Effect of soil particle size and extraction method on the oral bioaccessibility of arsenic.

Recent findings indicate that incidental ingestion of soil by humans primarily involves soil particles <150 µm, rather than <250 µm-sized fraction previously used for most oral bioaccessibility and bioavailability studies. It was postulated that a greater soil surface area in the finer fraction (<150 versus <250 µm) might increase oral bioaccessibility of arsenic (As) in soil. Bioaccessibility and concentrations of As were compared in <150 and <250 µm fractions of 18 soil samples from a variety of arsenic-contaminated sites. The two methods used to measure bioaccessibility were compared - EPA Method 1340 and the California Arsenic Bioaccessibility (CAB) method. Arsenic concentrations were nearly the same or higher in the <150 fraction compared with <250 µm. EPA Method 1340 and the CAB method presented significantly different bioaccessibility results, as well as estimated relative oral bioavailability (RBA) based upon algorithms specific to the methods, but there was no marked difference for <150 and <250 µm soil fractions within either method. When compared with RBA determined previously for these soil samples in vivo in non-human primates, EPA Method 1340 was generally more predictive than the CAB method. Data suggest that soil- or site-specific factors control bioaccessibility under either method and that the test method selected is more important than the particle size fraction (<150 or <250) in using these in vitro methods to predict As RBA for use in risk assessment.

Bioaccessibility of polychlorinated dioxins and furans in soil from a Superfund site.

Bioaccessibilities of PCDD/PCDF congeners contributing to cancer risk were determined in twelve soil samples from the American Creosote Works Superfund site in Florida. Based upon sample locations, congener profiles (i.e., the same dominant congeners), and total (Toxic Equivalent; TEQ) concentrations, each of these samples has PCDD/PCDF contamination reasonably attributable to the site. Bioaccessibility was determined using a 2-phase in vitro extraction method that included both simulated gastric and intestinal conditions of the human GI tract. Measured congener-specific bioaccessibility values ranged from 34.3 to 62.1%. There was no apparent relationship between the extent of chlorination of PCDD/PCDF congeners and their bioaccessibility. TEQ-weighted bioaccessibility values varied among individual soil samples, which is not unexpected based upon the literature. This variability could not be explained by differences in soil pH, composition, or organic carbon content. The average TEQ-weighted bioaccessibility value of 59% for the twelve samples was accepted as representing site-specific bioavailability of PCDD/PCDFs. This value is higher than most dioxin/furan bioaccessibility values reported in the literature and at the upper end of the range of relative oral bioavailability (RBA) values reported for PCDD/PCDFs from in vivo bioavailability studies. This study used a finer fraction of soil particles (<150 microns versus the more typical <250 microns) to better represent soil that is incidentally ingested. This finer fraction would be expected to have a greater surface area available for extraction of PCDD/PCDFs per unit mass, which might account for the greater than expected bioaccessibility.

Dermal absorption of benzo[a]pyrene into human skin from soil: Effect of artificial weathering, concentration, and exposure duration.

In vitro assessments of 14C-benzo[a]pyrene (BaP) absorption through human epidermis were conducted with the sub-63-µm fraction of four test soils containing different amounts of organic and black carbon. Soils were artificially weathered for eight weeks and applied to epidermis at nominal BaP concentrations of 3 and 10 mg/kg for 8 or 24 h. Experiments were also conducted at 24 h with unweathered soils and with BaP deposited onto skin from acetone at a comparable chemical load. For the weathered soils, absorption was independent of the amount of organic or black carbon, the mass in the receptor fluid was proportional to exposure duration but independent of concentration, and the mass recovered in the skin after washing was proportional to concentration and independent of exposure time. Results from the weathered and unweathered soils were similar except for the mass recovered in the washed skin, which was lower for the weathered soil only at the higher concentration. We hypothesize that chemical concentrations exceeded the BaP sorption capacity accessible within the artificial weathering timeframe for all soils tested, and that BaP mass in the washed skin was dominated by particles that were not removed by washing. Fluxes into and through skin from soils were lower by an order of magnitude than from acetone-deposited BaP.

Effects of Source and Concentration on Relative Oral Bioavailability of Benzo(a)pyrene from Soil.

