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  • More accuracy to the EROD measurements--the resorufin fluorescence differs between species and individuals.

More accuracy to the EROD measurements--the resorufin fluorescence differs between species and individuals.

Aquatic toxicology (Amsterdam, Netherlands) (2012-04-10)
Eeva-Riikka Vehniäinen, Eija Schultz, Heli Lehtivuori, Janne A Ihalainen, Aimo O J Oikari
ABSTRACT

Ethoxyresorufin-O-deethylase (EROD) activity is a biomarker of exposure to planar aromatic hydrocarbons, and it is often measured from the S9 fraction. The effect of the liver S9 fraction of seven boreal freshwater fish species on the fluorescence of resorufin was studied. The S9 fractions diminished resorufin fluorescence by 40-80%, and there were large differences between species. Thus, using a resorufin standard curve without the S9 fraction leads to a large underestimation of the EROD activity. Therefore a microwell plate EROD method was developed that takes into account the effect of each sample on resorufin fluorescence. At least two mechanisms were involved in the decrease of the fluorescence: opaqueness of the sample, and enzymes (DT-diaphorase and plausibly NADPH-CYP450 oxidoreductase) that reduce resorufin to a non-fluorescent form.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
DT Diaphorase (NQO1) human, lyophilized powder, recombinant, expressed in E. coli, ≥90% (SDS-PAGE)