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Measuring reactive species and oxidative damage in vivo and in cell culture: how should you do it and what do the results mean?
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Free radicals and other reactive species (RS) are thought to play an important role in many human diseases. Establishing their precise role requires the ability to measure them and the oxidative damage that they cause. This article first reviews what is meant by the terms free radical, RS, antioxidant, oxidative damage and oxidative stress. It then critically examines methods used to trap RS, including spin trapping and aromatic hydroxylation, with a particular emphasis on those methods applicable to human studies. Methods used to measure oxidative damage to DNA, lipids and proteins and methods used to detect RS in cell culture, especially the various fluorescent "probes" of RS, are also critically reviewed. The emphasis throughout is on the caution that is needed in applying these methods in view of possible errors and artifacts in interpreting the results.
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“Unfortunately, many of the assays in the sports science literature merely serve to create confusion as they are fundamentally ill suited to deciphering whether a compound with antioxidant activity is actually acting as an antioxidant.”
From New strategies in sport nutrition to increase exercise performance (2016) · cites this paper as [156]
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Metadata
- Authors
- Halliwell, B., & Whiteman, M.
- Affiliations
- National University of Singapore (SG)
- Venue
- British Journal of Pharmacology
- Publisher
- Wiley
- Published
- 2004-05
- Volume
- 142
- Issue
- 2
- Pages
- 231-255
- ISSN
- 0007-1188 (ISSN-L)
- 1476-5381
- PMC
- PMC1574951
- OpenAlex
- W1977913875
- Cited by
- 2,238
- References
- 385
- Citations by year
- 2026: 47 · 2025: 51 · 2024: 77 · 2023: 78 · 2022: 92 · 2021: 103 · 2020: 127 · 2019: 106 · 2018: 110 · 2017: 113 · 2016: 123 · 2015: 129 · 2014: 134 · 2013: 141 · 2012: 134
- FWCI
- 6.98
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- British Journal of Pharmacology (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- National University of Singapore (submittedVersion)
- Topics
- Health Sciences › Medicine › Biochemistry (medical) › Antioxidant Activity and Oxidative Stress
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Physical Sciences › Chemistry › Organic Chemistry › Free Radicals and Antioxidants
- Keywords
- reactive species, free radicals, oxidative stress, oxidative damage, antioxidants, spin trapping, aromatic hydroxylation, fluorescent probes, DNA damage, lipid peroxidation, protein oxidation, in vivo measurement, cell culture, measurement artifacts
- MeSH terms
- Animals, Electron Spin Resonance Spectroscopy, Humans, Predictive Value of Tests, Spectrometry, Fluorescence, Reactive Oxygen Species, Oxidative Stress, Cell Culture Techniques
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