Estimation of arsenic intake from drinking water and food (raw and cooked) in a rural village of northern Chile. Urine as a biomarker of recent exposure.

The aim of this study was to estimate both the contribution of drinking water and food (raw and cooked) to the total (t-As) and inorganic (i-As) arsenic intake and the exposure of inhabitants of Socaire, a rural village in Chile´s Antofagasta Region, by using urine as biomarker. The i-As intake from food and water was estimated using samples collected between November 2008 and September 2009. A 24-hour dietary recall questionnaire was given to 20 participants. Drinking water, food (raw and cooked) and urine samples were collected directly from the homes where the interviewees lived. The percentage of i-As/t-As in the drinking water that contributed to the total intake was variable (26.8-92.9). Cereals and vegetables are the food groups that contain higher concentrations of i-As. All of the participants interviewed exceeded the reference intake FAO/OMS (149.8 µg∙i-As·day⁻¹) by approximately nine times. The concentration of t-As in urine in each individual ranged from 78 to 459 ng·mL⁻¹. Estimated As intake from drinking water and food was not associated with total urinary As concentration. The results show that both drinking water and food substantially contribute to i-As intake and an increased exposure risk to adult residents in contaminated areas.

Mercury and selenium in fish and shellfish: occurrence, bioaccessibility and uptake by Caco-2 cells.

This study evaluates Hg and Se concentrations and bioaccessibility (element solubilised after simulated gastrointestinal digestion) in 16 raw seafood species consumed in Spain. The concentrations varied greatly (Hg, 3.8-1621 ng/g wet weight, ww; Se, 84-1817 ng/g ww). Only one sample of swordfish exceeded the Hg limit permitted in Spain (1mg/kg), and for this sample the Hg/Se molar ratio and Se Health Benefit Value food safety criteria also indicated the presence of a risk. Bioaccessibility of Hg (35-106%) and Se (17-125%) was very variable and the Hg/Se molar ratio in the bioaccessible fraction was less than one for all samples. Transport by Caco-2 cells, an intestinal epithelium model, was also evaluated from the swordfish bioaccessible fraction. Hg and Se transport from the food was less than 14%, and cell retention was much greater for Hg (49-69%) than Se (8-12%).

Content of selenium, total and inorganic arsenic and bioaccessibility of arsenic in children diets of Mexico.

BACKGROUND: The aim of this study was to evaluate the intake of selenium, total and inorganic arsenic and the bioaccessibility of arsenic in diets of Mexican schoolchildren using an in vitro method. RESULTS: Total and inorganic arsenic concentrations in the diets ranged from 0.05 to 1.15 mg kg(-1) dry weight (DW) and from 0.023 to 0.088 mg kg(-1) DW respectively, while selenium levels were between 0.08 and 0.20 mg kg(-1) DW. Daily intake of total and inorganic arsenic ranged from 0.15 to 10.49 µg day(-1) kg(-1) body weight (BW) and from 0.06 to 1.11 µg day(-1) kg(-1) BW respectively, while that of selenium varied from 0.29 to 2.63 µg day(-1) kg(-1) BW. Bioaccessibility of total and inorganic arsenic ranged from 4 to 97% (mean 44 ± 21.5%) and from 33 to 97% (mean 71 ± 22.5%) respectively. CONCLUSION: Inorganic arsenic levels in the diets were low. Therefore there appears to be a low risk of adverse effects resulting from excess inorganic arsenic intake from these diets.

Total and inorganic arsenic concentrations in different species of economically important algae harvested from coastal zones of Chile.

Chile is one of the major producers of phytocolloids extracted from seaweed. Multicellular algae are considered to be primary accumulators of arsenic. We analyzed 14 species of algae belonging to the groups Rhodophyceae (10), Phaeophyceae (3) and Chlorophyceae (1) from different coastal zones of Chile in 2003-2004. Dry ashing mineralization (for total As) and acid digestion (for inorganic As) together with quantification by flow injection hydride generation atomic absorption spectrometry (FI-HG-AAS) were employed. In general, total arsenic concentrations varied between 3.0 and 68 mg kg(-1), whereas inorganic arsenic concentrations ranged between 0.15 and 1.06 mg kg(-1). The algal species Durvillaea antarctica and Porphyra columbina, used for direct human consumption, did not have inorganic arsenic levels that represent a health risk to consumers. Among species used for phytocolloids production, such as Macrocystis piryfera, Gracilaria chilensis and Gigartina skottsbergii, observed levels of inorganic arsenic were greater than 1 mg kg(-1), the limit value established by the regulations of some countries. Among the 14 species of algae tested, inorganic arsenic levels were between 0.8% and 13% of the total arsenic concentrations; that is, arsenic present in these algae was found primarily as organic arsenic.

