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Cellular and Chemical Assays for Discovery of Novel Antioxidants in Marine Organisms

  • Tim HoferEmail author
  • Tonje Engevik Eriksen
  • Espen Hansen
  • Ingrid Varmedal
  • Ida-Johanne Jensen
  • Jeanette Hammer-Andersen
  • Ragnar Ludvig Olsen
Chapter
Part of the Oxidative Stress in Applied Basic Research and Clinical Practice book series (OXISTRESS)

Abstract

Low-molecular weight antioxidants receive increased interest both as protectors from disease and therapeutics, but are also used in other applications such as sun screens and food additives. Antioxidants are present in all organisms and are well characterized in terrestrial organisms whereas little knowledge exists for marine organisms. Only in Norwegian Arctic and sub-Arctic waters, thousands of species (invertebrates, algae, bacteria) are yet to be chemically characterized. Here we describe cell-based antioxidant assays (CAA and CLPAA in microplate format, and the Comet assay) measuring both hydrophilic and lipophilic antioxidant activities of single compounds and extract mixtures. Contrary to our chemical methods (ORAC and FRAP), cell-based assays assess cell membrane permeability and operate under more physiologically relevant conditions. A brief overview of our group’s work on finding novel antioxidants in marine organisms including testing activities of semipurified natural product extracts, identification of active compounds, purification of compounds in greater amounts, as well as how antioxidants can be tested in animals and humans is also described.

Keywords

Antioxidants Chemical assays Comet assay Marine organisms 

Abbreviations

ABAP

2,2′-azobis(2-amidinopropane)

C11-BODIPY

4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid

CAA

Cellular antioxidant activity

CLPAA

Cellular lipid peroxidation antioxidant activity

cumOOH

Cumene hydroperoxide

DCF

2′,7′-Dichlorofluorescein

DCFH

2′,7′-Dichlorofluorescin

DCFH-DA

2′,7′-Dichlorofluorescin diacetate

DMSO

Dimethyl sulfoxide

EGCG

Epigallocatechin gallate

FRAP

Ferric reducing antioxidant power

HepG2

Human hepatocellular liver carcinoma cell line

IC50

Half maximal inhibitory concentration

ORAC

Oxygen radical absorbance capacity

Notes

Acknowledgements

We thank Rudi Caeyers for help with graphical illustrations. We are grateful to the Norwegian Research Council and our industrial partner ABC Bioscience for financial support.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Tim Hofer
    • 1
    Email author
  • Tonje Engevik Eriksen
  • Espen Hansen
  • Ingrid Varmedal
  • Ida-Johanne Jensen
  • Jeanette Hammer-Andersen
  • Ragnar Ludvig Olsen
  1. 1.MabCent-SFI, University of TromsøTromsøNorway

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