Interpretation of toxicity test results may be hampered when doubt exists about the actual exposure concentration. Processes that are responsible for differences between the nominal and the actual concentration in aqueous test systems may include sorption, precipitation, volatilization, chemical and biological degradation, and uptake into biological or test tissue. In this study, the use of a poly(dimethylsiloxane) (PDMS) film containing the test compound is introduced as a versatile technique for partition controlled delivery of hydrophobic compounds to aqueous toxicity tests. Two methods developed produced preloaded films, having toxicant added to the PDMS prepolymer solution before film deposition and curing, and postloaded films, which are created by the addition of toxicant in a solvent to an already-polymerized PDMS film. Preloaded films were generally more easily prepared, may better accommodate larger molecules, and have a higher capacity than postloaded films. Postloaded films provided film-solution partition coefficients with higher precision and allowed for the use of films from stock and thus for a more portable technique. Chemical analysis showed that equilibrium between films and the aqueous solution was established within 7-10 min and was maintained for a suite of aromatic compounds (log Kow ranging from 2.8 to 6.1). The reliability of the film technique was demonstrated by application to the Microtox bacterial toxicity tests of solutions of polycyclic aromatic hydrocarbons (PAHs).
In bioassays, exposure concentrations of test compounds are usually expressed as nominal concentrations. As a result of various processes, such as adsorption, degradation, or uptake, the actual freely dissolved concentration of the test compound may differ from the nominal concentration. The goal of the present study was to develop a method to dose passively the freely dissolved fraction of organic chemicals in an in vitro bioassay with adherent cells. To this end, a polydimethylsiloxane (PDMS) film-based method was developed for a reporter gene assay for dioxin-like compounds in a rat liver cell line. Polydimethylsiloxane films loaded with test compounds ensure that the concentration during exposure is in equilibrium and that the ratio between the concentration on the film and the concentration in medium is constant. Benzo[k]fluoranthene (BkF) was used as a model compound to develop the passive dosing method in transwell plates, which was further tested with a complex mixture, i.e., an extract prepared from a contaminated sediment. A higher dioxin-like activity was found when extracts were dosed by passive dosing with PDMS than when directly added to medium. Comparison with analysis of the concentration of BkF in medium shows that passive dosing of individual chemicals may not be necessary if freely dissolved concentrations are known. Use of PDMS for passive dosing of complex samples may represent a more realistic method for exposure in in vitro bioassays.
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