HPLC Separation of Fluorescent Products of Lipid Peroxidation in Erythrocytes and Mitochondria

Abstract

Polyunsaturated fatty acids found in membrane phospholipids are readily oxidized by free radicals producing highly reactive aldehydes such as malondialdehyde. These aldehydes react with biological material to form fluorescent lipid peroxidation end products known as lipofuscin-like pigments. We studied fluorescent pigments from beef heart mitochondria incubated with malondialdehyde in vitro and from erythrocytes of patients with Alzheimer’s disease to evaluate their potential as markers of oxidative stress and to develop HPLC methods for their qualitative analysis. We used tridimensional fluorescence spectral arrays and synchronous fluorescence spectra in connection with HPLC separation with fluorescence detection. Stable fluorophores were found in both models and were successfully resolved into several distinctive fractions. This creates the basis for further characterization of this relatively less studied group of products of oxidative damage.