The objective of this study was to examine the influence of soil composition, PAH concentration, and source material type on PAH bioavailability using an approach capable of measuring uptake at low, environmentally relevant PAH concentrations (down to 1 ppm). Contaminated soil samples were constructed using PAHs from three source materials-solvent, soot, and fuel oil-to which 3H-benzo(a)pyrene (3H-BaP; total BaP concentrations of 1, 10, and 100 ppm) was added in a mixture of PAHs. The soils were weathered for 8 weeks using weekly wet-dry cycles. Each soil was administered as a single dose to rats, and blood samples were taken over 6 days. Relative oral bioavailability (RBA) of the BaP from soil was estimated by comparing the area under the curve (AUC) for 3H concentration versus time in blood with the AUC observed from the same PAH mixture dosed in a food matrix. The extent to which BaP RBA was diminished in soil versus food varied among the source materials, but little or no difference was observed among the soil types examined unless carbon amendments were added. These results suggest that the type of PAH source material can have a strong influence on PAH oral bioavailability.

Predicting oral relative bioavailability of arsenic in soil from in vitro bioaccessibility.

Several investigations have been conducted to develop in vitro bioaccessibility (IVBA) assays that reliably predict in vivo oral relative bioavailability (RBA) of arsenic (As). This study describes a meta-regression model relating soil As RBA and IVBA that is based upon data combined from previous investigations that examined the relationship between As IVBA and RBA when IVBA was determined using an extraction of soil in 0.4 M glycine at pH 1.5. Data used to develop the model included paired IVBA and RBA estimates for 83 soils from various types of sites such as mining, smelting, and pesticide or herbicide application. The following linear regression model accounted for 87% of the observed variance in RBA (R(2) = .87): RBA(%) = 0.79 × IVBA(%) + 3. This regression model is more robust than previously reported models because it includes a larger number of soil samples, and also accounts for variability in RBA and IVBA measurements made on samples collected from sites contaminated with different As sources and conducted in different labs that have utilized different experimental models for estimating RBA.

Oral Bioavailability, Bioaccessibility, and Dermal Absorption of PAHs from Soil-State of the Science.

This article reviews the state of the science regarding oral bioavailability, bioaccessibility, and dermal absorption of carcinogenic polycyclic aromatic hydrocarbons (cPAHs) in soil by humans, and discusses how chemical interactions may control the extent of absorption. Derived from natural and anthropomorphic origins, PAHs occur in a limited number of solid and fluid matrices (i.e., PAH sources) with defined physical characteristics and PAH compositions. Existing studies provide a strong basis for establishing that oral bioavailability of cPAHs from soil is less than from diet, and an assumption of 100% relative bioavailability likely overestimates exposure to cPAHs upon ingestion of PAH-contaminated soil. For both the oral bioavailability and dermal absorption studies, the aggregate data do not provide a broad understanding of how different PAH source materials, PAH concentrations, or soil chemistries influence the absorption of cPAHs from soil. This article summarizes the existing studies, identifies data gaps, and provides recommendations for the direction of future research to support new default or site-specific bioavailability adjustments for use in human health risk assessment.

Fish consumption as a driver of risk-management decisions and human health-based water quality criteria.

The use and interpretation of fish consumption surveys and interviews, the application of fish consumption rates for sediment evaluation and cleanup, and the development of human health water quality criteria (HH WQC) are complex and interrelated issues. The present article focuses on these issues using examples from the United States, although the issues may be relevant for other countries. Some key considerations include the fact that there are many types of fish consumption surveys (e.g., 24-h recall surveys, food frequency questionnaires, creel surveys), and these surveys have different advantages and limitations. Identification of target populations for protection, identification of the species and quantities of fish consumed, and determination of bioaccumulation assumptions are important factors when developing water quality and sediment screening levels and standards. Accounting for the cultural importance of fish consumption for some populations is an even more complex element. Discussions about HH WQC often focus only on the fish consumption rate and may not have broad public input. Some states are trying to change this through extensive public participation efforts and use of probabilistic approaches to derive HH WQC. Finally, there are limits to what WQC can achieve. Solutions beyond the establishment of WQC that target toxics reduction from other sources may provide the greatest improvements to water quality and reductions in human health risks in the future.

An in vitro method for estimation of arsenic relative bioavailability in soil.

