Use of Bioassay Methods to Evaluate Mutagenicity of Ambient Air Collected Near a Municipal Waste Combustor

An ambient air sampling study was conducted around a municipal waste combustor; a primary goal was to develop procedures and methods to evaluate the emissions of organic mutagens resulting from incomplete combustion of municipal waste. The products of Incomplete combustion from incineration include complex mixtures of organics, particularly polycycllc aromatic compounds, which are present after atmospheric dilution and cooling In emissions as semi-volatile or particle bound organic compounds. Combustion emissions are generally recognized as a potential cancer risk since they contain many carcinogenic and mutagenlc polycyclic aromatic hydrocarbons. Analyzing such a complex mixture for the presence of even a few selected chemicals is difficult and provides risk information on only a fraction of the chemicals present. Bioassay methods, however, may be directly applied to evaluate the mutagenic and potential carcinogenic activity of the complex organics from combustion emissions. The Salmonella (Ames) assay was used to determine the mutagenicity associated with particles from ambient air collected near a municipal waste combustor. Dose-response data was generated, and mutagenicity concentrations were calculated to demonstrate the utility of bioassay In assessing the potential Impact of emissions from municipal waste combustion. This phase of study quantified mutagenicity concentrations In ambient air but did not detect organic mutagens that could be attributed to Incinerator emissions.     

Investigating the sources of the mutagenic activity found in a river using the Salmonella assay and different water extraction procedures

In the routine São Paulo state (Brazil) surface water quality-monitoring program, which includes the Salmonella microsome mutagenicity assay as one of its parameters, a river where water is taken and treated for drinking water purposes has repeatedly shown mutagenic activity. A textile dyeing facility employing azo-type dyes was the only identifiable source of mutagenic compounds. We extracted the river and drinking water samples with XAD4 at neutral and acidic pH and with blue rayon, which selectively adsorbs polycyclic compounds. We tested the industrial effluent, raw, and treated water and sediment samples with YG1041 and YG1042 and compared the results with the TA98 and TA100 strains. The elevated mutagenicity detected with YG-strains suggested that nitroaromatics and/or aromatic amines were causing the mutagenicity detected in the samples analyzed. Positive responses for the blue rayon extracts indicated that mutagenic polycyclic compounds were present in the water samples analyzed. The mutagen or mixture of mutagens present in the effluent and water samples cause mainly frameshift mutations and are positive with and without metabolic activation. The Salmonella assay combined with different extraction procedures proved to be very useful in the identification of the origin of the pollution and in the identification of the classes of chemical compounds causing the mutagenic activity in the river analyzed.     

Effect of a base-catalyzed dechlorination process on the genotoxicity of PCB-contaminated soil

We evaluated the genotoxicity of dichloromethane (DCM) extracts of PCB-contaminated soil before and after the soil had been treated by a base-catalyzed dechlorination process. The treatment process involves heating a mixture of the soil, polyethylene glycol (or hydrocarbons with boiling points of 310–387°C), and sodium hydroxide to 250–350°C. Dechlorination reduced by >99% the PCB concentration of the soil, which was initially 2,200 ppm. The DCM extracts of both control and treated soils were not mutagenic in strain TA100 of Salmonella, but they were mutagenic in strain TA98. Based on results in strain TA98, the base-catalyzed dechlorination process reduced the mutagenic potency of the soil by approximately one-half. The DCM extracts of the soils before and after treatment were equally genotoxic in a prophage-induction assay in . , which detects some chlorinated organic carcinogens that are not detected by the Salmonella mutagenicity assay. These results suggest that treatment of PCB-contaminated soil by base-catalyzed dechlorination reduced the mutagenicity of the soil slightly.     