Influence of mercury bioaccessibility on exposure assessment associated with consumption of cooked predatory fish in Spain.

BACKGROUND: Predatory fish tend to accumulate high levels of mercury (Hg). Food safety assessment of these fish has been carried out on the raw product. However, the evaluation of the risk from Hg concentrations in raw fish might be modified if cooking and bioaccessibility (the contaminant fraction that solubilises from its matrix during gastrointestinal digestion and becomes available for intestinal absorption) were taken into account. Data on Hg bioaccessibility in raw predatory fish sold in Spain are scarce and no research on Hg bioaccessibility in cooked fish is available. The aim of the present study was to evaluate Hg bioaccessibility in various kinds of cooked predatory fish sold in Spain to estimate their health risk. RESULTS: Both Hg and bioaccessible Hg concentrations were analysed in raw and cooked fish (swordfish, tope shark, bonito and tuna). There were no changes in Hg concentrations during cooking. However, Hg bioaccessibility decreased significantly after cooking (42 ± 26% in raw fish and 26 ± 16% in cooked fish), thus reducing in swordfish and tope shark the Hg concentration to which the human organism would be exposed. CONCLUSION: In future, cooking and bioaccessibility should be considered in risk assessment of Hg concentrations in predatory fish.

Comparison of a static and a dynamic in vitro model to estimate the bioaccessibility of As, Cd, Pb and Hg from food reference materials Fucus sp. (IAEA-140/TM) and Lobster hepatopancreas (TORT-2).

Bioaccessibility, the fraction of an element solubilized during gastrointestinal digestion and available for absorption, is a factor that should be considered when evaluating the health risk of contaminants from food. Static and dynamic models that mimic human physiological conditions have been used to evaluate bioaccessibility. This preliminary study compares the bioaccessibility of arsenic (As), cadmium (Cd), lead (Pb) and mercury (Hg) in two food certified reference materials (CRMs) (seaweed: Fucus sp., IAEA-140/TM; Lobster hepatopancreas: TORT-2), using two in vitro gastrointestinal digestion methods: a static method (SM) and a dynamic multicompartment method (TIM-1). There are significant differences (p<0.05) between the bioaccessible values of As, Cd, Pb and Hg obtained by SM and TIM-1 in the two CRMs. The specific form in which the elements studied are present in the CRM may help to explain the bioaccessibility values obtained.

Characterization of the intestinal absorption of arsenate, monomethylarsonic acid, and dimethylarsinic acid using the Caco-2 cell line.

Many toxicological studies have been conducted with arsenic species in target organ cell lines. However, although epithelial gastrointestinal cells constitute the first barrier to the absorption of contaminants, studies using intestinal cells are scarce. The present study examines absorption through the intestinal epithelium of the pentavalent arsenic species most commonly found in foods [arsenate, As(V); monomethylarsonic acid, MMA(V); and dimethylarsinic acid, DMA(V)], using the Caco-2 cell line as a model. Different concentrations (1.3-667.6 microM) and culture conditions (media, pH, addition of phosphates, and treatment with ethylenediaminetetraacetic acid) were evaluated to characterize such transport. The apparent permeabilities indicate that the methylated species show low absorption, whereas As(V) is a compound with moderate absorption. The kinetic study shows only a saturable component for MMA(V) transport in the range of concentrations assayed. The existence of paracellular transport was shown for all of the species, with greater significance in the case of the methylated forms. As(V) absorption was inhibited by 10 mM phosphate, and a phosphate transporter therefore could take part in intestinal absorption. Acidification of the medium (pH 5.5) resulted in a marked increase in As(V) and DMA(V) permeability (4-8 times, respectively) but not in MMA(V) permeability. This makes it necessary to consider the possible existence of absorption in the proximal intestine and even in the stomach, where the environment is acidic; alternatively, an H(+)-dependent transporter may be involved. The results obtained constitute the basis for future research on the mechanisms involved in the intestinal absorption of arsenic and its species, a decisive step in relation to their toxic action.

