Liquid-gel partitioning using Lipidex in the determination of polychlorinated biohenyls,napthalenes, dibenzo-p-dioxins and dibenzofurans in blood plasma

A method was developed for the transfer of fat, polychlorinated biphenyls (PCBs), naphthalenes (PCNs), dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) from blood plasma into the lipophilic gel Lipidex 5000. Subsequent elution of the gel separated about 70% of the fat from the analytes. Different adsorbents and activated charcoal were applied for further purification of the sample and separation of analytes. Identification and determination of the chlorinated compounds were made by gas chromatography with electron-capture detection (GC-ECD) or gas chromatography-mass spectrometry (GC-MS). Recoveries were studied by addition of Halowax 1014 and different congeners of PCBs, PCNs, PCDDs and PCDFs to 50 ml of plasma. The mean recoveries of the individual compounds studied were 72–99%. By using the liquid-gel partitioning technique emulsions were avoided. Concentrations of lipids in plasma obtained by the present method agreed well with the concentrations obtained using liquid-liquid partitioning with chloroform-methanol.     

Risk Assessment of 2,3,7,8-Tetrachlorodibenzo-p-Dioxin Equivalents in Tissue Samples from Three Species in the Denver Metropolitan Area

Polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and polychlorinated biphenyls (PCBs) are possible contaminants of concern in areas with a history of organochlorine pesticide production or various other industrial chlo-rination processes. The Rocky Mountain Arsenal (RMA), a Superfund site near Denver, CO, has a history of various industrial processes that may have led to the release of these compounds and subsequent exposure and risk to surrounding biota. PCDDs, and PCDFs, and non- and mono-ortho-substituted PCBs cause a common set of toxic effects that are mediated through the aryl-hydrocarbon receptor (AhR). The total AhR-medi-ated activity of tissue samples collected from the RMA and surrounding reference areas was determined by both instrumental and bioanalytical techniques. Concentrations of dioxin equivalents were measured in carp eggs, great horned owl livers, and American kestrel eggs collected from the RMA and from off-post reference sites. Detected concentrations of dioxin equivalents were used to assess risk to exposed wildlife through the use of Toxicity Reference Values (TRVs) and Hazard Quotients (HQs). Carp and kestrel egg samples had small HQ values, while some great horned owls had relatively large HQ values, indicating exposure to and possible risks from dioxins or dioxin-like chemicals.     

Sorption/Desorption of Polychlorinated Dibenzo-P-Dioxins and Polychlorinated Dibenzo Furans (PCDDs/PCDFs) in the Presence of Cyclodextrins

The goal of this study was to investigate the usefulness of cyclodextrins (CDs) for the removal of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzo furans (PCDDs/PCDFs) in soil and water. Five CDs having different molecular cavities and active functional groups were selected and evaluated for their ability to include (trap) PCDDs/PCDFs in soil and water. For the soil experiments, CDs were added to the soil on day one and the concentrations of unbound PCDDs/PCDFs were monitored over a 28-day period. Parallel control experiments were conducted to assist in the process performance evaluation. The ability of CDs to remove PCDDs/PCDFs from the contaminated soil was dependent upon the type of CD used and constituents of PCDDs/PCDFs. Among the five CDs investigated, hydroxypropyl-β -cyclodextrin (HPBCD) gave the highest removal efficiency for all components of PCDDs/PCDFs. The removal efficiency of total PCDDs/PCDFs was 81% one day after application of CDs and then increased to 96% after 28 days. The α -cyclodextrin (ACD) and β -cyclodextrin (BCD) removed only 45% and 50% of the total PCDDs/PCDFs after 28 days, respectively, whereas hydroxypropyl-γ -cyclodextrin (HPGCD) removed 80% of the total PCDDs/PCDFs.     

