Arsenocholine Research Articles

Metabolome analysis revealed the critical role of betaine for arsenobetaine biosynthesis in the marine medaka (Oryzias melastigma)

Arsenobetaine (AsB), a non-toxic arsenic (As) compound found in marine fish, structurally resembles betaine (GB), a common methyl donor in organisms. This study investigates the potential role of GB in AsB synthesis in marine medaka (Oryzias melastigma) using metabolomic analysis. Dietary exposure to arsenate (As(V)) and varying GB concentrations (0.05% and 0.1% in diets) increased total As and AsB bioaccumulation, particularly in marine medaka muscle. Metabolomic analysis revealed that GB played a crucial role in promoting up-regulation in methylthioadenosine (MTA) by modulating the methionine cycle and down-regulation in glutathione (GSH) by modulating the glutathione cycle. Methionine metabolism and GSH, potentially binding again to exogenous GB, could synchronously produce more non-toxic AsB. Combining verification experiments of differential metabolites of Escherichia coli in vitro, GB, GSH, S-adenosylmethionine (SAM), and arsenocholine (AsC) entered methionine and glutathione metabolism pathways to generate more AsB. These findings underscore the GB's crucial regulatory role in modulating the synthesis of AsB. This study provides vital insights into the interplay between the structural analogs GB and AsB, offering specific strategies to enhance the detoxification mechanisms of marine fish in As-contaminated environments.

Speciation of arsenic in milk from cows fed seaweed.

Including seaweed in cattle feed has gained increased interest, but it is important to take into account that the concentration of toxic metals, especially arsenic, is high in seaweed. This study investigated the arsenic species in milk from seaweed-fed cows. Total arsenic in milk of control diets (9.3 ± 1.0 μg As kg-1, n = 4, dry mass) was significantly higher than seaweed-based diet (high-seaweed diet: 7.8 ± 0.4 μg As kg-1, P < 0.05, n = 4, dry mass; low-seaweed diet: 6.2 ± 1.0 μg As kg-1, P < 0.01, n = 4, dry mass). Arsenic speciation showed that the main species present were arsenobetaine (AB) and arsenate (As(V)) (37% and 24% of the total arsenic, respectively). Trace amounts of dimethylarsinic acid (DMA) and arsenocholine (AC) have also been detected in milk. Apart from arsenate being significantly lower (P < 0.001) in milk from seaweed-fed cows than in milk from the control group, other arsenic species showed no significant differences between groups. The lower total arsenic and arsenate in seaweed diet groups indicates a possible competition of uptake between arsenate and phosphate, and the presence of AC indicates that a reduction of AB occurred in the digestive tract. Feeding a seaweed blend (91% Ascophyllum nodosum and 9% Laminaria digitata) does not raise As-related safety concerns for milk. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Key genes for arsenobetaine synthesis in marine medaka (Oryzias melastigma) by transcriptomics

Marine fish undergo detoxification to overcome As stress, forming non-toxic metabolites arsenobetaine (AsB). Genes associated with AsB synthesis remain unknown. Therefore, in this study, we explored the key genes involved in the synthesis of AsB by transcriptomic analysis in marine medaka (Oryzias melastigma), and then screened candidate genes related to AsB synthesis. In the liver, 40 genes were up-regulated and 23 genes were down-regulated, whereas in muscle, 83 genes were up-regulated and 331 genes were down-regulated. We revealed that bhmt, mat2aa, and gstt1a can play a significant role in the glutathione and methionine metabolic pathway. These three genes can affect the conversion of arsenocholine (AsC) to AsB by the vitro gene transformation experiments of E. coli BL21(DE3). E. coli BL21-bhmt overexpressing bhmt resulted in more oxidation of precursor AsC to AsB. Furthermore, the AsB concentration was decreased after E. coli BL21 overexpressing mat2aa and gstt1a, which were down-regulated in marine medaka. Therefore, we concluded that bhmt, mat2aa, and gstt1a are involved in AsB synthesis. Overall, this is the first report on transcriptome screening and identification of key genes for AsB synthesis in marine medaka. We provided important insights to reveal the mystery of AsB synthesis in marine fish.

