Abstract
1. A method of assaying 14C in ketone bodies present in blood by using liquid-scintillation counting is described. 2. d(−)-β-Hydroxy[14C]butyrate is converted quantitatively into [14C]acetoacetate by means of a coupled oxidoreduction reaction involving NAD+, d(−)-β-hydroxybutyrate dehydrogenase and malic dehydrogenase in the presence of a high concentration of oxaloacetate. 3. [14C]Acetoacetate is decarboxylated to acetone and carbon dioxide which are trapped separately in a double-well flask and counted subsequently. 4. The method permits the determination of 14C activity in the individual ketone bodies and allows the activity in the carboxyl carbon atoms of acetoacetate or of d(−)-β-hydroxybutyrate to be assayed separately from the activity in the remainder of the molecule. 5. Recoveries of 14C-labelled ketone bodies added to blood approach 100% with good reproducibility in replicate analyses.
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Selected References
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- CHAIKOFF I. L., GOLDMAN D. S., BROWN G. W., Jr, DAUBEN W. G., GEE M. Acetoacetate formation in liver. I. From palmitic acid-1-C14, 5-C14 and 11-C14. J Biol Chem. 1951 May;190(1):229–240. [PubMed] [Google Scholar]
- CRANDALL D. I., GURIN S. Studies of acetoacetate formation with labeled carbon; experiments with pyruvate, acetate, and fatty acids in washed liver homogenates. J Biol Chem. 1949 Dec;181(2):829–843. [PubMed] [Google Scholar]
- HIRD F. J., SYMONS R. H. The metabolism of glucose and butyrate by the omasum of the sheep. Biochim Biophys Acta. 1959 Oct;35:422–434. doi: 10.1016/0006-3002(59)90392-0. [DOI] [PubMed] [Google Scholar]
- KATZ J., CHAIKOFF I. L. A chromatographic-radioautographic method for study of acetate utilization in animal tissues. J Biol Chem. 1954 Feb;206(2):887–900. [PubMed] [Google Scholar]
- WILLIAMSON D. H., MELLANBY J., KREBS H. A. Enzymic determination of D(-)-beta-hydroxybutyric acid and acetoacetic acid in blood. Biochem J. 1962 Jan;82:90–96. doi: 10.1042/bj0820090. [DOI] [PMC free article] [PubMed] [Google Scholar]