Identification of As accumulation plant species growing on highly contaminated soils.

Soils from the alluvial flats of the Turia River, Valencia, Spain, which were highly contaminated by decades of industrial activity, were surveyed for native plant species that could be candidates useful in phytoremediation. Concentrations of heavy metals and arsenic (As) in soils reached 25,000 mg Kg(-1) Pb, 12,000 mg Kg(-1) Zn, 70 mg Kg(-1) Cd, and 13500 mg Kg(-1) As. The predominant vegetation was collected and species identified. Soil samples and the corresponding plant shoots were analyzed to determine the amount of As accumulated by the various plant species. Several were able to tolerate more than 1000 mg Kg(-1) As in the soil. Bassia scoparia (Chenopodiaceae) survive in soil with 8375 mg Kg(-1) As. Arsenic accumulation in shoots of the various plant species investigated ranged from 0.1 to 107 mg Kg(-1) dw. Bassia scoparia (Chenopodiaceae), Inula viscosa (Asteraceae), Solanum nigrum (Solanaceae), and Hirschfeldia incana (Brassicaceae) had the highest values for As accumulation.

Total and inorganic arsenic concentrations in rice sold in Spain, effect of cooking, and risk assessments.

Rice can contain a relatively high amount of arsenic (As). We evaluated total and inorganic As concentrations in 39 samples of different types of rice sold in Spain. The analyses were performed in raw rice and in rice cooked by boiling to dryness in water spiked with As(V) (0.1-1 microg mL(-1)). In raw rice, inorganic As represented 27-93% of total As: total As = 0.188 +/- 0.078 microg g(-1) dry weight (dw); inorganic As = 0.114 +/- 0.046 microg g(-1) dw. After cooking, the rice retained between 45% and 107% of the As(V) added to the cooking water, and the inorganic As concentrations ranged between 0.428 microg g(-1) dw (0.1 microg mL(-1) in the cooking water) and 3.89 microg g(-1) dw (1.0 microg microL(-1) in the cooking water). For raw rice, the inorganic As intake of the Spanish population (16 g raw rice/day) remains below the tolerable daily intake (TDI) proposed by the WHO (2.1 microg inorganic As/day/kg body weight). In rice cooked with water contaminated with As(V), this cereal intake is sufficient to attain the TDI. The results reveal the need to consider the determination of inorganic As and the influence of cooking when evaluating the risks associated with the consumption of rice.

Total and inorganic arsenic in Antarctic macroalgae.

The Antarctic region offers unparalleled possibilities of investigating the natural distribution of metals and metalloids, such as arsenic. Total and inorganic As were analysed in nine species of Antarctic macroalgae collected during the 2002 summer season in the Potter Cove area at Jubany-Dallmann Station (South Shetland Islands, Argentinian Base). Total As was determined by inductively coupled plasma-optical emission spectrometry after microwave-assisted acid digestion. Inorganic As was determined by acid digestion, solvent extraction, flow injection-hydride generation-atomic absorption spectrometry. Total As ranged from 5.8 microg g(-1) dry weight (dw) (Myriogramme sp.) to 152 microg g(-1)dw (Himantothallus grandifolius). Total As concentrations were higher in Phaeophytes (mean+/-SD: 71+/-44 microg g(-1)dw) than in Rhodophytes (mean+/-SD: 15+/-11 microg g(-1)dw). Inorganic As ranged from 0.12 microg g(-1) (Myriogramme sp.) to 0.84 microg g(-1)dw (Phaeurus antarcticus). The percentage of inorganic As with respect to total As was 0.7 for Phaeophytes, but almost 4 times higher for Rhodophytes (2.6). The work discusses possible causes for the presence of As in marine organisms in that pristine environment.

Total arsenic, inorganic arsenic, lead and cadmium contents in edible seaweed sold in Spain.

Total arsenic, inorganic arsenic, lead and cadmium contents were determined in 112 samples of seaweed preparations sold in Spain (seaweed packed in plastic or cardboard box, seaweed in the form of tablets and concentrates, foods containing seaweed, and canned seaweed). The concentration ranges found, expressed in mg/kg, dry weight, were: total As (0.031-149), inorganic As (<0.014-117), Pb (<0.050-12.1) and Cd (<0.003-3.55). For all the contaminants there were failures to comply with legislated values. In particular, all the samples of Hizikia fusiforme exceeded the inorganic As limit established in some countries, and a considerable number of species exceeded the Cd limit set by international regulations. With respect to food safety, consumption of 3 g/day of the samples analysed could represent up to 15% of the respective Tolerable Daily Intakes (TDI) established by the WHO. The situation is especially alarming for intake of inorganic As from H. fusiforme, which can be three times the TDI established.

