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Methodological considerations for characterizing potential antioxidant actions of bioactive components in plant foods
1Department of Neuroinflammation, Faculty of Medicine, Division of Neuroinflammation and Psychological Medicine, Imperial College London, Charing Cross Hospital Campus, Fulham Palace Road, London W6 8RF, UK. o.aruoma@ic.ac.uk
Abstract:
The study of free radicals and antioxidants in biology is producing medical revolution that promises a new age of health and disease management. From prevention of the oxidative reactions in foods, pharmaceuticals and cosmetics to the role of reactive oxygen species (ROS) in chronic degenerative diseases including cancer, autoimmune, inflammatory, cardiovascular and neurodegenerative (e.g. Alzheimer's disease, Parkinson's disease, multiple sclerosis, Downs syndrome) and aging challenges continue to emerge from difficulties associated with methods used in evaluating antioxidant actions in vivo. Our interest presently is focused on development of neurodegeneration models based on the integrity of neuronal cells in the central nervous system and how they are protected by antioxidants when challenged by neurotoxins as well as Fenton chemistry models based on the profile of polyunsaturated fatty acids (PUFAs) for the assessment of antioxidant actions in vivo. Use continues to be made of several in vitro analytical tools to characterise the antioxidant propensity of bioactive compounds in plant foods and supplements. For example, the oxygen radical absorbance capacity (ORAC), ferric reducing antioxidant power (FRAP), total oxidant scavenging capacity (TOSC), the deoxyribose assay, assays involving oxidative DNA damage, assays involving reactive nitrogen intermediates (e.g. ONOO(-)), Trolox equivalent antioxidant capacity (TEAC) and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. There is need to agree governance on in vitro antioxidant methods based on an understanding of the mechanisms involved. Because some of the assays are done in non-physiological pH values, it is impossible to extrapolate the results to physiological environment. The consensus of opinion is that a mix of these tools should be used in assessing the antioxidant activities in vitro. The proof of bio-efficacy must emanate from application of reliable in vivo models where markers of baseline oxidative damage are examined from the standpoint of how they are affected by changes in diet or by antioxidant supplements.
Insights
Free radicals and antioxidants are revolutionizing health. Reliable in vivo models are crucial for evaluating antioxidant efficacy in preventing oxidative stress and neurodegenerative diseases.
Area of Science:
- Biochemistry
- Neuroscience
- Pharmacology
Background:
- Free radicals and reactive oxygen species (ROS) play critical roles in health and disease.
- Oxidative stress is implicated in aging and chronic degenerative conditions like cancer and neurodegenerative diseases.
- Current methods for evaluating antioxidant actions in vivo face significant challenges.
Purpose of the Study:
- To develop reliable neurodegeneration models for assessing antioxidant protection against neurotoxins.
- To establish Fenton chemistry models for evaluating antioxidant actions based on polyunsaturated fatty acid profiles.
- To address the need for standardized in vitro and in vivo methods for antioxidant efficacy assessment.
Main Methods:
- Development of neurodegeneration models utilizing neuronal cell integrity in the central nervous system.
- Utilizing Fenton chemistry models with polyunsaturated fatty acids (PUFAs) to assess antioxidant capacity.
- Review and critique of various in vitro antioxidant assays (e.g., ORAC, FRAP, DPPH) and their limitations.
Main Results:
- In vitro assays provide characterization of antioxidant propensity but have limitations due to non-physiological conditions.
- A consensus exists that a combination of in vitro methods is necessary for comprehensive assessment.
- The ultimate proof of bio-efficacy requires validation through reliable in vivo models.
Conclusions:
- There is a critical need for standardized governance of in vitro antioxidant assays.
- Extrapolation of in vitro results to physiological environments is often unreliable.
- Reliable in vivo models are essential for validating antioxidant efficacy and understanding their impact on oxidative damage.
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