DETERMINATION OF HYDROPEROXIDES WITH THE USE OF IMMOBILIZED GLUTATHIONE-PEROXIDASE IN A FLOW SYSTEM

Various hydroperoxides were determined in a flow system with the use of glutathione peroxidase immobilized on N-(2-aminoethyl)-3-aminopropyl glass. In this system, the glutathione diminished by the reaction with hydroperoxide was monitored through its reaction with 5,5'-dithio-bis-(2-nitrobenzoic acid). The optimum pH for this immobilized enzyme was found to be 7.8. The rates of three carrier solutions were examined to obtain a favorable peak. The calibration curves for hydrogen peroxide, tert-butyl hydroperoxide, cumene hydroperoxide, linoleate hydroperoxide and linolenate hydroperoxide showed high linearity over the sample concentration range from not more than 10 to 100 muM. The detection limits for the above hydroperoxides were in the range from 0.2 to 0.5 nmol. The determination of hydroperoxides could be performed in 4 min.