Evaluation of a sensitive HPLC method for the determination of malondialdehyde, and application of the method to different biological materials

Reactive oxygen species (ROS) important mediators of cell and tissue injury during inflammation, are produced by several types of inflammatory cells. The formation of ROS can be monitored by detection of lipid peroxidation pr oducts. The extremely broad spectrum of biological effects of aldehydic lip id peroxidation products has necessitated the development of a technique th at enables the sensitive routine quantitation of aldehydes formed in biolog ical materials. Malondialdehyde (MDA) is a by-product of enzymatic eicosano id formation and an end-product of nonenzymatic peroxidation of polyunsatur ated fatty acids with three or more bisallylic double bonds. The determinat ion of the thiobarbituric acid derivative of MDA (TBA-MDA) is a widely used method for estimating overall lipid peroxidation. We describe a rapid, iso cratic, simple, and sensitive high-performance liquid chromatographic (HPLC ) method with spectrofluorimetric detection for measurement of MDA-TBA in h uman biological samples such as plasma, urine, wound secretions, amniotic f luid, sputum and tissue samples. By use of this method, picomole quantities of MDA can be readily and specifically detected in different biological ma terials. Coefficients of variation of repeated MDA-TBA assays were 4.4% wit hin run and 6.9% from run to run. Reference values are given for a variety of human body fluids and for rat tissues.