Acute toxicity of 353-nonylphenol and its metabolites for zebrafish embryos

  • Ulrike KammannEmail author
  • Michael Vobach
  • Werner Wosniok
  • Andreas Schäffer
  • Andreas Telscher


Background, aim and scope

Nonylphenol (NP) can be detected in the aquatic environment all over the world. It is applied as a technical mixture of isomers of which 353-NP is the most relevant both in terms of abundance (about 20% of total mass) and endocrine potential. 353-NP is metabolised in sewage sludge. The aims of the present study were to determine and to compare the acute toxicity of t-NP, 353-NP and its metabolites as well as to discuss if the toxicity of 353-NP changes during degradation.

Materials and methods

353-NP and two of its metabolites were synthesised. The zebrafish embryo test was performed according to standard protocols. Several lethal and non-lethal endpoints during embryonal development were reported. NOEL, LOEL and EC50 were calculated.


All tested compounds caused lethal as well as non-lethal malformations during embryo development. 353-NP showed a higher toxicity (EC50 for lethal endpoints 6.7 mg/L) compared to its metabolites 4-(3.5-dimethyl-3-heptyl)-2-nitrophenol (EC50 13.3 mg/L) and 4-(3,5-dimethyl-3-heptyl)-2-bromophenol (EC50 27.1 mg/L).


In surface water, concentrations of NP are far below the NOEC identified by the zebrafish embryo test. However, in soils and sewage sludge, concentrations may reach or even exceed these concentrations. Therefore, sludge-treated sites close to surface waters should be analysed for NP and its metabolites in order to detect an unduly high contamination due to runoff events.


The results of the present study point out that the toxicity of 353-NP probably declines during metabolisation in water, sediment and soil, but does not vanish since the major metabolites exhibit a clear toxic potential for zebrafish embryos.

Recommendations and perspectives

Metabolites of environmental pollutants should be included in the ecotoxicological test strategy for a proper risk assessment.


