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UNIT 14.10 Assessment of Metabolic Stability Using the Rainbow Trout (Oncorhynchus mykiss) Liver S9 Fraction

  1. Karla Johanning1,
  2. Gregg Hancock2,
  3. Beate Escher3,
  4. Adebayo Adekola4,
  5. Mary Jo Bernhard5,
  6. Christina Cowan-Ellsberry6,
  7. Jeanne Domoradzki7,
  8. Scott Dyer5,
  9. Curtis Eickhoff8,
  10. Michelle Embry9,
  11. Susan Erhardt10,
  12. Patrick Fitzsimmons11,
  13. Marlies Halder12,
  14. James Hill13,
  15. Dustin Holden14,
  16. Rebecca Johnson15,
  17. Sibylle Rutishauser16,
  18. Helmut Segner17,
  19. Irvin Schultz18,
  20. John Nichols11

Published Online: 1 AUG 2012

DOI: 10.1002/0471140856.tx1410s53

Current Protocols in Toxicology

Current Protocols in Toxicology

How to Cite

Johanning, K., Hancock, G., Escher, B., Adekola, A., Bernhard, M. J., Cowan-Ellsberry, C., Domoradzki, J., Dyer, S., Eickhoff, C., Embry, M., Erhardt, S., Fitzsimmons, P., Halder, M., Hill, J., Holden, D., Johnson, R., Rutishauser, S., Segner, H., Schultz, I. and Nichols, J. 2012. Assessment of Metabolic Stability Using the Rainbow Trout (Oncorhynchus mykiss) Liver S9 Fraction. Current Protocols in Toxicology. 53:14.10:14.10.1–14.10.28.

Author Information

  1. 1

    KJohanning Consultancy, Pura Vida Connections LLC, Austin, Texas

  2. 2

    Waterborne Environmental Inc., Leesburg, Virginia

  3. 3

    National Research Centre for Environmental Toxicology (Entox), The University of Queensland, Brisbane, Australia

  4. 4

    BASF, Florham Park, New Jersey

  5. 5

    Miami Valley Laboratory, The Procter & Gamble Company, Cincinnati, Ohio

  6. 6

    CE2 Consulting, LLC, Cincinnati, Ohio

  7. 7

    Dow Corning Corporation, Auburn, Michigan

  8. 8

    MAXAM Analytics, Burnaby, British Columbia, Canada

  9. 9

    ILSI Health and Environmental Sciences Institute, Washington, D.C.

  10. 10

    Department of Entomology, Michigan State University, East Lansing, Michigan

  11. 11

    Mid-Continent Ecology Division, U.S. Environmental Protection Agency, Duluth, Minnesota

  12. 12

    European Commission, Joint Research Centre, Institute for Health and Consumer Protection, Validation of Alternative Methods, Ispra, Italy

  13. 13

    Spot on Sciences, Manor, Texas

  14. 14

    Department of Chemistry and Biochemistry, University of Texas at Austin, Austin, Texas

  15. 15

    Dell Inc., Round Rock, Texas

  16. 16

    Institute of Pharmacology and Toxicology, University of Zurich, Zurich, Switzerland

  17. 17

    Centre for Fish and Wildlife Health, University of Bern, Bern, Switzerland

  18. 18

    Marine Sciences Lab, Battelle Pacific Northwest National Laboratory, Sequim, Washington

Publication History

  1. Published Online: 1 AUG 2012


Standard protocols are given for assessing metabolic stability in rainbow trout using the liver S9 fraction. These protocols describe the isolation of S9 fractions from trout livers, evaluation of metabolic stability using a substrate depletion approach, and expression of the result as in vivo intrinsic clearance. Additional guidance is provided on the care and handling of test animals, design and interpretation of preliminary studies, and development of analytical methods. Although initially developed to predict metabolism impacts on chemical accumulation by fish, these procedures can be used to support a broad range of scientific and risk assessment activities including evaluation of emerging chemical contaminants and improved interpretation of toxicity testing results. These protocols have been designed for rainbow trout and can be adapted to other species as long as species-specific considerations are modified accordingly (e.g., fish maintenance and incubation mixture temperature). Rainbow trout is a cold-water species. Protocols for other species (e.g., carp, a warm-water species) can be developed based on these procedures as long as the specific considerations are taken into account. Curr. Protoc. Toxicol. 53:14.10.1-14.10.28. © 2012 by John Wiley & Sons, Inc.


  • liver S9 fraction;
  • rainbow trout;
  • metabolism;
  • in vitro assay;
  • fish;
  • metabolism;
  • CYP450;
  • phase I and II metabolism enzymes