Species composition and health risk assessment of arsenic in Agaricus blazei Murrill and Tricholoma matsutake from Yunnan Province, China

High concentrations of total arsenic (As) has been found in Agaricus blazei Murrill and Tricholoma matsutake samples. It is important to study the As species in A. blazei and T. matsutake for accurate health risk assessment. Total As was measured by inductively coupled plasma-mass spectrometry (ICP-MS). An analytical method was developed to determine As species in mushroom by using anion exchange and cation exchange chromatography coupled to ICP-MS. Total As contents ranged from 5.25 to 69.5 mg kg1. Arsenobetaine (AsB), inorganic arsenic (iAs), and monomethylarsenic acid (MMA) were the main form of As in A. blazei samples, which accounted for 42.7–88.8 %, 1.59–26.2 %, and 0.74–1.12 % of the extractable As, respectively. AsB, and dimethylarsenic acid (DMA) were the main form of As in T. matsutake samples, which accounted for 88.9–97.9 %, and 1.79–3.80 % of the extractable As, respectively. Arsenocholine (AsC), trimethylarsine oxide (TMAO), and tetramethylarsonium cation (TMA+) were present as well, but only at trace concentrations. The health risk index estimation results indicated that no particular risk of iAs exposure to consumers existed in T. matsutake. The iAs exposure in A. blazei can poses certain health risks.

Concentrations of blood and urinary arsenic species and their characteristics in general Korean population

Arsenic (As) exposure has been extensively studied by investigating As species (e.g., inorganic arsenic (iAs), monomethylarsonic acid (MMA), and dimethylarsinic acid (DMA)) in urine, yet recent research suggests that blood could be a possible biomarker of As exposure. These investigations, however, were conducted on iAs-contaminated areas, and evidence on populations exposed to low levels of iAs is limited. This study aimed to describe the levels and distributions of As species in urine and blood, as well as to estimate methylation efficiency and related factors in the Korean population. Biological samples were obtained by the Korean Ministry of Food and Drug Safety. A total of 2025 urine samples and 598 blood samples were utilized in this study. Six As species were measured using ultra-high-performance liquid chromatography with inductively coupled plasma mass spectrometry (UPLC-ICP-MS): As(V), As(III), MMA, DMA, arsenobetaine (AsB), and arsenocholine (AsC). Multiple linear regression models were used to examine the relationship between As species (concentrations and proportions) and covariates. AsB was the most prevalent species in urine and blood. The relative composition of iAs, MMA, DMA, and AsC in urine and blood differed significantly. Consumption of blue-backed fish was linked to higher levels of AsB in urine and blood. Type of drinking water and multigrain rice consumption were associated with increased iAs concentration in urine. Except for iAs, every species had correlations in urine and blood in both univariate and multivariate analyses. Adolescents and smokers presented a lower methylation efficiency (higher %MMA and lower %DMA in urine) and females presented a higher methylation efficiency (lower %iAs, %MMA, and higher %DMA in urine). In conclusion, blood iAs concentration cannot represent urinary iAs; nonetheless, different compositions of urine and blood might reflect distinct information about iAs exposure. Further investigations on exposure factors and health are needed using low-exposure groups.

Form and valence of arsenic in dry and fresh Cordyceps breeding products based on HPLC-ICP-MS and its risk assessment