Cytotoxic effect of As(III) in Caco-2 cells and evaluation of its human intestinal permeability.

Inorganic arsenic has been classified as a carcinogen for humans (Group I). However, its transit across the human intestinal epithelium has not been characterized. Using Caco-2 cells, the thiol-redox balance and apparent permeability coefficients (P(app)) for As(III) in the apical to basolateral (AP-BL) and basolateral to apical (BL-AP) direction were evaluated. After As(III) exposure, GSH-induced synthesis was observed, increasing the GSH/GSSG ratio by elevating the As(III) concentration. The AP-BL permeabilities decreased as the As(III) concentrations increased, indicating the existence of a mediated transport mechanism. The (BL-AP)/(AP-BL) permeability ratios were higher than unity, suggesting the existence of a secretion process.

Transformation of organoarsenical species by the microflora of freshwater crayfish.

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

Bioavailability of inorganic arsenic in cooked rice: practical aspects for human health risk assessments.

Arsenic is present in rice grain mainly as inorganic arsenic. Little is known about the effect of cooking on inorganic arsenic content in rice and its bioavailability. This study evaluated total arsenic and inorganic arsenic in rice cooked with arsenic-contaminated water, the bioaccessibility of As(III) and As(V) after simulated gastrointestinal digestion, and the extent of arsenic retention and transport by Caco-2 cells used as a model of intestinal epithelia. After cooking, inorganic arsenic contents increase significantly. After simulated gastrointestinal digestion, the bioaccessibility of inorganic arsenic reached 63-99%; As(V) was the main species found. In Caco-2 cells, arsenic retention, transport, and total uptake (retention + transport) varied between 0.6 and 6.4, 3.3 and 11.4, and 3.9 and 17.8%, respectively. These results show that in arsenic endemic areas with subsistence rice diets, the contribution of inorganic arsenic from cooked rice should be considered in assessments of arsenic health risk.

Arsenosugars in raw and cooked edible seaweed: characterization and bioaccessibility.

The aim of this study was to examine arsenic species contents in raw and cooked edible seaweed and the bioaccessibility (maximum soluble concentration in gastrointestinal medium) of arsenosugars (glycerol ribose, phosphate ribose, sulfonate ribose, and sulfate ribose). For the analysis, a new chromatographic separation was developed in anion exchange, coupled with thermooxidation-hydride generation-atomic fluorescence spectrometry. An in vitro digestion (pepsin, pH 2; pancreatin-bile extract, pH 7) was applied to estimate arsenosugar bioaccessibility. Cooking of Undaria pinnatifida and Porphyra sp. did not alter the arsenic species present in the methanol-water extract, but it produced a substantial increase (2 and 5 times) in the As(V) extracted from Hizikia fusiforme. In all of the seaweeds analyzed, arsenosugar bioaccessibility was high (>80%) and did not vary as a result of cooking. Arsenosugar degradation as a result of in vitro digestion was not observed.

An approach to As(III) and As(V) bioavailability studies with Caco-2 cells.

Foods and drinking water are the main sources of human exposure to inorganic arsenic [As(III) and As(V)]. After oral ingestion, the intestinal epithelium is the first barrier to absorption of these species. A human intestinal cell line (Caco-2) was used to evaluate cell retention and transport of As(III) (15.6-156.0 microM) and/or As(V) (15.4-170.6 microM). Cell monolayer integrity, cell viability, membrane damage and effects on cell metabolism were evaluated. Only the highest concentrations assayed [As(III): 156.0 microM; As(V): 170.6 microM] produced a cytotoxic effect with different cellular targets: As(III) altered the permeability of tight junctions, and As(V) caused uncoupling of the respiratory chain. Retention and transport of As(III) was more efficient than that of As(V). After 4h of exposure to As(III) or As(V), monolayer retention percentages varied between 0.87-2.28% and 0.14-0.39%, respectively. Transepithelial transport was greater for As(III) (5.82-7.71%) than for As(V) (not detectable-1.55%). The addition of As(III) and As(V) jointly produced a transport rate similar to that observed when they were added independently.