Relationship between Measures of Exposure to PCBs/Dioxins and Behavioral Effects in Recent Developmental Studies

This paper does not reflect official EPA policy. Epidemiological studies on the neurodevelopmental effects of exposure to PCBs initiated in the last decade have had the opportunity to take advantage of modern methodologies for the analysis of congeners of PCBs, dioxins, and related orga-nochlorine compounds. Each of these studies is a longitudinal prospective study, in which women were recruited during pregnancy and the children are being followed for at least several years after birth. The study from which the largest body of data has been published to date is being performed in the Netherlands, in which exposure to PCBs and related compounds is through the general food supply. Mother-infant pairs were recruited in two cities. Half of the infants were bottle-fed and half breast-fed in each city. Four PCB congeners (118, 138, 153, 180) were assessed in maternal and cord plasma, breast milk, and plasma of the child at 3.5 years. TEQ in breast milk was calculated based on PCDDs/PCDFs and dioxin-like PCBs. Various measures of in utero exposure were associated with suboptimal neurological status during infancy, whereas maternal plasma PCB concentration was associated with cognitive deficits (Kaufman scores) at 3.5 years of age. The child's concurrent plasma PCB levels and maternal PCB plasma levels independently predicted performance on various aspects of a vigilance task, and maternal and cord plasma levels predicted impairment of complex play behavior. Poor scores on behavioral ratings were associated with concurrent blood PCB concentrations in the child. A study in Oswego in Lake Ontario fish eaters includes mothers who never ate Great Lakes fish and mothers who consumed greater than 40 PCB-equivalent pounds of Lake Ontario fish over their lifetime. Sixty-eight PCB congeners were measured in cord blood. Suboptimal neurological status during infancy was associated with maternal fish consumption and highly chlorinated cord PCB levels, whereas deficits in short-term memory at 6 months and 1 year of age were associated with total chlorinated cord PCB levels. In a study in Germany of 171 mother-infant pairs, PCB congeners 138, 153, and 180 were measured in cord plasma and milk 2 weeks after birth; both measures are considered markers of in utero exposure. Suboptimal neurological status during infancy, decrements in Bayley scores at 30 months and Kaufman scores at 42 months were associated with PCBs in milk but not cord plasma. These studies, combined with data from previous studies, reveal a consistent relationship between PCB exposure and suboptimal neurological status during infancy, and cognitive deficits associated with in utero exposure. Data from the Dutch study revealed effects on other behavioral domains associated with concurrent (postnatal) exposure. Although it is not possible to identify specific congeners or groups of congeners that may be responsible for the neurotoxic effects observed in these studies, the TEQ approach was not particularly predictive for neurotoxic outcomes.     

Degradation of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans by the white rot fungus Phanerochaete sordida YK-624.

A method for the degradation of dioxins by white rot fungi was developed. Degradation of a mixture of 10 kinds of tetra- to octachlorodibenzo-p-dioxins (polychlorinated dibenzo-p-dioxins [PCDDs]) and tetra- to octachlorodibenzofurans (polychlorinated dibenzofurans [PCDFs]), which were chlorinated at 2-, 3-, 7-, and 8-positions of the molecules, by the white rot fungus Phanerochaete sordida YK-624 was studied in a stationary low-nitrogen medium. The percent degradation values of PCDDs and PCDFs were approximately 40 (tetra-chloro-) to 76% (hexachloro-) and 45 (tetrachloro-) to 70% (hexachloro-), respectively. Metabolites of 2,3,7,8-tetra- and octaCDD formed by P. sordida YK-624 included 4,5-dichlorocatechol and tetrachlorocatechol, respectively. These results suggest that white rot fungus is able to substantially degrade both PCDDs and PCDFs. This is the first report of the degradation of highly chlorinated PCDDs and PCDFs by a microorganism.     

Formation of reactive 1-nitropyrene metabolites by lung microsomes and isolated lung cells

