Fixed wavelength fluorescence to detect PAH metabolites in fish bile: Increased statistical power with an alternative dilution method

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Abstract
Polyaromatic hydrocarbons (PAHs) are a group of environmental toxicants mainly emitted from diffuse sources. Fixed wavelength fluorescence of fish bile is a simple screening method that allows environmental monitoring of PAH exposure to fish. One drawback with this method is that the results can be biased by the presence of a so-called inner filter effect. This effect can be reduced by dilution of the bile samples. However, bile samples differ in density and extensive dilution may cause increased measurement error. An alternative method is to adjust the dilution rate to the density of each bile sample. Here, both methods have been applied to bile samples collected from caged rainbow trout. The bile density was estimated using biliverdin. The results show that the variance within experimental groups is reduced when the dilution rate is adjusted to the bile density, resulting in increased statistical power to detect differences when applied for environmental monitoring.

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