Pretreatment Techniques for Arsenic Speciation Analysis in Biosamples

Introduction Coupled techniques such as HPLC–HG–AAS,HPLC–ICP–MS,HPLC–HG–AFS are widely used for arsenic speciation analysis in biosamples,during which pretreatment method usually plays a significant role.This

Arsenic species analysis in porewaters and sediments using hydride generation atomic fluorescence spectrometry

It was observed that the atomic fluorescence emission due to signal during the determination of As（Ⅲ） in the mixture of As（Ⅲ） and As（V） could has a 10% to 40% of fluorescence emission As（V）. Besides, interferes from heavy metals such as Pb（Ⅱ）, Cu（Ⅱ） can cause severe increase of the signals as compared to the insignificant effects caused by Cd（Ⅱ）, Zn（Ⅱ）, Mn（Ⅱ） and Fe（Ⅲ）. On the basis of further studies, the masking agent of 8-hydroxyquinoline was used as an efficient agent to eliminate interference of As（V） emission and the heavy metal of Cu^2＋ and Pb^2＋ in the measurements of arsenic species, After a series standard additions and CRM researches, a sensitive and interference-free analytical procedure was developed for the speciation of arsenic in samples of porewaters and sediments in Poyang Lake, China.

Arsenic Exposure and Association with Hepatitis E IgG Antibodies

Arsenic is a known toxic chemical, has immuno-modulatory properties, and can change the susceptibility of infection in humans. Acute hepatitis E is an infectious disease;it can be self-limiting, but in severe cases, can cause acute-on-chronic liver failure. The presence of IgG anti-HEV (hepatitis E IgG antibody) in blood represents a past hepatitis E infection in an individual. Very few studies have investigated the association between arsenic levels and hepatitis E seroconversion in humans. The primary objective of this study was to assess the relationship between total urinary arsenic and speciated urinary arsenic (urinary arsenous acid, urinary arsenic acid, urinary arsenobetaine, urinary arsenocholine, urinary dimethylarsinic acid, urinary monomethylarsonic acid) and the presence of IgG anti-HEV. The 2011-2012, 2013-2014, and 2015-2016 National Health and Nutrition Examination Survey (NHANES) III data sets were analyzed, with participants aged 20 years and older (n = 7061). We used weighted logistic regression to determine the association between total and speciated urinary arsenic concentrations and IgG anti-HEV. For each analyte considered, a separate weighted logistic regression model was fitted. Each of these models regressed log-transformed analyte levels on the log-odds of the presence of IgG Anti-HEV. To evaluate the relationships between the urinary arsenic measurements, pairwise Pearson correlation coefficients were determined for each of the urinary arsenic measurements. Of the human subjects included, 6628 (93.9%) were negative for IgG Anti-HEV while 433 (6.1%) were positive for IgG anti-HEV. Total urinary arsenic was associated with 1.161 times greater odds of IgG anti-HEV (95% CI: [1.035, 1.303]) for each unit increase in concentration on a log-scale. For speciated urinary arsenic measurements, the odds ratios and 95% CI’s were: arsenobetanine 1.168 [1.075, 1.270], arsenocholine 1.201 [0.944, 1.529], dimethylarsinic acid 1.183 [1.009, 1.386], monomethylacrsonic acid 1.095 [0.915, 1.310], aresnous acid 0.907 [0.762, 1.079], and arsenic acid 1.951 [0.905, 4.209]. Our analysis indicates that total urinary arsenic, arsenobetanine, and dimethylarsinic acid are significantly associated with the odds of the presence of IgG anti-HEV. Future prospective studies are required to evaluate the association between hepatitis E infection and arsenic exposures.

