Arsenic Relative Bioavailability Research Articles

Intra- and Interlaboratory Evaluation of an Assay of Soil Arsenic Relative Bioavailability in Mice.

Hand-to-mouth activity in children can be an important route for ingestion of soil and dust contaminated with inorganic arsenic. Estimating the relative bioavailability of arsenic present in these media is a critical element in assessing the risks associated with aggregate exposure to this toxic metalloid during their early life. Here, we evaluated the performance of a mouse assay for arsenic bioavailability in two laboratories using a suite of 10 soils. This approach allowed us to examine both intralaboratory and interlaboratory variations in assay performance. Use of a single vendor for preparation of all amended test diets and of a single laboratory for arsenic analysis of samples generated in the participating laboratories minimized contributions of these potential sources of variability in assay performance. Intralaboratory assay data showed that food and water intake and cumulative urine and feces production remained stable over several years. The stability of these measurements accounted for the reproducibility of estimates of arsenic bioavailability obtained from repeated intralaboratory assays using sodium arsenate or soils as the test material. Interlaboratory comparisons found that estimates of variables used to evaluate assay performance (recovery and urinary excretion factor) were similar in the two laboratories. For all soils, estimates of arsenic relative bioavailability obtained in the two laboratories were highly correlated (r2 = 0.94 and slope = 0.9) in a linear regression model. Overall, these findings show that this mouse assay for arsenic bioavailability provides reproducible estimates using a variety of test soils. This robust model may be adaptable for use in other laboratory settings.

Comparison of mouse and swine bioassays for determination of soil arsenic relative bioavailability

Evaluation of soil arsenic (As) relative bioavailability (RBA) is essential to accurately assess human exposure to As contaminated soils via the incidental ingestion pathway. A variety of animal bioassays have been developed to estimate As RBA in contaminated soils and dusts, with uncertainty regarding how physiological differences between animal models or differences in assay methodologies may influence As RBA estimates. This study compared As RBA estimates across 20 As pesticide and mine-impacted soils in two commonly utilized animal bioassays, a mouse model based on steady state urinary excretion factor (UEF) and two swine models based on UEF and area under the curve (AUC), to elucidate factors influencing As RBA estimates. Multiple regression analysis supported combining the RBAs from swine UEF and swine AUC assays into a single regression model. The weighted least squares regression model derived from these data was as follows: swine RBA (%) = 1.23·mouse RBA (%) - 0.80 (R2 = 0.50). The mean relative percent difference between swine and mouse RBAs was +13%. This study extends and confirms conclusions from previous smaller studies to indicate that mouse and swine As RBA assays yield similar RBA estimates when applied to the same soils, although there appears to be a trend for the swine assay to predict higher RBAs than the mouse UEF assay.

Arsenic Relative Bioavailability in Rice Using a Mouse Arsenic Urinary Excretion Bioassay and Its Application to Assess Human Health Risk.

A steady-state mouse model was developed to determine arsenic (As) relative bioavailability (RBA) in rice to refine As exposure in humans. Fifty-five rice samples from 15 provinces of China were analyzed for total As, with 11 cooked for As speciation and bioavailability assessment. Arsenic concentrations were 38-335 μg kg-1, averaging 133 μg kg-1, with AsIII being dominant (36-79%), followed by DMAV (18-58%) and AsV (0.5-16%). Following oral doses of individual As species to mice at low As exposure (2.5-15 μg As per mouse) over a 7-d period, strong linear correlations (R2 = 0.99) were observed between As urinary excretion and cumulative As intake, suggesting the suitability and sensitivity of the mouse bioassay to measure As-RBA in rice. Urinary excretion factor for DMAV (0.46) was less than inorganic As (0.63-0.69). As-RBA in cooked rice ranged from 13.2 ± 2.2% to 53.6 ± 11.1% (averaging 27.0 ± 12.2%) for DMAV and 26.2 ± 7.0% to 49.5 ± 4.7% (averaging 39.9 ± 8.3%) for inorganic As. Calculation of inorganic As intake based on total inorganic As in rice overestimated As exposure by 2.0-3.7 fold compared to that based on bioavailable inorganic As. For accurate assessment of the health risk associated with rice consumption, it is important to consider As bioavailability especially inorganic As in rice.

