Evaluation of lipid peroxidation of human erythrocyte hemolysates

Lipid peroxidation of human erythrocyte hemolysates was evaluated by determining thiobarbituric acid reactive substance by fluorospectrophotometer. Because of the non-linear relationship between malondialdehyde formation and hemoglobin concentration, a constant hemoglobin concentration is required to obtain reproducible results. Disruption of the hemolysate by sonication and the presence of formed elements are necessary to induce the reaction, the optimal pH of which was found to be limited to the region of 7.4. Either ferric or ferrous ions are essential for the reaction to proceed, but if both are present, their catalytic activity is abolished. The present method will be useful in the investigation of the mechanism of oxidative damage to erythrocytes under various pathological conditions.