Measurement of fractional lipid synthesis using deuterated water (2H2O) and mass isotopomer analysis

W. N P Lee, S. Bassilian, Z. Guo, D. Schoeller, J. Edmond, E. A. Bergner, L. O. Byerley

Research output: Contribution to journalArticle

96 Citations (Scopus)

Abstract

Fractional biosynthesis of palmitate, stearate, and cholesterol was determined with deuterated water (2H2O) using mass isotopomer analysis in Hep G2 and MCA sarcoma cells in culture. The method employed differs from previous ones in that the number of deuterium atoms from 2H2O incorporated into newly synthesized molecules was determined and not assumed. After correction for background natural abundances, the isotopomer distribution due to deuterium incorporation in fatty acids and cholesterol was shown to follow a simple binomial distribution depending on the deuterium enrichment in water (p) and the maximum number of deuterium atoms incorporated per molecule (N). Under a wide range of 2H2O enrichments, N could be determined to be 17 for palmitate, 20 for stearate, and 20 for cholesterol by regression analysis or from a series of consecutive mass isotopomer ratios. The fraction derived from de novo synthesis was given by the ratio of the observed to the theoretical deuterium enrichment, which is the product (N x p). The new synthesized fraction of palmitate and stearate by Hep G2 cells for the length of the experiment was found to be 77 and 65%, respectively. These values were confirmed by experiments with [U-13C]glucose as the precursor. In MCA sarcoma cells grown in lipid-poor medium, the average values for fractional synthesis of palmitate, stearate, and cholesterol were 70, 35, and 70%, respectively. This approach should be generally applicable to the simultaneous determination of fractional synthesis of a number of compounds with either deuterium or 13C tracers. Its application is only limited by the accuracy of mass spectrometric analysis.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Endocrinology and Metabolism
Volume266
Issue number3 29-3
StatePublished - 1994
Externally publishedYes

Fingerprint

Deuterium
Palmitates
Lipids
Water
Stearates
Sarcoma
Cholesterol
Binomial Distribution
Atoms
Molecules
Biosynthesis
Hep G2 Cells
Regression analysis
Fatty Acids
Cell Culture Techniques
Experiments
Cells
Regression Analysis
Glucose
cholesterol stearate

Keywords

  • binomial distribution
  • human hepatoma cell line Hep G2
  • MCA sarcoma cell line

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Physiology

Cite this

Lee, W. N. P., Bassilian, S., Guo, Z., Schoeller, D., Edmond, J., Bergner, E. A., & Byerley, L. O. (1994). Measurement of fractional lipid synthesis using deuterated water (2H2O) and mass isotopomer analysis. American Journal of Physiology - Endocrinology and Metabolism, 266(3 29-3).

Measurement of fractional lipid synthesis using deuterated water (2H2O) and mass isotopomer analysis. / Lee, W. N P; Bassilian, S.; Guo, Z.; Schoeller, D.; Edmond, J.; Bergner, E. A.; Byerley, L. O.

In: American Journal of Physiology - Endocrinology and Metabolism, Vol. 266, No. 3 29-3, 1994.

Research output: Contribution to journalArticle

Lee, WNP, Bassilian, S, Guo, Z, Schoeller, D, Edmond, J, Bergner, EA & Byerley, LO 1994, 'Measurement of fractional lipid synthesis using deuterated water (2H2O) and mass isotopomer analysis', American Journal of Physiology - Endocrinology and Metabolism, vol. 266, no. 3 29-3.
Lee, W. N P ; Bassilian, S. ; Guo, Z. ; Schoeller, D. ; Edmond, J. ; Bergner, E. A. ; Byerley, L. O. / Measurement of fractional lipid synthesis using deuterated water (2H2O) and mass isotopomer analysis. In: American Journal of Physiology - Endocrinology and Metabolism. 1994 ; Vol. 266, No. 3 29-3.
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