The metabolism and activation of 1-nitropyrene (1-NP) to reactive intermediates by lung microsomes and isolated lung cells was studied. Mutagenicity of 1-NP metabolites was assayed in Salmonella typhimurium TA98NR, a strain lacking a major component of nitroreductase activity. In the presence of NADPH, microsomes from rabbit, rat and hamster lung metabolized 1-NP to mutagenic products to a similar degree. Pretreatment with a mixture of polychlorinated biphenyls (PCB) decreased the formation of mutagenic metabolites by rabbit lung microsomes, but did not affect the production of mutagens by rat or hamster lung microsomes. ³H-1-NP was metabolized to covalently bound protein products at a rate of 82 and 10 pmol/mg by rabbit and hamster lung microsomes, respectively, whereas no binding was detected in rat lung microsomes. PCB-pretreatment increased covalent protein binding of ³ H-1-NP in lung microsomes from hamster and rat, but decreased the binding in rabbit lung microsomes. High performance liquid chromatography analysis indicated that ³H-1-NP was readily converted to ring-hydroxylated products by rabbit and hamster lung microsomes; the rate was much lower with rat lung microsomes. ³H-1-NP was activated to metabolites that covalently bound to protein in isolated rabbit lung cells, with the following rates being observed: Clara cells > lung digest > type II cells. In contrast, covalent protein binding in cells isolated from rat lung was very low. 1-NP was not activated to products mutagenic for S. typhimurium TA 98 N R when co-incubated with cells isolated either from rabbit or rat lung.Abbreviations 1-AP 1-aminopyrene - DMSO dimethyl sulfoxide - EGTA ethylene glycol-bis(ß-aminoethyl ether) - EM electron microscopy - HEPES N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid - HPBS HEPES-phosphate-buffered-saline - HPLC high performance liquid chromatography - NBT nitroblue tetrazolium - 1-NP 1-nitropyrene - 1-NP-4,5-diol trans-4,5-dihydro-4,5-dihydroxy-1-nitropyrene - 1-NP-9,10-diol trans-9,10-dihydro-9,10-dihydroxy-1-nitropyrene - 1-NP-4,5-oxide 1-nitropyrene-4,5-oxide - 1-NP-9,10-oxide 1-nitropyrene-9,10-oxide - 3-OH-1-NP 3-hydroxy-1-nitropyrene - 6-/8-OH-1-NP a mixture of 6- and 8-hydroxy-1-nitropyrene - PBS phosphate-buffered saline - PCB a mixture of polychlorinated biphenyls (Aroclor 1254) - TLC thin layer chromatography     

In vitro binding of estrogens by dietary fiber and the in vivo apparent digestibility tested in pigs.

Within the framework of experiments related to the association between dietary fiber and breast cancer an in vitro test system was used to study the binding of estrogens to various fibers (e.g. cholestyramin, lignin and cellulose) and fiber sources (e.g. wheat bran, cereals, seeds and legumes). Furthermore, the in vivo apparent digestibility of the different fiber sources was tested using a mobile nylon bag technique in intestine-cannulated pigs. Estradiol-17 beta (E2) bound more strongly to the various fibers than did estrone (E1), estriol or estrone-3-glucuronide. At increasing pH (greater than 7) binding of both E1 and E2 to wheat bran decreased significantly. Cholestyramine and lignin bound almost all estrogens present in the medium. Linseed (91%), oats (83%), barley chaff (88%) and wheat bran (82%) are other excellent binders of E2. Corn, rye and white wheat flour showed lower binding capacity with a relatively low affinity. Cereals with the highest percentage of lignin in the fiber (greater than 3%) were also the fiber sources with the lowest apparent digestibility. Estrogens bound with the highest affinity (relative to bovine serum albumin) to these fiber sources. Together with wheat bran and lignin, oats, linseed and soybean seem to be products with good perspectives for in vivo evaluation of the lowering effect of dietary fiber on estrogen exposure of estrogen-sensitive tissues.     

Fecal mutagens and Bacteroides fragilis levels in the feces of dimethylhydrazine-treated rats: influence of diet

In a study designed to investigate the effects of dietary synergisms on 1,2-dimethylhydrazine (DMH)-induced rat colon carcinogenesis, fecal pellets were examined for the presence of direct-acting fecal mutagens and levels of Bacteroides fragilis group organisms. Intraperitoneal injections of DMH at 10 mg/kg were given for 16 weeks (weeks 3-18) to 160 male F344 rats consuming 4 supplemental dietary factors in all possible combinations. The dietary factors examined were wheat bran (15%), cholesterol (1%), beef tallow (18%) and indole-3-carbinol (IC) (0.1%). Feces were collected 3, 10, 17, 24 and 31 weeks after commencing the dietary treatments and dichloromethane extracts were assayed using the Salmonella typhimurium TA100 without metabolic activation. The numbers of B. fragilis group organisms were enumerated in feces collected at the same time. Most feces samples were negative for mutagens but extracts from weeks 17-31 showed a significant mutagenic response from the IC factor in the diet. The fecal levels of B. fragilis were significantly increased by the inclusion of cholesterol in the diets. The B. fragilis counts and fecal mutagen production were not correlated (r = 0.09), although species of the B. fragilis group have been implicated in the production on human fecal mutagens.     

