Evaluation of phospholipid peroxidation as malondialdehyde during myocardial ischemia and reperfusion injury

Peroxidation of membrane phospholipid polyunsaturated fatty acids is considered a major mechanism of the damage occurring on reperfusion of the myocardium after a prolonged period of ischemia. The evidence in support of this mechanism of damage is based on tissue malondialdehyde quantitation by the thiobarbituric acid test (TBA test). In an attempt to verify this topic, we have subjected isolated and Langendorff-perfused rabbit hearts to a period of 60 min of severe ischemia plus 30 min of reperfusion. At appropriate time points, malondialdehyde was determined in the tissue by means of TBA test and directly by reversed-phase, high-pressure liquid chromatography (HPLC). We have found no correlation between the two compared assays. During reperfusion, there was the formation of non-lipid-related, malondialdehyde-like, TBA-reactive substance that leads to overestimation of the extent of lipid peroxidation. On the contrary, by direct HPLC quantitation, there was a decrease of tissue malondialdehyde during ischemia and during the early phases of reperfusion. Our results demonstrate that TBA test is not a reliable index of lipid peroxidation in organ systems.