4
Tools

This chapter describes the physical, chemical, and biological tools that have been used to evaluate bioavailability, and it assesses their scientific basis. In general, understanding contaminant bioavailability from soils and sediments requires studying the processes illustrated in Figure 1-1. A first-order need is to identify the contaminant of concern and determine its form, concentration, and distribution (which can correlate with understanding bioavailability process A). These characteristics can be inferred from the soil or sediment matrix or determined directly with operational or mechanistic measurements. Some analytical techniques like spectroscopy can directly address where and how a chemical is associated with sediment or soil, while techniques like extractions operationally address form. Biological tools typically consider entry of the contaminant into the living organism (D in Figure 1-1) without directly measuring processes A–C. Of course, processes A, B or C might be manipulated or measured by other means, with biological tools then being used to evaluate an organism’s responses to those manipulations or measurements. One class of biological tools addresses complex responses like toxicity (E in Figure 1-1), for which bioavailability is only one of several possible influences. This chapter does not discuss tools applicable to processes B and C, like fate and transport models, as there are numerous other reports dealing with fate and transport. Rather, the tests that are part of this chapter mainly deal with bioavailability processes A, D, and E; such tests usually assume a constant transport condition.



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Bioavailability of Contaminants in Soils and Sediments: Processes, Tools, and Applications 4 Tools This chapter describes the physical, chemical, and biological tools that have been used to evaluate bioavailability, and it assesses their scientific basis. In general, understanding contaminant bioavailability from soils and sediments requires studying the processes illustrated in Figure 1-1. A first-order need is to identify the contaminant of concern and determine its form, concentration, and distribution (which can correlate with understanding bioavailability process A). These characteristics can be inferred from the soil or sediment matrix or determined directly with operational or mechanistic measurements. Some analytical techniques like spectroscopy can directly address where and how a chemical is associated with sediment or soil, while techniques like extractions operationally address form. Biological tools typically consider entry of the contaminant into the living organism (D in Figure 1-1) without directly measuring processes A–C. Of course, processes A, B or C might be manipulated or measured by other means, with biological tools then being used to evaluate an organism’s responses to those manipulations or measurements. One class of biological tools addresses complex responses like toxicity (E in Figure 1-1), for which bioavailability is only one of several possible influences. This chapter does not discuss tools applicable to processes B and C, like fate and transport models, as there are numerous other reports dealing with fate and transport. Rather, the tests that are part of this chapter mainly deal with bioavailability processes A, D, and E; such tests usually assume a constant transport condition.

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Bioavailability of Contaminants in Soils and Sediments: Processes, Tools, and Applications SUMMARY TABLES In illustrating the range of physical, chemical, and biological approaches that have been used to evaluate bioavailability processes, this chapter reflects the existing state of knowledge. It is not meant to be an exhaustive list from which one can choose the ultimate tool, nor should it be read as a list of approved approaches for explicitly considering bioavailability. The state of the science is such that little consensus exists about optimal approaches. Among the tests reviewed here, some are appropriate for some situations, but most are not generally applicable to a wide spectrum of situations. Table 4-1 summarizes the characteristics of the tools covered in the chapter, including what process the tool studies, the approximate cost, and the status of the tool in terms of its future use. It is important to recognize that most tools are still in development and few are fully validated by a body of work relating their predictions to independent measures from nature. Almost all of the tools are broadly applicable to both soils and sediments. Where a test is specific to one or the other, it is mentioned in the description of that test, rather than in the table. Table 4-2 specifies some generic strengths and limitations of each method and thereby illustrates that every method has tradeoffs. The criteria used for Table 4-2 are: Application to the field. Some methods can be employed in complex natural settings (score 3), some can be used on materials collected from the field (score 2), and some require experimental manipulations such as contaminant spiking (score 1). Application to solid phase. A method that directly addresses processes in the solid phase of sediments or soils, such as a method that evaluates contaminant form in the solid, would score 3. In contrast, a method that requires measurement of the properties of an extract scores 1. A biological test that addresses the solid phase in situ scores higher (a field bioaccumulation survey) than a method that takes the solid phase out of context for the evaluation (a lab sediment bioassay), which scores higher than a test that uses an extract (pore water, Microtox or elutriate bioassay). Single vs. lumped processes. Methods that measure a single process are most likely to illustrate a specific mechanism at work. For example, some physical-chemical methods directly evaluate metal form, while other methods measure one mechanism instrumental to bioavailability such as initial biouptake. These score 3. Speciation can be inferred from some methods, as can biouptake from methods like whole organism bioaccumulation (score 2). Other methods that measure a mixture of processes are more operational and less mechanistic (score 1). For example, extractions remove contaminants from an unknown suite of forms without quantifying any processes. Biological methods like toxicity tests are influenced by biouptake plus other processes that influence toxicity.