Bioaerosols and Particle Release During Composting of Contaminated Sawmill Soil

Compost windrows for bioremediation of soil were built at a wood-preserving site contaminated with chlorophenols, polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs). Sampling of airborne particles during the mixing of the compost windrows found concentrations of PCDDs and PCDFs in different particle sizes. The congener distribution of PCDDs and PCDFs in the collected air particle fractions was similar to that in the compost windrows, and the level of PCDDs and PCDFs was 1000-fold higher than the atmospheric background values reported previously. Viable particle-sizing samplers and several selective growth media were used to enumerate bacteria and fungi in the airborne particles. From the collected air samples, 40 bacteria were isolated and identified. Among the isolated bacteria, 80% were Gram-positive and spore-forming. Two of the identified airborne bacteria, Pseudomonas aeruginosa and Bacillus cereus, may cause human disease and are classified in biological agent hazard group 2. The amounts of airborne fungi, molds, and yeasts were 1000 to 2000 colony-forming units (CFUs) per m³. The number of actinomycetes was up to 6-fold, and the number of bacteria was 2- to 20-fold compared to background values. The overall level of airborne bacteria (200 to 3500 CFUs per m³) was low compared to the level of bacteria (10⁵ to 10⁸ CFUs per m³) found when composting municipal waste.     

Effect of wheat bran on excretion of radioactively labeled estradiol-17 beta and estrone-glucuronide injected intravenously in male rats.

Urinary and fecal estrogen excretion were studied in male rats fed a non-fiber wheat starch diet (dietary fiber less than 1%; NF group; n = 4), a low-fiber wheat flour diet (dietary fiber 2%; LF group; n = 4) or a high-fiber wheat bran diet (dietary fiber 11.6%; HF group; n = 3). Short-term effects of the experimental diet on estrogen excretion were studied after i.v. injection of 5 microCi (0.185 MBq) of [14C]estradiol-17 beta (E2) into the tail vein of the rats fed the diets for 2 days. After 3 weeks on the experimental diets, the long-term effects were studied after injection of 5 microCi of [14C]E2 and 10 microCi of [3H]estrone-3-glucuronide (E1-gluc). The diet was found to affect estrogen excretion. The short-term effect indicated that rats fed the HF diet excreted a relatively large amount of labeled compounds in the feces during the first day after injection, while rats fed the NF or the LF diets excreted about half that amount over the same period. On the other hand, urinary excretion of labeled compounds was significantly higher in the NF and LF rats. The long-term effect resulted in steeper slopes (P less than 0.05) of the fecal excretion profiles of rats fed the HF diet as compared with rats fed the NF and LF diets, indicating an accelerated fecal excretion of labeled compounds in the HF rats. The kinetic profiles of 14C and 3H radioactivity in blood plasma indicated a fast decrease (t1/2 of less than 2 min) for both [14C]E2 and [3H]E1-gluc. It was concluded that, owing to the short-term effect of wheat bran intake, during the first 24 h after i.v. administration relatively large amounts of radioactively labeled compounds are excreted in feces of rats fed the HF diet. In contrast, excretion is lower in urine of these rats. When the microflora is adapted to the experimental diet the wheat bran diet still results in an accelerated fecal excretion of labeled compounds, which might be attributed to an interruption of the enterohepatic circulation of estrogens. This might result in lowered plasma and/or tissue estrogen levels and hence a decreased exposure of estrogen-sensitive tissue to estrogens, which might decrease risk on mammary (breast) cancer development.     

Effect of chitosan intake on fecal excretion of dioxins and polychlorinated biphenyls in healthy men

Six healthy male subjects were treated with 0 g, 1 g, 3 g, and 0 g of chitosan for the first, second, third, and fourth of four weeks, respectively. They were administered chitosan before breakfast on the second, third, and fourth days of the week, and fecal specimens were collected corresponding to the prescribed diet consumed for breakfast on the second day to breakfast on the fourth day. Fecal excretion of dioxins and polychlorinated biphenyls (PCBs) was promoted by intake of 3 g of chitosan (p=0.0589 and p<0.05 respectively), and was positively correlated with that of fat (p<0.01 for both). We found that chitosan intake increased the fecal excretion of dioxins and PCBs, as well as that of fat, suggesting that it might be useful for reducing the adverse effects of lipophilic endocrine-disrupting chemicals.     

