Accumulation of nonesterified fatty acids in the dog myocardium during coronary artery occlusion determined by a method using 9-anthryldiazomethane

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Abstract
The levels of nonesterified fatty acids (NEFA) in the myocardium during ischemia were determined by a simple method, which requires neither previous separation with thin-layer chromatography nor heating. After being extracted with Folch's solution, NEFA were subjected to fluorescent labeling with 9-anthryldiazomethane at room temperature, separation with high-pressure liquid chromatography, and then detection by a flow-through fluorometer. Calibration and validation studies revealed that this method was satisfactory. The left anterior descending coronary artery was completely occluded for 90 min in dogs anesthetized with pentobarbital. In the nonischemic myocardium, the levels of lauric, myristic, palmitic, palmitoleic, stearic, oleic, linoleic, and arachidonic acids were 5.31, 15.85, 47.06, 2.59, 23.92, 37.90, 38.69, and 3.99 nmol/g wet tissue, respectively, and those in the ischemic myocardium 5.67, 22.16, 75.94, 6.65, 42.67, 61.75, 70.06, and 13.39 nmol/g wet tissue, respectively, the total NEFA in the former being 175.3 and that in the latter 298.3 nmol/g wet tissue. The increase in myocardial NEFA during ischemia was significant except for lauric and myristic acids, and the increase in arachidonic acid was the greatest. The ratio of the level of arachidonic acid in the ischemic myocardium to that in the nonischemic myocardium was 335.6%.