Skip to main content
SpringerLink
Log in

Measurement of EROD Activity: Caution on Spectral Properties of Standards Used

  • Short Communication
  • Published:
Marine Biotechnology Aims and scope Submit manuscript

Abstract

The activity of the enzyme 7-ethoxy-resorufin-O-deethylase (EROD) has been extensively used in biomonitoring studies for more than a decade. Although the analytical procedure is simple, it is often poorly characterized. In this study spectral properties of particular standard compounds used to measure EROD activity (ethoxyresorufin and resorufin, standards from Molecular Probes) were tested in order to optimize excitation and emission wavelengths to be used in the fluorimetric assay of EROD activity. The optimal excitation wavelength for the detection of resorufin was 560  nm. At this wavelength the excitation represents only 37% of its maximum level for ethoxyresorufin, while it represents 86% for resorufin. This allows discrimination between the fluorescence emitted by both standards, favoring the formed resorufin. Our results demonstrate that any analytical work using spectrofluorometry to measure EROD activity should be preceded by precise determination of the spectral characteristics of each set of standards used.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure  1
Figure  2
Figure  3

Similar content being viewed by others

References

  • R.F. Addison A.J. Edwards (1988) ArticleTitleHepatic microsomal mono-oxygenase activity in flounder Platichthys flesus from polluted sites in Langesundfjord and from mesocosms experimentally dosed with diesel oil and copper Mar Ecol Prog Ser 46 51–54 Occurrence Handle1:CAS:528:DyaL1cXlt1Cjtrg%3D

    CAS  Google Scholar 

  • C.M. Brumley V.S. Haritos J.T. Ahokas D.A. Holdway (1995) ArticleTitleValidation of biomarkers of marine pollution exposure in sand flathead using Aroclor 1254 Aquat Toxicol 31 249–262 Occurrence Handle10.1016/0166-445X(94)00074-Z Occurrence Handle1:CAS:528:DyaK2MXkvVaisbc%3D

    Article  CAS  Google Scholar 

  • T. Burgeot G. Bocquéné P. Truquet L. Le Dean F. Galgani (1994) ArticleTitleInduction of EROD activity in red mullet (Mullus barbatus) along the French Mediterranean coasts Sci Total Environ 142 213–220 Occurrence Handle10.1016/0048-9697(94)90329-8 Occurrence Handle1:CAS:528:DyaK2cXjtlSmt7Y%3D

    Article  CAS  Google Scholar 

  • M. Burke R. Mayer (1974) ArticleTitleEthoxyresorufin: direct fluometric assay of a microsomal O-dealkylation which is preferentially inducible by 3-methylchloranthrene Drug Metab Dispos 2 583–588 Occurrence Handle1:CAS:528:DyaE2MXhtVOqtLw%3D Occurrence Handle4155680

    CAS  PubMed  Google Scholar 

  • M. Burke R. Mayer (1983) ArticleTitleDifferential effects of phenobarbitone and 3-methylcholanthrene induction on the hepatic microsomal metabolism and cytochrome P-450-binding of phenoxazone and a homologous series of its n-alkyl ethers (alkoxyresorufins)1 Chem Biol Interact 45 243–258 Occurrence Handle10.1016/0009-2797(83)90072-8 Occurrence Handle1:CAS:528:DyaL3sXlvVKqu7k%3D Occurrence Handle6883573

    Article  CAS  PubMed  Google Scholar 

  • M. Eggens F. Galgani (1992) ArticleTitleEthoxyresorufin-O-deethylase (EROD) activity in flatfish: fast determination with a fluorescence plate reader Mar Environ Res 33 213–221 Occurrence Handle10.1016/0141-1136(92)90149-G Occurrence Handle1:CAS:528:DyaK3sXitVen

    Article  CAS  Google Scholar 

  • M. Eggens A. Bergman D. Vethaak (1995) ArticleTitleSeasonal variation of hepatic EROD activity in flounder (Platichthys flesus) in the Dutch Wadden Sea Mar Environ Res 39 231–234 Occurrence Handle10.1016/0141-1136(94)00023-I Occurrence Handle1:CAS:528:DyaK2MXkvFeht7c%3D

    Article  CAS  Google Scholar 

  • F. Galgani J.F. Payne (1991) ArticleTitleBiological effects of contaminants: microplate method for measurement of ethoxyresorufin-O-deethylase (EROD) in fish. Palægade 2-4, Copenhagen, Denmark ICES Tech Mar Environ Sci . 13

    Google Scholar 

  • F. Galgani G. Bocquéné M. Luçon D. Grzebyk F. Letrouit D. Claisse (1991) ArticleTitleEROD measurements in fish from the northwest part of France Mar Pollut Bull 22 494–500 Occurrence Handle10.1016/0025-326X(91)90403-F Occurrence Handle1:CAS:528:DyaK38Xms1Kmtw%3D%3D

    Article  CAS  Google Scholar 

  • F. Galgani G. Bocquéné P. Truquet T. Burgeot J.F. Chiffoleau D. Claisse (1992) ArticleTitleMonitoring of pollutant biochemical effects on marine organisms of the French coasts Oceanol Acta 15 355–364 Occurrence Handle1:CAS:528:DyaK3sXoslWisA%3D%3D

