Detection of glutathione in whole blood after stabilization with N-ethylmaleimide. 2011

Daniela Giustarini, and Isabella Dalle-Donne, and Aldo Milzani, and Ranieri Rossi
Department of Evolutionary Biology, Laboratory of Pharmacology and Toxicology, University of Siena, 53100 Siena, Italy. giustarini@unisi.it

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.

UI MeSH Term Description Entries
D002851 Chromatography, High Pressure Liquid Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed. Chromatography, High Performance Liquid,Chromatography, High Speed Liquid,Chromatography, Liquid, High Pressure,HPLC,High Performance Liquid Chromatography,High-Performance Liquid Chromatography,UPLC,Ultra Performance Liquid Chromatography,Chromatography, High-Performance Liquid,High-Performance Liquid Chromatographies,Liquid Chromatography, High-Performance
D005033 Ethylmaleimide A sulfhydryl reagent that is widely used in experimental biochemical studies. N-Ethylmaleimide,N Ethylmaleimide
D005978 Glutathione A tripeptide with many roles in cells. It conjugates to drugs to make them more soluble for excretion, is a cofactor for some enzymes, is involved in protein disulfide bond rearrangement and reduces peroxides. Reduced Glutathione,gamma-L-Glu-L-Cys-Gly,gamma-L-Glutamyl-L-Cysteinylglycine,Glutathione, Reduced,gamma L Glu L Cys Gly,gamma L Glutamyl L Cysteinylglycine
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D057230 Limit of Detection Concentration or quantity that is derived from the smallest measure that can be detected with reasonable certainty for a given analytical procedure. Limits of Detection,Detection Limit,Detection Limits

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