Mutagens in feces from vegetarians and non-vegetarians

Mutagens in water extracts from feces of persons in 3 different diet groups were measured with the fluctuation test for weak mutagens using Salmonella typhimurium TA100 and TA98 as tester strains. The 3 diet groups were ovo-lacto vegetarians (N = 6), strict vegetarians (N = 11) and non-vegetarians (N = 12). All subjects were from the urban area of Vancouver, British Columbia, Canada. On TA100 ovo-lacto vegetarians and strict vegetarians had significantly lower levels of fecal mutagens than non-vegetarians (P less than or equal to 0.025 and P less than 0.010, resp.). The same pattern, although less significant, was obtained with TA98. Correlation studies between mutagenicity on TA100 and TA98 and between the pH of the fecal homogenate and mutagenicity indicate the presence of 2 or more major fecal mutagens.     

Cellulase–lignin interactions—The role of carbohydrate-binding module and pH in non-productive binding

Non-productive cellulase adsorption onto lignin is a major inhibitory mechanism preventing enzymatic hydrolysis of lignocellulosic feedstocks. Therefore, understanding of enzyme–lignin interactions is essential for the development of enzyme mixtures and processes for lignocellulose hydrolysis. We have studied cellulase–lignin interactions using model enzymes, Melanocarpus albomyces Cel45A endoglucanase (MaCel45A) and its fusions with native and mutated carbohydrate-binding modules (CBMs) from Trichoderma reesei Cel7A. Binding of MaCel45A to lignin was dependent on pH in the presence and absence of the CBM; at high pH, less enzyme bound to isolated lignins. Potentiometric titration of the lignin preparations showed that negatively charged groups were present in the lignin samples and that negative charge in the samples was increased with increasing pH. The results suggest that electrostatic interactions contributed to non-productive enzyme adsorption: Reduced enzyme binding at high pH was presumably due to repulsive electrostatic interactions between the enzymes and lignin. The CBM increased binding of MaCel45A to the isolated lignins only at high pH. Hydrophobic interactions are probably involved in CBM binding to lignin, because the same aromatic amino acids that are essential in CBM–cellulose interaction were also shown to contribute to lignin-binding.     

Fecal mutagens from subjects consuming a mixed-western diet

Because of potential significance of fecal mutagens (presumptive carcinogens) in the pathogenesis of colon cancer, feces from 99 healthy subjects from the New York metropolitan area were studied. The diet histories indicate that all participants were consuming a mixed-western diet which is high in total fat and low in fiber. Fecal samples that were incubated under anaerobic conditions at 37 degrees C for 96 h or frozen without incubation, were extracted with hexane: peroxide-free diethyl ether (1:1), partially purified on a silica Sep-pak cartridge and assayed for mutagenicity using the Salmonella typhimurium/mammalian microsome system. Aliquots of fecal samples incubated anaerobically showed a higher frequency of mutagenic activity (per cent samples showing activity) in strains TA98 and TA100 with and without microsomal (S9) activation. In addition, the mutagens requiring S9 activation, were more frequently inactivated when the fecal samples were frozen immediately after defecation and transported to the laboratory. Compared with hexane: ether, extraction of fecal samples with acetone increased the mutagenic activity mostly with TA98 with S9 activation. The HPLC fractionation of hexane: ether extract with methanol: water gradient using reverse phase C-18 column and UV detector at 254 nm indicated that the mutagenic activity (TA98 with S9 activation) is concentrated in several peaks. This is the first demonstration of HPLC profile of fecal samples that are active in TA98 with S9 activation. HPLC profile of fecal extracts and mutagenic activity of these extracts in strains TA98 and TA100 suggest the presence of several types of mutagens in the feces of healthy subjects consuming a high-fat, low-fiber mixed-western diet.     

