Useful agents for the study of glutathione metabolism in erythrocytes: organic hydroperoxides

t Butyl hydroperoxide and cumene hydroperoxide, both known to be substrates for glutathione peroxidase, were used to oxidize erythrocyte GSH. Addition of concentrations of hydroperoxides equimolar with respect to GSH in the erythrocytes or whole blood quantitatively oxidizes GSH in the erythrocytes with a half time of 4.5 sec at 37°C and about three times as long at 4°C. In the presence of glucose, normal erythrocytes regenerate all the GSH in about 25 min. However, glucose 6 phosphate dehydrogenase deficient erythrocytes failed to regenerate GSH. Treatment of erythrocytes with hydroperoxides does not affect erythrocyte survival in rabbits. Oxidation of erythrocyte GSH with equimolar concentrations of hydroperoxides does not lead to formation of mixed disulphides of haemoglobin and GSH. The hydroperoxides do not affect erythrocyte sulphides of haemoglobin and GSH. The hydroperoxides do not affect erythrocyte glycolytic and hexose monophosphate shunt pathway enzymes. Previous studies on transport of GSSG from erythrocytes were confirmed by using t butyl hydroperoxide to oxidize erythrocyte GSH.