Mutagenicity of complex mixtures in Salmonella typhimurium. A report of the International Programme on Chemical Safety's Collaborative Study on Complex Mixtures.
Explore the source record for details and available documents.
Mutagenicity Tests↗
SEARCH · PubMed Health
Results for “Complex Mixtures”
Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.
Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.
Salmonella mutagenicity of three complex mixtures assayed with the microsuspension technique. A WHO/IPCS/CSCM study.
In a collaborative study on complex mixtures, three complex mixtures and two pure compounds were assayed with the Salmonella microsuspension technique. The two pure compounds were benzo[a]pyrene (BaP) and 1-nitropyrene (1-NP). The three complex mixtures were standard reference materials (SRMs) from the U.S. National Institute of Standards and Technology, SRM 1649, SRM 1650 and SRM 1597. The two samples SRM 1649, an urban dust particulate matter, and SRM 1650, a diesel particulate matter, were sonicated with dichloromethane. Sample SRM 1597 was an extract of a coal tar sample with a complex mixture of polycyclic aromatic hydrocarbons. The microsuspension assay was performed with Salmonella strains TA98 and TA100 according to Kado et al. (1983) with minor modifications (Löfroth et al., 1988). The results showed that the microsuspension technique is a more sensitive assay than the plate incorporation method. Depending on sample, strain and metabolic condition the mutagenic responses were 3-37 times higher in the microsuspension assay than in the conventional plate incorporation assay. The microsuspension method is thus useful for environmental samples which are often available in only small amounts.
Air Pollutants↗
Emerging methodologies for assessment of complex mixtures: application of bioassays in the Integrated Air Cancer Project.
The assessment of complex mixtures of environmental pollutants requires new interdisciplinary strategies. Integration of bioassay methodologies into these strategies is an important tool that provides direct evidence of the toxicity of a mixture. Short-term genetic bioassays are now widely used in the cancer assessment of complex mixtures. New and emerging interdisciplinary methodologies for assessing complex mixtures using bioassays are illustrated by the U.S. EPA's Integrated Air Cancer Project (IACP). The goals of this project are to identify the major airborne carcinogens and their emission sources and to improve the methodology and data available for human exposure and risk assessment from airborne carcinogens. The research effort is focused primarily on characterizing the impact of complex mixtures of products of incomplete combustion, including gaseous, semi-volatile, and particle-bound organic species. Short-term genetic bioassays have been integrated into several analysis strategies, including environmental biomonitoring; bioassay-directed fractionation/characterization; transformation studies; source apportionment; and exposure, dosimetry, and risk assessment.
Air Pollutants↗
Dose-related differences in DNA adduct levels in rodent tissues following skin application of complex mixtures from air pollution sources.
Dose-related differences in the binding of DNA reactive intermediates for three environmentally important complex mixture particulate extracts and a well-studied carcinogen, benzo[a]pyrene (BaP), were examined in female C-57 mice following multiple topical treatments ranging from 1 to 120 mg/mouse. Particulate extracts from coke oven, coal soot and diesel exhaust were selected as model complex mixtures based on short-term mutagenicity assays, animal bioassays for carcinogenicity or epidemiological studies, where increased incidences of lung cancer in exposed populations were detected. Positive and negative control animals were treated with 1.2 mg BaP or acetone respectively. DNA was isolated from skin, lung and liver 24 h following the last application and analyzed for DNA adducts using the nuclease P1 version of the 32P-postlabeling assay. Each of the particulate extracts produced distinct patterns of DNA adducts. A diagonal zone of radioactivity, presumably representing multiple putative DNA adducts, was observed for coke-oven, coal-soot- and diesel-modified DNA samples. One adduct, common to all three complex-mixture-modified DNA samples, co-migrated with the major BaP adduct observed following treatment with BaP alone. Based on the BaP concentration for each of the extracts it seems unlikely that this adduct is derived from BaP alone. It is possible that an adduct is formed with chromatographic properties similar to the major BaP-derived adduct detected in mice treated with BaP alone. This adduct was detected in all tissues examined and represented approximately 12-34% of the total number of adducts detected within the diagonal radioactive zone for all coke-oven- and coal-soot-exposed tissues (skin, lung and liver). In contrast, this adduct represented 49-67% of the total radioactivity recovered from the diagonal zone of DNA isolated from lungs of animals exposed to diesel extract. The highest total number of adducts resulted from the metabolism of coke oven extract followed by coal soot and diesel treatments respectively. A dose-dependent increase in adduct formation was observed for all tissues in the diesel- and coal-soot-treatment mice. Liver and lung, but not skin, DNA adduct levels increased in a dose-dependent manner in the coke-oven-treated mice. The percentage of dose administered, detected as DNA adducts increased in all tissues as the dose decreased for all three complex mixtures. These data have important implications for risk assessment of these complex mixtures.
