METHOD AND REAGENT FOR DETERMINING UNSATURATED FATTY ACID HYDROPEROXIDE

摘要

PURPOSE:To determine N-methylindoxyl formed by incubating an unsaturated fatty acid hydroperoxide in a specimen with peroxygenase in the presence of N-methylindole as an oxygen acceptor. CONSTITUTION:To 0.1-1.0ml of a specimen, e.g., serum, maintained at 6.8- 8.0pH are added an oxygen acceptor and a peroxygenase and the mixture is heated at 370 deg.C for 10-40min. The reaction product is measured using a spectrophotometer with a blank solution as control. The same operation is conducted using the standard unsaturated fatty acid hydroperoxide, and the amount of an unsaturated fatty acid hydroperoxide in a specimen is obtained from a prepared calibration curve. The peroxygenase is prepared from a microsome fraction obtained from peas in its germination period. Indole, phenol and aniline derivatives are used as the oxygen acceptor, and a N-methylindole is the most suitable. The reaction product obtained from the above composition is stable, and so enables the measurement using a minute amount of the specimen.