A comparative study was conducted for the first time on the form and valence of arsenic in the dry and fresh Cordyceps breeding products to clarify the specific morphology and valence of arsenic in Cordyceps breeding products and its safety. Arsenic betai-ne(AsB), arsenite(AsⅢ), dimethyl arsenic(DMA), arsenocholine(AsC), monomethyl arsenic(MMA), and arsenate(AsⅤ) in the dry and fresh samples were investigated using a bionic extraction method combined with HPLC-ICP-MS. The HPLC separation was performed on a DioncxIonPac~(TM) AS7 anion exchange column with a mobile phase of 100 mmol·L~(-1) ammonium carbonate-water for gradient elution at room temperature and the flow rate of 0.8 mL·min~(-1). HPLC was coupled with ICP-MS for the determination. The arsenic content was characterized in combination with chemometrics. The health safety risk of inorganic arsenic in the samples was assessed using the margin of exposure(MOE). The results of methodological validation showed that the six arsenic compounds showed good linearity(R~2&gt;0.999) from 10 to 500 ng·mL~(-1), with precision RSDs of 1.8%-3.0%, recoveries(n=6) of 84.15%-98.28%, reproducibility RSDs of 6.4%-7.7%, and sample stability RSDs of 8.3%-14% within 10 h. Trace AsⅢ and AsⅤ were detected in 30 batches of dry and fresh Cordyceps breeding products, while arsenic compounds in other forms and valence were not detected. In the dry products, AsⅢ was 0.019-0.040 mg·kg~(-1) and AsV was 0.024-0.061 mg·kg~(-1), while in the fresh products, AsⅢ was 0.002 3-0.006 1 mg·kg~(-1) and AsⅤ was 0.008-0.016 mg·kg~(-1). The risk assessment results showed that the MOE of inorganic arsenic was much higher than 1 in both dry and fresh products, and the potential health safety risk of inorganic arsenic was low. The HPLC-ICP-MS method established in this study was efficient, rapid, accurate, and stable for the determination of six arsenic compounds in Cordyceps breeding products. The results of this study provide a basis for the safety and quality control of Cordyceps breeding products.

Green and Simple Extraction of Arsenic Species from Rice Flour Using a Novel Ultrasound-Assisted Enzymatic Hydrolysis Method

It is well established that arsenic (As) has many toxic compounds, and in particular, inorganic As (iAs) has been classified as a type-1 carcinogen. The measuring of As species in rice flour is of great importance since rice is a staple of the diet in many countries and a major contributor to As intake in the Asian diet. In this study, several solvents and techniques for the extraction of As species from rice flour samples prior to their analysis by HPLC-ICP-MS were investigated. The extraction methods were examined for their efficiency in extracting various arsenicals from a rice flour certified reference material, NMIJ-7532a, produced by the National Metrology Institute of Japan. Results show that ultrasound-assisted extraction at 60 °C for 1 h and then heating at 100 °C for 2.5 h in the oven using a thermostable α-amylase aqueous solution was highly effective in liberating the arsenic species. The recoveries of iAs and dimethylarsinic acid (DMA) in NMIJ-7532a were 99.7% ± 1.6% (n = 3) and 98.1% ± 2.3% (n = 3), respectively, in comparison with the certificated values. Thus, the proposed extraction method is a green procedure that does not use any acidic, basic, or organic solvents. Moreover, this extraction method could effectively maintain the integrity of the native arsenic species of As(III), As(V), monomethylarsonate (MMA), DMA, arsenobetaine (AsB) and arsenocholine (AsC). Under the optimum extraction, chromatography and ICP-MS conditions, the limits of detection (LOD) obtained were 0.47 ng g−1, 1.67 ng g−1 and 0.80 ng g−1 for As(III), As(V) and DMA, respectively, while the limits of quantification (LOQ) achieved were 1.51 ng g−1, 5.34 ng g−1 and 2.57 ng g−1 for As(III), As(V) and DMA, respectively. Subsequently, the proposed method was successfully applied to As speciation analysis for several rice flour samples collected from contaminated areas in China.