Effect of sample treatment on determination of arsenic （Ⅲ） and arsenic （Ⅴ） in aqueous and tissue samples by hydride generation atomic absorption spectrometry

The purpose of this procedure was to optimize and improve a method that used for the determination of arsenic （Ⅲ） and arsenic （Ⅴ） in biological and environmental samples. The method is based on hydride generation and atomic absorption spectrometry. For both As （Ⅲ） and As （Ⅴ） the parameters such as NaBH4, HCI concentration, and pH were optimized. Absorption signal of As （Ⅴ） was approximately 17% of As （Ⅲ） signal. Therefore, for estimation of As （Ⅲ） and As （Ⅴ） concentrations in various samples the difference between the absorbance obtained for arsenic, without and with previous treatment of samples with potassium iodide （KI）, can be applied. The calibration graphs were linear （r〉0.99）, and the detection limits of the method based on three times the standard deviation of the blank were 0.14 and 0.64 μL^-1 for As （Ⅲ） and As （Ⅴ）, respectively. The relative standard deviation （R.S.D.） of measurements was less than 10%. As a means of checking performance method, water samples were spiked with known concentrations of both As （Ⅲ） and As （Ⅴ）, and recovery above 94% was obtained. The proposed method was applied successfully to determine inorganic As （Ⅲ） and As （Ⅴ） in various environmental and total As in biological samples.

Removal of Arsenic from Chlamys farreri with Different Methods

Arsenic is commonly found in seafood,and the toxicity of different speciation varies widely.Therefore,there is an urgent need to reduce the content and analyze the speciation of arsenic to improve the safety of seafood.In this study,arsenic was removed from Chlamys farreri by adding ferric ions and sodium citrate or reducing the salinity of seawater.Then,the speciation of arsenic was analyzed.The results showed that the removal rates of arsenic were 20%,23%,24%and 18%when C.farreri individuals were treated with 0.1 g L^(−1)Fe^(3+),0.4 g L^(−1),0.6 g L^(−1)sodium citrate or decreasing seawater salinity to 17 within 48 h,respectively.Furthermore,it was detected that the arsenic speciation in the C.farreri was mainly polar arsenic,and among the polar arsenic was mainly arsenic betaine.These results indicated that the removal of arsenic can be achieved by adding the removal agent or changing the seawater salinity under the experimental conditions.The speciation of arsenic did not change during the removal process and it was mainly excreted in the speciation of arsenic betaine.

A novel colorimetric method for field arsenic speciation analysis

Accurate on-site determination of arsenic （As） concentration as well as its speciation presents a great environmental challenge especially to developing countries. To meet the need of routine field monitoring, we developed a rapid colorimetric method with a wide dynamic detection range and high precision. The novel application of KMnO4 and CHaN2S as effective As（III） oxidant and As（V） reductant, respectively, in the formation of molybdenum blue complexes enabled the differentiation of As（III） and As（V）. The detection limit of the method was 8 ~tg/L with a linear range （R2 = 0.998） of four orders of magnitude in total As concentrations. The As speciation in groundwater samples determined with the colorimetric method in the field were consistent with the results using the high performance liquid chromatography atomic fluorescence spectrometry, as evidenced by a linear correlation in paired analysis with a slope of 0.9990- 0.9997 （p 〈 0.0001, n = 28）. The recovery of 96%-116% for total As, 85%-122% for As（III）, and 88%-127% for As（V） were achieved for groundwater samples with a total As concentration range 100-800 μg/L. The colorimetric result showed that 3.61 g/L As（III） existed as the only As species in a real industrial wastewater, which was in good agreement with the HPLC-AFS result of 3.56 g/L As（Ⅲ）. No interference with the color development was observed in the presence of sulfate, phosphate, silicate, humic acid, and heavy metals from complex water matrix. This accurate, sensitive, and easy-to-use method is especially suitable for field As determination.

Effects of water management on arsenic and cadmium speciation and accumulation in an upland rice cultivar

Pot and field experiments were conducted to investigate the effects of water regimes on the speciation and accumulation of arsenic（As） and cadmium（Cd） in Brazilian upland rice growing in soils polluted with both As and Cd. In the pot experiment constant and intermittent flooding treatments gave 3–16 times higher As concentrations in soil solution than did aerobic conditions but Cd showed the opposite trend. Compared to arsenate, there were more marked changes in the arsenite concentrations in the soil solution as water management shifted, and therefore arsenite concentrations dominated the As speciation and bioavailability in the soil. In the field experiment As concentrations in the rice grains increased from 0.14 to 0.21 mg/kg while Cd concentrations decreased from 0.21 to 0.02 mg/kg with increasing irrigation ranging from aerobic to constantly flooding conditions. Among the various water regimes the conventional irrigation treatment produced the highest rice grain yield of 6.29 tons/ha. The As speciation analysis reveals that the accumulation of dimethylarsinic acid（from 11.3% to 61.7%） made a greater contribution to the increase in total As in brown rice in the intermittent and constant flooding treatments compared to the intermittent-aerobic treatment. Thus, water management exerted opposite effects on Cd and As speciation and bioavailability in the soil and consequently on their accumulation in the upland rice. Special care is required when irrigation regime methods are employed to mitigate the accumulation of metal（loid）s in the grain of rice grown in soils polluted with both As and Cd.

