In vitro biotransformation rates in fish liver S9: Effect of dosing techniques
In vitro biotransformation assays are currently being explored to improve estimates of bioconcentration factors of potentially bioaccumulative organic chemicals in fish. This study compares thin‐film and solvent‐delivery dosing techniques, as well as single vs. multiple chemical dosing for measuring biotransformation rates of selected polycyclic aromatic hydrocarbons in rainbow trout (Oncorhynchus mykiss) liver S9. 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 due to 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 (KOW = 105.60) and benzo[a]pyrene (KOW = 106.04) while there were no statistical differences in pyrene (KOW = 105.18) biotransformation rates between the two 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 super‐hydrophobic chemicals. Environ Toxicol Chem. © 2014 SETAC
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