Quantitative Measurement of Ascorbate and Glutathione by Spectrophotometry

Methods Mol Biol. 2022:2526:87-96. doi: 10.1007/978-1-0716-2469-2_6.

Abstract

Ascorbate and glutathione are key chemical antioxidants present at relatively high concentrations in plant cells. They are also reducing cofactors for enzymes that process hydrogen peroxide in the ascorbate-glutathione pathway. Due to these two related biochemical functions, the compounds form an interface between reactive oxygen species and sensitive cellular components. Therefore, their status can provide reliable and direct information on cell redox state, signaling, and plant health. While several methods exist for quantification of ascorbate and glutathione, simple enzyme-dependent assays allow them to be measured easily and inexpensively in common extracts. This chapter describes a protocol to measure total contents, as well as the major oxidized and reduced forms, of both compounds in plant tissues.

Keywords: Antioxidant; Ascorbate; Glutathione; H2O2; ROS; Redox state.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antioxidants / metabolism
  • Ascorbate Peroxidases / metabolism
  • Ascorbic Acid* / metabolism
  • Glutathione* / metabolism
  • Hydrogen Peroxide / metabolism
  • Oxidation-Reduction
  • Spectrophotometry

Substances

  • Antioxidants
  • Hydrogen Peroxide
  • Ascorbate Peroxidases
  • Glutathione
  • Ascorbic Acid