Arsenic Relative Bioavailability in Contaminated Soils: Comparison of Animal Models, Dosing Schemes, and Biological End Points.

Different animals and biomarkers have been used to measure the relative bioavailability of arsenic (As-RBA) in contaminated soils. However, there is a lack of As-RBA comparison based on different animals (i.e., swine and mouse) and biomarkers [area under blood As concentration curve (AUC) after a single gavaged dose vs steady-state As urinary excretion (SSUE) and As accumulation in liver or kidney after multiple doses via diet]. In this study, As-RBA in 12 As-contaminated soils with known As-RBA via swine blood AUC model were measured by mouse blood AUC, SSUE, and liver and kidney analyses. As-RBA ranges for the four mouse assays were 2.8-61%, 3.6-64%, 3.9-74%, and 3.4-61%. Compared to swine blood AUC assay (7.0-81%), though well correlated (R(2) = 0.83), the mouse blood AUC assay yielded lower values (2.8-61%). Similarly, strong correlations of As-RBA were observed between mouse blood AUC and mouse SSUE (R(2) = 0.86) and between urine, liver, and kidney (R(2) = 0.75-0.89), suggesting As-RBA was congruent among different animals and end points. Different animals and biomarkers had little impact on the outcome of in vivo assays to validate in vitro assays. On the basis of its simplicity, mouse liver or kidney assay following repeated doses of soil-amended diet is recommended for future As-RBA studies.

Predicting Arsenic Relative Bioavailability Using Multiple in Vitro Assays: Validation of in Vivo-in Vitro Correlations.

In this study, previously established arsenic (As) in vivo-in vitro correlations (IVIVC) were assessed for their validity using an independent data set comprising As relative bioavailability (RBA) and bioaccessibility values for 13 herbicide- and mine-impacted soils. The validation process established the correlation between As RBA (swine model) and bioaccessibility (five in vitro assays), determined whether correlations differed significantly from previous relationships and assessed model bias and error. The capacity of in vitro assays to predict As RBA was demonstrated by the strength of IVIVC; goodness of fit ranged from 0.53 (DIN-I) to 0.74 (UBM-I). When compared to previous IVIVC (Juhasz et al. Environ. Sci. Technol. 2009 , 43 , 9487 ; Juhasz et al. J. Hazard. Mater. 2011 , 197 , 161 ), there was no significant difference (P < 0.01) in the slope and y-intercept for IVG-G, UBM-G, and UBM-I indicating the consistency of these assays for predicting As RBA. However, variability in model bias and prediction error was observed with significantly lower (P < 0.01) error determined for IVG-G suggesting that As RBA predictions using IVG-G may be more robust compared to UBM-G and UBM-I. In contrast, differences in the slope and/or y-intercept were observed for SBRC-I, IVG-I, PBET-G, PBET-I, DIN-G, and DIN-I suggesting that these methodologies may not be suitable for predicting As RBA.

In vitro bioaccessibility and in vivo relative bioavailability in 12 contaminated soils: Method comparison and method development

