Estimation of toxicity of chemical mixtures through modeling of chemical interactions.

The Agency for Toxic Substances and Disease Registry (ATSDR), in collaboration with the Dutch Organization for Applied Scientific Research (TNO) Nutrition and Food Research Institute, is conducting studies to evaluate the role of chemical interactions in the expression of toxicity from low-level exposure to combinations of chemicals. The goal of this collaborative effort is to use a weight-of-evidence (WOE) approach to estimate joint toxicity of some simple chemical mixtures and to compare the estimations with test results from animal toxicity studies. The WOE approach uses individual chemical dose-response assessments and algorithms that incorporate various assumptions regarding potential chemical interactions. Qualitative evaluations were prepared for binary combinations of chemicals for the effect of butyl hydroxyanisole on di(2-ethylhexyl)phthalate, the effect of stannous chloride on Cd chloride (CdCl2), and the effect of CdCl2 on loperamide. Analyses of these evaluations and their comparison with the conclusions of laboratory animal experiments indicate that the WOE approach can be used to estimate qualitatively the joint toxicity of such simple mixtures. To further test the utility of the WOE approach, qualitative and semiquantitative evaluations were prepared for two chemical mixtures--one with similarly acting halogenated aliphatics (trichloroethylene, tetrachloroethylene, hexachloro-1,3-butadiene[HCBD], and 1,1,2-trichloro-3,3,3-trifluoropropene [TCTFP]) and the other with dissimilarly acting nephrotoxic components (mercuric chloride, lysinolalanine, D-limonene, and HCBD). These two sets of data were used to estimate the overall toxicities of the mixtures using the WOE algorithm for the mixture. The comparison of the results of the estimated toxicity with experimentally determined toxicity of the mixture of similarly acting nephrotoxicants demonstrated that the WOE approach correctly adjusted for the observed interactions in experimental animal studies. However, this was not true for the mixture of dissimilarly acting nephrotoxicants. This could be attributed to the fact that WOE evaluations are based on dose additivity that postulates that all chemicals in a given mixture act in the same way--by the same mechanism--and differ only in their potencies. In these cases the WOE approach evaluations, based on consideration of common mechanisms for simple chemical mixtures, can lead to better estimates of joint toxicity of chemical mixtures than the default assumption of dose additivity. The results also show that the WOE evaluations should be target-organ specific because none of the models tested could approximate the observed responses in organs other than the target organs in the laboratory animal studies.

Categorical regression of toxicity data: a case study using aldicarb.

Categorical regression is a mathematical tool that can be adapted to estimate potential health risk from chemical exposures. By regressing ordered categories of toxic severity or pathological staging on exposure dose, this method can estimate the likelihood of observing any of the categories of severity at any dose level. Depending on the nature of the available data, these estimates can take the form of incidence rates for any of the categories in an exposed population or the probability of a new study conducted at a specified dose level being classified as one of the categories. Categorical regression is illustrated using toxicity data on aldicarb. For aldicarb, the data fall into three different groups: human clinical studies, dietary exposures in experimental animals, and accidental human exposure by contaminated crops. The U.S. EPA has assessed this literature and developed a reference dose (RfD) of 0.001 mg/kg-day. The results of applying categorical regression to data from human clinical studies suggests a maximum likelihood risk estimate of adverse effects of 0.008% at a 10-fold higher dose than the RfD when blood cholinesterase inhibition is not considered as an adverse effect. When blood cholinesterase inhibition of 20% or more is considered as an adverse effect, a maximum likelihood risk estimate of adverse effects is 0.1% at a dose 10-fold higher than the RfD.

Bioconcentration and biokinetics of heavy metals in the earthworm.

This study examines the steady state and non-steady state kinetics of five metals, cadmium, copper, lead, nickel, and zinc in earthworms. The steady state kinetics are based on field studies in which worms from contaminated and uncontaminated sites were collected and measurements were made of concentrations in the earthworms and soils. For each of the metals, evidence suggests that bioconcentration depends on the metal concentrations in the soil; bioconcentration is greater at lower soil concentrations. The studies of non-steady state kinetics involve uptake and elimination experiments in which worms are transferred from an uncontaminated soil to a contaminated soil (uptake studies) or from a contaminated soil to an uncontaminated soil (elimination studies). The voiding time is shown to be an important experimental variable in determining the measured levels of metal in earthworms because experimental measurements are usually made on a worm-soil complex (i.e. the soft tissue of the worm and the soil in the gut of the worm). Thus, for metals that are bioconcentrated in worm tissue, increasing the voiding period increases the concentration of the metal in the worm-soil complex. Conversely, for metals that are not bioconcentrated, increasing the voiding time leads to a decrease in concentrations in the worm-soil complex.

Methods of assessing dermal absorption with emphasis on uptake from contaminated vegetation.

