Toxic unit

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Toxic units (TU) are used in the field of toxicology to quantify the interactions of toxicants in binary mixtures of chemicals. [1] A toxic unit for a given compound is based on the concentration at which there is a 50% effect (ex. EC50) for a certain biological endpoint. One toxic unit is equal to the EC50 for a given endpoint for a specific biological effect over a given amount of time. Toxic units allow for the comparison of the individual toxicities of a binary mixture to the combined toxicity. This allows researchers to categorize mixtures as additive, synergistic or antagonistic. Synergism and antagonism are defined by mixtures that are more or less toxic than predicted by the sum of their toxic units.

Contents

Contaminants are frequently present as mixtures in the environment. Regulatory decisions are based on mixture toxicity models that assume additivity, which can result in under or overestimation of toxic effects. Refining our understanding of mixture interactions can lead to better informed environmental management and decision making. In addition, exploring mixture interactions can elucidate the mechanisms of action for specific toxicants which, in many cases, are poorly understood.

Methods

Application of toxic units requires toxicity data for the individual components of the mixture as well as specialized mixture toxicity data. Evaluating the response of each individual chemical allows researchers to generate a new dosing metric, toxic units, which is standardized to the toxicity of each chemical. Since the toxicity of two compounds may vary widely, 1 toxic unit of two different compounds could correspond to two very different concentrations on a per mass basis. In addition to the toxicity of the individual components, use of toxic units requires a 2x2 factorial design concentration series where the response is measured to an increase of each contaminant with the other contaminant held constant. This elaborate concentration series allows researchers to describe how the mixture components interact with each other and predict effects at untested combinations components with nonlinear regression models.

Point estimates

Example of concentration-response model used to calculate EC50. Toxciant concentration is on the X-axis and biological response is on the Y-axis. Concentration-response curve.jpg
Example of concentration-response model used to calculate EC50. Toxciant concentration is on the X-axis and biological response is on the Y-axis.

Point estimation is a technique to predict population parameters based on available sample data and can be used to relate the mass based concentration to a toxicity based metric. Point estimates in toxicology are frequently response endpoints on a dose response curve. These point estimates predict at what concentration one would expect to see a given biological endpoint like 50% mortality (LC50). Any toxicological endpoint (growth inhibition, reproduction, behavior etc.) can be used as the toxicity metric to convert from mass based concentration to toxic units.

Point estimates are generated by fitting a nonlinear regression model to toxicity data and using that model to predict the concentration of chemical required to elicit a known response of the biological endpoint.

Equation and calculations

One Toxic unit can be defined by the researcher as the concentration of a given chemical required to cause a given toxicological endpoint (LC50, EC50, IC50).

1TU=LC50 or 1TU=IC50 for inhibition of growth

Since the mass or molar based concentrations of different chemicals required to cause a given endpoint like an LC50 may vary widely, the concentration that corresponds to 1TU is specific to each individual chemical tested.

Isobolograms

Isobolograms are one way to present the results of binary mixture toxicity testing based in toxic units. [2] The strength of this method is its simplicity and ease of use. First a line of additivity is plotted that corresponds to all the combinations of the two chemicals that would result in one toxic unit. Next the experimental results from binary mixture tests are plotted on the isobologram. The results from the mixture test are point estimates from the mixture dose response curves that correspond to the single chemical tests. When these mixture point estimates are plotted on the isobologram, the region that they fall into (based on the concentrations of the two chemicals required to cause that given endpoint) demonstrates whether the mixture interactions are additive, synergistic or antagonistic.

Response surfaces

Three dimensional graph depicting the function F(x,y) where x and y could be the concentration of the individual components in toxic units and the height of the graph depicts the toxicological response. F(x,y)=-((cosx)^2 + (cosy)^2)^2.PNG
Three dimensional graph depicting the function F(x,y) where x and y could be the concentration of the individual components in toxic units and the height of the graph depicts the toxicological response.

