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Estimation of total phenolic content and other oxidation substrates in plant tissues using Folin–Ciocalteu reagent

Abstract

Non-structural phenolic compounds perform a variety of functions in plants, including acting as antioxidants. We describe a microplate-adapted colorimetric total phenolics assay that utilizes Folin–Ciocalteu (F–C) reagent. The F–C assay relies on the transfer of electrons in alkaline medium from phenolic compounds to phosphomolybdic/phosphotungstic acid complexes, which are determined spectroscopically at 765 nm. Although the electron transfer reaction is not specific for phenolic compounds, the extraction procedure eliminates approximately 85% of ascorbic acid and other potentially interfering compounds. This assay is performed in microcentrifuge tubes and assessed in a 96-well plate reader. At least 64 samples can be processed in 1 d.

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Figure 1: Example of a gallic acid standard curve.

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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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Correspondence to Elizabeth A Ainsworth.

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Ainsworth, E., Gillespie, K. Estimation of total phenolic content and other oxidation substrates in plant tissues using Folin–Ciocalteu reagent. Nat Protoc 2, 875–877 (2007). https://doi.org/10.1038/nprot.2007.102

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