This report summarizes the results of a study to develop an in vitro bioaccessibility (IVBA) extraction technique for estimating the relative bioavailability (RBA) of arsenic (As) in soil. The study was implemented in several steps. In step 1, key variables in the extraction protocol were identified. In step 2, 21 different extraction conditions were tested on 12 different soils with reliable RBA values measured in swine or monkeys to identify which yielded useful in vivo-in vitro correlations (IVIVC). In step 3, three extraction conditions were evaluated using 39 different test soils to make a final selection of the best IVIVC. In step 4, the within- and between-lab reproducibility of the extraction method was examined. The optimum IVIVC model for swine utilized a pH 1.5 IVBA extraction fluid, with an R (2) value of .723. For monkeys, the optimum IVIVC model was obtained using a pH 7 IVBA extraction fluid that contained phosphate, with an R (2) value of .755. Within-lab precision of IVBA results was typically less than 3%, with an average of 0.8% for all 4 labs. Between-lab variation in mean IVBA values was generally less than 7%, with an overall average of 3%. The principal advantages of this IVBA method compared to other in vitro methods described in the literature are that (1) the fluids and extraction conditions are simple, (2) the results have been calibrated against a larger data set than any other method, and (3) the method has been demonstrated to be reproducible both within and between labs.

Variability of bioaccessibility results using seventeen different methods on a standard reference material, NIST 2710.

Bioaccessibility is a measurement of a substance's solubility in the human gastro-intestinal system, and is often used in the risk assessment of soils. The present study was designed to determine the variability among laboratories using different methods to measure the bioaccessibility of 24 inorganic contaminants in one standardized soil sample, the standard reference material NIST 2710. Fourteen laboratories used a total of 17 bioaccessibility extraction methods. The variability between methods was assessed by calculating the reproducibility relative standard deviations (RSDs), where reproducibility is the sum of within-laboratory and between-laboratory variability. Whereas within-laboratory repeatability was usually better than (<) 15% for most elements, reproducibility RSDs were much higher, indicating more variability, although for many elements they were comparable to typical uncertainties (e.g., 30% in commercial laboratories). For five trace elements of interest, reproducibility RSDs were: arsenic (As), 22-44%; cadmium (Cd), 11-41%; Cu, 15-30%; lead (Pb), 45-83%; and Zn, 18-56%. Only one method variable, pH, was found to correlate significantly with bioaccessibility for aluminum (Al), Cd, copper (Cu), manganese (Mn), Pb and zinc (Zn) but other method variables could not be examined systematically because of the study design. When bioaccessibility results were directly compared with bioavailability results for As (swine and mouse) and Pb (swine), four methods returned results within uncertainty ranges for both elements: two that were defined as simpler (gastric phase only, limited chemicals) and two were more complex (gastric + intestinal phases, with a mixture of chemicals).

Selective soil particle adherence to hands: implications for understanding oral exposure to soil contaminants.

Over the last 30 years, there has been extensive research designed to quantify the extent of oral bioavailability and bioaccessibility of organic and inorganic contaminants in soil. One aspect of this research is the soil particle size selected to represent environmental exposures, which may affect study results and comparability across studies. Different research groups have studied soil particle sizes ranging from <45 µm to <2000 µm. This article reviews the historical and technical considerations that pertain to the selection of an appropriate particle size fraction for evaluating the relative oral bioavailability of chemicals from soil, which include (1) how the resultant data will be used in human health risk assessment, (2) soil fractions historically used in oral bioavailability studies, (3) studies of soil adherence to human hands, (4) the distribution of contaminants in soils as a function of particle size, and (5) the effect of differential bioavailability as a function of soil particle size and geochemical matrix. These factors are first discussed from a general perspective, applicable to all contaminants in soil, and then more specifically for polycyclic aromatic hydrocarbons (PAHs) in soil. Based on this review, a specific soil particle size of <150 µm is recommended for future studies on the oral bioavailability and bioaccessibility of PAHs in soil.

Importance of considering the framework principles in risk assessment for metals.

The recent EPA Framework for Metals Risk Assessment provides the opportunity for contextual risk assessment for sites impacted by metals (such as the depicted Dauntless Mine in Colorado).

Dermal absorption of arsenic from soils as measured in the rhesus monkey.