Adenosine Triphosphate↗
Statistical aspects of experimental studies with complex mixtures.
Some formal concepts are introduced relating to the statistical design and analysis of experiments with complex mixtures. Aspects relating to the chemical analysis of complex mixtures, identification of their major components, continuous monitoring of samples or related issues are not addressed. As a surrogate for experimental studies in general, the discussion is oriented towards the situation of long-term animal experiments. When complex mixtures of unknown components are to be tested, samples collected under a standardized protocol must be applied to the different exposure groups in such a way that the experimental groups differ only in the dose of the exposure they receive. If the composition of a mixture is known, selected components may be combined in experimental testing in order to study the nature of their joint effect. Careful consideration guided by a priori knowledge about the individual compounds' dose-response relationships is required to design and analyse such experiments. There is an apparent need to avoid confusion by unifying the terminology in this field.
Animals↗
Comparative antimutagenicity of 5 compounds against 5 mutagenic complex mixtures in Salmonella typhimurium strain TA98.
Using the Ames Salmonella/microsome assay, we compared the antimutagenic activities of chlorophyllin, retinol, beta-carotene, vitamin C, and vitamin E against solvent extracts of coal dust, diesel emission particles, airborne particles, fried beef, and tobacco snuff. The results show that chlorophyllin inhibited 69% of the mutagenic activity of tobacco snuff and over 90% of that of the other 4 complex mixtures. Retinol inhibited 29-48% of the mutagenic activity of all 5 complex mixtures. beta-Carotene, vitamin C, and vitamin E inhibited, if any, less than 39% of the activity of the complex mixtures studied. Vitamin C enhanced the mutagenicity of airborne particles. These results indicate that for these dietary and environmental complex mixtures chlorophyllin is a more effective antimutagen than retinol, beta-carotene, vitamin C, and vitamin E.
Air↗
Application of modified Salmonella/microsome prescreen to petroleum-derived complex mixtures and polynuclear aromatic hydrocarbons (PAH)
In some cases, the Salmonella mutagenicity assay may fail to predict the carcinogenic potential of PAH (and of complex mixtures containing PAH) because of nonoptimal in vitro metabolic activation parameters. In this study, 7 petroleum-derived complex mixtures, as well as a number of individual PAH which were representative constituents of such mixtures, were tested in a Salmonella prescreen using quadrant plates with rat or hamster S9 at concentrations approximately 2-8 times those used in the standard assay. Some PAH (perylene, quinoline, benzo[b]chrysene, phenanthrene, anthracene) were optimally activated to mutagens by S9 at 400 microliters/plate. Rat S9 was similar to hamster S9 for most tested PAH, but anthracene and quinoline mutagenicity was enhanced by hamster S9. All 7 complex mixtures were more mutagenic with 200-400 microliters/plate S9; rat was generally slightly more efficient than hamster. Modifying this assay to include a prescreen using a range of S9 concentrations (and perhaps from species other than rat) may improve prediction of the potential carcinogenicity of complex petroleum-derived mixtures.