Design of a compact dilution sampler for stationary combustion sources

The dilution sampling method simulates the rapid cooling and dilution processes after hot flue gas have left the stack. This allows gases or vapors to nucleate both homogeneously and heterogeneously, and to condense on preexisting particles in processes analogous to those that occur in the ambient environment. Using this method the authors can collect filterable particulate matter (PM) and condensible PM, that is, primary PM, simultaneously. In order to make this method more suitable for field investigation, a compact dilution sampler was developed. The sampler enhances mixing of dilution air with the stack gas, and thus shortens the length of the mixing section. The design decreases the nominal flow rate through the aging section, and accordingly reduces the size of the residence chamber. The decreased size of the sampler is suitable for field test. Sampling gas is pressured into the residence chamber, and air pressure in the chamber is micro-positive. Uncollected redundant gas is automatically discharged through unused sampling ports, which keeps the unit stable. Performance evaluation tests demonstrate that the design is reasonable. The sampler has been applied to characterize PM emissions from various combustion sources in China.     

Source contributions to the mutagenicity of urban particulate air pollution

Using organic compounds as tracers, a chemical mass balance model was employed to investigate the relationship between the mutagenicity of the urban organic aerosol sources and the mutagenicity of the atmospheric samples. The fine particle organic mass concentration present in the 1993 annual average Los Angeles-area composite sample was apportioned among eight emission source types. The largest source contributions to fine particulate organic compound mass concentration were identified as smoke from meat cooking, diesel-powered vehicle exhaust, wood smoke, and paved road dust. However, the largest source contributions to the mutagenicity of the atmospheric sample were natural gas combustion and diesel-powered vehicles. In both the human cell and bacterial assay systems, the combined mutagenicity of the composite of primary source effluents predicted to be present in the atmosphere was statistically indistinguishable from the mutagenicity of the actual atmospheric sample composite. Known primary emissions sources appear to be capable of emitting mutagenic organic matter to the urban atmosphere in amounts sufficient to account for the observed mutagenicity of the ambient samples. The error bounds on this analysis, however, are wide enough to admit to the possible importance of additional mutagenic organics that are formed by atmospheric reaction (e.g., 2-nitrofluoranthene has been identified as an important human cell mutagen in the atmospheric composite studied here, accounting for approximately 1% of the total sample mutagenic potency).     

The influence of application rate on the bacterial mutagenicity of soil amended with municipal sewage sludge

The mutagenic potential of two soils amended with a municipal sewage sludge at two application rates was monitored over a 2-year period using Salmonella/microsome mutagenicity assay. Samples were collected from undisturbed monolith lysimeters of Weswood sandy clay (Fluventic Ustochrept) and Padina sandy loam (Grossarenic Paleustalf) amended with dried sewage sludge at 50 and 100 Mg/ha. Soil samples were collected and sequentially extracted with methylene chloride and methanol. The residues from these extracts were tested for mutagenicity at five doses with and without metabolic activation in Salmonella strain TA98. In general, the mutagenic potential of the amended soils of both application rates for the first 8 weeks following sludge application increased and then slowly decreased. The maximum mutagenic response observed in the soil extracts was 222 revertants at a dose of 10 mg of residue. This response was induced by the methanol extract from the Weswood soil collected 56 days after the application of 50 Mg/ha sewage sludge as compared to the 100 Mg/ha application which induced 202 revertants/mg. The mutagenicity of all fractions extracted from the sludge-amended soil at both application rates collected 717 days after application were not appreciably different from extracts from the unamended soils. The data indicate that chemicals that were mutagenic in bacteria persist in the soil and that at the higher application rates, as much as 2 years may be required for the mutagenic potential of the soil to return to background levels.     

Urban-Center CO Air Quality Projections

The distribution of mutagenic activity and nitroaromatic components of polycyclic organic matter (POM) in ambient air at industrial, urban, suburban, rural, and remote sites was studied using organic extracts from high volume aerosol samples. Direct-acting mutagens including 1-nitropyrene (1-NP), dinitropyrenes (DNP), and hydroxynitropyrenes (HNP) were measured by high performance liquid chromatography while the mutagenicity was determined in the Salmonella bioassay with strain TA-98. Benzo(a)pyrene (BaP), one of the possible precursors of nitroaromatic compounds in POM, was also measured. In comparing samples from a range of sites, TSP and the concentration of BaP per mass of particulate matter decreased, as expected, at greater distances from urban and industrial combustion sources. However, the concentrations of polar nitroaromatic POM compounds per mass of particles were higher at a remote site than in nonindustrial urban and suburban areas. The mutagenicity in particulate matter extracts from the remote area was predominantly (>90 percent) in the very polar fractions. There were also high atmospheric levels of nitroaromatic compounds and mutagenicity in heavily industrialized areas. These observations may reflect the influences of source emissions, atmospheric transformations of POM compounds, and ther atmospheric processes on the composition of ambient suspended particulate matter.     

