Measurement of reduced, oxidized and total ascorbate content in plants

Kelly M. Gillespie; Elizabeth A. Ainsworth

doi:10.1038/nprot.2007.101

Measurement of reduced, oxidized and total ascorbate content in plants

Kelly M. Gillespie, Elizabeth A. Ainsworth

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Abstract

Ascorbate is one of the major antioxidant metabolites in plant tissues. This protocol describes a microplate-adapted colorimetric ascorbate assay, in which ferric ion is reduced by ascorbate to the ferrous ion. The ferrous ion reacts with α-ά-bipyridl to form a complex with characteristic absorbance at 525 nm. With the chemical reduction of any dehydroascorbate (DHA) in a sample, total ascorbate can be assayed using the α-ά-bipyridl method, and DHA can be estimated by subtracting the reduced portion from the total ascorbate pool. The assay is performed in microcentrifuge tubes and assessed in a 96-well plate reader. Reduced ascorbate, DHA and total ascorbate of at least 64 experimental samples can be analyzed easily in 1 d.

Original language	English (US)
Pages (from-to)	871-874
Number of pages	4
Journal	Nature Protocols
Volume	2
Issue number	4
DOIs	https://doi.org/10.1038/nprot.2007.101
State	Published - Apr 2007

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)

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10.1038/nprot.2007.101

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title = "Measurement of reduced, oxidized and total ascorbate content in plants",

abstract = "Ascorbate is one of the major antioxidant metabolites in plant tissues. This protocol describes a microplate-adapted colorimetric ascorbate assay, in which ferric ion is reduced by ascorbate to the ferrous ion. The ferrous ion reacts with α-ά-bipyridl to form a complex with characteristic absorbance at 525 nm. With the chemical reduction of any dehydroascorbate (DHA) in a sample, total ascorbate can be assayed using the α-ά-bipyridl method, and DHA can be estimated by subtracting the reduced portion from the total ascorbate pool. The assay is performed in microcentrifuge tubes and assessed in a 96-well plate reader. Reduced ascorbate, DHA and total ascorbate of at least 64 experimental samples can be analyzed easily in 1 d.",

author = "Gillespie, {Kelly M.} and Ainsworth, {Elizabeth A.}",

note = "Funding Information: ACKNOWLEDGMENTS This research was supported by the Office of Science (BER), U.S. Department of Energy, Grant no. DE-FG02-04ER63849.",

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AU - Gillespie, Kelly M.

AU - Ainsworth, Elizabeth A.

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N2 - Ascorbate is one of the major antioxidant metabolites in plant tissues. This protocol describes a microplate-adapted colorimetric ascorbate assay, in which ferric ion is reduced by ascorbate to the ferrous ion. The ferrous ion reacts with α-ά-bipyridl to form a complex with characteristic absorbance at 525 nm. With the chemical reduction of any dehydroascorbate (DHA) in a sample, total ascorbate can be assayed using the α-ά-bipyridl method, and DHA can be estimated by subtracting the reduced portion from the total ascorbate pool. The assay is performed in microcentrifuge tubes and assessed in a 96-well plate reader. Reduced ascorbate, DHA and total ascorbate of at least 64 experimental samples can be analyzed easily in 1 d.

AB - Ascorbate is one of the major antioxidant metabolites in plant tissues. This protocol describes a microplate-adapted colorimetric ascorbate assay, in which ferric ion is reduced by ascorbate to the ferrous ion. The ferrous ion reacts with α-ά-bipyridl to form a complex with characteristic absorbance at 525 nm. With the chemical reduction of any dehydroascorbate (DHA) in a sample, total ascorbate can be assayed using the α-ά-bipyridl method, and DHA can be estimated by subtracting the reduced portion from the total ascorbate pool. The assay is performed in microcentrifuge tubes and assessed in a 96-well plate reader. Reduced ascorbate, DHA and total ascorbate of at least 64 experimental samples can be analyzed easily in 1 d.

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Measurement of reduced, oxidized and total ascorbate content in plants

Abstract

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