Cited by 156Antioxidant Activity and Oxidative StressFree Radicals and AntioxidantsPhytochemicals and Antioxidant Activities

A method to measure antioxidant activity in organic media: Application to lipophilic vitamins

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Cano, A., Acosta, M., Arnao, M.B.·2000·Redox Report

Abstract

The 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) radical (ABTS(.+)) can be generated by the enzymatic system formed by hydrogen peroxide and horseradish peroxidase in an organic medium. The ABTS radical is easily generated in acidified ethanol medium in about 100 s with a stability of 1.7 x 10(-3) (-deltaabs/min) monitored at 730 nm. Other organic solvents, such as methanol or acetone, have lower radical generation times but the radical is less stable. The addition of Trolox or a lipophilic antioxidant such as alpha-tocopherol or beta-carotene produces a decrease in absorbance that can be used to estimate antioxidant capacity. Using a spectrophotometric end-point method and microplate-reader equipment, we have developed a method that estimates the antioxidant activity of different lipophilic vitamins. The use of Trolox as an antioxidant standard led to a limit of detection of 0.08 nmoles and limit of quantitation of 0.28 nmoles, while similar values were obtained for alpha-tocopherol and beta-carotene. The relative antioxidant activity values obtained by different antioxidants showed that alpha-tocopherol has a similar antioxidant potential to Trolox and that beta-carotene has 2.6 times the antioxidant potential of Trolox. In our opinion, this method can be useful for estimating the antioxidant activity in lipophilic samples and as a complement to other methods that measure antioxidant activity in aqueous media.

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Research works printed but not linked: 3

  1. Arnao MB, Cano A, Acosta M. Total antioxidant activity in plant material and its interest in food technology. Rec Res Devel Agric Food Chem 1998; 2: 893Ð905.
  2. Tatsumi K, Wada S, Ichikawa H. Removal of chlorophenols from wastewater by immobilized horseradish peroxidase. Biotech Bioeng 1996; 51: 126Ð130.
  3. Campos AM, Lissi EA. Evaluation of the antioxidant capacity of herbal teas by a procedure based on the bleaching of ABTS radical cations. Bol Soc Chil Qu'm 1995; 40: 375Ð381.

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Authors
Cano, A., Acosta, M., & Arnao, M. B.
Affiliations
Universidad de Murcia (ES)
Venue
Redox Report
Published
2000-12
Volume
5
Issue
6
Pages
365-370
ISSN
  • 1351-0002 (ISSN-L)
  • 1743-2928
OpenAlex
W2004853818
Cited by
156
References
24
Citations by year
2026: 1 · 2025: 7 · 2024: 11 · 2023: 12 · 2022: 10 · 2021: 13 · 2020: 14 · 2019: 5 · 2018: 3 · 2017: 7 · 2016: 5 · 2015: 9 · 2014: 6 · 2013: 8 · 2012: 6
FWCI
1.37
Type
journal article
Language
en
Open access
Locations
  • Redox Report (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Health Sciences › Medicine › Biochemistry › Antioxidant Activity and Oxidative Stress
  • Physical Sciences › Chemistry › Organic Chemistry › Free Radicals and Antioxidants
  • Health Sciences › Medicine › Biochemistry › Phytochemicals and Antioxidant Activities
Keywords
antioxidant activity, ABTS radical, organic media, lipophilic vitamins, alpha-tocopherol, beta-carotene, Trolox, horseradish peroxidase, hydrogen peroxide, spectrophotometric assay, detection limit
MeSH terms
Ethanol, Antioxidants, Calibration, Chromans, Free Radicals, Horseradish Peroxidase, Indicators and Reagents, Sensitivity and Specificity, Solvents, Spectrophotometry, Sulfonic Acids, Vitamin E, Vitamins, Reproducibility of Results, beta Carotene, Benzothiazoles

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