The contribution of azo dyes to the mutagenic activity of the Cristais River

To verify whether dyes emitted within the discharge of a dye processing plant were contributing to the mutagenicity repeatedly found in the Cristais River, Sao Paulo, Brazil, we chemically characterized the following mutagenic samples: the treated industrial effluent, raw and treated water, and the sludge produced by a Drinking Water Treatment Plant (DWTP) located approximately 6 km from the industrial discharge. Considering that 20% of the dyes used for coloring activities might be lost to wastewaters and knowing that several dyes have mutagenic activity, we decided to analyze the samples for the presence of dyes. Thin layer chromatographic analysis indicated the presence of three prevalent dyes in all samples, except for the drinking water. This combination of dyes corresponded to a commercial product used by the industry, and it tested positive in the Salmonella assay. The structures of the dye components were determined using proton magnetic resonance and mass spectrometric (MS) methods, and the dyes were tested for mutagenicity. The blue component was identified as the C.I. Disperse Blue 373, the violet as C.I. Disperse Violet 93, and the orange as C.I. Disperse Orange 37. The dyes showed mutagenic responses of 6300, 4600, and 280 revertants/microg for YG1041 with S9 respectively. A bioassay-directed fractionation/chemical analysis showed that the C.I. Disperse Blue 373 contributed 55% of the mutagenic activity of the DWTP sludge. We showed that these dyes contributed to the mutagenic activity found in the Cristais River environmental samples analyzed and are indirectly affecting the quality of the related drinking water. Therefore, we believe that this type of discharge should be more thoroughly characterized chemically and toxicologically. Additionally, human and ecological risks associated with the release of dye processing plant effluents should be more fully investigated, especially where the resultant water is taken for human consumption.     

Application of semipermeable membrane device (SPMD) to assess air genotoxicity in an occupational environment

Semipermeable membrane device (SPMD) is a passive sampler that sequesters lipophilic contaminants, mimicking the bioconcentration in the fatty tissue of organisms. This study was designed to assess the use of SPMD and biological tests (Comet assay and Ames test) for air monitoring. For this purpose an occupational environment with expected polycyclic aromatic hydrocarbons (PAHs) contamination (coke plant) was selected for a case study. The SPMDs were deployed in five occupational contaminated sites and in a control site. The SPMD dialysates were chemically analysed and examined for in vitro DNA-damaging activity in human cells (Jurkat) by Comet assay and for mutagenicity with the Ames test (TA98 strain, w/o S9). Total suspended particulates were also collected and analysed (GC–MS). No biological effect of SPMD extract was revealed in the control site. On the other hand, air samples collected with SPMDs within the coke plant showed variable degrees of genotoxic and mutagenic activity. The highest effects were associated with the highest PAH level recovered in the SPMDs extracts and in particulate samples.Results obtained support the sensitivity of biological tests associated to SPMD sampling for evaluating the health risk of potentially contaminated work environments highlighting the usefulness of SPMDs for environmental air quality monitoring.     

