Search

A number of individual metabolites of aromatic compounds were found in the hydrolyzed bile of five English sole (Parophrys vetulus) captured from polluted sites in Puget Sound, Washington. Metabolites of fluorene, dibenzofuran, phenanthrene, fluoranthene, pyrene and benzo-[a]pyrene (BaP) were tentatively identified by gas chromatography/mass spectrometry (GC/MS) or by high-pressure liquid chromatography (HPLC) with fluorescence detection. Identifications of these metabolites were confirmed by comparing their fluorescence excitation/emission and mass spectra to those obtained from metabolites produced after other English sole from a reference site were injected with individual parent aromatic compounds. High concentrations of many of the individual metabolites quantitated (up to 300,000 ng/g) were found in the hydrolyzed bile of sole sampled from polluted sites. None of these metabolites were found in bile of sole sampled from a relatively clean (reference) site in Puget Sound.

Nonradiometric techniques were used to identify and quantitate two aromatic hydrocarbons and several of their metabolites in biological samples. High-performance liquid chromatographic separation combined with ultraviolet fluorescence detection provided a sensitive and selective method of trace analysis. Metabolites of benzo[ a ]pyrene and 2,6-dimethylnaphthalene were prepared by in vitro incubation of these substrates, singly and together, with liver microsomes of coho salmon ( Oncorhynchus kisutch ). Individual metabolites were separated by high-performance liquid chromatography and quantitated at fluorescence excitation and emission wavelengths characteristic of the corresponding parent hydrocarbon. Future refinement of these techniques may allow the determination of more complex mixtures of xenobiotics and their metabolites in marine organisms.

High prevalences of idiopathic hepatic lesions, including neoplasms (e.g., hepatocellular carcinomas, cholangiocellular carcinomas) (27%, 20 of 75 fish) and foci of cellular alteration (putative preneoplastic lesions) (44%, 33 of 75 fish), were found in English sole (Parophrys vetulus) exposed to creosote-contaminated sediments in Eagle Harbor, Puget Sound, WA. Sediments from the contaminated region of the harbor contained particularly high concentrations of aromatic hydrocarbons (e.g., benzo(a)pyrene and benz(a)anthracene), and a variety of nitrogen-containing aromatic compounds (e.g., carbazole and acridine). The composition of the aromatic compounds was characteristic of creosote. Dramatically lower concentrations of aromatic compounds were found in sediments from a reference site in which the bottom-dwelling fish examined were free of detectable neoplastic or preneoplastic hepatic lesions. Food organisms in the stomachs of the English sole from Eagle Harbor contained substantially higher concentrations of aromatic hydrocarbons than comparable organisms from the reference site. The concentrations of individual aromatic hydrocarbons in muscle and liver from the Eagle Harbor fish were low; however, high concentrations of metabolites of aromatic compounds were present in the bile. The findings strongly suggest an association between exposure to creosote and the prevalence of hepatic lesions, including neoplasms, in the bottom-dwelling fish, and furthermore support the putativemore » role of aromatic hydrocarbons in liver carcinogenesis in fish.« less

High prevalences of idiopathic liver lesions, including 7.5% hepatic neoplasms (e.g., hepatocellular and cholangiocellular carcinomas) and 16.7% foci of cellular alteration (putative preneoplastic hepatic lesions), were found in English sole (Parophrys vetulus) from waters near Mukilteo, a small community on Puget Sound in Washington State. Sediments from the sampling sites contained particularly high concentrations of aromatic hydrocarbons. Substantially lower concentrations of these compounds were found in sediments from another Puget Sound (reference) site in which fish showed no evidence of hepatic neoplasms or foci of cellular alteration. Stomach contents from the fish at Mukilteo contained substantially higher concentrations of the chemicals than did stomach contents of fish from the relatively uncontaminated site. High concentrations of metabolites of aromatic compounds were measured in the bile of fish from Mukilteo. These findings support previously observed relationships between sediment chemicals (e.g., aromatic hydrocarbons) and high prevalences of liver lesions in English sole from Puget Sound. In addition, a dietary route of uptake by English sole of environmental chemicals, including known carcinogens, was documented.

Metabolites of several two- and three-ring aromatci hydrocarbons (AHs) have been found in lives of English sole exposed to No. 2 fuel oil. Four metabolites of the C2H5-naphthalenes, six of the C3H7-naphthalenes and one each of fluorene, phenanthrene and anthracene have been partially characterized and their concentrations, which ranged from 50 to 1100 ng/g, were determined. Metabolites were separated from the liver matrix using an automated extractor/concentrator.The resulting extract was then purified by high-performance liquid chromatography, and the metabolites were characterized and quantitated by gas chromatography—mass spectrometry.

