Arsenic Compounds in Higher Fungi

In 50 mushroom species (56 samples) from Slovenia, Switzerland, Brazil, Sweden, The Netherlands and USA, total arsenic was determined by radiochemical neutron activation analysis (RNAA). Arsenic concentrations ranged from 0.1 to 30 μg g⁻¹ (dry mass). Arsenic compounds were determined in methanol extracts from the mushrooms by HPLC–ICP–MS. The aim of the study was not only to quantify arsenic compounds in mushrooms but also to uncover trends relating the methylating ability of a mushroom to its taxonomic or evolutionary status. The main arsenic compound found in many mushrooms (various puffballs, Agaricales and Aphyllophorales) was arsenobetaine. Arsenate [As(V)] was the main arsenic species in Laccaria fraterna and Entoloma rhodopolium and arsenite [As(III)] in Tricholoma sulphureum. A mixture of arsenite and arsenate was present in Amanita caesarea. Dimethylarsinic acid (DMA) and methylarsonic acid were present in many mushrooms, but generally as minor components. In Laccaria laccata, Leucocoprinus badhamii and Volvariella volvacea, DMA was the major metabolite. Arsenocholine (AC) and the tetramethylarsonium ion were present in a few species, generally at low concentrations, except for Sparassis crispa, in which AC was the main compound. Tri- methylarsine oxide was not found in any of the mushrooms. In some species small amounts of unknown compounds were also present. The possible taxonomic significance of the metabolite patterns and the predominance of arsenobetaine in more advanced fungal types are discussed. © 1997 John Wiley & Sons, Ltd.     

Arsenobetaine and other arsenic species in mushrooms

Arsenic species in arsenic accumulating mush- rooms (Sarcosphaera coronaria, Laccaria amethystina, Sarcodon imbricatum, Entoloma lividum, Agaricus haemorrhoidaius, Agaricus placomyces, Lycoperdon perlatum) were determined. HPLC/ICP MS and ion-exchange chromatogra- phy–instrumental neutron activation analysis (NAA) combinations were used. The remarkable accumulator Sarcosphaera coronaria (up to 2000 mg As kg^?1 dry wt) contained only methylarsonic acid, Entoloma lividum only arsenite and arsenate. In Laccaria amethystina dimethylarsinic acid was the major arsenic compound. Sarcodon imbricatum and the two Agaricus sp. were found to contain arsenobetaine as the major arsenic species, a form which had previously been found only in marine biota. Its identification was confirmed by electron impact MS.     

Arsenic biomethylation by the microorganism apiotrichum humicola in the presence of l-methionine-methyl-d3

The microorganism Apiotrichum humicola (previously known as Candida humicola) grown in the presence of either arsenate, arsenite, methylarsonic acid or dimethylarsinic acid, produces arsenic-containing metabolites in the growth medium. When L-methionine-methyl-d₃ is added to the cultures, the CD₃ label is incorporated intact into the metabolites to a considerable extent to form deuterated dimethylarsenic and trimethyl-arsenic species, indicating that S-adenosylmethionine, or some related sulphonium compound, is involved in the biological methylation. Conclusive evidence of CD₃ incorporation in the arsenicals found in the growth medium was provided by using a specially developed hydride generation-gas chromatography-mass spectrometry system (HG–GC–MS).     

Behaviour of organoarsenic compounds in contact with natural zeolites

Batch experiments were conducted on aqueous solutions containing arsenite, arsenobetaine, methylarsonic acid or phenylarsonic acid in contact with natural zeolites to examine their interaction. The concentration of the arsenic species in the liquid phase at equilibrium before and after contact was measured by means of liquid chromatography coupled with inductively coupled plasma mass spectrometry detection. Clinoptilolites completely removed arsenobetaine from the solution and the resulting amounts of dimethylarsinic acid were detected. The methylarsonic acid maximum concentration diminution was reached at a mass—to volume V value of m/V = 0.2. Phenylarsonic acid solution decreased its concentration 75% after treatment with clinoptilolites. Untreated mordenites in contact with arsenite solutions led to the formation of arsenate, whereas acid‐washed mordenites practically removed arsenobetaine and were less effective for methylarsonic acid. To show the incompatibility of molecular dimensions with the zeolite windows, the molecular parameters of surface area, molecular volume, molecular length, and the width and depth of arsenite, arsenate and a series of ten organic arsenic compounds were calculated. Since sorption onto the external zeolite surface rather than a sieve process defined the interaction, an acid‐catalysed reaction mechanism is proposed to explain the transformation results. Copyright © 2001 John Wiley & Sons, Ltd.     