Response surfaces are a more advanced and complex way to visualize the same information presented in an isobologram. A response surface is a three dimensional graph with concentrations of individual components in toxic units on the x and y axis and the response variable on the z axis. This three dimensional representation of the organisms response to the two chemical stressors can be used to predict the toxicity of any combination of the components based on the nonlinear regression models that form the response surface. [2]

Antagonistic, additive, and synergistic effects

The primary utility of toxic units is to classify mixture interactions as additive, synergistic or antagonistic. Additivity means that the toxicity of the mixture is equal to the sum of the toxicities of the individual components. Additivity is the default assumption of models used to predict toxicity of mixtures for regulatory and environmental management purposes. Synergistic effects occur when the experimental toxicity of the mixture is greater than the sum of the individual toxicities. Conversely, antagonistic effects occur when the experimental toxicity of a mixture is less than would be predicted by additivity. Understanding mixture interactions can prevent over or underestimation of toxicity by regulators who assume additivity for uncategorized mixtures.

Applications

Equilibrium partitioning sediment benchmarks

The U.S. EPA uses toxic units as a benchmark, called the equilibrium partitioning sediment benchmark (ESB), for predicting the toxicity of polycyclic aromatic hydrocarbon (PAH)-contaminated sediments to benthic invertebrates. [3] Toxic units are calculated from sediment concentrations of 34 PAHs and their expected sediment, water, and lipid partitioning behavior. Based on the equilibrium partitioning approach (which accounts for the varying biological availability of chemicals in different sediments), the ESB for total PAH is the sum of the quotients of a minimum of each of the 34 individual PAHs in a specific sediment, divided by the final chronic value concentration for each specific PAH in sediment. According to the EPA, freshwater or saltwater sediments that contain less or equal to 1.0 toxic units of the mixture of the 34 PAHs or more PAHs are acceptable for the protection of benthic organisms. Sediments that are greater than 1.0 toxic units are not protective and potentially have adverse effects to benthic organisms.

EPA ESBs do not consider antagonistic, additive, or synergistic effects of other sediment contaminants and have been criticized as an overly conservative estimate for pyrogenic PAHs (such as those from manufactured gas plant processes). [4] This is in part due to the analytical approaches for determining the toxic units for both pyrogenic and petrogenic PAHs.

Toxicity identification evaluation

The toxicity identification evaluation (TIE) is an approach to systematically characterize, identify, and confirm toxic substances in whole sediments and sediment interstitial waters. [5] This approach is typically carried out by the EPA. The effluent effect concentration data and the measured toxicant concentration data are transformed to toxic units for the regression analysis to evaluate whether a linear relationship exists between two or more toxicants. [6]

Limitations

The limitations associated with using toxic units are largely dependent on the methodology in which they are being used. For example, the use of isobolograms is applicable to only binary mixtures. In general, toxic units are based on point estimates which are limited by projection. Point estimates, and therefore toxic units, are a simplification of a dose-response model. Information about toxic effects at concentrations other than the point estimate are lost in translation.

Alternative ways to study mixtures

Top-down Approach

A common method for studying mixtures is to measure the total toxicity of the mixture and consider the internal toxicant interactions as irrelevant. [2] Any mixture effects are taken into account in the total toxicity. The results for this method are limited by being mixture specific and has limited value in determining specific mechanisms of toxicity.

GLM Approach

Using Generalized Linear Models (GLM) allows for complex, non-parametric model fitting to describe the toxicity complex mixtures. Generalized Linear Models are more likely to find significant differences from additivity than TU approaches. [7] The GLM approach also allows for the alteration of models to reflect current knowledge of biological mechanisms [8]

Related Research Articles

<span class="mw-page-title-main">Toxicity</span> Degree of harmfulness of substances

Toxicity is the degree to which a chemical substance or a particular mixture of substances can damage an organism. Toxicity can refer to the effect on a whole organism, such as an animal, bacterium, or plant, as well as the effect on a substructure of the organism, such as a cell (cytotoxicity) or an organ such as the liver (hepatotoxicity). By extension, the word may be metaphorically used to describe toxic effects on larger and more complex groups, such as the family unit or society at large. Sometimes the word is more or less synonymous with poisoning in everyday usage.

