Micro-determination of Lipoperoxide in the Mouse Myocardium by Thiobarbituric Acid Fluorophotometry1)

Abstract

The analytical conditions or fluorometric assay of lipoperoxide in the myocardium of mice were investigated in detail. It was found that the reaction of lipoperoxide with thiobarbituric acid (TBA) depended on the acidity of the reaction medium, not on the kind of acid used. The best result was obtained when 2.0 m acetate buffer (pH 3.6) solution was used. In extraction of the fluorescent substance from the reaction medium after the reaction of lipoperoxide with TBA, the addition of one-fifteenth volume of pyridine to n-butanol was suitable to remove the turbidity of the reaction medium. Calibration plots gave a good linear relationship in the range of 0.1–1.0 nmol of malondialdehyde. The TBA aqueous solution was found to decompose and therefore we used the solution within 2 weeks after preparation. This method was used to assay lipoperoxide in the myocardium of mice which had been treated with adriamycin (ADR). Lipoperoxide level in the myocardium was increased from 250.7 nmol/g wet tissue to 674.5 nmol/g wet tissue on the 4th day after ADR administration. © 1981, The Pharmaceutical Society of Japan. All rights reserved.