Comparison of mild extraction procedures for determination of arsenic compounds in different parts of pepper plants (Capsicum annum,L.)

Eight extraction agents (water, methanol–water mixtures in various ratios, methanol, a 20 mmol l^?1 ammonium phosphate buffer, and a methanol–phosphate buffer) were tested for the extraction of arsenic compounds from fruits, stems + leaves, and roots of pepper plants grown on soil containing 17.2 mg kg^?1 of total arsenic. The arsenic compounds in the extracts were determined using high‐performance liquid chromatography–hydride generation inductively coupled plasma mass spectrometry. Whereas pure water was the most effective extraction agent for fruits (87 ± 3.3% extraction yield) and roots (96 ± 0.6% extraction yield), the 20 mM ammonium phosphate buffer at pH 6 extracted about 50% of the arsenic from stems + leaves. Decreasing extractability of the arsenic compounds was observed with increasing methanol concentrations for all parts of the pepper plant. In pepper fruits, arsenic(III), arsenic(V), and dimethylarsinic acid (DMA) were present (25%, 37%, and 39% respectively of the extractable arsenic). Arsenic(V) was the major compound in stems + leaves and roots (63% and 53% respectively), followed by arsenic(III) representing 33% and 42% respectively, and small amounts (not exceeding 5%) of DMA and methylarsonic acid were also detected. Hence, for a quantitative extraction of arsenic compounds from different plant tissues the extractant has to be optimized individually. Copyright © 2005 John Wiley & Sons, Ltd.     

Metabolism of arsenic compounds by the blue mussel mytilus edulis after accumulation from seawater spiked with arsenic compounds

Blue mussels (Mytilus edulis) were exposed to 100 μg As dm^?3 in the form of arsenite, arsenate, methylarsonic acid, dimethylarsinic acid, arsenobetaine, arsenocholine, trimethylarsine oxide, tetramethylarsonium iodide or dimethyl-(2-hydroxyethyl)arsine oxide in seawater for 10 days. The seawater was renewed and spiked with the arsenic compounds daily. Analyses of water samples taken 24 h after spiking showed that arsenobetaine and arsenocholine had been converted to trimethylarsine oxide, whereas trimethylarsine oxide and tetramethylarsonium iodide were unchanged. Arsenobetaine was accumulated by mussels most efficienty, followed in efficiency by arsenocholine and tetramethylarsonium iodide. None of the other arsenic compounds was significantly accumulated by the mussels. Extraction of mussel tissues with methanol revealed that control mussels contained arsenobetaine, a dimethyl-(5-ribosyl)arsine oxide and an additional arsenic compound, possibly dimethylarsinic acid. Mussels exposed to arsenobetaine contained almost all their experimentally accumulated arsenic as arsenobetaine, and mussels exposed to tetramethylarsonium iodide contained it as the tetramethylarsonium compound. Mussels exposed to arsenocholine had arsenobetaine as the major arsenic compound and glycerylphosphorylarsenocholine as a minor arsenic compound in their tissues. The results show that arsenobetaine and arsenocholine are efficiently accumulated from seawater by blue mussels and that in both cases the accumulated arsenic is present in the tissues as arsenobetaine. Consequently arsenobetaine and/or arsenocholine present at very low concentrations in seawater may be responsible for the presence of arsenobetaine in M. edulis and probably also among other marine animals. The quantity of arsenobetaine accumulated by the mussels decreases with increasing concentrations of betaine. HPLC-ICP-MS was found to be very powerful for the investigation of the metabolism of arsenic compounds in biological systems.     