Arsenate and phosphate interaction in Saccharomyces cerevisiae

In the present study, arsenate（As（V）） and phosphate（P（V）） interactions were investigated in growth, uptake and RNA content in yeast（Saccharomyces cerevisiae）. Yeast grew slowly with As（V） concentrations increasing in the medium. However, the maximal population density was almost the same among different As（V） treatments. It was in the late log phase that yeast growth was aug- mented by low As（V）, which was maybe due to the fact that methionine metabolism was stressed by vitamin B6 deprivation, so As（V） treatments did not affect maximal population density. However, with P （V） concentrations increasing, the maximal population density increased. Therefore, the maximal population density was determined by P （V） concentrations in the medium but not by As （V） concentrations in the medium. Ycflp（a tonoplast transpor） transports As（GS）3 into the vacuole, but arsenic（As） remaining in the thalli was 1.27% with As（V） exposure for 60 h, from which it can be speculated that the percentage of As transported into vacuole should be lower than 1.27%. However, the percentage of As pumped out of cell was 71.49% with As （V） exposure for 68 h. Although two pathways （extrusion and sequestration） were involved in As detoxification in yeast, the extrusion pathway played a major role in As detoxification. RNA content was the highest in the early-log phase and was reduced by As（V）.

Tannic acid and saponin for removing arsenic from brownfield soils: Mobilization, distribution and speciation

Plant biosurfactants were used for the first time to remove As and co-existing metals from brownfield soils. Tannic acid （TA）, a polyphenol, and saponin （SAP）, a glycoside were tested. The soil washing experiments were performed in batch conditions at constant biosurfactant concentration （3%）. Both biosurfactants differed in natural pH, surface tension, critical micelle concentration and content of functional groups. After a single washing, TA （pH 3.44） more efficiently mobilized As than SAP （pH 5.44）. When both biosurfactants were used at the same pH （SAP adjusted to 3.44）, arsenic mobilization was improved by triple washing. The process efficiency for TA and SAP was similar, and depending on the soil sample, ranged between 50%-64%. Arsenic mobilization by TA and SAP resulted mainly from decomposition of Fe arsenates, followed by Fe3＋ complexation with biosurfactants. Arsenic was efficiently released from reducible and partially from residual fractions. In all soils, As（V） was almost completely removed, whereas content of As（III） was decreased by 37%-73%. SAP and TA might be used potentially to remove As from contaminated soils.

Effects of root oxidation ability and P on As mobility and bioavailability in rice

The effects of root oxidization ability and P fertilization on As mobility in soils, and subsequently As uptake, translocation and speciation in rice plants were investigated. Results show that root oxidation significantly influences As mobility in rhizoshphere.Genotype TD71 with higher radial oxygen loss(ROL) induces more Fe plaque formation and sequesters more As and P in iron plaque and rhizoshphere soil, leading to the reduction of As accumulation in rice plants. Additionally, P addition mobilizes As in soil solution, and increases As accumulation in rice plants. Arsenic speciation results show that the majority of As species in husks detected is inorganic As, accounting for 82%-93% of the total As, while in grains the majority of As is inorganic As and dimethyl arsenic(DMA), with DMA accounting for 33%-64% of the total As. The fraction of inorganic As decreases while fraction of DMA increases, with increasing As and P concentrations. The study further elucidates the mechanisms involved in effects of ROL on As tolerance and accumulation in rice.

