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Updated: Jun 2, 2026

Rapid Quantification of Oxidized and Reduced Forms of Glutathione Using Ortho -phthalaldehyde in Cultured Mammalian Cells In Vitro
Published on: June 28, 2024
Detection of glutathione in whole blood after stabilization with N-ethylmaleimide
Daniela Giustarini1, Isabella Dalle-Donne, Aldo Milzani
1Department of Evolutionary Biology, Laboratory of Pharmacology and Toxicology, University of Siena, 53100 Siena, Italy. giustarini@unisi.it
Abstract:
The measurement of glutathione is a challenging task in that, during sample manipulation, a large percentage of this compound can be artificially oxidized. Here a high-performance liquid chromatography (HPLC) method to measure reduced glutathione in blood, based on the analysis of its conjugate with N-ethylmaleimide, has been developed and validated. The method is linear in the concentration range 0.1-2 mM, and the lower limit of quantification is 0.05 mM. Blood samples treated with N-ethylmaleimide are stable at -20°C for 90 days. This method has simplicity and rapidity as its main advantages; therefore, it is particularly suitable for large-scale clinical studies.
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