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Bioavailability of Contaminants in Soils and Sediments: Processes, Tools, and Applications TABLE 4-1 Characteristics of Tools for Measuring Bioavailability Tool Process Studieda Costb Statusc Physical/chemical characterization of the solid phase General characteristics • Organic carbon content • Particle/grain size • pH • CEC • Humic and fulvic acid content Chemical characteristics of the solid (except particle size which is a physical characteristic). $ Standard protocols available Specific structures • Characterization of carbonaceous and other solid phases using NMR, petrography, EA, IR/FTIR Molecular characteristics of solid substrate. $$ to $$$ Currently research grade for contaminated site application Specific forms of contaminant bound to solids • XRD and SEM • XAS • μL2MS • SIMS • NMR • EPR • XPS Association and dissociation processes, including the roles of surface morphology, oxidation state, and compound or element location. XRD, SEM—$$ All others—$$$ XRD, SEM—Standard protocols available; all others are research grade Extraction of soils and sediments for inorganic contaminants Extracts that change the solid phase • Conventional • Sequential • TCLP, SPLP Dissociation from the solid phase. Sequential extracts attempt to differentiate between forms of elements associated with different components or phases of the particle. $ Some extracts in use and in regulations and thus standardized, but sequential extracts at research stage or in development Passive approaches • Passive extracts • Pore water measurements with ASV or ion-specific electrode • Exchangeable resins Passive extracts determine dissociation from the solid phase. ASV and electrodes measure pore water concentrations. Exchange resins measure dissociation from the solid phase and physical flux to aqueous phase. $(but need ICP-MS for exchangeable resins) Research grade, no standard protocols developed; exchangeable resins better developed for sediments

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Bioavailability of Contaminants in Soils and Sediments: Processes, Tools, and Applications Tool Process Studieda Costb Statusc In vitro tests to mimic human intake for both organics and inorganics Dissociation from the solid phase that mimics physiological fluids. $ to $$ Variable: validated for lead, but in various stages of development for others Extraction and other tests of soils and sediments for organic contaminants Fluid-phase extractions • Mild solvents • SWE • Supercritical CO2 extraction • PTD Dissociation from the solid phase. $ Mild solvents have standardized protocols; supercritical CO2 and SWE are in development Solid phase and membrane-based extractions • Tenax • C-18 • SPME • SPMD • DGT Dissociation from the solid phase and physical flux to aqueous phase by capturing desorbed contaminant on highly sorptive matrix or gel device. $ Standard protocols for using these methods for measuring contaminants in water; for soils and sediments, all of these techniques are in development Other desorption tests • Gas purge • Desorption kinetics and activation energy Dissociation from the solid phase. $$ to $$$$ In development Normalizations Organic and inorganic correlations • Ratios and models • AVS/SEM • EqP EqP and AVS/SEM assume reactants control dissociation from the solid phase; other ratios are determined empirically from regressions in field data. $ Research grade; varies with evaluator Biological approaches to measuring uptake Assimilation efficiency Biological uptake across the gut wall. $$ Research grade Mineralization/assimilation assays for microorganisms Integrated measure of bacterial uptake and metabolic degradation. $ to $$ Research grade