Concentration and speciation of arsenic in an insect feeding on the leaves of Pteris vittata

The development of an effective and green bioinsecticide is a research hotspot.This study demonstrated the possibility of using an arsenic(As)hyperaccumulator as a bioinsecticide.When the As concentration in the Pteris vittata fronds exceeded 138 mg kg^(–1),the larva of the hawk moth(Theretra clotho)displayed apparent preference to lower-As-concentration P.vittata fronds.The As concentration in the larva body was as high as 850 mg kg^(–1).Such high concentration of As in the larva body might have been the case that T.clotho lacks a process to exclude As.The larval frass showed an As concentration of only 1%–4%of that in the larva body.The predominant As species in the larva body and frass was As(III)-SH.The percentage of As(III)-SH was slightly higher in the frass than that in the larval body.Chelation with thiols may be a universal detoxification mechanism for As in both plants and insects.In general,the adoption of P.vittata as a bioinsecticide should be feasible.However,the exact processes to achieve this goal still need further study.The mechanism of different animals to detoxify As is another interesting research topic.

Alcohol consumption promotes arsenic absorption but reduces tissue arsenic accumulation in mice

Alcohol consumption alters gut microflora and damages intestinal tight junction barriers,which may affect arsenic(As)oral bioavailability.In this study,mice were exposed to arsenate in the diet(6μg/g)over a 3-week period and gavaged daily with Chinese liquor(0.05 or 0.10 mL per mouse per day).Following ingestion,78.0%and 72.9%of the total As intake was absorbed and excreted via urine when co-exposed with liquor at daily doses of 0.05 or 0.10 mL,significantly greater than when As was supplied alone(44.7%).Alcohol co-exposure significantly altered gut microbiota but did not significantly alter As biotransformation in the intestinal tract or tissue.Significantly lower relative mRNA expression was observed for genes encoding for tight junctions in the ileum of liquor co-exposed mice,contributing to greater As bioavailability attributable to enhanced As absorption via the intestinal paracellular pathway.However,As concentration in the liver,kidney,and intestinal tissue of liquortreated mice was decreased by 24.4%-42.6%,27.5%-38.1%,and 28.1%-48.9%compared to control mice.This was likely due to greater renal glomerular filtration rate induced by alcohol,as suggested by significantly lower expression of genes encoding for renal tight junctions.In addition,in mice gavaged daily with 0.05 mL liquor,the serum antidiuretic hormone level was significantly lower than control mice(2.83±0.59 vs.5.40±1.10 pg/mL),suggesting the diuretic function of alcohol consumption,which may facilitate As elimination via urine.These results highlight that alcohol consumption has a significant impact on the bioavailability and accumulation of As.

Arsenic biotransformation and an arsenite Sadenosylmethionine methyltransferase in plankton

Arsenic（As）is a well-recognized toxicant and carcinogen.Chronic exposure to inorganic arsenic causes a range of human cancers（e.g.,skin,bladder,and lung）and increases the risk of developing diabetes,hypertension,and cardiovascular and neurological diseases.The prevalence of arsenic species and the severity of their health effects continue to drive and demand for extensive research（Carlin et al.,2016）.

Methylated and thiolated arsenic species for environmental and health research——A review on synthesis and characterization

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 following14 trivalent（Ⅲ） and pentavalent（） arsenic compounds： monomethylarsonous acid（MMA~Ⅲ）, dicysteinylmethyldithioarsenite（MMA~Ⅲ（Cys）_2）, monomethylarsonic acid（MMA~Ⅴ）,monomethylmonothioarsonic acid（MMMTAⅤ） 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-DMAⅤ, 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.

