Skip to main content
SpringerLink
Log in

A Method for Evaluation of Antioxidant Activity Based on Inhibition of Free Radical-Induced Erythrocyte Hemolysis

  • Protocol
  • First Online:
Advanced Protocols in Oxidative Stress II

Part of the book series: Methods in Molecular Biology ((MIMB,volume 594))

Abstract

There are many in vitro methods for evaluating antioxidant activity. In this chapter, we describe an operationally simple cell-based assay, oxidative hemolysis inhibition assay (OxHLIA). OxHLIA is based on inhibition of free radical-induced membrane damage in erythrocytes by antioxidants. The advantage of this method is that it uses peroxyl radicals as pro-oxidants and erythrocytes as oxidizable targets so that the results obtained reflect biologically relevant radical-scavenging activity and microlocalization of antioxidants. We also present here a comparison of OxHLIA with other common methods (DPPH, ABTS•+, and ORAC assays).

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Aruoma OI (2003) Methodological considerations for characterizing potential antioxidant actions of bioactive components in plant foods. Mutat Res 523–524:9–20

    PubMed  Google Scholar 

  2. Prior RL, Wu X, Schaich K (2005) Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements. J Agric Food Chem 53:4290–4302

    Article  PubMed  CAS  Google Scholar 

  3. Pérez-Jiménez J, Arranz S, Tabernero M, Díaz-Rubio ME, Serrano J, Goñi I, Saura-Calixto F (2008) Updated methodology to determine antioxidant capacity in plant foods, oils and beverages: extraction, measurement and expression of results. Food Res Intern 41:274–285

    Article  Google Scholar 

  4. Magalhães LM, Segundo MA, Reis S, Lima JL (2008) Methodological aspects about in vitro evaluation of antioxidant properties. Anal Chim Acta 613:1–19

    Article  PubMed  Google Scholar 

  5. Brand-Williams W, Cuvelier ME, Berset C (1995) Use of a free radical method to evaluate antioxidant activity. Lebenson Wiss Technol 28:25–30

    CAS  Google Scholar 

  6. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C (1999) Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med 26:1231–1237

    Article  PubMed  CAS  Google Scholar 

  7. Prior RL, Hoang H, Gu L, Wu X, Bacchiocca M, Howard L, Hampsch-Woodill M, Huang D, Ou B, Jacob R (2003) Assays for hydrophilic and lipophilic antioxidant capacity (oxygen radical absorbance capacity (ORACFL)) of plasma   and other biological and food samples. J Agric Food Chem 51:3273–3279

    Article  PubMed  CAS  Google Scholar 

  8. Niki E, Komuro E, Takahashi M, Urano S, Ito E, Terao K (1988) Oxidative hemolysis of erythrocytes and its inhibition by free radical scavengers. J Biol Chem 263:19809–19814

    PubMed  CAS  Google Scholar 

  9. Sato Y, Kamo S, Takahashi T, Suzuki Y (1995) Mechanism of free radical-induced hemolysis of human erythrocytes: hemolysis by water-soluble radical initiator. Biochemistry 34:8940–8949

    Article  PubMed  CAS  Google Scholar 

  10. Takebayashi J, Kaji H, Ichiyama K, Makino K, Gohda E, Yamamoto I, Tai A (2007) Inhibition of free radical-induced erythrocyte hemolysis by 2-O-substituted ascorbic acid derivatives. Free Radic Biol Med 43:1156–1164

    Article  PubMed  CAS  Google Scholar 

  11. Niki E, Noguchi N (2000) Evaluation of antioxidant capacity. What capacity is being measured by which method? IUBMB Life 2000:323–329

    Google Scholar 

  12. Frankel EN, Meyer AS (2000) The problems of using one-dimensional methods to evaluate multifunctional food and biological antioxidants. J Sci Food Agric 80:1925–1941

    Article  CAS  Google Scholar 

  13. Niki E (1990) Free radical initiators as source of water- or lipid-soluble peroxyl radicals. Methods Enzymol 186:100–108

    Article  PubMed  CAS  Google Scholar 

  14. Yoshida H, Yanai H, Namiki Y, Fukatsu-Sasaki K, Furutani N, Tada N (2006) Neuroprotective effects of edaravone: a novel free radical scavenger in cerebrovascular injury. CNS Drug Rev 12:9–20

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was partly supported by a Grant-in-Aid for Young Scientists (B) (No. 17780103) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.

Author information

Authors and Affiliations

  1. Food Function and Labeling Program, Incorporated Administrative Agency, National Institute of Health and Nutrition, Tokyo, Japan

    Jun Takebayashi & Jianbin Chen

  2. Prefectural University of Hiroshima, Hiroshima , Japan

    Akihiro Tai (Faculty of Life and Environmental Sciences)

Authors
  1. Jun Takebayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jianbin Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Akihiro Tai
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jun Takebayashi .

Editor information

Editors and Affiliations

  1. Golden Leaf Rd. 215, Mars Hill, 28754, USA

    Donald Armstrong

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Takebayashi, J., Chen, J., Tai, A. (2010). A Method for Evaluation of Antioxidant Activity Based on Inhibition of Free Radical-Induced Erythrocyte Hemolysis. In: Armstrong, D. (eds) Advanced Protocols in Oxidative Stress II. Methods in Molecular Biology, vol 594. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-411-1_20

Download citation

  • DOI: https://doi.org/10.1007/978-1-60761-411-1_20

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-410-4

  • Online ISBN: 978-1-60761-411-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation