More accuracy to the EROD measurements鈥擳he resorufin fluorescence differs between species and individuals

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• 作者：Eeva-Riikka Vehniä ; inen^a ; ^{eeva-riikka.vehniainen@jyu.fi} ; ^{eevariikka@gmail.com} ; Eija Schultz^b ; Heli Lehtivuori^a ; ^c ; Janne A. Ihalainen^a ; ^c ; Aimo O.J. Oikari^a
• 关键词：EROD activity ; Resorufin ; Fluorescence ; Ethoxyresorufin-O-deethylase ; S9 fraction
• 刊名：Aquatic Toxicology
• 出版年：2012
• 期刊代码：19_0166445x
• 类别：abs
• 出版时间：15 July, 2012
• 卷：116-117
• 期：Complete
• 页码：102-108
• 文件大小：548 K

摘要

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.