Danio rerio Degradation Metabolites Nonylphenol Toxicity 


  1. Amacher SL, Draper BW, Summers BR, Kimmel CB (2002) The zebrafish T-box genes no tail and spadetail are required for development of trunk and tail mesoderm and medial floor plate. Development 129:3311–1123Google Scholar
  2. Braunbeck T, Böttcher M, Hollert H, Kosmehl T, Lammer E, Leist E, Rudolf M, Seitz N (2005) Towards an alternative for the acute fish LC50 test in chemical assessment: the fish embryo toxicity test goes multi-species—an update. Altex-Altern Tierexp 22:87–102Google Scholar
  3. DIN (2001) DIN 38 415-T6, German standard methods for the examination of water, waste water and sludge—Subanimal testing (group T)—Part 6: Determination of the non-acute-poisonous effect of waste water to fish eggs by dilution limits. German Standardisation Organisation, Beuth Vertrieb GmbH, BerlinGoogle Scholar
  4. Hollert H, Keiter S, König N, Rudolf M, Ulrich M, Braunbeck T (2003) A new sediment contact assay to assess particle-bound pollutants using zebrafish (Danio rerio) embryos. J Soil Sediment 3(3):197–207CrossRefGoogle Scholar
  5. Hoyt PR, Doktycz MJ, Beattie KL, Greenley MS (2003) DNA microarray detect 4-nonylphenol induced alterations in gene expression during zebrafish early development. Ecotoxicol 12:469–474CrossRefGoogle Scholar
  6. ISO (2007) ISO 15088:2007 Water quality—determination of the acute toxicity of waste water to zebrafish eggs (Danio rerio)Google Scholar
  7. Isobe T, Takada H (2004) Determination of degradation products of alkylphenol polyethoxylates in municipal wastewaters and rivers in Tokyo, Japan. Environ Toxicol Chem 23:599–605CrossRefGoogle Scholar
  8. Johnson AJM, Jürgens M (2003) Endocrine active industrial chemicals: release and occurrence in the environment. Pure Appl Chem 75:1895–1904CrossRefGoogle Scholar
  9. Kammann U, Biselli S, Huhnerfuss H, Reineke N, Theobald N, Vobach M, Wosniok W (2004) Genotoxic and teratogenic potential of marine sediment extracts investigated with comet assay and zebrafish test. Environ Pollut 132(2):279–287CrossRefGoogle Scholar
  10. Kammann U, Biselli S, Reineke N, Wosniok W, Danischewski D, Hühnerfuss H, Kinder A, Sierts-Herrmann A, Theobald N, Vahl HH, Vobach M, Westendorf J, Steinhart H (2005) Bioassay-directed fractionation of organic extracts of marine surface sediments from the North and Baltic Sea—Part II: Results of the biotest battery and development of a biotest index. J Soil Sediment 5(4):225–232CrossRefGoogle Scholar
  11. Kammann U, Vobach M, Wosniok W (2006) Toxic effects of brominated indoles and phenols to zebrafish embryos. Arch Environ Con Tox 51(1):97–102CrossRefGoogle Scholar
  12. Kannan K, Keith TL, Naylor CG, Staples CA, Snyder SA, Giesy JP (2003) Nonylphenol and nonylphenol ethoxylates in fish, sediment, and water from the Kalamazoo River, Michigan. Arch Environ Con Tox 44:77–82CrossRefGoogle Scholar
  13. Kawahata H, Ohta H, Inoue M, Suzuki A (2004) Endocrine disrupter nonylphenol and bisphenol A contamination in Okinawa and Ishigaki Islands, Japan—within coral reefs and adjacent river mouths. Chemosphere 55(11):1519–1527CrossRefGoogle Scholar
  14. Kelly SA, Di Giulio RT (2000) Developmental toxicity of estrogenic alkylphenols in killifish (Fundulus heteroclitus). Environ Toxicol Chem 19:2564–2570CrossRefGoogle Scholar
  15. Kim YS, Katase T, Horii Y, Yamashita N, Makino M, Uchijama T, Fujimoto Y, Inoue T (2005) Estrogen equivalent concentration of individual isomer-specific 4-nonylphenol in Ariake sea water, Japan. Mar Pollut Bull 51:850–856CrossRefGoogle Scholar
  16. Kolpin DW, Furlong ET, Meyer MT, Thurmann EM, Zaugg SD, Barber LB, Buxton HT (2002) Pharmaceuticals, hormones, and other organic wastewater contaminants in U.S. streams, 1999–2000: a national reconnaissance. Environ Sci Technol 36:1202–1211CrossRefGoogle Scholar
  17. Kosmehl T, Hallare AV, Reifferscheid G, Manz W, Braunbeck T, Hollert H (2006) A novel contact assay for testing genotoxicity of chemicals and whole sediments in zebrafish embryos. Environ Toxicol Chem 25:2097–2106CrossRefGoogle Scholar
  18. Lavado R, Thibaut R, Raldua D, Martin R, Porte C (2004) First evidence of endocrine disruption in feral carp from the Ebro River. Toxicol Appl Pharm 196(2):247–57CrossRefGoogle Scholar
  19. Lee H, Peart TE, Chan J, Gris G (2004) Occurrence of endocrine-disrupting chemicals in sewage and sludge samples in Toronto, Canada. Water Qual Res J Can 39:57–63Google Scholar
  20. Nagel R (2002) DarT: the embryo test with the zebrafish Danio rerio—a general model in ecotoxicology and toxicology. Altex-Altern Tierexp 19(Suppl 1):38–48Google Scholar
  21. OECD (2006) Organisation for Economic Co-operation and development, Environment Directorate (OECD)—Current approaches in the statistical analysis of ecotoxicity data: a guidance to application. OECD Series on Testing and Assessment, Number 54, Paris 2006Google Scholar
  22. Petrovic M, Diaz A, Ventura F, Barcelo D (2001) Simultaneous determination of halogenated derivatives of alkylphenol ethoxylates and their metabolites in sludges, river sediments, and surface, drinking, and wastewaters by liquid chromatography–mass spectrometry. Anal Chem 73(24):5886–5895CrossRefGoogle Scholar
  23. Preuss TG, Ratte HT (2007) Ecotoxicological characterisation of nonylphenol isomers. Umweltwiss Schadst Forsch 19(4):227–233CrossRefGoogle Scholar
  24. Preuss TG, Gehrhardt J, Schirmer K, Coors A, Rubach M, Russ A, Joned PD, Giesy JP, Ratte T (2006) Nonylphenol isomers differ in estrogenic activity. Environ Sci Technol 40(16):5147–5153CrossRefGoogle Scholar
  25. Russ AS, Vinken R, Schuphan I, Schmidt B (2005) Synthesis of branched paranonylphenol isomers: occurrence and quantification in two commercial mixtures. Chemosphere 60:1624–1635CrossRefGoogle Scholar
  26. SAS Institute Inc. (2004) SAS OnlineDoc® 9.1.3. Cary: SAS Institute Inc.Google Scholar
  27. Scholz S, Fischer S, Gündel U, Küster E, Luckenbach T, Voelker D (2008) The zebrafish embryo model in environmental risk assessment—applications beyond acute toxicity testing. Environ Sci Pollut Res 15:394–404CrossRefGoogle Scholar
  28. Shao B, Hu J, Yang M (2003) Nonylphenol ethoxylates and their biodegradation intermediates in water and sludge of a sewage treatment plant. B Environ Contam Tox 70:527–532CrossRefGoogle Scholar
  29. Stachel B, Jantzen E, Knoth W, Kruger F, Lepom P, Oetken M, Reincke H, Sawal G, Schwartz R, Uhlig S (2005) The Elbe flood in August 2002—organic contaminants in sediment samples taken after the flood event. J Environ Sci Heal A 40:265–287CrossRefGoogle Scholar
  30. Telscher MJH, Schuller U, Schmidt B, Schäffer A (2005) Occurrence of a nitro metabolite of a defined nonylphenol isomer in soil/sewage sludge mixtures. Environ Sci Technol 39(20):7896–7900CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Ulrike Kammann
    • 1
    Email author
  • Michael Vobach
    • 1
  • Werner Wosniok
    • 2
  • Andreas Schäffer
    • 3
    • 4
  • Andreas Telscher
    • 3
  1. 1.Johann Heinrich von Thünen-Institute (vTI), Federal Research Institute for Rural Areas, Forests and Fisheries, Institute for Fishery EcologyHamburgGermany
  2. 2.Institute of StatisticsUniversity of BremenBremenGermany
  3. 3.RWTH Aachen, Institute of Environmental Biology and ChemodynamicsAachenGermany
  4. 4.Fraunhofer Institute of Molecular Biology and Applied EcologySchmallenbergGermany

Personalised recommendations