Quantitation of Total And Reduced Glutathione In Erythrocytes By High Performance Liquid Chromatography]

Glutathione is the main non-protein thiol compound present in mammalian cells inmillimolar level. Glutathione which is a tripeptide functions as a principle reducingagent in biological systems. The most important function of this tripeptide is tomaintain intracellular redox status and to protect cells from reactive oxygen species.It is also involved in physiological processes such as synthesis of DNA andtransport of amino acids. This study was carried out to optimize the most suitableconditions for quantitation of total and reduced glutathione in erythrocytes by HighPerformance Liquid Chromatography. In our study, the most suitable compositionof acetonitrile+acetate buffer and the flow rate for the ideal elution of glutathionewere determined by evaluating the number of theoretical plate, capacity factor, peakasymmetry and resolution values, indicating the system and method performance.In addition, intra and inter assay reproducibility and recovery studies were done.The results obtained from this study provided an improved method for glutathionedetermination by High Performance Liquid Chromatography. Since this method isfaster than the most commonly used methods and it also permits better reproducibilityand high sensitivity, thus it is suggested that this method can be used confidentlyto determine the amount of glutathione in biologic samples.