Animals↗
Genotoxicity of complex mixtures: strategies for the identification and comparative assessment of airborne mutagens and carcinogens from combustion sources.
Two strategies for assessment of the toxicity of complex mixtures are described and illustrated with examples from genotoxicity studies of complex combustion mixtures. The first, a strategy for identifying biologically active compounds or compound classes in complex mixtures, is called bioassay-directed fractionation and characterization. The identification and assessment of mutagens and potential carcinogens in complex mixtures has been significantly advanced by the use of short-term genetic bioassays. Bioassay-directed fractionation coupled with new analytical characterization methods has provided the tools needed to more efficiently identify potential carcinogens in complex combustion emissions and urban air samples. These studies have shown that a significant portion of the mutagenicity in combustion emissions and urban air is found in fractions more polar than polynuclear aromatic hydrocarbons (PAHs). A second strategy, the comparative potency method, provides an approach to evaluating the relative toxicities of a series of mixtures. The comparative mutagenicity and carcinogenicity of a series of combustion emissions has been assessed using dose-response studies in bacteria, mammalian cells, and rodents. Comparative mutagenic and tumorigenic emission rates or emission factors provide a means to directly compare the relative hazard of the sources. This data base has also been used to develop a comparative risk assessment methodology for combustion emissions.
Air Pollutants↗
In vitro mutagenicity of water contaminants in complex mixtures.
Trihalomethanes, Carbon tetrachloride and trichloroethylene were tested in single, binary and multi-complex mixtures using standard tester strains TA1535, TA1537, TA98 and TA100 of Salmonella typhimurium with and without addition of an in vitro metabolizing fraction S-9. Chloroform (CHCl3) was found to be mutagenic in all strains without S-9 activation. However, when tested with Bromoform (15%), which was nonmutagenic singly, the combined effect of the mixture was nonmutagenic. CCl4 was a direct mutagen (without S-9) in all strains except TA 1535. When combined with 85% CHCl3, only strains TA1535 and TA1537 were mutagenic. When tested with mammalian activation (S-9), CCl4 was mutagenic in all strains. However, when tested with CHCl3 (CHCl3 and CCl4-85:15), the mutagenic capability was lost. With or without S-9 Activation multi-complex mixture of CHCl3, CCl4 and TCE (85:8:7) was mutagenic for a narrow range of doses in all strains.
Carbon Tetrachloride↗
Validation of the SOS/Umu test with mutagenic complex mixtures.
The SOS/Umu test, a rapid system for detecting genotoxic agents by monitoring SOS responses, was evaluated with the extracts of 8 mutagenic complex mixtures (airborne particles, coal dust, tobacco snuff, fried beef, fried shredded pork, airborne particles from polyurethane plants). In this system, the SOS function induced by genotoxicants is detected by a colorimetric measurement of beta-galactosidase in tester cells carrying a umuC-lacZ-fused gene on the plasmid. Results from the study show that a higher beta-galactosidase activity was found when the enzyme substrate and treated cells were added simultaneously into the enzyme reaction mixture and post-treatment dilution (10 X dilution with fresh medium) and incubation (for 2 h) were incorporated. The post-treatment dilution is necessary to reduce a possible false positive due to the color of test substances. The extracts of all mutagenic complex mixtures tested were found to induce dose-related SOS responses, indicating that the SOS/Umu test is potentially useful for the detection of mutagenic complex environmental mixtures.
Air Pollutants↗
Evaluation of cyano bonded phases for the fractionation of bioactive complex mixtures.
