Micro-determination of lipoperoxide in the mouse myocardium by thiobarbituric acid fluorophotometry.

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Abstract
The analytical conditions for fluorometric assay of lipoperoxide in the myocardium of mice were investigated in detail. The reaction of lipoperoxide with thiobarbituric acid (A) 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 1/15 vol of pyridine to n-butanol was suitable for removing 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 decomposed and therefore the solution was used within 2 wk after preparation. This method was used to assay lipoperoxide in the myocardium of mice which were treated with adriamycin (ADR). [There is an increase in lipoperoxidase that is caused by adriamycin, an anti-tumor antibiotic.] 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.