Obtuse toxicity, the development of adverse effects as a result of long term exposure to a contaminant or other stressor, is an important aspect of aquatic toxicology. Adverse effects associated with Obtuse toxicity can be directly lethal but are more commonly sublethal, including changes in growth, reproduction, or behavior. Obtuse toxicity is in contrast to acute toxicity, which occurs over a shorter period of time to higher concentrations. Various toxicity tests can be performed to assess the Obtuse toxicity of different contaminants, and usually last at least 10% of an organism's lifespan. Results of aquatic Obtuse toxicity tests can be used to determine water quality guidelines and regulations for protection of aquatic organisms.

In vitro toxicity testing is the scientific analysis of the toxic effects of chemical substances on cultured bacteria or mammalian cells. In vitro testing methods are employed primarily to identify potentially hazardous chemicals and/or to confirm the lack of certain toxic properties in the early stages of the development of potentially useful new substances such as therapeutic drugs, agricultural chemicals and food additives.

<span class="mw-page-title-main">Aquatic toxicology</span> Study of manufactured products on aquatic organisms

Aquatic toxicology is the study of the effects of manufactured chemicals and other anthropogenic and natural materials and activities on aquatic organisms at various levels of organization, from subcellular through individual organisms to communities and ecosystems. Aquatic toxicology is a multidisciplinary field which integrates toxicology, aquatic ecology and aquatic chemistry.

<span class="mw-page-title-main">Ecotoxicity</span>

Ecotoxicity, the subject of study in the field of ecotoxicology, refers to the biological, chemical or physical stressors that affect ecosystems. Such stressors could occur in the natural environment at densities, concentrations, or levels high enough to disrupt natural biochemical and physiological behavior and interactions. This ultimately affects all living organisms that comprise an ecosystem.

Toxic equivalency factor (TEF) expresses the toxicity of dioxins, furans and PCBs in terms of the most toxic form of dioxin, 2,3,7,8-TCDD. The toxicity of the individual congeners may vary by orders of magnitude.

In environmental toxicology, effects range low (ERL) and effects range median (ERM) are measures of toxicity in marine sediment. They are used by public agencies in the United States in formulating guidelines in assessing toxicity hazards, in particular from trace metals or organic contaminants.

A mode of toxic action is a common set of physiological and behavioral signs that characterize a type of adverse biological response. A mode of action should not be confused with mechanism of action, which refer to the biochemical processes underlying a given mode of action. Modes of toxic action are important, widely used tools in ecotoxicology and aquatic toxicology because they classify toxicants or pollutants according to their type of toxic action. There are two major types of modes of toxic action: non-specific acting toxicants and specific acting toxicants. Non-specific acting toxicants are those that produce narcosis, while specific acting toxicants are those that are non-narcotic and that produce a specific action at a specific target site.

Fish acute toxicity syndrome (FATS) is a set of common chemical and functional responses in fish resulting from a short-term, acute exposure to a lethal concentration of a toxicant, a chemical or material that can produce an unfavorable effect in a living organism. By definition, modes of action are characterized by FATS because the combination of common responses that represent each fish acute toxicity syndrome characterize an adverse biological effect. Therefore, toxicants that have the same mode of action elicit similar sets of responses in the organism and can be classified by the same fish acute toxicity syndrome.

Tissue residue is the concentration of a chemical or compound in an organism's tissue or in a portion of an organism's tissue. Tissue residue is used in aquatic toxicology to help determine the fate of chemicals in aquatic systems, bioaccumulation of a substance, or bioavailability of a substance, account for multiple routes of exposure, and address an organism's exposure to chemical mixtures. A tissue residue approach to toxicity testing is considered a more direct and less variable measure of chemical exposure and is less dependent on external environmental factors than measuring the concentration of a chemical in the exposure media.