Dilution-based emissions sampling from stationary sources: Part 1--Compact sampler methodology and performance

This paper presents the design and performance of a compact dilution sampler (CDS) for characterizing fine particle emissions from stationary sources. The sampler is described, along with the methodology adopted for its use. Dilution sampling has a number of advantages, including source emissions that are measured under conditions simulating stack gas entry and mixing in the ambient atmosphere. This is particularly important for characterizing the semivolatile species in effluents as a part of particulate emissions. The CDS characteristics and performance are given, along with sampling methodology. The CDS was compared with a reference dilution sampler. The results indicate that the two designs are comparable for tests on gas-fired units and a diesel electrical generator. The performance data indicate that lower detection limits can be achieved relative to current regulatory methods for particulate emissions. Test data for the fine particulate matter (PM2.5) emissions are provided for comparison with U.S. Environment Protection Agency (EPA) Conditional Test Method 040 for filterable particulate matter (FPM) and the EPA Method 202 for condensable particulate matter. This comparison showed important differences between methods, depending on whether a comparison is done between in situ FPM determinations or the sum of such values with condensable PM from liquid filled impingers chilled in an ice bath. These differences are interpretable in the light of semivolatile material present in the stack effluent and, in some cases, differences in detection and quantification limits. Determination of emissions from combustors using liquid fuels can be readily achieved using 1-hr sampling with the CDS. Emissions from gasfired combustors are very low, requiring careful attention to sample volumes. Sampling volumes corresponding with 6-hr operation were used for the combined mass and broad chemical speciation. Particular attention to dilution sampler operation with clean dilution air also is essential for gas-fired sources.     

Compositions of Particles from Selected Sources in Philadelphia for Receptor Modeling Applications

As a part of a receptor model study of the Philadelphia, PA atmosphere, particulate samples were collected from seven air pollution sources in the area: two oil-fired power plants, a coal-fired power plant, a fluidized catalytic cracker, a refuse incinerator, a secondary aluminum smelter and an antimony ore roaster. Samples were collected In two size fractions with a dilution source sampler connected to a modified dichotomous sampler. Masses of collected material were determined gravlmetrlcally. Samples were analyzed for elements by x-ray fluorescence followed by Instrumental neutron activation analysis of some samples. Other samples were analyzed by chemical methods for volatile and nonvolatile carbon, SO₄ ^2? and NH₄ ⁺. Data are presented for up to 46 elements and species on fine (<2.5-μm aerodynamic equivalent diameter) and coarse (2.5 μm < diam < 7-10 μm) particles from each source. Although the data were collected for use in Philadelphia, they should be of value for receptor modeling of other areas having similar sources. The most unexpected results were the large amounts of rare earth elements on particles from the catalytic cracker (e.g., 0.31 percent La In fine fraction) and the oil-fired power plants (120 and 420 ppm La in fine fraction). Substantial amounts of primary SO₄ ^2? are released from the oil-fired plants, the SO₄ ^2? concentrations accounting for 40-45 percent of the fine particulate mass.     

Determination of mutagenic potential and organic contaminants of Great Lakes drinking water

Extracts of organic compounds were obtained, using XAD-2 macroreticular resin, from drinking water supplies in 12 Great Lakes municipalities. The extracts were tested for mutagenic potential using the Salmonella/mammalian-microsome assay and analysed for organochlorine pesticides, polyaromatic hydrocarbons, organophosphorous pesticides and trialkyl-arylphosphates. Grab samples of drinking water were also analysed for volatile organic compounds. Dose-related increases in mutagenicity were found in extracts from 11 of the drinking water supplies.     

Changes in toxicity and genotoxicity of industrial sewage sludge samples containing nitro- and amino-aromatic compounds following treatment in bioreactors with different oxygen regimes

Goals, Scope and Background

From 2005, deposition of organic waste will be banned in Sweden. Likewise, in Germany and Austria, similar bans are being planned, and further countries will probably follow. Thus, there is a need to develop new methods and to refine established techniques for sludge management in the whole of the European Union. For this end, there is also an urgent need for appropriate ecotoxicological approaches to elucidate and assess the hazard potential of sewage sludge. Therefore, the present study was designed to assess the capacity of various established sludge treatment methods using different oxygen regimes to degrade recalcitrant nitro-substituted organic compounds and reduce their toxicity. Sewage sludge samples from a wastewater treatment plant in Sweden (Cambrex Karlskoga AB, industrial area Björkborn) receiving wastewater from industries manufacturing pharmaceutical substances, chemical intermediates and explosives were processed with different sludge treatment methods. Among other treatment methods, bioreactors (for anaerobic and aerobic sludge treatment) were used. In the present investigation, a battery ofin vitro bioassays was employed to compare the cytotoxic and genotoxic potentials of different fractions of sludge samples in order to elucidate whether the treatments were suitable to reduce the toxicity of the sludge.

