Determination of glutathione in biological material by high pressure liquid chromatography

A rapid and selective determination of reduced glutathione in biological material is described, based on its conjugation with 1-chloro-2,4-dinitrobenzene. The reaction product has a UV absorption maximum at 340 nm and is analysed by reversed phase high pressure liquid chromatography. Linear calibration graphs were obtained in the concentration range between 50 microM and 2 mM glutathione in standard solutions and in biological material (rat liver and bacterial homogenates). The detection limit is about 2 microM glutathione when using 20 microliters injection samples.