Simultaneous measurement of foliar glutathione, gamma-glutamylcysteine, and amino acids by high-performance liquid chromatography: comparison with two other assay methods for glutathione

Anal Biochem. 1998 Nov 1;264(1):98-110. doi: 10.1006/abio.1998.2794.

Abstract

The recent production of transformed plants with enhanced capacity for glutathione synthesis has highlighted the interactions between foliar glutathione and turnover of free amino acid pools. The development of a convenient method for simultaneous measurement of glutathione, gamma-glutamylcysteine, and 16 amino acids is reported. This method utilizes derivatization of compounds with o-phthalaldehyde in the presence of 2-mercaptoethanol followed by separation using reversed-phase high-performance liquid chromatography. Eluted compounds are detected fluorimetrically. The method was tested using untransformed poplars and poplars in which foliar thiol contents have been enhanced by overexpression of gamma-glutamylcysteine synthetase. Foliar contents of glutathione determined by this method were comparable to those measured in common extracts by two other techniques.

Publication types

  • Comparative Study

MeSH terms

  • Amino Acids / analysis*
  • Chromatography, High Pressure Liquid / methods*
  • Dipeptides / analysis*
  • Fluorometry
  • Glutathione / analysis*
  • Mercaptoethanol
  • Plant Leaves / chemistry*
  • Sulfhydryl Compounds / analysis
  • Trees
  • o-Phthalaldehyde

Substances

  • Amino Acids
  • Dipeptides
  • Sulfhydryl Compounds
  • Mercaptoethanol
  • o-Phthalaldehyde
  • Glutathione
  • gamma-glutamylcysteine