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In vitro biotransformation assays are currently being explored to improve estimates of bioconcentration factors of potentially bioaccumulative organic chemicals in fish. The present study compares thin-film and solvent-delivery dosing techniques as well as single versus multiple chemical dosing for measuring biotransformation rates of selected polycyclic aromatic hydrocarbons in rainbow trout (Oncorhynchus mykiss) liver S9. The findings show that biotransformation rates of very hydrophobic substances can be accurately measured in thin-film sorbent-dosing assays from concentration-time profiles in the incubation medium but not from those in the sorbent phase because of low chemical film-to-incubation-medium mass-transfer rates at the incubation temperature of 13.5 °C required for trout liver assays. Biotransformation rates determined by thin-film dosing were greater than those determined by solvent-delivery dosing for chrysene (octanol-water partition coefficient [KOW ] =10(5.60) ) and benzo[a]pyrene (KOW  =10(6.04) ), whereas there were no statistical differences in pyrene (KOW  =10(5.18) ) biotransformation rates between the 2 methods. In sorbent delivery-based assays, simultaneous multiple-chemical dosing produced biotransformation rates that were not statistically different from those measured in single-chemical dosing experiments for pyrene and benzo[a]pyrene but not for chrysene. In solvent-delivery experiments, multiple-chemical dosing produced biotransformation rates that were much smaller than those in single-chemical dosing experiments for all test chemicals. While thin-film sorbent-phase and solvent delivery-based dosing methods are both suitable methods for measuring biotransformation rates of substances of intermediate hydrophobicity, thin-film sorbent-phase dosing may be more suitable for superhydrophobic chemicals., (© 2014 SETAC.)

The present study examines concentrations and risks of polycyclic aromatic hydrocarbons (PAHs), nitro-PAHs (NPAHs), steranes, and hopanes in lake trout collected in Lake Michigan. A total of 74 fish were collected in 2 seasons at 3 offshore sites. The total PAH concentration (Σ9 PAH) in whole fish ranged from 223 pg/g to 1704 pg/g wet weight, and PAH concentrations and profiles were similar across season, site, and sex. The total NPAH (Σ9 NPAH) concentrations ranged from 0.2 pg/g to 31 pg/g wet weight, and carcinogenic compounds, including 1-nitropyrene and 6-nitrochrysene, were detected. In the fall, NPAH concentrations were low at the Illinois site (0.2-0.5 pg/g wet wt), and site profiles differed considerably; in the spring, concentrations and profiles were similar across sites, possibly reflecting changes in fish behavior. In the fall, the total sterane (Σ5 Sterane) and total hopane (Σ2 Hopane) levels reached 808 pg/g and 141 pg/g wet weight, respectively, but concentrations in the spring were 10 times lower. Concentrations in eggs (fall only) were on the same order of magnitude as those in whole fish. These results demonstrate the presence of target semivolatile organic compounds in a top predator fish, and are consistent with PAH biodilution observed previously. Using the available toxicity information for PAHs and NPAHs, the expected cancer risk from consumption of lake trout sampled are low. However, NPAHs contributed a significant portion of the toxic equivalencies in some samples. The present study provides the first measurements of NPAHs in freshwater fish, and results suggest that additional assessment is warranted., (© 2014 SETAC.)

Accurate estimates for the oral bioavailability of hydrophobic contaminants bound to solid matrices are challenging to obtain because of sorption to organic matter. The purpose of this research was to measure the bioavailability of [14C]chrysene sorbed to soil using an in vitro model of gastrointestinal digestion and absorption to a surrogate intestinal membrane, ethylene vinyl acetate (EVA) thin film. The [14C]chrysene moved rapidly from soil into the aqueous compartment and reached steady state within 2 h. Equilibrium was reached in the EVA film within 32 h. Aging the spiked soil for 6 or 12 months had no effect on chrysene mobilization. This was supported by the finding that the data best fit a one-compartment model. Despite significant decreases in [14C]chrysene mobilization when water or nonneutralized gastrointestinal fluids were used in place of the complete medium, the equilibrium concentration of [14C]chrysene in EVA film remained the same in all conditions. Thus, the driving force for uptake was the fugacity gradient between the aqueous phase and the EVA film. Cultured human enterocytes (human colorectal carcinoma cell line [Caco-2 cells]) had a higher lipid-normalized fugacity capacity than EVA film, but the elimination rate constants were the same, suggesting that the rate was controlled by the resistance of the unstirred aqueous layer at the membrane-water interface.

Chrysene is one of the basic polycyclic aromatic hydrocarbons that are toxic environmental pollutants. The photoproducts of 6-aminochrysene (6AC) include 5,6-chrysenequinone (5,6-CQ) along with some minor products. In this study, cytotoxicity and genotoxicity of 6AC and 5,6-CQ to a human skin cell line, HaCaT, were measured with the fluorescein diacetate uptake (FDA) test and comet assay, respectively, in the presence or absence of ultraviolet A (UVA) irradiation. The FDA test result showed that HaCaT cell viability decreased dose dependently after exposure to UVA irradiation in both 6AC (0, 0.1, 0.5, 1, 5, 10, 50 microM) and 5,6-CQ (0, 0.05, 0.25, 0.5, 2.5, 5, 25 microM) groups, with the 6AC group having lower cell viability at the same substrate concentrations; therefore, 6AC was more cytotoxic. Results of the comet assay showed that the extent of DNA damage was also dose dependent after the combined UVA and 6AC treatment (0, 0.05, 0.1, 0.5, 1 microM), although no DNA damage was detectable in the 6AC group without UVA irradiation. In addition, no DNA damage was found in the 5,6-CQ group with or without UVA irradiation. Our study indicated that 5,6-CQ, the major photoproduct of 6AC, was less photocytotoxic than the parent compound and was not photogenotoxic to HaCaT cells under the experimental conditions.

Polycyclic aromatic hydrocarbons (PAHs) are major components of wastes from municipal gas plants and many wood preservatives. Soil contaminated with these wastes is a potential threat to human health because of the carcinogenicity of many PAHs. This study follows the fate of two four-ring PAHs, pyrene and chrysene, in three matrices: an adapted soil (obtained from a site contaminated with PAHs for more than 75 years), an uncontaminated soil (with and without an inoculum of adapted soil), and sand mixed with an inoculum of adapted soil. Radiolabeled pyrene, chrysene, and salicylic acid (a metabolite of PAH biodegradation) were used to trace the mineralization, transformation, extractability, and formation of an unextractable residual over time. Linear approximations of the rates of these processes were made. High-performance liquid chromatography (HPLC) analysis of extracts from inoculated soil showed the transient formation of two known metabolites: 1-hydroxypyrene (from pyrene) and 1-hydroxy-2-naphthoic acid (from chrysene). The amount of extractable label diminished steadily over the course of the study in systems that were not inhibited with sodium azide, whereas the amount of extractable label remained relatively constant in inhibited systems. Correspondingly, the amount of nonextractable residual label generally increased during each incubation in uninhibited systems, whereas the amount of this residual label remained relatively constant in inhibited systems. In contrast, the rate and extent of mineralization varied widely across matrix types. This suggests that alterations of the PAH that impact extractability and residual formation are common, in contrast to mineralization, which was apparently limited to adapted communities.