    CAS  Google Scholar 

  • D. Grzebyk F. Galgani (1991) ArticleTitleMeasurement of the effect of organic pollution on marine organisms: rapid determinantion of EROD induction using plate reader Aquatic Living Resour 4 53–59

    Google Scholar 

  • A.J. Gunther R.B. Spies J. Stegeman B. Woodin D. Carney J. Oakden L. Hain (1997) ArticleTitleEROD activity in fish is an independent measure of contaminated-induced mortality of invertebrates in sediment bioassays Mar Environ Res 44 41–49 Occurrence Handle10.1016/S0141-1136(96)00101-8 Occurrence Handle1:CAS:528:DyaK2sXisVKqtrg%3D

    Article  CAS  Google Scholar 

  • S. Hewitt H. Fenet C. Casellas (1998) ArticleTitleInduction of EROD activity in European eel (Anguilla anguilla) by different polychlorobiphenyls Wat Sci Technol 38 245–252 Occurrence Handle10.1016/S0273-1223(98)00625-8 Occurrence Handle1:CAS:528:DyaK1MXjtVWgug%3D%3D

    Article  CAS  Google Scholar 

  • D.A. Holdway S.E. Brennan J.T. Ahokas (1994) ArticleTitleUse of hepatic MFO and blood enzyme biomarkers in sand flathead (Platycephalus bassensis) as indicators of pollution in Port Phillip Bay, Australia Mar Pollut Bull 28 683–695 Occurrence Handle10.1016/0025-326X(94)90304-2 Occurrence Handle1:CAS:528:DyaK2MXisFWntr4%3D

    Article  CAS  Google Scholar 

  • S.W. Kennedy S.P. Jones (1994) ArticleTitleSimultaneous measurement of cytochrome P450IA catalytic activity and total protein concentration with a fluorescent plate reader Anal Biochem 222 217–223 Occurrence Handle10.1006/abio.1994.1476 Occurrence Handle1:CAS:528:DyaK2cXmslCqs70%3D Occurrence Handle7856852

    Article  CAS  PubMed  Google Scholar 

  • K.R. Munkittrick M.R. Heuvel ParticleVan Den M.R. Metner W.L. Lockhart J.J. Stegeman (1993) ArticleTitleInterlaboratory comparison and optimisation of hepatic microsomal ethoxyresorufin O-deethylase in white sucker Catostomas commersoni exposed to bleached Kraft pulp mill effluent Environ Toxicol Chem 12 1273–1283 Occurrence Handle1:CAS:528:DyaK3sXkvVyns7c%3D

    CAS  Google Scholar 

  • H.M. Sleiderink I. Oostigh A. Goksøyr J.P. Boon (1995) ArticleTitleSensitivity of cytochrome P450 1A induction in dab (Limanda limanda) of different age and sex as a biomarker for environmental contaminants in the southern North Sea Arch Environ Contam Toxicol 28 423–430 Occurrence Handle1:CAS:528:DyaK2MXltFGntr0%3D Occurrence Handle7755396

    CAS  PubMed  Google Scholar 

  • R. Stagg A. McIntosh (1998) ArticleTitleBiological effects of contaminants: determination of CYPlA-dependent mono-oxygenase activity in dab by fluorimetric measurement of EROD activity ICES Tech Mar Environ Sci . 23

    Google Scholar 

  • N. Sulaiman S. George M.D. Burke (1991) ArticleTitleAssessment of sublethal pollutant impact on flounders in an industrialised estuary using hepatic biochemical indices Mar Ecol Prog Ser 68 207–212 Occurrence Handle1:CAS:528:DyaK3MXktlGhsbk%3D

    CAS  Google Scholar 

Download references

Acknowledgments

We thank the team of the Organic Chemistry Laboratory (ULB) for providing access to analytical facilities and for insightful advice. Ph.D. and M.W. are, respectively, Research Associate and Honorary Research Associate of the National Fund for Scientific Research (NFSR, Belgium). G.C. and B.D. were holders of a FRIA doctoral grant. This work was supported by the SSTC covenant MN-11-30 (Prime Minister Services, Belgium). This is a contribution of the Centre interuniversitaire de Biologie marine (CIBIM). The IAEA Marine Environment Laboratory operates under a bipartite agreement between the International Atomic Energy Agency and the Government of the Principality of Monaco.

Author information

Authors and Affiliations

  1. Laboratoire de Biologie & Environnement Marins, EA 3168-Université de La Rochelle, Institut de la Mer & du Littoral, Port des Minimes, F-17000, La Rochelle, France

    G. Radenac

  2. Laboratoire de Biologie Marine, Université Libre de Bruxelles, Brussels, Belgium

    G. Coteur, B. Danis & Ph. Dubois

  3. International Atomic Energy Agency, Marine Environment Laboratory, Principality of Monaco

    M. Warnau

Authors
  1. G. Radenac
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G. Coteur
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. B. Danis
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ph. Dubois
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M. Warnau
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to G. Radenac.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Radenac, G., Coteur, G., Danis, B. et al. Measurement of EROD Activity: Caution on Spectral Properties of Standards Used. Mar. Biotechnol. 6, 307–311 (2004). https://doi.org/10.1007/s10126-004-3014-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10126-004-3014-4

Keywords:

Search

Navigation