Immunotoxicity of Organic Arsenic Compounds in Marine Animals

In the present study, we demonstrated for the first time the immunotoxic effects of organic arsenic compounds in marine animals, namely arsenocholine [AsCho; trimethyl(2-hydroxyethyl)arsonium cation], arsenobetaine [AsBe; the trimethyl(carboxymethyl)arsonium zwitterion] and the tetramethylarsonium ion (TetMA), to murine principal immune effector cells (macrophages and lymphocytes), comparing them with the effects of inorganic arsenicals in vitro . Inorganic arsenicals (arsenite and arsenate) showed strong cytotoxicity to both macrophages and lymphocytes. The concentration of arsenite that reduced the number of surviving cells to 50% of that in untreated controls (IC₅₀) was 3–5 μmol dm⁻³, and the cytotoxicity of arsenate (IC₅₀=100 μ-1 m mol dm⁻³) was lower than that of arsenite. Compared with these findings, trimethylarsenic compounds in marine animals, AsCho and AsBe, were less toxic even at a concentration over 10 mmol dm⁻³ to both macrophages and lymphocytes; however, TetMA had weak, but significant, cytotoxicity to these cells (IC₅₀ was about 6 mmol dm⁻³).     

The separation of dimethylarsinic acid,methylarsonous acid,methylarsonic acid,arsenate and dimethylarsinous acid on the Hamilton PRP‐X100 anion‐exchange column

In order to separate the potential arsenite metabolites methylarsonous acid and dimethylarsinous acid from arsenite, arsenate, methylarsonic acid and dimethylarsinic acid, the pH‐dependent retention behaviour of all six arsenic compounds was studied on a Hamilton PRP‐X100 anion‐exchange column with 30 mM phosphate buffers (pH 5, 6, 7, 8 and 9) containing 20% (v/v) methanol as mobile phase and employing an inductively coupled plasma atomic emission spectrometer (ICP–AES) as the arsenic‐specific detector. Baseline separation of dimethylarsinic acid, methylarsonous acid, methylarsonic acid, arsenate and dimethylarsinous acid was achieved with a 30 mmol dm⁻³ phosphate buffer (pH 5)–methanol mixture (80:20, v/v) in 25 min. Arsenite is not baseline‐separated from dimethylarsinic acid under these conditions. Copyright © 1999 John Wiley & Sons, Ltd.     

The Cumulation of Chromium and Arsenic Species in Fish (Poecilia reticulata)

Abstract

Chromium-51 and arsenic-74 were used for the investigation of the uptake and the release of different chromium and arsenic species in fish. It has been found that only trimethylarsine can be rapidly taken up directly from water. The release of chromium(III), consumed by fish in food, is very rapid: about 99.9% of chromium is released within a few days. The same results were obtained with chromium(III) acetylacetonate or chromium(III) ethylenediaminotetraacetate. About 95% of arsenic acid, methylarsonic acid, dimethylarsinic acid or arsenic(III) diethyldithiocarbamate are released within a few days whereas the remaining arsenic is released with the biological half time 35 ± 5 days.     

Effects of Arsenic Compounds on Proliferation and Nitric Oxide Synthesis in C3H 10T 1/2 Murine Fibroblasts

Exposure to arsenic, either through chronic consumption of contaminated water or inhalation, is associated with increased risk of cancer, yet the mechanism by which arsenicals promote neoplastic change remains undefined. The carcinogenic process involves the formation of heritable genetic changes in the DNA of normal cells and this process may be enhanced by environmental agents that increase cellular proliferation, increase DNA damage and decrease the ability to repair damage or cause immunosuppression. We describe the inhibition of cellular proliferation of C3H 10T1/2 murine fibroblasts in the presence of 1.0 μM arsenate or arsenite; yet cacodylic acid had no significant effect on cell growth in culture at this concentration. Both arsenate and cacodylate, at micromolar concentrations, slightly stimulated cell growth and cell density when cells were treated with interferon-γ/lipopolysaccharide (IFN-γ/LPS). At 1 μM , arsenate and cacodylate also slightly increased IFN-γ/LPS-induced nitric oxide (NO) synthesis in this cell line, consistent with the increase in cell number observed, whereas 1 μM arsenite significantly increased NO production on a per-cell basis. In contrast, arsenite significantly inhibited NO synthesis at concentrations above 10 μM arsenite as, to a lesser extent, did arsenate and cacodylate. These results suggest that ingestion of arsenicals could alter cellular generation of NO and interfere with its associated physiological functions. © 1997 by John Wiley & Sons, Ltd.     