Dermal absorption may be an important route of exposure in several exposure scenarios for workers and the general public. Because criteria (e.g. RfDs or MRLs) for chemical exposures are usually expressed in units of mg/kg/day, risk assessments often attempt to convert dermal exposure data to units of mg/kg/day absorbed dose. For some types of dermal exposure involving direct and continuous contact with liquids, Fick's first law may be used. In other cases, such as those involving spills onto the skin surface or dermal exposure to contaminated vegetation, the applicability of Fick's first law is limited. This analysis focuses on a method for estimating absorbed dose from dermal contact with contaminated vegetation or other surfaces. The method involves two steps: estimating the transfer rate from contaminated vegetation to the skin surface, and estimating the extent of absorption from the skin surface into the body. A generic equation can be derived for estimating the transfer rate (TR) from dislodgeable foliar residues (DFR): logTR = 1.09 logDFR + 0.05. Given the surface area of the exposed skin and the duration of contact, this equation can be used to estimate the amount of chemical deposited on the surface of the skin. This equation is based on data from eight different studies using 16 different pesticides. Excluding one outlier (Vinclozolin), the squared correlation coefficient for this equation is 0.78, and the model is significant at p < 0.00001. Data from a series of studies by Feldmann and Maibach (1969, 1970, 1974) are used to estimate dermal absorption. These studies were selected as the most relevant for risk assessment because most of the experimental subjects are human and because of the nature of dermal exposures was closely related to many exposure scenarios used in risk assessments. For all 47 compounds included in this series of studies, there were no significant correlations between commonly available physical or chemical properties and dermal absorption. For those compounds with a K0/w > 1.85, however, the average daily absorption rate (AR) over a five-day postexposure period can be estimated from the molecular weight: logAR = -0.04MW + 1.5. The squared correlation coefficient for this equation is 0.68, and the model is significant at p < 0.00001. The usefulness of this approach is evaluated using a study by Harris and Solomon (1992) in which the absorption of 2,4-D from contaminated turf was measured in a group of volunteers. The estimated absorbed dose using the equations above is very close to the measured values.

A weight-of-evidence approach for assessing interactions in chemical mixtures.

The risk assessment process must encompass all available toxicological data and scientific evidence on the plausible toxicities of a chemical or chemical mixture. As an extension to the approaches used to conduct risk assessments on chemical mixtures, a preliminary scheme, analogous to the IARC classification of carcinogens, is proposed to express the weight of evidence for the interactions in binary mixtures. This scheme is based on composite representation of all the toxicological evidence from animal bioassays and human data, pharmacokinetics studies, metabolism studies, and structure activity relationships. In addition, factors such as the relevance of route, duration and sequence of exposure, toxicological significance of interactions and the quality of in vivo and in vitro data are taken into consideration. The scheme yields an alphanumeric classification that can be used for qualitative risk assessment, and has the potential, as demonstrated in this paper, for quantitative application to site-specific risk assessments. Furthermore, the scheme can be used to estimate interactions or form hypotheses concerning binary interactions. It is flexible and allows all pertinent information to be incorporated in a methodical and consistent manner. Research is needed to identify interaction patterns for simultaneous and sequential exposure scenarios of chemical pollutants in order that this scheme may be developed further and its usefulness and limitations may be tested.

Comparative toxicity of ten organic chemicals to four earthworm species.

Ten organic chemicals were tested for toxicity to four earthworm species: Allolobophora tuberculata, Eisenia fetida, Eudrilus eugeniae and Perionyx excavatus, using the European Economic Community's (EEC) earthworm artificial soil and contact testing procedure. The phenols were the most toxic chemicals tested, followed by the amine, substituted benzenes, halogenated aliphatic hydrocarbon, polycyclic aromatic hydrocarbon and phthalate as the least toxic chemical tested. Correlations among species within each type of test for a given chemical were extremely high, suggesting that the selection of earthworm test species does not markedly affect the assessment of a chemical's toxicity. The correlation between the two tests was low for all test species. The contact test LC50 for a given chemical cannot be directly correlated to an artificial soil test LC50 for the same earthworm species.

Ranking chemicals based on chronic toxicity data.

During the past 3 years, EPA's ECAO/Cincinnati has developed a method to rank chemicals based on chronic toxicity data. This ranking system reflects two primary attributes of every chemical: the minimum effective dose and the type of effect elicited at that dose. The purpose for developing this chronic toxicity ranking system was to provide the EPA with the technical background required to adjust the RQs of hazardous substances designated in Section 101(14) of CERCLA or "Superfund." This approach may have applications to other areas of interest to the EPA and other regulatory agencies where ranking of chemicals based on chronic toxicity is desired.

Antimutagenic effect of rat small intestine in vitro on 1,1,2,3-tetrachloropropene and 1,1,2,3,3-pentachloropropene.

The direct mutagenic activity of 1,1,2,3- tetrachloropropene and 1,1,2,3,3- pentachloropropene in Salmonella typhimurium was measured before and after incubation in the presence of intact segments of rat small intestine in vitro. The number of revertants in tester strain TA100 was reduced by about 95% when these chloropropenes were exposed to rat small intestine for 10-30 min immediately prior to determination of mutagenicity. The existence of an intestine-mediated detoxication reaction was postulated, and was supported by observations that incubation with the chloropropenes for 30 min caused a 48-68% depletion of intestinal glutathione in vitro. Although addition of glutathione to the chloropropenes reduced mutagenicity, the amount of tissue glutathione consumed during incubation of mutagen with intestinal segments is probably insufficient to account for the detoxication. Additional metabolic reactions and/or non-specific protein binding may occur in the intact intestine which contribute to the antimutagenic effect. These initial results support the existence of an effective detoxication mechanism by the small intestine which is likely to reduce the absorption of direct-acting mutagens and other electrophiles.