Regulatory agencies have relied on dermal absorption data for soluble forms of arsenic as the technical basis for specific absorption values that are used to calculate exposure to arsenic in weathered soil. These evaluations indicate that percutaneous absorption of arsenic from soil ranges from 3.2 to 4.5% of the dermally applied dose, based on studies of arsenic freshly mixed with soil. When this value is incorporated into risk assessments and combined with other assumptions about dermal exposures to soil, the conclusion is often that dermal exposure to arsenic from soil may contribute significantly to overall exposure to arsenic in soil. Prior characterization research has indicated that the solubility of arsenic in soil varies, depending on the provenance of the soil, the source of the arsenic, and the chemical interaction of arsenic with other minerals present within the soil matrix. Weathering produces forms of arsenic that are more tightly bound within the soil and less available for absorption. Our research expands on prior in vivo studies to provide insights into the potential for dermal absorption of arsenic from the more environmentally relevant substrate of soil. Specifically, two soils with very high concentrations of arsenic were evaluated under two levels of skin hydration. One soil, containing 1400 mg/kg arsenic, was collected adjacent to a pesticide production facility in New York. The other soil, containing 1230 mg/kg arsenic, was collected from a residential area with a history of application of arsenical pesticides. Although the results of this research are constrained by the small study size dictated by the selection of an animal research model using monkeys, the statistical power was optimized by using a "crossover" study design, wherein each animal could serve as its own comparison control. No other models (animal or in vitro) were deemed adequate for studying the dermal absorption of soil arsenic. Our results show dermal absorption of soluble arsenic in solution to be 4.8 +/- 5.5%, which is similar to results reported earlier for arsenic in solution (and used by regulatory agencies in recommendations regarding dermal absorption of arsenic). Conversely, absorption following application of arsenic in the soil matrices resulted in mean estimated arsenic absorption of 0.5% or less for all soils, and all individual estimates were less than 1%. More specifically, following application of arsenic-bearing soils to the abdomens of monkeys, urinary arsenic excretion could not be readily distinguished from background. This was true across all five soil-dosing trials, including application of the two dry soils and three trials with wet soil. These findings are consistent with our understanding of the environmental chemistry of arsenic, wherein arsenic can be present in soils in complexed mineral forms. This research addresses an important component involved in estimating the true contribution of percutaneous exposures to arsenic in soil relative to exposures via ingestion. Our findings suggest that dermal absorption of arsenic from soil is truly negligible, and that EPA's current default assumption of 3% dermal absorption of arsenic from soils results in significant overestimates of exposure.

Relative oral bioavailability of arsenic from contaminated soils measured in the cynomolgus monkey.

A number of studies have found that gastrointestinal absorption of arsenic from soil is limited, indicating that a relative oral bioavailability (RBA) adjustment is warranted when calculating risks from exposure to arsenic-contaminated soil. However, few studies of arsenic bioavailability from soil have been conducted in animal models with phylogenetic similarity to humans, such as nonhuman primates. We report here the results of a study in which the RBA of arsenic in soil from a variety of types of contaminated sites was measured in male cynomolgus monkeys. A single oral dose of each contaminated soil was administered to five adult male cynomolgus monkeys by gavage, and the extent of oral absorption was evaluated through measurement of arsenic recovery in urine and feces. Urinary recovery of arsenic following doses of contaminated soil was compared with urinary recovery following oral administration of sodium arsenate in water in order to determine the RBA of each soil. RBA of arsenic in 14 soil samples from 12 different sites ranged from 0.05 to 0.31 (5-31%), with most RBA values in the 0.1-0.2 (10-20%) range. The RBA values were found to be inversely related to the amount of arsenic present with iron sulfate. No other significant correlations were observed between RBA and arsenic mineralogic phases in the test soils. The lack of clear relationships between arsenic mineralogy and RBA measured in vivo suggests that gastrointestinal absorption of arsenic from soil may be more complex than originally thought, and subject to factors other than simple dissolution behavior.

Chemical speciation and bioaccessibility of arsenic and chromium in chromated copper arsenate-treated wood and soils.