Simple and reliable determination of total arsenic and its species in seafood by ICP-MS and HPLC-ICP-MS

Quantitative, rapid, selective and sensitive methods for the determination of total arsenic (tAs) and six arsenic compounds (arsenite (As(III)), arsenate (As(V)), arsenobetaine (AsB), arsenocholine (AsC), monomethylarsonic acid (MMA), dimethylarsonic acid (DMA)) in seafood were developed. The measurement of the tAs concentration was performed using quadrupole inductively-coupled plasma mass spectrometry (ICP-MS). Microwave-assisted extraction was used for the isolation of arsenic species. The separation and quantification of analysed compounds were performed by ion-exchange chromatography coupled with ICP-MS in one chromatographic run using ammonium carbonate-based buffers, which has little effect on ICP-MS sensitivity compared to commonly used phosphate buffers. The results of validation and proficiency tests confirmed the reliability, robustness, and applicability of the developed procedures to various types of matrices. The proposed methods are relatively simple, time- and cost-efficient, therefore could be used to routinely analyse tAs content and arsenic species in different types of seafood at trace and ultra-trace levels.

Rare, Protein-Altering Variants in AS3MT and Arsenic Metabolism Efficiency: A Multi-Population Association Study.

Background:Common genetic variation in the arsenic methyltransferase (AS3MT) gene region is known to be associated with arsenic metabolism efficiency (AME), measured as the percentage of dimethylarsinic acid (DMA%) in the urine. Rare, protein-altering variants in AS3MT could have even larger effects on AME, but their contribution to AME has not been investigated.Objectives:We estimated the impact of rare, protein-coding variation in AS3MT on AME using a multi-population approach to facilitate the discovery of population-specific and shared causal rare variants.Methods:We generated targeted DNA sequencing data for the coding regions of AS3MT for three arsenic-exposed cohorts with existing data on arsenic species measured in urine: Health Effects of Arsenic Longitudinal Study (HEALS, ), Strong Heart Study (SHS, ), and New Hampshire Skin Cancer Study (NHSCS, ). We assessed the collective effects of rare (allele frequency ), protein-altering AS3MT variants on DMA%, using multiple approaches, including a test of the association between rare allele carrier status (yes/no) and DMA% using linear regression (adjusted for common variants in 10q24.32 region, age, sex, and population structure).Results:We identified 23 carriers of rare-protein-altering AS3MT variant across all cohorts (13 in HEALS and 5 in both SHS and NHSCS), including 6 carriers of predicted loss-of-function variants. DMA% was 6–10% lower in carriers compared with noncarriers in HEALS [ (95% CI: , )], SHS [ (95% CI: , )], and NHSCS [ (95% CI: , )]. In meta-analyses across cohorts, DMA% was 8.7% lower in carriers [ (95% CI: , )].Discussion:Rare, protein-altering variants in AS3MT were associated with lower mean DMA%, an indicator of reduced AME. Although a small percentage of the population (0.5–0.7%) carry these variants, they are associated with a 6–10% decrease in DMA% that is consistent across multiple ancestral and environmental backgrounds. https://doi.org/10.1289/EHP8152

Arsenic speciation in rice samples for trace level determination by high performance liquid chromatography-inductively coupled plasma-mass spectrometry

Six arsenic species, namely arsenite (AsIII), arsenate (AsV), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), arsenobetaine (AsB) and arsenocholine (AsC) were speciated using a combination of high-performance liquid chromatography and inductively coupled plasma mass spectrometry (HPLC–ICP-MS). Under optimum chromatographic conditions, six arsenic species were well separated, and the performance of the combined system (HPLC–ICP-MS) for the species was determined. The limits of detection were calculated in the range of 0.14–0.29 ng/mL, and the corresponding quantification limits ranged between 0.45 and 0.97 ng mL−1 for the species. Spike recovery experiments performed on rice samples were used to validate the method’s applicability to complex matrices. The recovery results calculated ranged between 93 and 109%, validating the method’s applicability. Triplicate measurements for all spiked samples recorded percent relative standard deviation values below 10%.