Calorimetric Investigation of Adsorption of Organic Compounds by Active Carbons

Enthalpies of wetting of two active carbons and one brown coal coke by several pure liquids (mainly homologous series) were measured at 298.15 K with two quasi-isothermal microcalorimeters. Different measuring cells with cavities of about 0.5 and 15 cm³ were used. The advantage of the larger measuring cell with three independent 15 cm³ cavities is the higher mass of active carbon, leading to a high reproducibility with standard deviations below 1% and a reduced measuring time. Experiments were carried out with n-alkanes, l-alkanols, cycloalkanes and isomers thereof, i.e. 2-propanol, methylcyclohexane, 2,2,4-trimethylpentane (isooctane and 2-ethyl-1-hexanol, and water. Like the polarity, the size and the three-dimensional expansion of the molecules, the energetic and geometric heterogeneity of the adsorbent influences the enthalpy of wetting. This revised version was published online in August 2006 with corrections to the Cover Date.     

Induction of mitotic arrest and aneuploidy by organic arsenic compounds in human lymphocytes

Inorganic arsenic is methylated in the mammalian body to methylarsonic acid (MMA), dimethylarsinic acid (DMA) and trimethylarsine oxide (TMA). To achieve a more precise understanding of arsenic carcinogenicity, we examined the genotoxic effects of organic arsenic compounds on human lymphocytes by assessing induction of mitotic arrest, sister chromatid exchange (SCE) and aneuploidy. MMA, DMA and TMA arrested mitosis, DMA induced hyperdiploid cells, and DMA and TMA induced tetraploid cells. Of the three arsenic metabolites tested, DMA had the strongest effects on cell mitosis and aneuploidy induction. DMA arrested mitosis and induced c-mitosis significantly. These results suggest that DMA arrests mitosis and induces aneuploidy through spindle disruptions similar to those observed with known spindle poisons, such as colchicine or vinblastine. Since aneuploidy has been thought to be associated with tumor induction or neoplastic transformation, induction of aneuploidy by organic metabolites of arsenic may play a major role in arsenic carcinogenesis in humans. © 1997 John Wiley & Sons, Ltd.     

The lon-chromatographic behavior of arsenite,arsenate, methylarsonic acid and dimethylarsinic acid on the hamilton PRP-X100 anion-exchange column

The HPLC separation of arsenite, arsenate, methylarsonic acid and dimethylarsinic acid has been studied in the past but not in a systematic manner. The dependence of the retention times of these arsenic compounds on the pH of the mobile phase, on the concentration and the chemical composition of buffer solutions (phosphate, acetate, potassium hydrogen phthalate) and on the presence of sodium sulfate or nickel sulfate in the mobile phase was investigated using a Hamilton PRP-X100 anion-exchange column. With a flame atomic absorption detector and arsenic concentrations of at least 10 mg dm^?3 all investigated mobile phases will separate the four arsenic compounds at appropriate pH values in the range 4–8. The shortest analysis time (?3 min) was achieved with a 0.006 mol dm^?3 potassium hydrogen phthalate mobile phase at pH 4, the longest (?10 min) with 0.006 mol dm^?3 sodium sulfate at pH 5.9 at a flow rate of 1.5 cm³ min^?1. With a graphite furnace atomic absorption detector at the required, much lower, flow rate of ?0.2 cm³ min^?1 acceptable separations were achievable only with the pH 6 phosphate buffer (0.03 mol dm^?3) and the nickel sulfate solution (0.005 mol dm^?3) as the mobile phase. To become detectable approximately 100 ng arsenic from each arsenic compound (100 μl injection) must be chromatographed with the phosphate buffer, and approximately 10 ng with the nickel sulfate solution.     

