Optimization of Ethoxyresorufin‐O‐deethylase Determination in the Microsomes of Earthworms and Its Induction by Polycyclic Aromatic Hydrocarbons

Publisher： John Wiley & Sons Inc

E-ISSN： 1863-0669|42|8|1121-1125

ISSN： 1863-0650

Source： CLEAN - SOIL, AIR, WATER (ELECTRONIC), Vol.42, Iss.8, 2014-08, pp. : 1121-1125

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Abstract

In this study, a fluorometric method was established to measure the ethoxyresorufin‐O‐dealkylase (EROD) activity in the microsomes of earthworm after a series of parameter modification (including excitation wavelengths; kinds, pH, and volumes of buffer solutions; kinds and volumes of sensitizers). Earthworms were exposed to increasing concentrations of benzo[a]pyrene (B[a]P) (0.009–90 µg cm⁻²) in filter papers to validate the modified method and to find out the possible effects of B[a]P to EROD activity. Results showed that 560 nm was the optimal excitation wavelength for detecting resorufin. The linear range was 0.05–20 pmol mL⁻¹ with the correlation coefficient of 0.9996. The detection limit was 0.045 pmol mL⁻¹ and the relative standard deviation was <1.3%. Based on the established method, we also found that EROD activity in earthworms was significantly induced by B[a]P at 0.009 and 0.09 µg cm⁻², suggesting that the EROD activity in earthworms could be served as a sensitive biomarker in the monitoring of soil contamination within low dosage range.