Risks associated with the consumption of inorganic and organic arsenic

It is known that the chronic intake of arsenic (As) leads to the development of multisystem pathologies. In the case of high levels of As consumption, the risks of negative effects of exposure to inorganic and methylated forms persist for decades. The International Agency for Research on Cancer (IARC) has established that all inorganic forms of As are absolute carcinogens (group 1). Methylated forms of As are classified as substances possibly carcinogenic to humans (group 2b). Not metabolized in the human body forms of organic As are not carcinogenic to humans (group 3). The aim - assessment of health risks of various forms of As contained in food, including seafood. Material and methods. The scientific data regarding the risks associated with As contamination of food obtained by using PubMed, Web of Science, Google Scholar databases, legislative and regulatory acts of the Eurasian Economic Union, the Russian Federation, the Codex Alimentarius Commission, the European Union and other coun tries have been analyzed. Results. The analysis of the available data showed that the degree of As toxicity decreased in the following order: glutathione of dimethylarsonic acid (DMAIIIGl) > methylarsic acid (MMAIII) > dimethylarsic acid (DMAIII) > arsenic hydrocarbons (AsHC) > arsenite (AsIII) > arsenate (AsV) > trimethylarsine (TMAIII) > methylarsonic acid (MMAV) > dimethylarsonic acid (DMAV) > DMAIII-sugar glyceride > DMAV-sugar glyceride > thio compounds of DMAV > arsenosugarsIII > arsenosugarsV > tetramethylarsonium chloride (TETPA) > trimethylarsine oxide (TMAO), arsenocholine (AsC) > arsenobetaine (AB). Consequently, the toxicity of some methylated and organic forms of As (for example, DMAIIIGl, AsHC) may be higher than that of its inorganic forms. It is known that As is found in foods mainly in organic forms. When ingested with food, As organic forms are metabolized and, thus, could cause a number of negative effects in the organism. Conclusion. High levels of organic and inorganic forms of As in food, including seafood, could have a negative impact on the health of the population, which makes it necessary to conduct additional assessments of the health risks of various forms of As entering the body. The insufficient amount of data on the toxicity of organic forms of As indicates the impossibility of separate setting of safety maximum levels for organic and inorganic forms of As in foods.

Occurrence, speciation analysis and health risk assessment of arsenic in Chinese mitten crabs (Eriocheir sinensis) collected from China

Arsenic (As) is a major hazardous element in natural environments, and arsenic pollution is becoming an issue of concern worldwide. The more toxic inorganic arsenic, such as trivalent arsenite As(Ⅲ) and pentavalent arsenate As(Ⅴ), which normally can transform to organic arsenic compounds, such as arsenobetaine (AsB), arsenocholine (AsC), monomethyl arsenic acid (MMA), dimethyl arsenic acid (DMA), trimethyl arsenic acid (TMA), arsenosugars and arsenolipids. In this study, a total of 2130 individuals of Chinese mitten crabs were collected from seven locations and mixed to 71 samples. Total arsenic and six major species (AsC, AsB, DMA, MMA, As (Ⅲ), and As (Ⅴ)) were determined by inductively coupled plasma mass spectrometry (ICP-MS) and high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS), respectively. The target hazard quotient (THQ) was utilized to evaluate the human health risk. The total arsenic concentration and totals for the six arsenic forms are 0.25–1.66 mg/kg and 0.05–1.19 mg/kg wet mass, respectively. Except for HH and LH, the six arsenic species concentrations accounted for more than 50.0 % of the total arsenic. The less toxic AsB is the most predominant in crabs, comprising 50.0 %–90.0 % of the sum of six types of arsenic. The more toxic inorganic arsenic only range from 0.01 to 0.21 mg/kg wet mass, much less than the limit content of 0.50 mg/kg inorganic As in crustacea. The THQ values of inorganic arsenic through the consumption of Chinese mitten crabs are estimated, and the values are all less than 1, indicating that intaking Chinese mitten crabs collected from China will not cause an appreciable hazard risk to human health.