Visible and near-infrared spectroscopy as a technique for screening the inorganic arsenic content in the red crayfish (procambarus clarkii Girard).

The potential of near-infrared spectroscopy (NIRS) for screening the inorganic arsenic (i-As) content in the red crayfish (Procambarus clarkii Girard 1852) was assessed. Sixty-two samples belonging to this species were freeze-dried and scanned by NIRS. The i-As contents of the samples were obtained by acid digestion-solvent extraction followed by hydride generation atomic absorption spectrometry and were regressed against different spectral transformations by modified partial least-squares (MPLS) regression. Second derivative transformation equations of the raw optical data, previously standardized by applying standard normal variate and de-trending algorithms, resulted in a coefficient of determination in the cross-validation (1-VR) of 0.84, indicative of equations of good quantitative information. The standard error of cross-validation to standard deviation ratio, shown by the second derivative equation, was similar to those obtained for other trace metal calibrations reported in NIRS reflectance. Spectral information related to chromophores and lipids of the red crayfish tissues, and also the plant matter contained in their stomachs, were the main organic components used by MPLS for modeling the selected prediction equation. This pioneering use of NIRS to predict the i-As content in red crayfish represents an important savings in time and cost of analysis.

Contribution of water, bread, and vegetables (raw and cooked) to dietary intake of inorganic arsenic in a rural village of Northern Chile.

Total and inorganic As contents of cooked vegetables obtained from an arsenic endemic area of Chile were analyzed. Inorganic As intake from those foods, bread, and water was estimated. The study was performed in two different periods, in which the water used by the population for drinking and cooking purposes contained 0.572 (first period) or 0.041 microg mL(-)(1) (second period). In the first period, the FAO/WHO reference intake was exceeded by all of the persons interviewed. In the second period, the reference intake was exceeded by all of the persons interviewed ages 13-15. The foods studied contributed 4% (first period) or 25% (second period) of the inorganic As intake. The results show the contribution of food to inorganic As intake and the risk to which those ages 15 or younger are exposed.

Heavy metals and arsenic uptake by wild vegetation in the Guadiamar river area after the toxic spill of the Aznalcóllar mine.

On 25 April 1998, approximately 4.5 hm(3) of pyritic sludge, containing 5000 mg of As kg(-1) among other pollutants, was spilled into the Agrio and Guadiamar rivers and the surrounding agricultural areas (Aznalcóllar, Seville, Southern Spain). Many trace metals such as Pb, Cu, Zn, Cd, Tl, Sb and As reached the Doñana National Park, the largest wetland area in Europe, affecting soils, different plant and animal species. In order to recuperate the affected lands by employing plants capable of accumulating high levels of contaminants in shoots, periodical field surveys have been made to identify the metal-tolerant species that are spontaneously growing in the polluted soils, and are able to uptake one or various of the contaminants. Among the 99 different plant species studied, Anchusa azurea, Beta vulgaris, Chamaemelum fuscatum, Convolvulus arvensis, Cynodon dactylon, Diplotaxis virgata, Erodium aethiopicum, Lavatera cretica, Malva nicaeensis, Silybum marianum and, above all, Amaranthus blitoides highlight as the most promising to be used in the remediation of the affected area.

Organoarsenical species contents in fresh and processed seafood products.

A study was carried out to determine organic species of arsenic in the main varieties of seafood consumed in the Basque country (Spain). The concentrations of arsenobetaine (AB), dimethylarsinic acid (DMA), monomethylarsonic acid (MMA), arsenocholine (AC), and tetramethylarsonium ion (TMA(+)) in 64 samples corresponding to different food items are presented. The study provides information about a possible distribution pattern of organoarsenical species in seafood products. AB was detected in all of the samples [0.3-104.1 microg g(-1) dry weight (dw)]. DMA was detected in all of the samples except squid and salted cod (0.027-1.757 microg g(-1) dw). MMA was detected only in certain fatty fish (0.004-0.028 microg g(-1) dw) and bivalves (0.031-0.047 microg g(-1) dw). AC was only present in some samples of lean fish (0.014-0.089 microg g(-1) dw), and TMA(+) was detected only in anchovy (0.039-0.169 microg g(-1) dw) and crustaceans (0.044-0.966 microg g(-1) dw).