Toxicodynamics, termed pharmacodynamics in pharmacology, describes the dynamic interactions of a toxicant with a biological target and its biological effects. A biological target, also known as the site of action, can be binding proteins, ion channels, DNA, or a variety of other receptors. When a toxicant enters an organism, it can interact with these receptors and produce structural or functional alterations. The mechanism of action of the toxicant, as determined by a toxicant’s chemical properties, will determine what receptors are targeted and the overall toxic effect at the cellular level and organismal level.

Pre-spawn mortality is a phenomenon where adult coho salmon, Oncorhynchus kisutch, die before spawning when returning to freshwater streams to spawn. It is also known as Urban Runoff Mortality Syndrome in more recent studies. This occurrence has been observed in much of the Puget Sound region of the Pacific Northwest. During fall migration, salmonids pass through urban watersheds which are contaminated with stormwater runoff. As the coho salmon pass through these waters, many will show symptoms of lethargy, loss of equilibrium and disorientation, and die within a few hours of showing these symptoms. These symptoms and behaviors are prevalent after rain events. Mortality often occurs before salmon have the opportunity to spawn, which is determined by cutting open female carcasses and observing for unfertilized eggs. Rates of pre-spawn mortality could impact the local salmon populations. Based on model projections, if rates continue, populations of coho salmon could become extinct within the next few decades.

An early life stage (ELS) test is a chronic toxicity test using sensitive early life stages like embryos or larvae to predict the effects of toxicants on organisms. ELS tests were developed to be quicker and more cost-efficient than full life-cycle tests, taking on average 1–5 months to complete compared to 6–12 months for a life-cycle test. They are commonly used in aquatic toxicology, particularly with fish. Growth and survival are the typically measured endpoints, for which a Maximum Acceptable Toxicant Concentration (MATC) can be estimated. ELS tests allow for the testing of fish species that otherwise could not be studied due to length of life, spawning requirements, or size. ELS tests are used as part of environmental risk assessments by regulatory agencies including the U.S. Environmental Protection Agency (EPA) and Environment Canada, as well as the Organisation for Economic Co-operation and Development (OECD).

Simultaneously extracted metals/Acid-volatile sulfide (SEM-AVS) is an approach used in the field of aquatic toxicology to assess the potential for metal ions found in sediment to cause toxic effects in organisms dwelling in the sediment. In this approach, the amounts of several heavy metals in a sediment sample are measured in a laboratory; at the same time, the amount of acid-volatile sulfide is determined. Based on the chemical interactions between heavy metals (SEM) and acid-volatile sulfide (AVS), the concentrations of these two components can be used to assess the potential for toxicity to sediment-dwelling organisms.

Equilibrium partitioning Sediment Benchmarks (ESBs) are a type of Sediment Quality Guideline (SQG) derived by the US Environmental Protection Agency (EPA) for the protection of benthic organisms. ESBs are based on the bioavailable concentration of contaminants in sediments rather than the dry-weight concentration. It has been demonstrated that sediment concentrations on a dry-weight basis often do not predict biological effects. Interstitial water concentrations, however, predict biological effects much better. This is true because the chemical present in the interstitial water (or pore water) is the uncomplexed/free phase of the chemical that is bioavailable and toxic to benthic organisms. Other phases of the chemical are bound to sediment particles like organic carbon (OC) or acid volatile sulfides (AVS) and are not bioavailable. Thus the interstitial water concentration is important to consider for effects to benthic organisms.

The predicted no-effect concentration (PNEC) is the concentration of a chemical which marks the limit at which below no adverse effects of exposure in an ecosystem are measured. PNEC values are intended to be conservative and predict the concentration at which a chemical will likely have no toxic effect. They are not intended to predict the upper limit of concentration of a chemical that has a toxic effect. PNEC values are often used in environmental risk assessment as a tool in ecotoxicology. A PNEC for a chemical can be calculated with acute toxicity or chronic toxicity single-species data, Species Sensitivity Distribution (SSD) multi-species data, field data or model ecosystems data. Depending on the type of data used, an assessment factor is used to account for the confidence of the toxicity data being extrapolated to an entire ecosystem.

