Analysis of GSH and GSSG after derivatization with N-ethylmaleimide

  title={Analysis of GSH and GSSG after derivatization with N-ethylmaleimide},
  author={D. Giustarini and I. Dalle-donne and A. Milzani and P. Fanti and R. Rossi},
  journal={Nature Protocols},
  • D. Giustarini, I. Dalle-donne, +2 authors R. Rossi
  • Published 2013
  • Chemistry, Medicine
  • Nature Protocols
  • This protocol describes a procedure for determining glutathione (GSH) and glutathione disulfide (GSSG) concentrations in blood and other tissues. Artifactual oxidation to GSSG of 5–15% of the GSH found in a sample can occur during deproteination of biological samples with any of the commonly used acids, with consequent marked overestimation of GSSG. This can be prevented by derivatizing GSH with the alkylating agent N-ethylmaleimide (NEM) to form GS-NEM before acid deproteination, followed by… CONTINUE READING
    147 Citations

    Paper Mentions

    Interventional Clinical Trial
    The study is done to find out whether the combined use of the nutritional supplements N-acetylcysteine and Siliphos (milk thistle extract) corrects the shedding of urine protein… Expand
    ConditionsDiabetic Nephropathy, Oxidative Stress, Proteinuria
    Interventional Clinical Trial
    The investigators hypothesized that acute consumption of Fuji apple juice (AJ) could increase the antioxidant status and/or decrease the oxidative stress (OS) biomarkers, without… Expand
    ConditionsOxidative Stress
    A direct comparison of methods used to measure oxidized glutathione in biological samples: 2-vinylpyridine and N-ethylmaleimide
    • 28
    A sensitive method for the analysis of glutathione in porcine hepatocytes
    • 7
    The determination of hepatic glutathione at tissue and subcellular level.
    • 10
    • Highly Influenced
    Pitfalls in the analysis of the physiological antioxidant glutathione (GSH) and its disulfide (GSSG) in biological samples: An elephant in the room.
    • D. Giustarini, D. Tsikas, +4 authors R. Rossi
    • Chemistry, Medicine
    • Journal of chromatography. B, Analytical technologies in the biomedical and life sciences
    • 2016
    • 56
    Comparison of N-ethyl maleimide and N-(1-phenylethyl) maleimide for derivatization of biological thiols using liquid chromatography-mass spectrometry
    • 3
    • Highly Influenced


    Blood glutathione disulfide: in vivo factor or in vitro artifact?
    • 210
    • PDF
    Protein glutathionylation in erythrocytes.
    • 59
    • PDF
    An improved HPLC measurement for GSH and GSSG in human blood.
    • 120