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Rapid measurement of total antioxidant capacity in plants

Nature Protocols volume 2, pages 867–870 (2007)Cite this article

Abstract

There is growing interest in measuring the antioxidant status of plant tissues. This protocol describes the oxygen radical absorbance capacity (ORAC) assay, which measures antioxidant inhibition of peroxyl radical–induced oxidations and is a measure of total antioxidant capacity. The assay is performed in a microplate and is assessed with a 96-well multi-detection plate reader. Total antioxidant capacity of 64 experimental samples can easily be analyzed in 1 d. This assay is presented along with rapid assays for total phenolic content and total ascorbate content. Overall, these assays provide a general diagnostic tool of the antioxidant capacity in leaf tissue extracts.

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Figure 1: Typical fluorescence curves and trolox standard curves for the oxygen radical absorbance capacity (ORAC) assay.
Figure 2: Relationship between total antioxidant capacity (oxygen radical absorbance capacity, ORAC) and phenolic content and total ascorbate content in field-grown soybean leaves.

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Acknowledgements

This research was supported by the Office of Science (BER), U.S. Department of Energy, Grant no. DE-FG02-04ER63849.

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Authors and Affiliations

  1. Program in Physiological and Molecular Plant Biology and Institute for Genomic Biology, University of Illinois Urbana-Champaign, 147 ERML, 1201 W. Gregory Drive, Urbana, 61801, Illinois, USA

    Kelly M Gillespie & Elizabeth A Ainsworth

  2. USDA ARS Photosynthesis Research Unit, Urbana, 61801, Illinois, USA

    June M Chae & Elizabeth A Ainsworth

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Correspondence to Elizabeth A Ainsworth.

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Gillespie, K., Chae, J. & Ainsworth, E. Rapid measurement of total antioxidant capacity in plants. Nat Protoc 2, 867–870 (2007). https://doi.org/10.1038/nprot.2007.100

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