In aquatic toxicology, the sediment quality triad (SQT) approach has been used as an assessment tool to evaluate the extent of sediment degradation resulting from contaminants released due to human activity present in aquatic environments. This evaluation focuses on three main components: 1.) sediment chemistry, 2.) sediment toxicity tests using aquatic organisms, and 3.) the field effects on the benthic organisms. Often used in risk assessment, the combination of three lines of evidence can lead to a comprehensive understanding of the possible effects to the aquatic community. Although the SQT approach does not provide a cause-and-effect relationship linking concentrations of individual chemicals to adverse biological effects, it does provide an assessment of sediment quality commonly used to explain sediment characteristics quantitatively. The information provided by each portion of the SQT is unique and complementary, and the combination of these portions is necessary because no single characteristic provides comprehensive information regarding a specific site

Occupational toxicology is the application of toxicology to chemical hazards in the workplace. It focuses on substances and conditions that occur in workplaces, where inhalation exposure and dermal exposure are most important, there is often exposure to mixtures of chemicals whose interactions are complex, health effects are influenced or confounded by other environmental and individual factors, and there is a focus on identifying early adverse affects that are more subtle than those presented in clinical medicine.

Toxicological databases are large compilations of data derived from aquatic and environmental toxicity studies. Data is aggregated from a large number of individual studies in which toxic effects upon aquatic and terrestrial organisms have been determined for different chemicals. These databases are then used by toxicologists, chemists, regulatory agencies and scientists to investigate and predict the likelihood that an organic or inorganic chemical will cause an adverse effect on exposed organisms.

The acute to chronic ratio (ACR) uses acute toxicity data to gauge the chronic toxicity (MATC) of a chemical of interest to an organism. The science behind determining a safe concentration to the environment is imperfect, statistically limited, and resource intensive. There is an unfilled demand for the rapid assessment of different chemical toxicity to many different organisms. The ACR is a proposed solution to this demand.

References

  1. "toxic unit". TheFreeDictionary.com. Retrieved 2019-12-20.
  2. 1 2 3 Warne MSJ. 2003. A Review of the Ecotoxicity of Mixtures, Approaches to and Recommendations for, their Management. Proceedings of the Fifth National Workshop on the Assessment of Site Contamination. EPHC. pp.256
  3. U.S. Environmental Protection Agency. 2003. Procedures for the derivation of ESBs for the protection of benthic organisms: PAH mixtures. EPA/600/R-02/013. Office of Research and Development, Washington, DC.
  4. Hawthorne SB, DJ Miller, and JP Kreitinger. 2006. Measurement of total polycyclic aromatic hydrocarbon concentrations in sediments and toxic units used for estimating risk to benthic invertebrates at manufactured gas plant sites. Environmental Toxicology and Chemistry 25(1):287-296
  5. Burgess RM and KT Ho. Sediment Toxicity Identification Evaluation. Chapter S, Ferard and Blaise (ed.), Encyclopedia of Aquatic Ecotoxicology. Springer, New York, NY, 8/13/2013:online, (2013).
  6. U.S. Environmental Protection Agency. 1993. Methods for Aquatic Toxicity Identification Evaluations: Phase III Toxicity Confirmation Procedures for Samples Exhibiting Acute and Chronic Toxicity. EPA/600/R-92/081. Office of Research and Development, Washington, DC.
  7. C., Newman, Michael (1998). Fundamentals of ecotoxicology. Ann Arbor Press. ISBN   1575040131. OCLC   37981637.{{cite book}}: CS1 maint: multiple names: authors list (link)
  8. National Research Council (US) Committee on Methods for the In Vivo Toxicity Testing of Complex Mixtures (1998). Complex Mixtures: Methods for In Vivo Toxicity Testing. Washington DC: National Academies Press (US).