An automated extractor—concentrator was used to extract metabolites of naphthalene, 2,6-dimethylnaphthalene, and benzo[ a ]pyrene from serum, bile and liver homogenate of rainbow trout ( Samlo gairdneri ). The extracts were analyzed by reversed-phase high-performance liquid chromatography (HPLC) with fluorescence detection. Recoveries of naphthalene and 2,6-dimethylnaphthalene metabolites from all matrices were generally greater than 90%; however, the recoveries of benzo[ a ]pyrene metabolites from serum ranged from 37–99%. In addition, conjugated metabolites of polycyclic aromatic hydrocarbons (PAHs) were separated from non-conjugated metabolites and parent PAHs by using two diol columns with normal-phase HPLC. The extraction and separation techniques were also applied to isolated metabolites in samples from fish fed 2,6-dimethylnaphthalene.

High concentrations of chemicals have been found in sediments from urban areas of Puget Sound. Hundreds of organic chemicals (including certain aromatic hydrocarbons [AHs] and various chlorinated compounds) were identified. Statistical methods were used to evaluate possible relationships between the chemistry data and fish diseases. Positive correlations were found between the frequencies of liver neoplasms (e. g., hepatocellular carcinoma) and other liver lesions in English sole (Parophrys vetulus) and concentrations of AHs in sediment; such correlations were not found with chlorinated hydrocarbons. Strong evidence was also obtained to show that many organic chemicals in sediment are bioavailable to bottom-dwelling fish. Stomach contents (consisting mainly of benthic invertebrates) from English sole had concentrations of a number of AHs similar to those in the sediment from which the fish were taken. In these same fish, metabolites of many aromatic compounds were found in bile using a procedure combining HPLC with fluorescence detection. Further, the concentrations of certain xenobiotic metabolites in bile were correlated positively with the occurrence of liver neoplasms in English sole.

1. An h.p.l.c.-fluorescence technique was used to estimate relative concentrations of metabolites of xenobiotics in bile of 103 English sole (Parophrys vetulus) from both polluted and minimally polluted (reference) sites in Puget Sound, WA.2. Fish from polluted sites had concentrations of xenobiotics in bile with naphthalene-, phenanthrene- and benzo[a]pyrene-like fluorescence that averaged 9, 14 and 19 times, respectively, those of fish from reference sites.3. Within a polluted site, fish with liver lesions had significantly higher bile concentrations of xenobiotics with benzo[a]pyrene-like fluorescence than did fish without liver lesions.4. Individual metabolites of fluorene, phenanthrene, anthracene, biphenyl and dimethylnaphthalene were determined by g.I.e.-mass spectrometry in extracts of hydro-Is sed bile of three English sole from polluted waterways; concentrations ranged from 90 to 19000 ng/g, wet wt. Other xenobiotics were tentatively identified, but not quantified.

Rainbow trout (Salmo gairdneri) were exposed to orally administered (/sup 3/H)naphthalene. Another group received naphthyl glucuronic acid and naphthyl sulfate via iv injection. Brain, liver, and blood were assayed for the parent compound and/or total metabolites. Individual naphthalene derivatives were determined by high-performance liquid chromatography (hplc) using either radiometric or on-line fluorimetric detection systems. Naphthalene concentrations in brain (8.2 pmol/mg dry wt at 16 hr after feeding) approximated those found at the same time in liver (7.4 pmol/mg dry wt). A nonconjugated naphthalene derivative, 1,2-dihydro-1,2-dihydroxynaphthalene, also accumulated in brain (0.041 pmol/mg dry wt after 16 hr), although to a lesser degree than in liver (0.10 pmol/mg dry wt after 16 hr). Conjugated naphthalene derivatives, 1-naphthyl sulfate and 1-naphthyl glucuronic acid, although present in liver and blood, were largely excluded from the brain. Low naphthalene hydroxylase activity (

We report a rapid, largely automated high-performance liquid chromatographic (HPLC) method, which uses an HPLC column packed with alkylnitrile-substituted secondary alkylamine (aminocyano) bonded phase, to isolate coprostanol from interfering compounds in sediment extracts. Coprostanol is then quantitated, as the trimethylsilyl ether, by gas chromatography with flame ionization detection. Results from using the HPLC method to analyze a sediment reference material for coprostanol were statistically comparable to a previously used gravity-flow column method. We also report the coprostanol concentrations in several sediment samples from the California coast which reflect a range of sewage contamination (62-15,000 ng/g).

Bottomfish and sediments were collected from 18 sites including a number of reference sites on the U.S. West Coast during the first year of the multiyear National Benthic Surveillance Project. Sediments and fish were analyzed for toxic chemicals, and fish were examined for diseases. Wide variations in the concentrations of aromatic hydrocarbons (AH), polychlorinated biphenyls (PCB), DDT and related compounds, and selected metals in sediments were found within locations ( i .e., embayments in the vicinity of major cities) and between locations. Concentrations of chemicals in tissues were generally less variable than those in sediments, apparently reflecting the ability of fish to integrate chemical sources over large geographical areas. Prevalences of diseases (i.e., fin erosion and liver lesions) were usually highest in fish exposed to the highest levels of environmental contaminants.