Previous studies have established in vivo–in vitro correlations (IVIVC) between arsenic (As) relative bioavailability (RBA) and bioaccessibility in contaminated soils. However, their ability to predict As-RBA in soils outside the models is unclear. In this study, As bioaccessibility and As-RBA in 12 As-contaminated soils (22.2–4172mgkg−1 As) were measured using five assays (SBRC, IVG, DIN, PBET, and UBM) and a mouse blood model. Arsenic RBA in the soils ranged from 6.38±2.80% to 73.1±17.7% with soils containing higher extractable Fe showing lower values. Arsenic bioaccessibility varied within and between assays. Arsenic bioaccessibility was used as input values into established IVIVC to predict As-RBA in soils. There were significant differences between predicted and measured As-RBA for the 12 soils, illustrating the inability of established IVIVC to predict As-RBA in those contaminated soils. Therefore, a new IVIVC was established by correlating measured As-RBA and As bioaccessibility for the 12 soils. The strength of the predictive models varied from r2=0.50 for PBET to r2=0.83 for IVG, with IVG assay providing the best prediction of As-RBA. When IVIVC were compared to those of Juhasz et al. (2014a), slopes of the relationships were significantly higher possibly due to different As-RBA measurements. Our research showed that IVG has potential to measure As bioavailability in contaminated soils from China though UBM and SBRC assays were also suitable. More research is needed to verify their suitability to predict As-RBA in soils for refining health risk assessment.

Validation of the predictive capabilities of the Sbrc-G in vitro assay for estimating arsenic relative bioavailability in contaminated soils.

A number of bioaccessibility methodologies have the potential to act as surrogate measures of arsenic (As) relative bioavailability (RBA), however, validation of the in vivo-in vitro relationship is yet to be established. Validation is important for human health risk assessment in order to ensure robust models for predicting As RBA for refining exposure via incidental soil ingestion. In this study, 13 As-contaminated soils were assessed for As RBA (in vivo swine model) and As bioaccessibility (Solubility Bioaccessibility Research Consortium gastric phase extraction; SBRC-G). In vivo and in vitro data were used to assess the validity of the As RBA-bioaccessibility correlation previously described by Juhasz et al. (2009). Arsenic RBA and bioaccessibility in the 13 soils ranged from 6.8±2.4% to 70.5±6.8% and 5.7±0.3% to 78.4±0.8% respectively with a strong linear relationship (R2=0.75) between in vivo and in vitro assays. When the As in vivo-in vitro correlation was compared that of Juhasz et al. (2009), there was no significant difference (P>0.05) indicating that the relationship between As RBA and As bioaccessibility was consistent thereby demonstrating its validation. For these data, a grouped linear regression model was developed (R2=0.82) with a slope and y-intercept of 0.84 and 3.56 respectively. A number of cross validation methodologies (2-fold, repeat random subsampling, leave one out) were utilized to determine the performance of the linear regression model. Residuals and prediction errors ranged from 5.4 to 9.4 and 6.9-12.2 respectively illustrating the capacity of the SBRC-G to accurately predict As RBA in contaminated soil.

Measurement of Arsenic Relative Bioavailability in Swine

This study describes a method for measuring the relative oral bioavailability (RBA) of arsenic (As) in soil and other soil-like media using young swine as the animal model. Groups of animals are exposed to site soil or sodium arsenate orally for 12 d. Forty-eight-hour urine samples were collected from each animal on d 6–7, 8–9, and 10–11 and were analyzed for total As. The urinary excretion fraction (UEF) for each group was estimated by plotting the mass of As excreted in urine by each animal as a function of the dose administered, and then fitting a linear model to the data using simultaneous weighted linear regression. The RBA of a test material is calculated as the ratio of the UEF value for the test material divided by the UEF of the reference material. Uncertainty around the RBA estimate is calculated using Fieller's theorem. Application of this method to a series of test soils indicates that RBA values for As can range from 18 to 52%. This wide variability supports the conclusion that there may be important differences in RBA between sites, and that use of a site-specific RBA value is likely to increase the accuracy of risk estimates for exposure to As in soil.