Methods

In order to investigate the cytotoxicity of the extracts of treated and untreated sludge samples, the acute cytotoxicity test with the permanent cell line RTL-W1 was used. Genotoxicity was tested by means of the comet assay (single cell gel electro-phoresis) with RTL-W1cells, and mutagenicity was assessed with the Ames test using the Salmonella typhimurium strains TA98, TA98NR and TA100. Sludge toxicity was tested in different fractions of organic extracts produced by acetone and hexane extractions. The subsequent clean-up procedure (silica gel chro-matography and elution with hexane and dichloromethane) resulted in two fractions, a lipophilic hexane-fraction and a semi-lipophilic dichloromethane-fraction. For the genotoxicity and mutagenicity tests, these fractions were reunited at equal ratios.

Results and Discussion

The acute cytotoxicity test with RTL-Wl cells revealed a high cytotoxic potential for the semi-li-pophilic DM-fractions of all sludge samples with NR₅₀ values (= effective concentration for 50% cell death in the neutral red test) from 8.9 up to 20 mg sludge d.w./ml medium. A low cytotoxic potential for the hexane fractions of the untreated sludge samples (NR₅₀ 400 to < 400 mg sludge d.w./ml medium) was observed, whereas the hexane fractions of the treated sludge samples showed elevated cytotoxicity increasing further with treatment in the bioreactors. The comet assay indicated that three out of eight of the reunited fractions had a significant genotoxic potential. Whereas the genotoxic potential of one sample treated anaerobically was very high with an induction factor of 11.6, a similar sample (taken from the same anaerobic reactor four months later) and one untreated sample showed lower potentials. The samples treated in another anaerobic bioreactor as well as the samples treated aerobically showed no genotoxic potential. Results indicate that aerobic treatment was basically adequate for reducing the genotoxicity of the sludge, whereas anaerobic treatment was only partly useful for reduction of genotoxicity. The Ames test revealed a very high mutagenic potential for the reunited fractions of the untreated sludge samples with strain TA98 (maximum induction factors (IF_max up to 45) and a relatively high potential for one of the samples treated aerobically (S2, IF_max = 18 (TA98, S9-)), thus documenting the suitability of both anaerobic and aerobic treatments to reduce the mutagenicity of the samples, however, with the aerobic treatment being less effective. Conclusions. Overall, none of the microbiological treatments for wastewater sludge in bioreactors was found to be ideal for general toxicity reduction of the sludge samples. Whereas cytotoxicity of the sludge increased or levelled off in most cases following either treatment, genotoxicity both increased or decreased after anaerobic treatment, depending on the specific sample. However, mutagenicity could generally be reduced by anaerobic treatment and, to a lesser degree, by aerobic treatment. Recommendationsand Perspectives. The complex modification of the diverse damage potentials of sludge sample extracts by use of anin vitro biotest battery following treatment for toxicity reduction in bioreactors showed that considerations of different toxicological endpoints is essential for an adequate hazard assessment. Whereas in the case of cytotoxicity reduction, the reactors proved ineffective, mutagenicity could be reduced significantly at least in some cases in this case study.

     

