6533b860fe1ef96bd12c38e1
RESEARCH PRODUCT
[21] Assay of blood glutathione oxidation during physical exercise
Miguel AsensiJuan SastreJose ViñaLester Packersubject
chemistry.chemical_compoundSulfosalicylic acidBiochemistryAutoxidationChemistryGlutathione reductasemedicineGlutathione disulfidePerchloric acidGlutathioneTrichloroacetic acidmedicine.disease_causeOxidative stressdescription
Publisher Summary This chapter describes a method to accurately measure glutathione (GSH) and glutathione disulfide (GSSG) in the blood of humans and animals that perform physical exercise. With this method, it is possible to assess the oxidative stress associated with physical exercise by measuring changes in the blood GSH/GSSG ratio. Glutathione measurement is performed by a modification of the glutathione S -transferase method of Brigelius et al. This is based on the conjugation of chlorodinitrobenzene with GSH catalyzed by glutathione S -transferase. The adduct formed, S-(2,4-dinitrophenyl)glutathione, exhibits a maximum of absorbance at 340 nm. The precipitation of proteins is carried out by acid treatment. Various acids can be used as deproteinizing agents: perchloric acid (PCA), trichloroacetic acid (TCA), metaphosphoric acid, sulfosalicylic acid, or picric acid. Perchloric acid causes the autoxidation of GSH during sample processing; this is especially important when assaying blood samples. The measurement of both GSH and GSSG in blood is essential as an index of oxidative stress in several physiological and pathological situations, because blood glutathione levels reflect glutathione status in other less accessible tissues.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 1995-01-01 |