Cyanopropyl bonded-phase sorbents are investigated for the liquid chromatographic (LC) fractionation of complex mixtures with the goal of extending routine sample fractionation to samples containing highly polar biologically active components. Separations based both on gravity-flow column chromatography and on high performance liquid chromatography (HPLC) are evaluated for efficiency and resolving power. Typical gravity-flow column separations of neutral to moderately polar mixtures are found to be less effective than those employing traditional sorbents; however, HPLC methods could be called upon to provide the resolving power necessary for satisfactory separations of these mixtures. For mixtures of polar reference compounds, cyano bonded phases provide good separations and are very efficient, permitting the elution of components too polar to be recovered from traditional sorbents. A scheme combining gravity-flow chromatography with HPLC is developed for the fractionation of complex mixtures into compound classes. It is based on the chromatographic behavior of reference compounds covering a wide polarity range utilizing cyanopropyl sorbents exclusively. Results are presented for the fractionation of two air particulate reference samples containing highly polar components.
Absorption↗
Epidemiologic approaches for assessing health risks from complex mixtures in indoor air.
Indoor air may be contaminated by diverse gaseous and particulate pollutants that may adversely affect health. As a basis for controlling adverse health effects of indoor air pollution, the presence of a hazard needs to be confirmed, and the quantitative relationship between exposure and response needs to be described. Toxicological, clinical, and epidemiological studies represent complementary approaches for obtaining the requisite evidence. The assessment of the effects of complex mixtures poses a difficult challenge for epidemiologists. Understanding the effects of exposure may require accurate assessment of concentrations and personal exposures to multiple agents and analytical approaches that can identify independent effects of single agents and the synergistic or antagonistic effects that may occur in mixtures. The array of epidemiological study designs for this task includes descriptive studies, cohort studies, and case-control studies, each having potential advantages and disadvantages for studying complex mixtures. This presentation considers issues related to exposure assessment and study design for addressing the effects of complex mixtures in indoor air.
Air Pollution, Indoor↗
Design and implementation of a collaborative study of the mutagenicity of complex mixtures in Salmonella typhimurium.
In 1987, the International Programme on Chemical Safety (IPCS) in collaboration with the U.S. Environmental Protection Agency (U.S. EPA) and the U.S. National Institute of Standards and Technology (U.S. NIST) initiated an international collaborative study of the mutagenicity of complex environmental mixtures in the Ames Salmonella typhimurium mutation assay. The objectives of this study were: (1) to estimate the inter- and intra-laboratory variability associated with the extraction of mixtures for bioassay, (2) to estimate the inter- and intra-laboratory variability associated with the Salmonella typhimurium bioassay when applied to complex mixtures, and (3) to determine whether standard reference complex mixtures would be useful in mutagenicity studies and to evaluate whether reference or certified mutagenicity values determined from this collaborative study should be reported. The complex mixtures used in this study were selected from standard reference materials (SRMs) which had previously been issued by the U.S. NIST as SRM 1597 (coal tar), SRM 1649 (diesel particulate matter) and SRM 1650 (urban air particulate matter) with certified values for polycyclic aromatic hydrocarbons. These SRM complex mixtures are available to scientists as reference standards for analytical chemistry research and are under consideration as SRMs for mutagenicity studies of complex environmental mixtures. This paper briefly describes the final study design, protocol, selection of the complex mixtures, and implementation of this international study.
Laboratories↗
Approaches for assessing health risks from complex mixtures in indoor air: a panel overview.
Critical to a more definitive human health assessment of the potential health risks from exposure to complex mixtures in indoor air is the need for a more definitive clinical measure and etiology of the health effects of complex mixtures. This panel overview highlights six of the eight presentations of the conference panel discussion and features a number of the major topical areas of indoor air concern. W. G. Meggs assessed clinical research priorities with primary focus on the role of volatile organic chemicals in human health, recognizing the areas where definitive data are lacking. By recognizing many types of chemical sensitivity, it may be possible to design studies that can illuminate the mechanisms by which chemical exposure may cause disease. The critically important topic of multiple chemical sensitivity was discussed by N. A. Ashford, who identified four high risk groups and defined the demographics of these groups. P. A. Schulte addressed the issue of biological markers of susceptibility with specific considerations of both methodological and societal aspects that may be operative in the ability to detect innate or inborne differences between individuals and populations. Three case studies were reviewed. H. Anderson discussed the past and present priorities from a public health perspective, focusing on those issues dealing with exposures to environmental tobacco smoke and formaldehyde off-gassing from materials used in mobile home construction. J. J. Osborne described several case studies involving wood smoke exposure to children, with emphasis on the significantly greater occurrence of chronic respiratory symptoms and acute chest illness for children from homes heated with woodburning stoves.(ABSTRACT TRUNCATED AT 250 WORDS)
Air Pollution, Indoor↗
A further characterization of Drosophila cuticular monoenes using a mass spectrometry method to localize double bonds in complex mixtures.