Arsenic species in electronic cigarettes:Determination and potential health risk

Human exposure to contaminants from electronic cigarettes(e-cigarettes)and the associated health effects are poorly understood.There has been no report on the speciation of arsenic in e-liquid(solution used for e-cigarettes)and aerosols.We report here determination of arsenic species in e-liquids and aerosols generated from vaping the e-liquid.Seventeen e-liquid samples of major brands,purchased from local and online stores in Canada and China,were analyzed for arsenic species using high-performance liquid chromatography and inductively coupled plasma mass spectrometry.Aerosols condensed from vaping the eliquids were also analyzed and compared for arsenic species.Six arsenic species were detected,including inorganic arsenate(iAsⅤ),arsenite(iAsⅢ),monomethylarsonic acid(MMA),and three new arsenic species not reported previously.In e-liquids,iAsⅢwas detected in 59%,iAsⅤin 94%,and MMA in 47%of the samples.In the condensate of aerosols from vaping the e-liquids,iAsⅢwas detected in 100%,iAsv in 88%,and MMA in 13%of the samples.Inorganic arsenic species were predominant in e-liquids and aerosols of e-cigarettes.The concentration of iAsⅢin the condensate of aerosols(median 3.27μg/kg)was significantly higher than that in the e-liquid(median 1.08μg/kg)samples.The concentration of inorganic arsenic in the vaping air was approximately 3.4μg/m3,which approaches to the permissible exposure limit(10μg/m3)set by the United States Occupational Safety and Health Administration(OSHA).According to the Environmental Protection Agency’s unit risk factor(4.3×10-3 perμg/m3)for inhalation exposure to inorganic arsenic in the air,the estimated excess lung cancer risk from lifetime exposure to inorganic arsenic in the ecigarette vaping air(3.4μg/m3),assuming e-cigarette vaping at 1%of the time,is as high as1.5×10-4.These results raise health concerns over the exposure to arsenic from electronic cigarettes.

Oxidation state specific analysis of arsenic species in tissues of wild-type and arsenic(+3 oxidation state)methyltransferase-knockout mice

Arsenic methyltransferase（As3mt） catalyzes the conversion of inorganic arsenic（i As） to its methylated metabolites, including toxic methylarsonite（MAs~Ⅲ） and dimethylarsinite（DMAs~Ⅲ）. Knockout（KO） of As3 mt was shown to reduce the capacity to methylate i As in mice. However, no data are available on the oxidation states of As species in tissues of these mice. Here, we compare the oxidation states of As species in tissues of male C57BL/6 As3mt-KO and wild-type（WT） mice exposed to arsenite（iA s~Ⅲ） in drinking water. WT mice were exposed to50 mg/L As and As3mt-KO mice that cannot tolerate 50 mg/L As were exposed to 0, 15, 20, 25 or30 mg/L As. iA s~Ⅲaccounted for 53% to 74% of total As in liver, pancreas, adipose, lung, heart, and kidney of As3mt-KO mice; tri- and pentavalent methylated arsenicals did not exceed 10% of total As. Tissues of WT mice retained iA s and methylated arsenicals： iA s~Ⅲ, MAs~Ⅲand DMAs~Ⅲ represented 55%‐68% of the total As in the liver, pancreas, and brain. High levels of methylated species, particularly MAs~Ⅲ, were found in the intestine of WT, but not As3mt-KO mice,suggesting that intestinal bacteria are not a major source of methylated As. Blood of WT mice contained significantly higher levels of As than blood of As3mt-KO mice. This study is the first to determine oxidation states of As species in tissues of As3mt-KO mice. Results will help to design studies using WT and As3mt-KO mice to examine the role of iA s methylation in adverse effects of iA s exposure.

SERS detection of arsenic in water:A review

Arsenic（As） is one of the most toxic contaminants found in the environment. Development of novel detection methods for As species in water with the potential for field use has been an urgent need in recent years. In past decades, surface-enhanced Raman scattering（SERS）has gained a reputation as one of the most sensitive spectroscopic methods for chemical and biomolecular sensing. The SERS technique has emerged as an extremely promising solution for in-situ detection of arsenic species in the field, particularly when coupled with portable/handheld Raman spectrometers. In this article, the recent advances in SERS analysis of arsenic species in water media are reviewed, and the potential of this technique for fast screening and field testing of arsenic-contaminated environmental water samples is discussed. The problems that remain in the field are also discussed and an outlook for the future is featured at the end of the article.

