Summary
In order to avoid the use of radioactive tracers for the determination of human ketone body turnover, we have developed a method using a primed-continuous infusion of 13C-labelled acetoacetate or D-β-hydroxybutyrate. Determination of the mole percent enrichment of blood acetoacetate and D-β-hydroxybutyrate was performed by gas chromatography/mass spectrometry. In the post-absorptive state, the mean total ketone body appearance rate, determined in four subjects, was 3.74 μmol · kg−1 · min−1 using [3,4-13 C2] acetoacetate and 2.76 μmol · kg−1 · min−1 using [3-13C]D-β-hydroxybutyrate, values in agreement with those reported in studies with 14C-labelled tracers. In order to evaluate the usefulness of the method for determination of ketone body kinetics in non steady-state conditions, we infused four subjects with natural sodium acetoacetate and calculated the isotopically determined total ketone body appearance rate using a single compartment model (volume of distribution 0.201/kg; functional pool fraction: 1). During the tests with [3,4-13C2]-acetoacetate, the actual infusion rates of natural acetoacetate were 7.3±0.3, 14.6±0.8, 21.9±1.2 and 10.9 ± 0.6 μ mol · kg−1 · min−1 whereas the corresponding isotopically determined total ketone body appearance rates were respectively 9.2±1.0, 16.3±0.7, 23.1±1.1 and 10.7±0.8 μmol· kg−1 · min−1. During the tests with [3-13C]D-β-hydroxybutyrate, the actual infusion rates were 8.4 ± 0.5, 16.8 ± 0.9, 25.2 ±1.4 and 12.6±0.8 μmol · kg−1 · min−1, and the isotopically determined appearance rates respectively 11.1±0.7, 16.7±0.7, 25.0±1.1 and 11.1 ± 0.7 μmol · kg−1 · min−1. Thus, using either tracer we found a good agreement between acetoacetate infusion rate and tracer-determined appearance rate of ketone bodies. In conclusion, the present method may be used to determine human ketone body kinetics under steady-state and non steady-state conditions.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Robinson AM, Williamson DH (1980) Physiological role of ketone bodies as substrates and signals in mammalian tissues. Physiol Rev 60: 143–187
Balasse EO, Neef MA (1975) Inhibition of ketogenesis by ketone bodies in fasting humans. Metabolism 24: 999–1007
Balasse EO (1979) Kinetics of ketone body metabolism in fasting humans. Metabolism 28: 41–50
Keller U, Sonnenberg GE, Stauffacher W (1981) Validation of a tracer technique to determine non steady state ketone body turnover rates in man. Am J Physiol 240: E 253-E 262
Miles JM, Haymond MW, Rizza RA, Gerich JE (1980) Determination of 14C radioactivity in ketone bodies: a new, simplified method and its validation. J Lipid Res 21: 646–650
Miles JM, Rizza RA, Haymond MW, Gerich JE (1980) Effects of acute insulin deficiency on glucose and ketone body turnover in man. Evidence for the primacy of overproduction of glucose and ketone bodies in the genesis of diabetic ketoacidosis. Diabetes 29: 926–930
Reichard GA, Owen OE, Haff AC, Paul P, Bortz WM (1974) Ketone body production and oxidation in fasting obese humans. J Clin Invest 53: 508–515
Keller U, Cherrington AD, Liljenquist JE (1978) Ketone body turnover and net hepatic ketone production in fasted and in diabetic dogs. Am J Physiol 235: E 238-E 247
Miles JM, Schwenk F, McClean KL, Haymond MW (1984) Determination of ketone body turnover in vivo with stable isotopes utilizing gas chromatography/mass spectrometry. Anal Biochem 141:110–115
Sonnenberg GE, Keller U (1982) Sampling of arterialized heated hand venous blood as a non-invasive technique for the study of ketone body kinetics in man. Metabolism 31: 1–5
Beylot M, Rion JP, Bienvenu F, Mornex R (1980) Increased ketonaemia in hyperthyroidism. Evidence for a beta-adrenergic mechanism. Diabetologia 19: 505–510
Hales CM, Randle PJ (1963) Immunoassay of insulin with insulin antibody precipitate. Biochem J 88: 137–148
Harris J, Faloona GR, Unger RH (1979) Glucagon. In: Jaffe BM, Behrman HR (eds) Methods of hormone radioimmunoassay. Academic Press, New York, pp 643–671
Okabe H, Uji Y, Nagashima K, Noha A (1980) Enzymic determination of free fatty acids in serum. Clin Chem 26: 11–15
Miles JE, Haymond MW, Nissen SL, Gerich JE (1983) Effects of free fatty acids availability, glucagon excess, and insulin deficiency on ketone body production in post-absorptive man. J Clin Invest 71:1554–1561
Fery F, Balasse EO (1983) Ketone body turnover during and after exercise in overnight fasted and starved humans. Am J Physiol 245: E 318-E 325
Sonnenberg GE, Stauffacher W, Keller U (1982) Failure of glucagon to stimulate ketone body production during acute insudeficiency or insulin replacement in man. Diabetologia 23: 94–100
Balasse EO, Fery F (1983) Aspects of ketone body metabolism in normal and diabetic subjects at rest and during exercise. In: Crepaldi G, Lefebvre PJ, Galton DJ (eds) Diabetes, obesity and hyperlipidemias, vol II. Academic Press, London, pp 249–255
Miles JE, Haymond MW, Gerich JE (1981) Suppression of glucose production and stimulation of insulin secretion by physiological concentrations of ketone bodies in man. J Clin Endocrinol Metab 52: 34–37
Barton RN (1980) Isotopic study of ketone body kinetics: invalidity of calculations based upon specific activity of total ketone bodies. Metabolism 29: 392–394
Cobelli C, Nosadini R, Toffolo G, McCulloch A, Avogaro A, Tiengo A, Alberti KGGM (1982) Model of the kinetics of ketone bodies in humans. Am J Physiol 243: R 7-R 17
Miles J, Schwenk W, Gerich J, Haymond M (1984) Are conventional isotopic methods for determination of in vivo ketone body kinetics valid? 7th International Congress of Endocrinology, July 1–7, Quebec City, Canada (Abstract)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Beylot, M., Beaufrère, B., Normand, S. et al. Determination of human ketone body kinetics using stable-isotope labelled tracers. Diabetologia 29, 90–96 (1986). https://doi.org/10.1007/BF00456116
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00456116