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Development and application of organic reagents for analysis. IX. A sensitive fluorometric method for the determination of glutathione peroxidase activity.

A fluorometric method for the determination of rat liver glutathione peroxidase (GSH-Px) activity has been developed, based on the determination of the concentration of unreacted reduced glutathione in the assay mixture with N-{p-[2-(6-dimethylamino)benzofuranyl]phenyl}maleimide as a fluorogenic reagent. This method is very sensitive and can assay GSH-Px activity in 50 μg wet weight of rat liver. A linear relationship was obtained between GSH-Px activity and amount of sample over the range of 10-100 μg protein. GSH-Px activities obtained by this method correlated well with those obtained by the conventional colorimetric method with 5, 5'-dithiobis(2-nitrobenzoic acid).