This research compares the As and Cr chemistry of dislodgeable residues from chromated copper arsenate (CCA)-treated wood collected by two different techniques (directly from the board surface either by rubbing with a soft bristle brush or by rinsing from human hands after contact with CCA-treated wood) and demonstrates that these materials are equivalent in terms of both the chemical form and bonding of As and Cr and in terms of the As leaching behavior. This finding links the extensive chemical characterization and bioavailability testing that has been done previously on the brush-removed residue to a material that is derived from human skin contact with CCA-treated wood. Additionally, this research characterizes the arsenic present in biological fluids (sweat and simulated gastric fluid) following contact with these residues. The data demonstrate that in biological fluids the arsenic is present primarily as free arsenate ions. Arsenic-containing soils were also extracted into human sweat to evaluate the potential for arsenic dissolution from soils at the skin surface. For soils from field sites, only a small fraction of the total arsenic is soluble in sweat. Based on comparisons to reference materials that have been used for in vivo dermal absorption studies, these findings suggest that the actual relative bioavailability via dermal absorption of As from CCA residues and soil may be well below the current default value of 3% used by U.S. EPA.

Percutaneous absorption of arsenic from environmental media.

Current knowledge of percutaneous absorption of arsenic is based on studies of rhesus monkeys using soluble arsenic in aqueous solution, and soluble arsenic mixed with soil (Wester et al., 1993). These studies produced mean dermal absorption rates in the range of 2.0-6.4% of the applied dose. Subsequently, questions arose as to whether these results represent arsenic absorption from environmental media. Factors such as chemical interactions, the presence of other metals, and the effects of weathering on environmental media all can affect the nature of arsenic and its potential for percutaneous absorption. Therefore, research specific to more relevant matrices is important. The focus of this effort is to outline study design considerations, including particle size, application rates, means of ensuring skin contact and appropriate statistical evaluation of the data. Appropriate reference groups are also important. The potential for background exposure to arsenic in the diet possibly obscuring a signal from a dermally applied dose of arsenic will also be addressed. We conclude that there are likely to be many site- or sample-specific factors that will control the absorption of arsenic, and matrix-specific analyses may be required to understand the degree of percutaneous absorption.

Chemical structure of arsenic and chromium in CCA-treated wood: implications of environmental weathering.

Chromated copper arsenate (CCA) has been used to treat lumber for over 60 years to increase the expected lifetime of CCA-treated wood. Because of the toxicity of the arsenic and chromium used in CCA treatment, regulatory and public attention has become focused on the potential risks from this exposure source. In particular, exposure of children to arsenic from CCA-treated wood used in decks and play sets has received considerable attention. X-ray Absorption Spectroscopy (XAS) was used to evaluate the chemical structure of As and Cr in three samples of CCA-treated materials: newly treated wood, aged wood (5 years as decking), and dislodgeable residue from aged (1-4 years as decking) CCA-treated wood. The form of the Cr and As in CCA-treated material is the same in fresh and aged samples, and between treated wood and dislodged residue. In all cases, the dominant oxidation state of the two elements is As(V) and Cr(III), and the local chemical environment of the two elements is best represented as a Cr/As cluster consisting of a Cr dimer bridged by an As(V) oxyanion. Long-term stability of the As/Cr cluster is suggested by its persistence from the new wood through the aged wood and the dislodgeable residue.

In vivo percutaneous absorption of arsenic from water and CCA-treated wood residue.

This study was conducted to evaluate the dermal absorption of arsenic from residues present on the surface of wood preserved with chromated copper arsenate (CCA). The research reported herein used methods parallel to those of earlier research on the dermal absorption of radiolabeled arsenic (R. C. Wester et al., 1993, Fund. Appl. Toxicol. 20, 336-340), with modifications to allow use of environmental matrices that are not radiolabeled. These modifications include the surface area of application and dietary intake of arsenic, thus maximizing the potential for detection of dermally absorbed arsenic in exposed animals above diet-associated background levels of exposure. Two forms of arsenic were administered in this work. The first, arsenic in solution, was applied to the skin of monkeys to calibrate the model against prior absorption research and to serve as the basis of comparison for absorption of arsenic from CCA-treated wood residues. The second substrate was residue that resides on the surface of CCA-treated wood. Results from this research indicate that this study methodology can be used to evaluate dermally absorbed arsenic without the use of a radiolabel. Urinary excretion of arsenic above background levels can be measured following application of soluble arsenic, and absorption rates (0.6-4.4% absorption) are consistent with prior research using the more sensitive, radiolabeled technique. Additionally, the results show that arsenic is poorly absorbed from CCA-treated wood residues (i.e., does not result in urinary arsenic excretion above background levels).