Predicting Arsenic Relative Bioavailability in Contaminated Soils Using Meta Analysis and Relative Bioavailability–Bioaccessibility Regression Models

A number of in vitro assays are available for the determination of arsenic (As) bioaccessibility and prediction of As relative bioavailability (RBA) to quantify exposure for site-specific risk assessment. These data are usually considered in isolation; however, meta analysis may provide predictive capabilities for source-specific As bioaccessibility and RBA. The objectives of this study were to predict As RBA using previously published in vivo/in vitro correlations and to assess the influence of As sources on As RBA independent of geographical location. Data representing 351 soils (classified based on As source) and 514 independent bioaccessibility values were retrieved from the literature for comparison. Arsenic RBA was predicted using published in vivo/in vitro regression models, and 90th and 95th percentiles were determined for each As source classification and in vitro methodology. Differences in predicted mean As RBA were observed among soils contaminated from different As sources and within source materials when various in vitro methodologies were utilized. However, when in vitro data were standardized by transforming SBRC intestinal, IVG, and PBET data to SBRC gastric phase values (through linear regression models), predicted As RBA values for As sources followed the order CCA posts ≥ herbicide/pesticide > mining/smelting > gossan soils with 95th percentiles for predicted As RBA of 78.0, 78.4, 67.0, and 23.7%, respectively.

Measurement of arsenic bioavailability in soil using a primate model.

Several studies have shown limited absorption of arsenic from soils. This has led to increased interest in including measurements of arsenic relative bioavailability from soils in the calculation of risks to human health posed by arsenic-contaminated sites. Most of the information in the literature regarding arsenic bioavailability from soils comes from studies of mining and smelter sites in the western United States. It is unclear whether these observations are relevant to other types of arsenic-contaminated sites. In order to obtain information regarding arsenic bioavailability for other types of sites, relative bioavailability of arsenic from selected soil samples was measured in a primate model. Sodium arsenate was administered to five male Cebus apella monkeys by the intravenous and oral routes, and blood, urine, and feces were collected. Pharmacokinetic behavior of arsenic after intravenous administration and the fractions of dose excreted in urine and feces after both intravenous and oral doses were consistent with previous observations in humans. Soil samples from five waste sites in Florida (one from an electrical substation, one from a wood preservative treatment site, two from pesticide sites, and one from a cattle-dip vat site) were dried and sieved. Soil doses were prepared from these samples and administered orally to the monkeys. Relative bioavailability was assessed based on urinary excretion of arsenic following the soil dose compared with excretion following an oral dose of arsenic in solution. Differences in bioavailability were observed for different sites, with relative bioavailability ranging from 10.7 +/- 4.9% (mean +/- standard deviation) to 24.7 +/- 3.2% for the five soil samples. These observations, coupled with data in the literature, suggest limited oral bioavailability of arsenic in soils from a variety of types of arsenic-contaminated sites.

An Arsenic Exposure Model: Probabilistic Validation Using Empirical Data

This paper describes development of a multi-pathway arsenic exposure model. The model uses information on arsenic concentrations in food, water, soil, and dust, combined with estimates of intake and medium-specific absorption. Urinary arsenic is predicted assuming that 60% of absorbed arsenic is excreted in urine under steady state conditions. Fecal arsenic is predicted assuming all unabsorbed arsenic is excreted in feces. We applied this model at a former copper smelter site. Site specific distributions were available for the following parameters: soil and dust arsenic concentration (geometric mean approximately 100 to 200 ppm and 50 to 100 ppm, respectively); the combined childhood soil and dust ingestion rate (geometric mean of 20 mg/d); soil and dust arsenic relative bioavailability (geometric mean 0.20 and 0.28, respectively); exposure duration; water arsenic concentration; air arsenic concentration; and total arsenic in food. Monte Carlo simulation was used to predict daily arsenic uptake and excretion in urine and feces for children. Predicted urine arsenic levels were less than measured levels (73% to 88% of measured values, depending on region of site). On the other hand, predicted fecal arsenic levels exceeded measured levels by a factor of 1.7 to 4.6. We were able to improve the correspondence between predicted and measured arsenic excretion rates by decreasing the assumed value of the combined soil and dust ingestion rate, and increasing the assumed bioavailability of arsenic in soil and dust.