Exposure to polychlorinated biphenyls causes endothelial cell dysfunction

Environmental chemicals, such as polychlorinated biphenyls (PCBs), may be atherogenic by disrupting normal functions of the vascular endothelium. To investigate this hypothesis, porcine pulmonary artery-derived endothelial cells were exposed to 3,3′,4,4′-tetrachlorobiphenyl (PCB 77), 2,3,4,4′,5-pentachlorobiphenyl (PCB 114), or 2,2′,4,4′,5,5′-hexachlorobiphenyl (PCB 153) for up to 24 hours. These PCBs were selected for their varying binding avidities with the aryl hydrocarbon (Ah) receptor and differences in their induction of cytochrome P450. PCB 77 and PCB 114 significantly disrupted, in a dose-dependent manner, endothelial barrier function by allowing an increase in albumin transfer across endothelial monolayers. These PCBs also contributed markedly to cellular oxidative stress, as measured by 2,7-dichlorofluorescin (DCF) fluorescence and lipid hydroperoxides, and caused a significant increase in intracellular calcium ([Ca²⁺]_i) levels. Enhanced oxidative stress and [Ca²⁺]_i in PCB 77- and PCB 114-treated cells were accompanied by increased activity and content of cytochrome P450 1A and by a decrease in the vitamin E content in the culture medium. In contrast to the effects of PCB 77 and PCB 114, cell exposure to PCB 153 had no effect on cellular oxidation, [Ca²⁺]_i, or endothelial barrier function. These results suggest that certain PCBs may play a role in the development of atherosclerosis by causing endothelial cell dysfunction and a decrease in the barrier function of the vascular endothelium. It is possible that interaction of PCBs with the Ah receptor and activation of the cytochrome P450 1A subfamily are involved in this pathology. © 1995 John Wiley & Sons, Inc.     

Health Risk of PCBs and DDTs in Seafood from Southern Iran

Levels of polychlorinated biphenyls (PCBs) congeners and dichlorodiphenyltrichloroethane and its metabolites (DDTs) were measured in 18 species of fish, crab, shrimp, and bivalve samples collected from the northern region of the Persian Gulf in Iran. The levels of ∑PCBs varied from 259.92 ± 31.04 ng/g to 1648.88 ± 176.96 ng/g in lipid weight. CB118 showed the highest concentration. ∑DDT variations ranged from ND to 570.45 ± 806.74 ng/g in lipid weight. Health risk was assessed by estimating both dietary intakes and screening values (SVs). Daily intake levels were much lower than the Food and Agriculture Organization (FAO)/World Health Organization (WHO) standards for PCBs and DDTs. Based on an average bodyweight of 73 kg and consumption rates of 0.055 kg/person. day, the SVs for the carcinogen and non-carcinogen effects of PCB and DDT were calculated. Average concentrations of PCBs and DDTs showed that the levels of PCBs exceeded the established SV for carcinogens. This finding suggests the need to enhance risk management regarding seafood consumption through public advisory.     

Effect of stepwise microhydration on the guanidinium···π interaction

Polychlorinated biphenyls (PCBs) are potentially hazardous to the environment because of their chemical stability and biological toxicity. In this study, we identified the binding mode of a representative PCB180 to human serum albumin (HSA) using fluorescence and molecular dynamics (MD) simulation methods. PCB180 bound exactly at subdomain IIIA of HSA based on the fluorescence study along with site marker displacement experiments. PCB180 also induced conformational changes that were governed mainly by hydrophobic forces. MD studies and free energy calculations also made important contributions to the understanding of the effects of an HSA-PCB180 system on conformational changes. The simulations on binding behavior proved that PCB180 was located only in subdomain IIIA. Hydrophobic interactions dominated the mode of binding behavior. The results obtained using the two methods correlated well with each other. Our findings provide a framework for elucidating the mechanisms of PCB180-HSA binding, and may also help in further research on the transportation, distribution, and toxicity effects of PCBs when introduced into human blood serum.     

Fecapentaene concentration and mutagenicity in 718 North American stool samples

The fecapentaenes (FP) are the predominant fecal mutagens identified to date, but they have not been shown to be carcinogenic. Epidemiologists looking for other fecal mutagens that may be related to colorectal cancer must disentangle from their investigations the pervasive mutagenic effect of the fecapentaenes. As a first step to studying the epidemiology of fecal mutagenicity independent of fecapentaenes, we compared FP measurements and Salmonella mutagenicity assay results for 718 acetone-extracted stool samples collected from a variety of subjects in the Washington DC metropolitan areas. In this large group, 50% of mutagenic samples contained elevated fecapentaenes. Specifically, three-quarters of the samples mutagenic in TA100 contained high FP levels. In contrast, mutagenicity in TA98 was not generally explainable by fecapentaenes, suggesting that non-fecapentaene TA98 mutagenicity should be one focus of future efforts to uncover colorectal carcinogens of etiologic importance.