The mutagenic potentials of tap water samples in Shanghai

The tap water samples were collected from the users' ends in several areas of Shanghai, which is located in Taihu Lake basin, Eastern China. Source water samples were also collected from two municipal source water facilities at the same time. Samples were assayed by three different short-term mutagenicity test systems: Salmonella/microsome assay (Ames test), the Arabinose resistance test (Ara test) and the SOS/umu test. The data showed that two source water samples did not display direct mutagenic potentials. Two tap water samples from city north, which were directly from Yangtze River, were also not mutagenic. Water samples from city south and middle which used source water originating from Taihu Lake were proved to be contaminated with mutagenic potentials by three different assay techniques. The boiled water displayed an even stronger mutagenic potential compared to its original tap water. The molecular mechanism of mutagenicity was associated with a reading frame-shifting potential. GC-MS analysis of tap water extracts from city middle and corresponding source water was compared. Qualitatively similar spectra were observed except for the peaks of three chlorinated aromatic hydrocarbon compounds, which existed only in the tap water. Since the water source has been polluted, raw water was heavily chlorinated in order to sterilize. More toxic compounds, including mutagens, might form during the multi-chlorination. Caution about the possibility of elevated cancer risks in the population that consumes heavily chlorinated water should be kept in mind. A cohort study in the residents of Shanghai is required.     

Mutagenicity evaluation of industrial sludge from common effluent treatment plant

Sludge from common effluent treatment plant (CETP) receiving effluents from textile industries at Mandia Road, Pali, was analyzed to assess the level of mutagenicity. Mutagenicity assay using Salmonella typhimurium tester strains TA 98 and TA 100 gave positive results, thus suggesting presence of genotoxic contaminants in the samples investigated. Further, mutagenic activity of chemical sludge was found to be lesser than that of biological sludge. This result is very surprising and unexpected as it is indicating that some mutagenic compounds are either being formed or certain promutagenic compounds are being converted into stable mutagenic metabolites during the biological treatment of the wastewater effluents. There have been no previous reports giving similar or contrary results. Most of the previous studies have reported effects of single combined sludge.     

Chemical and biological profiles of sediments as indicators of sources of contamination in Hamilton Harbour. Part II: bioassay-directed fractionation using the Ames Salmonella/microsome assay

Bottom sediment and suspended sediment samples from Hamilton Harbour (western Lake Ontario) and from a major tributary were profiled using a bioassay-directed fractionation approach. Sample extracts were fractionated using an alumina/Sephadex gel clean-up procedure to afford non-polar aromatic fractions which were characterized using chemical analyses and the Ames/microsome bacterial assay in Salmonella typhimurium strains YG1025 with the addition of oxidative metabolism (S9), and YG1024 without S9. Non-polar aromatic fractions of selected samples were separated by normal phase HPLC into 1-min fractions which were subjected to bioassay analyses. The bioassays using strain YG1025+S9, a TA100-type strain, were performed to assess genotoxicity arising from the presence of polycyclic aromatic hydrocarbons (PAH). Fractions which exhibited mutagenic activity contained PAH with molecular masses of 252, 276 and 278 amu; these fractions contained over 80% of the genotoxicity attributable to PAH. Individual compounds identified using Gas Chromatography-Mass Spectrometry analyses in these active fractions included benzo[a]pyrene, indeno[cd]pyrene and dibenz[a,h]anthracene. The YG1025+S9 mutagenic activity profiles were similar for all samples. Mutagenic activity profiles generated using strain YG1024-S9, a TA98-type strain sensitive to compounds characteristic of mobile source emissions, were very different. The mutagenic activities in strain YG1024-S9 were greatest for harbour-suspended sediment samples collected from sites impacted by a major tributary. Suspended sediments collected near areas known to contain high levels of coal tar-contamination in the bottom sediments contained higher levels of genotoxic PAH than suspended sediments collected from other areas of the harbour.     

长江、嘉陵江(重庆段)源水有机提取物的致突变活性及其季节变化

为了确定并比较重庆主城区段长江、嘉陵江源水有机提取物的致突变性及其季节变化规律，分别于春、夏、冬季采用GDX－120大孔树脂，对位于城区上游、城区中段、城区下游以长江、嘉陵江源水的5个水厂的进厂水进行了有机物的浓缩提取。提取物的致突变活性采用经典的Ames试验平板掺入法评估，测试菌株为TA98及TA100，同时做加与不加S9的比较。结果显示，嘉陵江及长江源水的有机提取物均有不同程度的致突变活性。嘉陵江源水明显大于长江源水，城区中段源水明显大于上游段及下游段源水。多数断面显示平水期致突变活较为显著并且移码型致突变性大于碱基置换型致密变性。研究结果提示，城市污染源已导致长江、嘉陵江源水具备致突变活性，控制两江沿岸的各种水污染源已成为当务之急。     