Analysis of Organoarsenic Compounds Concentration in Fish with the Newly Developed LC-MS/MS Method

Most fish contain some kinds of organoarsenic compounds. To assess the health risk for the chronic effects due to intake of these compounds, it is necessary to quantify the concentration of each chemical form, since the toxicity is difference depending on the form. We developed and validated the LC-MS/MS method to determine the concentration of monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), trimethylarsine oxide (TMAO), tetramethylarsonium (TeMA), arsenobetaine (AB), and arsenocholine (AC) in fish. Using this method, we quantified the concentration of each organoarsenic compounds and total arsenic in 50 fish samples from across 10 groups. Total arsenic concentration ranged from 0.53 to 25 mg/kg in all samples, except for in thread-sail filefish where the concentration ranged from 8.3 to 25 mg/kg. With the exception of sardines, in all samples AB was found at the highest level in relation to the total arsenic concentration. In sardines, the concentration of DMA was higher than that of AB, accounting for 16 to 24% of total arsenic. In red sea bream, concentrations of total arsenic, AB, and AC in farm-raised fish were lower than those in wild-caught fish.

Ultrasonic treatment of arsenic species: Transformation kinetics analysis

A study on the transformation kinetics of arsenite (As(III)), arsenocholine (AsC) and arsenobetaine (AsB) was carried out in aqueous solution by ultrasonic treatment. High performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS) was used for this research under the condition of simulated ultrasound-assisted extraction. The results showed that the transformation of arsenic species accorded with the 1st-order kinetic equation at different ultrasonic water bath temperatures (0–80 °C) and ultrasonic powers (150–750 W). At the ultrasonic power of 300 W, the reaction rate constants of arsenic species at different ultrasonic water bath temperatures were as follows: k20 °C > k40 °C > k0 °C > k60 °C > k80 °C. At the ultrasonic water bath temperature of 60 °C, the reaction rate constants of arsenic species at different ultrasonic powers were as follows: k600 W > k450 W > k750 W > k300 W > k150 W. The study also provides a theoretical basis for the optimization of ultrasound-assisted extraction method without changing arsenic species.

Children exposure to inorganic and organic arsenic metabolites: A cohort study in Northeast Italy

The aim of this study was to provide urinary levels of total arsenic (TAs) and As species as arsenobetaine (AsB), arsenocholine (AsC), inorganic As (i.e., [As(III)+As(V)]), methylarsonic acid (MMA) and dimethylarsinic acid (DMA) in 7 year-old-children (n = 200) enrolled in the Northern Adriatic Cohort II (NACII), a prospective cohort in a coastal area of Northeast Italy. TAs was determined by sector field-inductively coupled plasma mass spectrometry (SF-ICP-MS) and AsB, AsC, As(III), As(V), MMA and DMA by ion chromatography coupled to ICP-MS (IC-ICP-MS). The geometric mean (GM) for TAs was 12.9 μg/L and for [iAs + MMA + DMA] was 4.26 μg/L. The species AsB (GM of 5.09 μg/L) and DMA (GM of 3.20 μg/L) had the greatest percentage contribution to TAs levels; a greater percentage contribution from AsB is seen at TAs >10 μg/L and from DMA at TAs <10 μg/L. Urinary [iAs + MMA] levels were positively associated with [iAs + MMA + DMA] and DMA with AsB levels. Fish, shellfish and crustaceans consumption increased the AsB and TAs levels, while rice intake, mothers’ education level and selenium (Se) concentration influenced the DMA concentration. Children have a high capacity to metabolize and detoxify the iAs because of the higher secondary methylation index (ratio DMA/MMA) with respect to primary methylation index (ratio MMA/iAs). In addition, the median level of [iAs + MMA + DMA] in the whole population of children was lower than the Biomonitoring Equivalent (BE) value for non-cancer endpoints. Also the Margin of Safety (MOS) value based on the population median was greater than 1, thus the exposure to the toxicologically relevant As species was not likely to be of concern.

Bioaccessibility of arsenic from gastropod along the Xiangjiang River: Assessing human health risks using an in vitro digestion model.

