When [1-14C], [U-14C], and [16-14C]palmitate were oxidized by isolated rat hepatocytes, there was a differential distribution of label as a percent of total oxidized products, such that 14CO2 from [1-14C] greater than [U-14C] greater than [16-14C]palmitate and acid-soluble radioactivity from [16-14C] greater than [U-14C] greater than [1-14C]palmitate. The oxidation of [2,3-14C]succinate to 14CO2 by isolated hepatocytes was only 9.1% of that from [1,4-14C]succinate, demonstrating that the differences in distribution of labeled products are in part due to less 14CO2 production from label in the even carbon positions entering the citric acid cycle. Apparent total ketone body production from [16-14C]palmitate was markedly higher than [1-14C] and [U-14C]palmitate. In addition, the 14C-acetone:14CO2 ratio derived from decarboxylation of labeled acetoacetate from [1-14C]palmitate was less than 1 and positively correlated to the rate of fatty acid oxidation in hepatocytes. These findings indicate that the known preferential incorporation of the omega-C2 unit of fatty acids into 14C-ketone bodies also contributed to the differential distribution of labeled products and that this contribution was greatest at the lower rates of fatty acid oxidation. In isolated mitochondria, the distribution of label to 14CO2 and acid-soluble radioactivity from [1-14C], [U-14C] and [16-14C]palmitate was qualitatively similar to that seen with hepatocytes.(ABSTRACT TRUNCATED AT 250 WORDS)