High Performance Liquid Chromatography of the Hydroperoxides Produced by Lipoxygenases

Abstract

Separation of 13-hydroperoxylinoleic acid or 13-hydroperoxylinolenic acid from linoleic acid or linolenic acid, respectively, was carried out easily and quickly by high performance liquid chromatography on porous polymer gel (TSK-Gel LS-140) using n-hexane/ethanol as an eluent. An eluent containing a large amount of n-hexane (96%) made possible the separation of 9- and 13-hydroperoxylinoleic acids. These methods were applicable for analyses of the products obtained by the incubation of soybean lipoxygenase-1 [linoleate: oxygen oxidoreductase, EC 1.13.11.12] with linoleic acid or 13-hydroperoxylinoleic acid.