges were 0.1–500 ng for As(III) and MMA (as arsenic), and 0.1–800 ng for DMA and As(V) (as arsenic). Arsenobetaine (AsB) was also determined by introducing an on-line photo-oxidation step after the chromatographic separation. In this case the limits of detection and linear ranges for the different species studied were similar to the values obtained previously for As(V). The technique was tested with a human urine reference material and a volunteer's sample. © 1998 John Wiley & Sons, Ltd.     

Simultaneous determination of arsenic species by ion chromatography-inductively coupled plasma mass spectrometry

Six arsenic species, arsenite, arsenate, monomethylarsonic acid, dimethylarsinic acid, arsenobetaine and arsenocholine, were separated by coupled column ion chromatography using carbonate and nitric acid as eluents, and were detected by inductively coupled plasma mass spectrometry. Coupling of an anion column with a cation column made the simultaneous determination of both the cationjic and the anionic arsenic species possible by ion chromatography. Extremely low detection limits, below 0.2 μg/1 (as arsenic), were obtained for all the species studied.     

Determination of arsenic species and arsenosugars in marine samples by HPLC–ICP–MS

Arsenic-speciation analysis in marine samples was performed by high-pressure liquid chromatography (HPLC) with ICP–MS detection. Separation of eight arsenic species—As^III, MMA, DMA, As^V, AB, TMAO, AC and TeMAs⁺—was achieved on a C₁₈ column with isocratic elution (pH 3.0), under which conditions As^III and MMA co-eluted. The entire separation was accomplished in 15 min. The HPLC–ICP–MS detection limits for the eight arsenic species were in the range 0.03–0.23 μg L⁻¹ based on 3σ for the blank response (n=5). The precision was calculated to be 2.4–8.0% (RSD) for the eight species. The method was successfully applied to several marine samples, e.g. oysters, fish, shrimps, and marine algae. Low-power microwave digestion was employed for extraction of arsenic from seafood products; ultrasonic extraction was employed for the extraction of arsenic from seaweeds. Separation of arsenosugars was achieved on an anion-exchange column. Concentrations of arsenosugars 2, 3, and 4 in marine algae were in the range 0.18–9.59 μg g⁻¹. This paper was presented at the European Winter Conference 2005     

HPLC-ICP-MS测定植物样品中6种砷形态化合物

通过优化色谱分离、样品前处理条件,同时对比了电感耦合等离子体质谱的标准模式(STD)、碰撞模式(KED)、氧气反应模式(Oxygen-DRC)、甲烷反应模式(Methane-DRC)的检测结果,建立了一种有效分离植物样品中砷甜菜碱(AsB)、二甲基砷酸(DMA)、亚砷酸(As(Ⅲ))、砷胆碱(AsC)、一甲基砷酸(MM...     

Arsenic metabolism in seaweed-eating sheep from Northern Scotland

Cation exchange and anion exchange liquid chromatography were coupled to an ICP-MS and optimised for the separation of 13 different arsenic species in body fluids (arsenite, arsenate, dimethylarsinic acid (DMAA), monomethylarsonic acid (MMAA), trimethylarsine oxide (TMAO), tetramethylarsonium ion (TMA), arsenobetaine (AsB), arsenocholine (AsC), dimethylarsinoyl ethanol (DMAE) and four common dimethylarsinoylribosides (arsenosugars). The arsenic species were determined in seaweed extracts and in the urine and blood serum of seaweed-eating sheep from Northern Scotland. The sheep eat 2–4 kg of seaweed daily which is washed ashore on the most northern Island of Orkney. The urine, blood and wool of 20 North Ronaldsay sheep and kidney, liver and muscle from 11 sheep were sampled and analysed for their arsenic species. In addition five Dorset Finn sheep, which lived entirely on grass, were used as a control group. The sheep have a body burden of approximately 45–90 mg arsenic daily. Since the metabolism of arsenic species varies with the arsenite and arsenate being the most toxic, and organoarsenic compounds such as arsenobetaine the least toxic compounds, the determination of the arsenic species in the diet and their body fluids are important. The major arsenic species in their diet are arsenoribosides. The major metabolite excreted into urine and blood is DMAA (95 ± 4.1%) with minor amounts of MMAA, riboside X, TMA and an unidentified species. The occurrence of MMAA is assumed to be a precursor of the exposure to inorganic arsenic, since demethylation of dimethylated or trimethylated organoarsenic compounds is not known (max. MMAA concentration 259 μg/L). The concentrations in the urine (3179 ± 2667 μg/L) and blood (44 ± 19 μg/kg) are at least two orders of magnitude higher than the level of arsenic in the urine of the control sheep or literature levels of blood for the unexposed sheep. The tissue samples (liver: 292 ± 99 μg/kg, kidney: 565 ± 193 μg/kg, muscle: 680 ± 224 μg/kg) and wool samples (10 470 ± 5690 μg/kg) show elevated levels which are also 100 times higher than the levels for the unexposed sheep. Received: 29 February 2000 / Revised: 26 April 2000 / Accepted: 1 May 2000     