Positions of double bonds of olefins of complex mixtures, such as those of Drosophila cuticular hydrocarbons, have been determined using a simple method combining methoxymercuration-demercuration of extracts and analysis by gas chromatography-mass spectrometry using ammonia for chemical ionization. Chemical similarities between cuticular monoenes of both sexes of Drosophila simulans and males of Drosophila melanogaster are specified: the major isomer is always in position 7.
Alkenes↗
Toxicological investigations in the semiconductor industry: I. Studies on the acute oral toxicity of a complex mixture of waste products from the aluminium plasma etching process.
In dry etching processes--one of the sources of potential exposure to toxic wastes in the semiconductor industry--complex mixtures of inorganic and organic compounds arise from reactions between feed stock gases (BCl3/Cl2), top layers (aluminium photoresist), and the carrier gas (N2). Two different fractions of the complex mixture--one an ethanolic solution (ES) and the other an insoluble liquid residue (LR)--were examined for acute oral toxicity in rats. Analytical data showed that the ethanol soluble fraction contained mainly inorganic compounds, whereas the residue contained various halogenated hydrocarbons. Neither death nor behavioral changes occurred after oral administration and observation up to 23 days. ES caused a lower mean arterial blood pressure in both sexes, increased P-R-intervals in male rats, and caused some mild biochemical and hematological alterations and changes in relative organ weights compared to the control groups. Exposure to LR influenced food and water intake, and caused a significant decrease in body weights, signs of polyurie, as well as changes in various relative organ weights and biochemical and hematological parameters. The blood pressure of the male animals fell and the heart rates of both sexes decreased.
Aluminum↗
Interactive and multi-sensory analysis of complex mixtures by an automated gas chromatography system.
Methods are presented for increased human-computer interactions in the analysis of metabolic profiles of urinary organic acids. Complex mixtures of trimethylsilylated derivatives are separated by capillary gas-liquid chromatography on a 25-m bonded-phase, fused-silica column. Retention times and areas, determined by a printing integrator, are transferred to a personal computer along with the digitized output of the flame-ionization detector. Post-run computer analysis gives enhanced visual graphic displays with software-driven zooming, scrolling, peak identification and quantitation. In addition, auditory (musical) forms can be produced that increase the scope of human interaction. This novel approach catalyzes simultaneous use of computer calculations and human intuition, optimizing their combined cognitive powers for accurate qualitative and quantitative analyses of these complex mixtures.
Acids↗
Specific high performance liquid chromatographic determination of the molecular weight and concentration of hyaluronic acid in complex mixtures by labelled hyaluronate binding proteins.
A simple High performance liquid chromatographic (HPLC) method for the specific determination of the molecular weight and concentration of hyaluronic acid (HA) in complex mixtures has been developed. Hyaluronate-binding proteins isolated from bovine cartilage labelled by 125I or fluoresceinisothiocyanate were used as specific markers. The specific binding affinities of the markers were compared and were found to have association constants of 1.6 x 10(7) M-1 and 1.2 x 10(7) M-1 respectively. The HA levels and molecular weight distributions can be easily determined in the range 10-500 ng/mL in complex mixtures by the use of markers, molecular sieving HPLC columns and appropriate detectors. It has been demonstrated clearly that the method is useful for the highly specific determination of the parameters in complex biological samples such as serum and synovial fluids and is recommended for clinical applications.
Animals↗