Abstract
Glutathione is a tripeptide natural product characterized by a non-canonical peptide bond with an amide moiety linking the nitrogen of cysteine to the γ-carboxyl of glutamate, and is found ubiquitously in nature, in animals, plants and microorganisms. One of the most abundant biological matrices is represented by erythrocytes, being glutathione the only sulfur-containing mechanism for the red blood cell oxidative protection. Several analytical methods for glutathione determination from different samples are described in the literature and most of these methods are based on the use of high-performance liquid chromatography. HPLC equipment is not available in all the biochemical laboratories, and, moreover, displays lot of economic and ecological limitations, including organic solvent consumption and time-consuming analysis. Here, an organic-free high-throughput fluorometric methodology for the analysis of total glutathione in erythrocytes is reported, avoiding the use of time-consuming and not-sustainable techniques.
Highlights
Erythrocytes are the blood cells with one of the most important roles for living beings, which is oxygen transport to all the districts of the organism
GSH is a tripeptide (γ-L-glutamyl-L-cysteinylglycine) characterized by a non-canonical peptide bond, with an amide moiety linking the nitrogen of cysteine to the carboxyl in the γ-position of the glutamic acid, and is found ubiquitously in nature, animals, plants and microorganisms
Glutathione levels are much higher in erythrocytes than in plasma and are often difficult to measure because of the presence of ferric ions (Fe3+), which accelerate the oxidation of GSH to GSSG [2]
Summary
Erythrocytes are the blood cells with one of the most important roles for living beings, which is oxygen transport to all the districts of the organism. They are free of nuclei, and are unable to perform the transcription process and protein synthesis. There are several methodologies, mostly HPLC-based, for the determination of GSH in different biological samples (GSH, GSSG, total GSH) [3,4,5,6,7]. The most common methods for HPLC determination of GSH utilize pre-column derivatization of the molecule to produce a chromophore of fluorophore moieties. A lot of derivatizing agents, with different chemical, physical and spectroscopic features, are described in the literature [8,9,10]
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