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Bioavailability of Contaminants in Soils and Sediments: Processes, Tools, and Applications Tool Process Studieda Costb Statusc Bioassays: cell cultures and isolated organs/tissues Biological uptake into cell or organ. $$ Research grade Bioassays: whole organism bioaccumulation • Plants • Invertebrates • Fish • Birds and mammals Biological uptake into whole organism.Various endpoints are measured, including tissue, blood,etc. Plants—$ to $$ Earthworm test—$$ Mammals—$$$ Standard protocols forplants, invertebrates, and birds; research grade when plants and other animals used as surrogates Field survey: whole organism bioaccumulation Biological uptake into whole organism in field. $$ Research grade Biological approaches to measuring organismal response and toxicity Reporter systems Integrated measure of dissociation from the solid phase,bacterial uptake, and effect on gene expression and subsequent events. $$$ Research grade Biomarkers Integrated measure of uptake and response at a subcellular level. $ to $$$$ (gene expression) In development Toxicity tests: spiked • Plant • Invertebrate • Fish • Mammal, bird Integrated measure of uptake and toxic effects. $ to $$ Standard protocols available for fresh and saltwater sediments Toxicity tests: site- specific materials Site-specific integrated measure of uptake and toxic effects. $ to $$$$ Standard protocol available Microbial community bioassays Integrated measure of uptake, toxic effects, and community interactions. $$ Research grade Ecosystem level mesocosms Integrated measure of many processes including ecosystem level processes like food web transfer. $$$$$ Standard protocols available

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Bioavailability of Contaminants in Soils and Sediments: Processes, Tools, and Applications Tool Process Studieda Costb Statusc Environmental exposure studies Integrated measure of many processes including measurable effects in humans. $$$$$ Research grade aProcess Studied: How does this tool address the physical, chemical or biological processes that influence bioavailability? bCost: $ to $$$$$: Costs in approximate order of magnitude, with $ equal to $100s. cStatus: standardized protocol, research grade, or in development. Immediacy or relevance to entry of contaminant into living cell (biouptake). Entry of a contaminant into a living cell across a biological membrane is the process most immediately relevant to determining bioavailability. Some biological methods involve direct determination of transport or biouptake (score 3). Some measure many processes including biouptake, or a process tangential to biouptake like toxicity, or they mimic biouptake as with certain extractions (score 2). Some physical-chemical methods are unrelated to biouptake (score 1). Ability to generalize. Although site-specific tests are essential to managing an individual site, methods that allow predictions (or development of predictive capabilities) without measuring all processes are ultimately a desirable approach. Methods that are predictive, like some models or some tests that determine a mechanism that can be unambiguously compared from site-to-site, score highest in this category (score 3). Methods that are predictive but not yet of proven reliability score 2. Approaches that are of value at a site but do little to explain how the bioavailability processes at that site are comparable to other sites score 1. Relevance to regulation. The relevance of a method to the pressing concerns at a site has led to the use of certain tests for regulatory purposes (like toxicity tests or direct evaluations of human health). Also, methods that are simple and practical to employ, or methods that yield a single value, are most likely to have been applied in the regulatory setting. Thus, methods that managers or decision-makers can interpret or have interpreted as directly relevant to their needs score 3. Methods that have seldom been used in a regulatory setting or have limited potential for such use score 1. Usefulness as a research tool. Relevance as a research tool is just as important as relevance to regulation because of the great need for better understanding the processes that govern bioavailability. Methods that are of use in explaining processes in specific circumstances or in mechanistic detail score highest (score 3), even if they are of limited use in applications. Methods that are of use in a correlative fashion in experimental studies score 2. Methods of limited use in research score 1.

