Determination of Human Skin Surface Lipid Peroxides by Chemiluminescence-HPLC

Abstract

The peroxidation of human skin surface lipids was investigated by the CL-HPLC (chemiluminescence-high performance liquid chromatography) system, using a reversed phased HPLC column and hydroperoxide-specific luminescent reagent consisting of cytochrome c and luminol. Squalene hydroperoxide was detected as the most prominent chemiluminescent peak in oxidized skin surface lipids, and the retention time was identical with that of hydroperoxide of the autoxidized squalene. The chemiluminescent peak disappeared by sodium borohydride treatment. Skin surface lipids oxidized by natural exposure to sunlight were analyzed quntitatively and squalene hydroperoxide formation was confirmed.