囊泡状二氧化硅的合成及油气吸附性能

以正硅酸四甲酯(TMOS)为硅源,P123(EO20PO70EO20)为表面活性剂,在p H=6的磷酸缓冲体系中制备了囊泡状二氧化硅材料.利用乙醇萃取脱除模板剂P123,电镜观测结果表明所得二氧化硅具有大孔囊泡结构,N2吸附结果表明其具有高比表面积和大孔容.通过Boehm滴定法确定了硅羟基数量与吸水率呈正相关.用囊泡状二氧化硅材料与商业化活性炭(AC)和硅胶(SG)对水蒸气、正己烷和油气进行静态吸附.在自建的动态正己烷吸附装置上用对囊泡状二氧化硅材料和商业化AC和SG对正己烷进行动态吸附.吸附结果表明,囊泡状二氧化硅材料的静/动态吸附容量和稳定性都远高于商业化活性炭和硅胶.     

Inorganic and Organic Arsenic Speciation in Seawaters by IEC-HG-AAS

Abstract

Arsenic (III), arsenic (V), monomethylarsonic acid (MMA), and dimethylarsinic acid (DMA) were separated by ion-exchange chromatography. The elution sequence was as follows: 1.5 M ammonia followed by 0.12 M hydrochloric acid yielding As(III) 1 M hydrochloric acid followed by water yielding first As(V) and then MMA. Detection was by hydride generation-atomic absorption spectrometry. No interference was noted from 13 metallic ions.

The method provides a good linearity, precision, accuracy, and sensitivity. It has been applied to the speciation of arsenic in seawaters from the North West coast of Spain (1200 Km).     

Arsenic Compounds in Zoo- and Phyto-plankton of Marine Origin

Major water-soluble arsenic compounds accumulated in some zoo- and phyto-plankton were identified. Zooplankton were collected at sampling stations in the Sea of Japan by a Norpac net towed from 600 m depth to the surface. Phytoplankton were cultivated under axenic conditions. Water-soluble arsenic compounds were extracted repeatedly from plankton tissues by aqueous methanol. The arsenic compounds in the extracts were analyzed by HPLC–ICP/MS. Among zooplankton analyzed in the present study, two carnivorous species, i.e. Amphipoda ( Themisto sp.) and Sagittoidea ( Sagitta sp.), contained arsenobetaine as the dominant arsenic species. Arsenobetaine was the major species in Euphausiacea ( Euphausia sp.), also. The most abundant arsenic compound in the herbivorous Copepoda species ( Calanus sp.), on the other hand, was an arsenic-containing ribofuranoside with a sulfate ester group, and arsenobetaine was only a minor component. Phytoplankton contained arsenic-containing ribofuranosides apparently in a species-speific manner. The arsenic compounds in zooplankton seem to reflect their feeding habit; i.e. carnivorous species eating zooplankton or other small animals accumulate arsenobetaine, while herbivorous ones eating phytoplankton accumulate arsenic-containing ribofuranosides as major arsenic compounds.     

二氧化硅气凝胶对挥发性有机污染物的吸附性能研究

以TEOS为前驱体,乙醇为溶剂,氢氟酸为催化剂,一步法合成了常规二氧化硅气凝胶。经乙醇超临界干燥后,通过SEM,FTIR和N2吸脱附分析仪等仪器对二氧化硅气凝胶样品进行表征,以更好地了解吸附机理与性质的关系。结果表明,样品的比表面积高达519 m2/g,孔体积为1.9 cm3/g,平均孔径为15.15 nm,是一种优良的吸附材料。制备的样品用作测试甲苯、对二甲苯和苯三种挥发性有机化合物的吸附效果。结果表明,二氧化硅气凝胶对三种污染物都具有很高的吸附量,其高吸附能力归因于气凝胶的三维纳米网络结构。样品对甲苯,对二甲苯和苯的最大吸附能力分别为1422.8 mg/g,707.4 mg/g和1299.4 mg/g。综上所述,二氧化硅气凝胶是一种很有前景的处理挥发性有机化合物的吸附剂,具有优异的吸附性能。     