Determination of arsenite, arsenate and monomethylarsonic acid in aqueous samples by gas chromatography of their 2,3-dimercaptopropanol (bal) complexes

Procedures are described for the determination of arsenite, arsenate and monomethylarsonic acid in aqueous samples. The arsenicals (after reduction of arsenic to the tervalent state) readily react with 2,3-dimercaptopropanol (BAL) to yield their BAL complexes. The products are extracted with benzene and introduced into a gas Chromatograph equipped with a flame-photometric detector for sulphur. One aliquot of sample is treated with stannous chloride solution and potassium iodide solution to reduce arsenate and monomethylarsonic acid, then BAL is added and the complexes are extracted with benzene. The extract is analysed for total inorganic As plus monomethylarsonic acid. Magnesia mixture and phosphate solution are added to another aliquot to remove arsenate by co-precipitation with magnesium ammonium phosphate. The precipitate is filtered off and arsenite determined in the filtrate. The detection limits are 0.02 ng of As for arsenate and arsenite and 0.04 ng of As for monomethylarsonic acid.     

Speciation of arsenic compounds in fish and oyster tissues by capillary electrophoresis-inductively coupled plasma-mass spectrometry

A capillary electrophoresis-inductively coupled plasma-mass spectrometric (CE-ICP-MS) method for the speciation of six arsenic compounds, namely arsenite [As(III)], arsenate [As(V)], monomethylarsonic acid, dimethylarsinic acid, arsenobetaine and arsenocholine is described. The separation has been achieved on a 70 cm length x 75 microm ID fused-silica capillary. The electrophoretic buffer used was 15 mM Tris (pH 9.0) containing 15 mM sodium dodecyl sulfate (SDS), while the applied voltage was set at +22 kV. The arsenic species in biological tissues were extracted into 80% v/v methanol-water mixture, put in a closed centrifuge tube and kept in a water bath, using microwaves at 80 degrees C for 3 min. The extraction efficiencies of individual arsenic species added to the sample at 0.5 microg As/g level were between 96% and 107%, except for As(III), for which it was 89% and 77% for oyster and fish samples, respectively. The detection limits of the species studied were in the range 0.3-0.5 ng As/mL. The procedure has been applied for the speciation analysis of two reference materials, namely dogfish muscle tissue (NRCC DORM-2) and oyster tissue (NIST SRM 1566a), and two real-world samples.     

微波辅助提取-液相色谱-氢化物发生-原子荧光光谱法分析沉积物中砷的形态

采用微波辅助提取-液相色谱-氢化物发生-原子荧光光谱法(LC-HG-AFS)联用技术分析了太湖沉积物中砷的形态[亚砷酸(As(III))、二甲基砷酸钠(DMA)、一甲基砷酸二钠(MMA)和砷酸As(V)]。测得沉积物中以无机砷为主,且以As(V)居多。选定以1mol/L的磷酸和0.1mol/L抗坏血酸为提取液,在微波辅助萃取(功率为60W,时间12min)下,萃取率达79.84%～91.57%,回收率在94.78%～107.6%之间。4种砷的形态在0～160μg/L之间时线性良好,检测限为0.6～2.3μg/L,相对标准偏差RSD为1.62%～2.20%。方法具有简便、快速、灵敏的特点。