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Bioavailability of Contaminants in Soils and Sediments: Processes, Tools, and Applications TABLE 4-2 Rankings of Bioavailability Tools According to Seven Criteria Technique Application to the Field Application to Solid Phase Single vs. Lumped Processes Physical/chemical characterization of the solid phase General characteristics • Organic carbon content • Particle/grain size • pH • CEC • HA/FA 2 Can test field samples in the laboratory. 3 Directly relevant to solid phase in situ; necessary to understand solid phase reactions. 2 Measures are the outcome of lumped processes, but can be used to interpret single processes. Specific structures • Characterization of carbonaceous and other solid phases using NMR, petrography, EA, IR/FTIR. 2 Can test field samples in the laboratory. 3 Directly measures the solid phase. 2 Determines nature of the phase but not contaminant-phase interactions. Specific forms of contaminant bound to solids • XRD and SEM • XAS • μL2MS • SIMS • NMR • EPR • XPS 2 Some methods hard to use on natural particles. Detection limits of equipment can cause problems in natural settings. 3 Directly applicable to solid phase. 3 Uniquely suited to identify mechanisms of association. Extraction of soils and sediments for inorganic contaminants Extracts that change the solid phase • Conventional • Sequential • TCLP, SPLP 2 Can extract field soils and sediments, but must remove from field for test. 2 Concentration extracted is qualitatively or operationally related to associations (form) in the solid phase. 1 Operational measure that lumps different association/dissociation processes.

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Bioavailability of Contaminants in Soils and Sediments: Processes, Tools, and Applications Immediately Relevant to Entry into Living Cell Ability to Generalize Relevance to Regulation Usefulness as a Research Tool   1 Characteristics alone are not predictive of biouptake, but are necessary for inferences about other measures and models. 2 Leads to generalization, but by themselves such measures are not predictive of bioavailability processes. 2 Regulators sometimes use such information in normalizations. 3 Essential to understanding contaminant form and links to biouptake in situ. 1 Characteristics alone are not predictive of biouptake, but are necessary for inferences about other measures and models. 2 Leads to generalization, but by themselves such measures are not predictive of bioavailability processes. 1 Seldom used for soil/sediment criteria. May be useful eventually. 3 Potential for contributing to mechanistic understanding. 1 Requires inference about link between specific form and biouptake. 2 Will eventually be essential to generalizing about bioavailability processes. 1 Complicated and consequently of limited use in regulatory environment. 3 Potential to understand what controls bioavailability processes. 2 Extracted concentrations are linked to biouptake by correlation. Best developed for use in particular conditions (e.g., restricted soil series; nutrient deficiency). 2 Generalizations are correlative and some are useful in the appropriate context. 2 Some extracts are in regulatory guidelines, mainly for use as screening tool (e.g., TCLP). Used where groundwater is focal point. Not for sequential extracts. 2 Better accepted for soils. Contentious for use in sediments. Relationships are correlative rather than mechanistic.

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Bioavailability of Contaminants in Soils and Sediments: Processes, Tools, and Applications Technique Application to the Field Application to Solid Phase Single vs. Lumped Processes Passive approaches • Passive extracts • ASV • Pore water measurements with ASV or ion-specific electrode • Exchangeable resins 2 Extracts miss in situ influences because you must remove materials from field setting. In situ pore water measurements are difficult to make and thus limited. 2 Passive extracts mimic solid phase exchange reactions, at equilibrium. Measures in pore water determine actual outcome of solid phase reactions and dissolved speciation. 1 Extracts and resins are operational measures that lump different association/dissociation processes. Pore water concentrations are the outcome of several processes. In vitro tests to mimic human intake for both organics and inorganics 2 Can use field soils and sediments, but must remove from field for test. 2 Extract the solid phase with simulated physiological fluid. 1 Operational measure that lumps multiple processes. Extraction and other tests of soils and sediments for organic contaminants Fluid-phase extractions • Mild solvents • SWE • Supercritical CO2 extraction • PTD 2 Can extract field sediments, but must remove from field for test. 2 Extracts mimic solid phase exchange reactions. 1 Operational measure that lumps multiple processes. Solid phase and membrane-based extractions • Tenax • C-18 • SPME • SPMD • DGT 2–3 Can use field soils and sediments, but must remove from field for test. May be able to use SPME, SPMD, DGT in situ. 3 Directly applicable to the solid phase or slurry. 1 Operational measure that lumps multiple processes. Other desorption tests • Gas purge • Desorption kinetics and activation energy 2 Can use field samples, but difficult to sustain in field setting. 3 Directly applicable to the solid phase or slurry. 2 Single vs. lumped processes can be decoupled by careful experimental design and working with component materials.