The bioaccessibility of total arsenic (tAs) and arsenic species in Bellamya aeruginosa collected from Xiangjiang River was evaluated using an in vitro digestion model, to assess the potential health risks to local residents. The tAs concentrations in gastropod samples ranged from 1.98 to 6.33mgkg-1 (mean 3.79±1.60mgkg-1). Five arsenic species including arsenite [As(III)], arsenate [As(V)], dimethylarsinic acid (DMA), arsenobetaine (AsB), and arsenocholine (AsC) were detected. Inorganic arsenic (iAs) concentrations, which were about a half of organic arsenic (oAs), were higher than the maximum permissible limit (≤0.50mgkg-1 in aquatic products). Bioaccessible concentrations of tAs in digestive juices were found to be decreased in the order: intestinal phase>gastric phase>salivary phase. As(III) and AsC were the predominant species, but AsB was not detectable in all digestive juices. Bioaccessible iAs concentrations, which were close to the level of bioaccessible oAs, were not significantly different among three digestive juices, but also above 0.50mgkg-1. Accordingly, bioaccessibility of tAs was highest in intestinal phase (48%), then in gastric phase (40%), and lowest in salivary phase (33%). Bioaccessibility of As(III) was close to 100%, and bioaccessibility of iAs was much higher than that of oAs. The mean values of target hazard quotient (THQ) and bioaccessible THQ were 0.80 and 0.70, respectively. The probability of experiencing non-carcinogenic effects was reduced to 18% down from 22% as considering iAs bioaccessibility. The mean values of carcinogenic risk (CR) and bioaccessible CR were higher than the acceptable value (1×10-4). Gastropod consumption from sampling sites may cause a potential carcinogenic risk.

An improved rapid analytical method for the arsenic speciation analysis of marine environmental samples using high-performance liquid chromatography/inductively coupled plasma mass spectrometry.

Arsenic contamination in marine environments is a serious issue because some arsenicals are very toxic, increasing the health risks associated with the consumption of marine products. This study describes the development of an improved rapid method for the quantification of arsenic species, including arsenite (AsIII), arsenate (AsV), arsenocholine (AsC), arsenobetaine (AsB), dimethylarsinic acid (DMA), and monomethyl arsonic acid (MMA), in seaweed, sediment, and seawater samples using high-performance liquid chromatography/inductively coupled plasma-mass spectrometry (HPLC/ICP-MS). ICP-MS based on dynamic reaction cells was used to eliminate spectral interference. Ammonium nitrate- and phosphate-based eluents were used as the mobile phases for HPLC analysis, leading to shorter overall retention time (6 min) and improved peak separation. Arsenicals were extracted with a 1% HNO3 solution that required no clean-up process and exhibited reasonable sensitivity and peak resolution. The optimized method was verified by applying it to hijiki seaweed certified reference material (CRM, NMIJ 7405-a) and to spiked blank samples of sediment and seawater. The proposed method measured the concentration of AsV in the CRM as 9.6 ± 0.6 μg/kg dry weight (dw), which is close to the certified concentration (10.1 ± 0.5 μg/kg dw). The recovery of the six arsenicals was 87-113% for the sediment and 99-101% for the seawater. In the analysis of real samples, AsV was the most abundant arsenical in hijiki and gulfweed, whereas AsB was dominant in other seaweed species. The two inorganic arsenicals (AsIII and AsV) and AsV were the most dominant in the sediment and seawater samples, respectively.

Effect of Ultrasonic Treatment on Transformations of Arsenic Species in Edible Mushrooms

