On the fluorometric assay of circulating lipoperoxides

The methodology for the measurement of circulating thiobarbituric acid (TBA)-reactive products of lipid peroxidation in human plasma by means of fluorometry has been reinvestigated. The lipid precipitation of plasma with phosphotungstic acid, which step is laid down in the standard assay method, strongly increases the apparent TBA-reactivity. However, subsequent washing of the precipitate may reduce the apparent levels of lipoperoxide to values close to those obtained by using untreated plasma. Also, short-time storage of plasma at either 4 degrees C or -20 degrees C induces large day to day variations in assay results. This phenomenon may be prevented almost completely by the addition of glutathione plus EDTA. It is proposed to use untreated plasma and to perform the assay immediately after collection of plasma or to add both glutathione and EDTA before storage.