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Bioavailability of Contaminants in Soils and Sediments: Processes, Tools, and Applications Immediately Relevant to Entry into Living Cell Ability to Generalize Relevance to Regulation Usefulness as a Research Tool 2 Extracted concentrations are linked to biouptake by correlation. Pore water concentrations are linked by inference that unassociated form is taken up; most useful for plant uptake. 2 The best methods (like DGT) correlate with bioavailability, but there is uncertain reliability of generalizations. 3 Used in some instances as trigger values for soils. Some sediment guidelines use porewater concentrations. 2 Used in research, although relationships are correlative rather than mechanistic. 2 Infers that what can be extracted will be taken up by organism (biomimetic). 1 Site-by-site test. Limited for generalization. 3 Simplicity makes it attractive to regulators. 2 Operational aspects limit use in research.   2 Infers that what can be extracted will be taken up by organism (biomimetic). 1 Reliability of generalizations is unproven. 1 Regulators seldom use such information for soil/sediment criteria; may be useful eventually. 2 Operational aspects limit use in research. 2 Biomimetic but still an inferential link to biouptake. 2 Reliability of generalizations about bioavailability is unproven; work in progress. 1 Regulators seldom use such information for soil/sediment criteria; may be useful eventually. 3 Potential to measure processes important to biouptake (e.g., can get rates of release). 1 Inferential link to biouptake by correlation or mechanistic model. 3 Generalizations possible with careful experimentation on component materials from different sites. 2 Potential to reveal the relationship between aqueous and solid phase concentrations and soil quality criteria. 3 Can lead to greater mechanistic understanding.

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Bioavailability of Contaminants in Soils and Sediments: Processes, Tools, and Applications Technique Application to the Field Application to Solid Phase Single vs. Lumped Processes Normalizations Organic and inorganic correlations • Ratios and models • AVS/SEM • EqP 2 Extracted contaminant normalized to in situ conditions. Difficult to mimic field setting. 2 Designed to describe associations with the solid phase that are relevant to biouptake (e.g., those that control exchange). 2 Ratioing assumes specific processes are described, and infers that they define biouptake. Biological approaches to measuring uptake Assimilation efficiency 1 Can use natural samples, but requires spiking and loss of in situ influences. 2 Direct intake from solid. Allows inferences about natural solids that are ingested. 3 Mechanistic. Determination of single process (biouptake). Mineralization/assimilation assays for microorganisms 1 Requires sample removal, and sometimes spiking. 1 Requires contaminant transfer to aqueous phase. 1 Measures the composite effect of several processes. Bioassays: cell cultures and isolated organs/tissues 2 Can use field soils and sediments, but must remove from field for test. 1–2 Some techniques can use solid phase material while others require extracts. 3 Mechanistic. Determination of single process (biouptake). Bioassays: whole organism bioaccumulation • Plants • Invertebrates • Fish • Birds • Mammals (all exposure routes) 1–2 Can use field soils and sediments, but must remove from field for test. May have to spike dermal tests. 2 Solid phase materials can be tested directly. 2 Whole organism bioaccumulation integrates influences of several biological processes, but is indicative of biouptake.

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