A study on the transformations of six arsenic species (arsenocholine (AsC), arsenobetaine (AsB), dimethylarsinic acid (DMA), monomethylarsonic acid (MMA), arsenite (As(III)), and arsenate (As(V))) was carried out in aqueous solution by ultrasonic treatment. The transformations of arsenic species increased with time. As the ultrasonic power or temperature was increased, transformations of arsenic species were at first enhanced and then decreased. AsC, AsB, As(III) were the most unstable at 600 W and 20 °C. DMA and MMA were stable; AsC was partially transformed to AsB, trimethylarsinic oxide (TMAO), and the more toxic tetramethylarsonium ion (TMA+); AsB was partially transformed to TMAO; and As(III) was easily oxidized to As(V). The reduction of As(V) to As(III) was only observed when the temperature was over 60 °C. The ultrasound treatment conditions with relatively stable arsenic species were 150 to 300 W, 40 to 80 °C, and 0 to 10 min. The relatively stable (150 W, 10 min, 80 °C) and unstable (600 W, 60 min, 20 °C) ultrasonic condition were applied to mushrooms and the transformations of AsC, AsB, and As(III) were observed. Currently, ultrasound-assisted extraction methods have only focused on the extraction efficiency of arsenic species and ignored their transformations. This study may serve as a guide for improving extraction efficiency without minimal transformation of the arsenic forms to reflect the actual contents in edible mushrooms. The study also provides a theoretical basis for making an accurate risk evaluation of human exposure by taking account of possible arsenic transformations during processing prior to consumption.

Arsenic Species Analysis at Trace Level by High Performance Liquid Chromatography with Inductively Coupled Plasma Mass Spectrometry

A sensitive and accurate simultaneous continuous analysis for six arsenic species including arsenobetaine (AsB), arsenocholine (AsC), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), arsenite (AsIII), and arsenate (AsV) has been developed by high performance liquid chromatography with inductively coupled plasma mass spectrometry (HPLC-ICP-MS). An anion-exchange column of Hamilton PRP-X100 (Switzerland) was applied for separation of the six arsenic species with gradient elution of 1.25 mmol/L Na2HPO4 and 11.0 mmol/L KH2PO4 as the mobile phase A and 2.5 mmol/L Na2HPO4 and 22.0 mmol/L KH2PO4 as the mobile phase B. The linearity ranges for AsB, AsC, MMA, DMA, AsIII, and AsV were between 0.5 and 50.0 μg/L, and the detection limits of the six arsenic species were all within 0.01–0.35 ng/L. The relative standard deviations (RSDs) were within 2.26–3.68% and the recovery rates of samples ranged from 95 to 103%. The proposed method was applied for the arsenic speciation analysis of sediment pore-water samples, which were taken from the supernatant after centrifugation and filtration.

Matrix-induced transformation of arsenic species in seafoods

The paper presents a study on the matrix-induced transformation of arsenic (As) species spiked into seafoods. Sixteen arsenicals were individually spiked into samples of finfish, crustaceans and molluscs. The spiked samples were subjected to hot water extraction at 90 °C, and extracts were analyzed by high pressure liquid chromatography (HPLC) interfaced with inductively coupled plasma mass spectrometry (ICP-MS). Arsenate (As5+), arsenobetaine (AsB), arsenocholine (AsC), dimethylarsinate (DMA), monomethylarsonate (MMA), tetramethylarsonium ion (TMA) and trimethylarsoniopropionate (TMAP) remained intact in all the matrices. Whereas arsenite (As3+), dimethylarsinoyl acetate (DMAA), dimethylarsinoyl ethanol (DMAE), dimethylarsinoyl propionate (DMAP), trimethylarsine oxide (TMAO) and glycerol-, sulfonate-, sulfate- and phosphate-arsinoylribosides (arsenosugars 328, 392, 408 and 482, respectively) were transformed to other forms in most finfish and crustaceans. The transformation of the arsenicals was discovered to be induced by matrix thiols. While As3+ was bound to sulfhydryl groups, DMAA, DMAE, DMAP, TMAO and arsenosugars 392, 408 and 482 were thiolated through conversion of their arsinoyl (As=O) functionalities to arsinothioyl (As=S). The newly formed arsinothioyl compounds were characterized by HPLC-ICP-MS and electrospray ionization high-resolution tandem mass spectrometry (ESI-HR-MS/MS) paired with HPLC. The observed matrix-induced transformation of the arsenic species could be prevented by treating the samples (prior to spiking) with a thiol-selective blocking agent, N-ethylmaleimide (NEM).