Photooxidation of arsenic(Ⅲ) to arsenic(Ⅴ) on the surface of kaolinite clay

As one of the most toxic heavy metals, the oxidation of inorganic arsenic has drawn great attention among environmental scientists. However, little has been reported on the solar photochemical behavior of arsenic species on top-soil. In the present work, the influencing factors（p H, relative humidity（RH）, humic acid（HA）, trisodium citrate, and additional iron ions） and the contributions of reactive oxygen species（ROS, mainly HO^- and HO2^-/O2^-） to photooxidation of As（Ⅲ） to As（Ⅴ） on kaolinite surfaces under UV irradiation（λ = 365 nm）were investigated. Results showed that lower p H facilitated photooxidation, and the photooxidation efficiency increased with the increase of RH and trisodium citrate.Promotion or inhibition of As（Ⅲ） photooxidation by HA was observed at low or high dosages, respectively. Additional iron ions greatly promoted the photooxidation, but excessive amounts of Fe^2＋competed with As（Ⅲ） for oxidation by ROS. Experiments on scavengers indicated that the HOUradical was the predominant oxidant in this system.Experiments on actual soil surfaces proved the occurrence of As（Ⅲ） photooxidation in real topsoil. This work demonstrates that the photooxidation process of As（Ⅲ） on the soil surface should be taken into account when studying the fate of arsenic in natural soil newly polluted with acidic wastewater containing As（Ⅲ）.

Experimental study on the uptake and effects of arsenic originated from poultry litter on the growth of Brassica napus in greenhouse pot cultivation and health risk assessment

Organoarsenics are widely used as growth promoters in poultry industry,resulting in arsenic(As)accumulation in poultry litter.A greenhouse pot study was implemented to investigate the fate of arsenic originated from poultry litter and their effects on the growth of Brassica napus(oilseed rape),and assess their potential health risks.Five poultry litter application rates(0,5%,10%,20%and 40%)were used,dividing into two groups:one for soil incubation(SI)and the other for plant cultivation(PC).Experimental results indicated that the total arsenic for composted poultry litter was(10.94±0.23)mg/kg,As(V)and As(III)decreased while methylated arsenic increased after 21 d in SI and PC treatments.Seed germination rates were negatively correlated with monomethylarsenic acid(MMA,R2=0.63,p<0.05).The length and biomass of roots and shoots were significantly inhibited by poultry litter,but plant length of 5%treatments was slightly stimulated.Within an average weekly intake of 0.5 kg Brassica napuss,the risk quotient(RQ)values induced from roots nearly all surpassed the acceptable limit(1),were two orders magnitude higher than shoots.According to the potential risk to order,child exhibited the highest risk,adolescent ranked secondly,and adult exhibited the lowest risk.Hence,people should better avoid intake Brassica napus roots to reduce arsenic potential risk.

The role of phosphorus in the metabolism of arsenate by a freshwater green alga, Chlorella vulgaris

A freshwater microalga, Chlorella vulgaris, was grown in the presence of varying phosphate concentrations（〈 10–500 μg/L P） and environmentally realistic concentrations of arsenate（As（Ⅴ））（5–50 μg/L As）. Arsenic speciation in the culture medium and total cellular arsenic were measured using AEC-ICP-MS and ICP-DRC-MS, respectively, to determine arsenic biotransformation and uptake in the various phosphorus scenarios. At high phosphate concentration in the culture medium, 〉 100 μg/L P, the uptake and biotransformation of As（Ⅴ） was minimal and dimethylarsonate（DMAs（Ⅴ）） was the dominant metabolite excreted by C. vulgaris, albeit at relatively low concentrations. At common environmental P concentrations, 0–50 μg/L P, the uptake and biotransformation of As（Ⅴ） increased. At these higher As-uptake levels, arsenite（As（Ⅲ）） was the predominant metabolite excreted from the cell. The concentrations of As（Ⅲ） in these low P conditions were much higher than the concentrations of methylated arsenicals observed at the various P concentrations studied. The switchover threshold between the（small） methylation and（large） reduction of As（Ⅴ） occurred around a cellular As concentration of 1 fg/cell. The observed nearly quantitative conversion of As（Ⅴ） to As（Ⅲ） under low phosphate conditions indicates the importance of As（Ⅴ） bio-reduction at common freshwater P concentrations. These findings on the influence of phosphorus on arsenic uptake, accumulation and excretion are discussed in relation to previously published research. The impact that the two scenarios of As（Ⅴ） metabolism, As（Ⅲ） excretion at high As（Ⅴ）-uptake and methylarsenical excretion at low As（Ⅴ）-uptake, have on freshwater arsenic speciation is discussed.