Fluorometric Optimized Determination of Total Glutathione in Erythrocytes

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.

Keywords

• glutathione
• <i>ortho</i>-phthalaldehyde
• erythrocytes
• spectrofluorometry