Association of the mutagenicity of airborne particles with the direct emission from combustion processes investigated in Osaka, Japan

The association of the direct-acting mutagenicity of soluble organic fraction of airborne particles toward Salmonella typhimurium YG1024 strain with the direct emission was investigated at a roadside and at a residential area in Osaka, Japan. The direct-acting mutagenicity was evaluated as mutagenic activity per unit volume of ambient air (rev m⁻³) and/or that per airborne particulate weight collected on a filter (rev mg⁻¹). The annual or diurnal changes of the mutagenicity of airborne particles at the residential site showed similar patterns to those of some gaseous pollutants such as NO₂ and SO₂, which were emitted from combustion processes. This result indicates that the mutagenicity is mainly attributable to the primary emissions. From the analysis of the relationship between the wind sector and the mutagenic intensity, rev m⁻³ and rev mg⁻¹ values were strongly affected by the emissions from the fixed sources and from the mobile sources, respectively. The rev m⁻³ value and concentration of 1-nitropyrene (1-NP) in unit per m³ at the roadside were a factor of 2.6 and 2.8 higher than those at the residential site, respectively, but the rev mg⁻¹ value and concentration of 1-NP in unit per mg at the roadside were substantially comparable to those at the residential area. These observations suggest that the characteristics of the airborne particles can be attributed to the automotive emissions even at the suburban area.     

Mutagenic nitrated benzo[a]pyrene derivatives in the reaction product of benzo[a]pyrene in NO2-air in the presence of O3 or under photoirradiation

In order to clarify the contribution of nitrated products to the direct-mutagenic activity of products of the reactions of benzo[a]pyrene in NO2-air under various conditions, heterogeneous reactions of BaP deposited on filter in the air containing 10 ppm of NO2 have been conducted in dark or under photoirradiation. The reaction products have been analyzed by gas chromatography and mutagenicity of the products fractionated by preparative HPLC was assayed for Salmonella typhimurium strains TA98 and YG1024 in the absence of S9 mix. 3,6-dinitrobenzo[a]pyrene and 1,3-dinitrobenzo[a]pyrene, which are strong direct-acting mutagens, largely contributed to the total direct-acting mutagenicity of the dark reaction products in NO2-air. On the other hand, both the dark reaction in the presence of O3 and the photoreaction in NO2-air resulted in the formation of much smaller amounts of nitrobenzo[a]pyrenes than that observed in the dark reaction in the absence of O3. These results show that the contribution of other direct-acting mutagens to the total direct-acting mutagenicity of the products in these reactions should be considered. Benzo[a]pyrene lactones were identified in a highly mutagenic fraction of the products of the dark reaction in the presence of O3 and photoreaction and a nitrobenzo[a]pyrene lactone was also identified in a highly mutagenic fraction of the dark reaction products in the presence of O3. Nitrated oxygenated benzo[a]pyrene derivatives such as nitrobenzo[a]pyrene lactone were considered to largely contribute to direct-acting mutagenicity of the products of the dark reaction in the presence of O3 and photoreaction.     

Toxicity and Mutagenicity of Component Classes of Oils Isolated from Soils at Petroleum- and Creosote-Contaminated Sites

Abstract

Microtox and Ames bioassays were employed to assess acute toxicity and mutagenicity of water soluble components of class–fractionated oils extracted from one creosote–and four petroleum–contaminated soils. Microtox results revealed that potential acute toxicity resides mainly in the polar class fractions at three sites and indicated potential synergistic and antagonistic effects between compounds in the total extracts at two sites. Ames Salmonella/microsome testing indicated that the polyaromatic fractions at two sites exhibit weak mutagenicity with enzymatic activation, while the polar fractions at two sites are weakly mutagenic without enzyme activation. Further chemical characterization of the polar and polyaromatic fractions is required to fully assess the potential of health and ecological risks at the creosote–and petroleum–contaminated sites exhibiting these toxic responses.