A high-performance liquid chromatography system with an immobilized enzyme reactor for detection of hydrophilic organic peroxides.

A short reactor column containing horseradish peroxidase immobilized on controlled-pore glass can replace the continuous flow of a solution of the enzyme in the detection of hydroperoxides in an HPLC system, thereby allowing the elimination of one of the three pumps previously required. The immobilized enzyme catalyzes the oxidation by hydroperoxides of (p-hydroxyphenyl)acetic acid to a fluorescent biphenyl derivative, which is the species actually detected. The simplified HPLC system is optimized for the analyste of H2O2 and a number of alkyl and 1-hydroxyalkyl hydroperoxides. The detection limit of the H2O2 analysis is 5 × 10-8 M (34 pg in a 20-μL sample), and the response is linear down to at least 10-7 M