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Assessment of antioxidant activity by using different in vitro methods
K Schlesier1, M Harwat, V Böhm
1Friedrich-Schiller-University Jena, Institute of Nutrition, Germany.
Free Radical Research
|May 10, 2002
Summary
This study compared six antioxidant activity assays using gallic acid, uric acid, ascorbic acid, and Trolox. Black currant juice demonstrated the highest antioxidant capacity, and using multiple assays is recommended for accurate results.
Area of Science:
- Food Science and Technology
- Biochemistry
- Analytical Chemistry
Background:
- Antioxidant activity measurement is crucial for assessing the health benefits of foods and biological samples.
- Various in vitro assays exist, but their comparability and suitability for different antioxidants and matrices remain a challenge.
- Understanding the mechanisms and limitations of these assays is essential for accurate interpretation of results.
Purpose of the Study:
- To evaluate and compare six common antioxidant activity assays: TEAC, TRAP, DPPH, DMPD, PCL, and FRAP.
- To assess the antioxidant capacity of four reference compounds (gallic acid, uric acid, ascorbic acid, Trolox) and common beverages (tea, juices).
- To highlight the differences in methodologies, reaction mechanisms, and sensitivity ranges of the evaluated assays.
Main Methods:
- Comparison of six antioxidant assays (TEAC I-III, TRAP, DPPH, DMPD, PCL, FRAP) using four antioxidants: gallic acid, uric acid, ascorbic acid, and Trolox.
- Application of these assays to various beverages including tea, apple juice, tomato juice, and black currant juice.
- Categorization of assays based on oxidizing reagents (organic radicals vs. metal ions) and reaction mechanisms (delay in oxidation vs. radical/ion reduction).
Main Results:
- Gallic acid exhibited the strongest antioxidant activity across most assays, except for DMPD.
- Black currant juice consistently showed the highest antioxidant activity among the tested beverages.
- Assay sensitivity varied, with PCL detecting activity in the nanomolar range while others operated in the micromolar range.
- Significant differences in results were observed between assays, underscoring the need for method selection based on the sample matrix and antioxidant of interest.
Conclusions:
- No single assay is sufficient for comprehensive antioxidant activity assessment; using at least two complementary methods is strongly recommended.
- The choice of assay significantly impacts the measured antioxidant capacity, necessitating careful consideration of the assay's mechanism and oxidizing agent.
- In vitro antioxidant assays provide valuable insights into the protective potential of plant-derived compounds and food products.