One-Step Preparation of Chitosan-Based Magnetic Adsorbent and Its Application to the Adsorption of Inorganic Arsenic in Water

Chitosan is a kind of biodegradable natural polysaccharide, and it is a very promising adsorber material for removing metal ions from aqueous solutions. In this study, chitosan-based magnetic adsorbent CMC@Fe₃O₄ was synthesized by a one-step method using carboxymethyl chitosan (CMC) and ferric salts under relatively mild conditions. The Fe₃O₄ microspheres were formed and the core–shell structure of CMC@Fe₃O₄ was synthesized in the meantime, which was well characterized via SEM/TEM, XRD, VSM, FT-IR, thermo gravimetric analysis (TGA), XPS, size distribution, and zeta potential. The effects of initial arsenic concentration, pH, temperature, contact time, and ionic strength on adsorption quantity of inorganic arsenic was studied through batch adsorption experiments. The magnetic adsorbent CMC@Fe₃O₄ displayed satisfactory adsorption performance for arsenic in water samples, up to 20.1 mg/g. The optimal conditions of the adsorption process were pH 3.0, 30−50 °C, and a reaction time of 15 min. The adsorption process can be well described by pseudo-second-order kinetic model, suggesting that chemisorption was main rate-controlling step. The Langmuir adsorption model provided much higher correlation coefficient than that of Freundlich adsorption model, indicating that the adsorption behavior is monolayer adsorption on the surface of the magnetic adsorbents. The above results have demonstrated that chitosan-based magnetic adsorbent CMC@Fe₃O₄ is suitable for the removal of inorganic arsenic in water.     

Formation of arsenobetaine from arsenocholine by micro-organisms occurring in sediments

As one of the experiments to pursue marine circulation of arsenic, we studied microbiological conversion of arsenocholine to arsenobetaine, because arsenocholine may be a precursor of arsenobetaine in these ecosystems. Two culture media, 1/5 ZoBell 2216E and an aqueous solution of inorganic salts, were used in this in vitro study. To each medium (25 cm³) were added synthetic arsenocholine (0.2%) and about 1 g of the sediment, and they were aerobically incubated at 25°C in the dark. These conversion experiments were performed in May and July 1990. In both seasons, two or three metabolites were derived in each mixture. These metabolites were purified using cation-exchange chromatography. Their structures were confirmed as arsenobetaine, trimethylarsine oxide and dimethylarsinic acid by high-performance liquid chromatography, thin-layer chromatography, FAB mass spectrometry and a combination of gas-chromatographic separation with hydride generation followed by a cold-trap technique and selected-ion monitoring mass spectrometric analysis. From this and other evidence it is concluded that, in the arsenic cycle in these marine ecosystems, as recently postulated by us, the pathway arsenocholine → arsenobetaine → trimethylarsine oxide → dimethylarsinic acid → methanearsonic acid → inorganic arsenic can be carried out by micro-organisms alone.     

Uptake and degradation of arsenobetaine by the microorganisms occurring in sediments

We have reported the degradation of arsenobetaine [(CH₃)₃As⁺CH₂COO^?] to inorganic arsenic by microorganisms from various marine origins such as sediments. However, there was no information as to the fate of the ingested arsenobetaine within the body of the microorganisms before excretion. In this study, arsenobetaine and sediments were added to two culture media (1/5 Zobell 2216E and a solution of inorganic salts) and aerobically incubated at 25°C in the dark. Despite the degradation and complete disappearance of arsenobetaine from the filtrates of the incubation mixtures, the major arsenic compound from the microorganisms harvested from the mixtures was identified by HPLC as arsenobetaine throughout the incubation period. The presence of arsenobetaine was further confirmed by TLC and fast atom bombardment mass spectrometry (FAB MS). A minor arsenical also present in the incubated microorganisms, dimethylarsinic acid, was detected.