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A method to measure antioxidant activity in organic media: application to lipophilic vitamins
Open accessAbstract
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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“These observations were probably due to limitations of the DPPH method that include restricted reactivity towards antioxidants and solubility in aqueous and organic solvents [74].”
From Comparison of Techniques and Solvents on the Antimicrobial and Antioxidant Potential of Extracts from Acacia dealbata and Olea europaea (2020) · cites this paper as [74]
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Analytical Methods Used in Determining Antioxidant Activity: A Review
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Self-citationABTS/PP Decolorization Assay of Antioxidant Capacity Reaction Pathways
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Metadata
- Authors
- Cano, A., Acosta, M., & Arnao, M. B.
- Affiliations
- Universidad de Murcia (ES)
- Venue
- Redox Report
- Publisher
- Informa UK Limited
- 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
- bronze
- doi.org
- Locations
- Redox Report (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Biochemistry (medical) › Antioxidant Activity and Oxidative Stress
- Physical Sciences › Chemistry › Organic Chemistry › Free Radicals and Antioxidants
- Health Sciences › Medicine › Biochemistry (medical) › 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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