Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1981 Nov;68(5):1197–1205. doi: 10.1104/pp.68.5.1197

Gas Chromatography-Mass Spectrometry of N- Heptafluorobutyryl Isobutyl Esters of Amino Acids in the Analysis of the Kinetics of [15N]H4+ Assimilation in Lemna minor L

David Rhodes 1, Ann C Myers 1, Gene Jamieson 1
PMCID: PMC426068  PMID: 16662074

Abstract

Rapid, sensitive, and selective methods for the determination of the 15N abundance of amino acids in isotopic tracer experiments with plant tissues are described and discussed. Methodology has been directly tested in an analysis of the kinetics of [15N]H4+ assimilation in Lemna minor L. The techniques utilize gas chromatography-mass spectrometry selected ion monitoring of major fragments containing the N moiety of N-heptafluorobutyryl isobutyl esters of amino acids. The ratio of selected ion pairs at the characteristic retention time of each amino acid derivative can be used to calculate 15N abundance with an accuracy of ±1 atom% excess 15N using samples containing as little as 30 picomoles of individual amino acids. Up to 11 individual amino acid derivatives can be selectively monitored in a single chromatogram of 30 minutes. It is suggested that these techniques will be useful in situations where the small quantities of N available for analysis have hitherto hindered the use of 15N-labeled precursors.

Full text

PDF
1200

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bauer A., Joy K. W., Urquhart A. A. Amino Acid metabolism of pea leaves: labeling studies on utilization of amides. Plant Physiol. 1977 May;59(5):920–924. doi: 10.1104/pp.59.5.920. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bauer A., Urquhart A. A., Joy K. W. Amino Acid metabolism of pea leaves: diurnal changes and amino Acid synthesis from N-nitrate. Plant Physiol. 1977 May;59(5):915–919. doi: 10.1104/pp.59.5.915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Kano H., Yoneyama T., Kumazawa K. Emission spectrometric 15-N analysis of the amino acids and amides in plant tissues separated by thin layer chromatography. Anal Biochem. 1975 Jul;67(1):327–331. doi: 10.1016/0003-2697(75)90300-0. [DOI] [PubMed] [Google Scholar]
  4. MacKenzie S. L., Tenaschuk D. Gas-liquid chromatography of N-heptafluorobutyryl isobutyl esters of amino acids. J Chromatogr. 1974 Oct 9;97(1):19–24. doi: 10.1016/s0021-9673(01)97579-x. [DOI] [PubMed] [Google Scholar]
  5. Pate J. S., Atkins C. A., Herridge D. F., Layzell D. B. Synthesis, Storage, and Utilization of Amino Compounds in White Lupin (Lupinus albus L.). Plant Physiol. 1981 Jan;67(1):37–42. doi: 10.1104/pp.67.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. SIMS A. P., FOLKES B. F. A KINETIC STUDY OF THE ASSIMILATION OF (15N)-AMMONIA AND THE SYNTHESIS OF AMINO ACIDS IN AN EXPONENTIALLY GROWING CULTURE OF CANDIDA UTILIS. Proc R Soc Lond B Biol Sci. 1964 Feb 18;159:479–502. doi: 10.1098/rspb.1964.0015. [DOI] [PubMed] [Google Scholar]
  7. Sims A. P., Ferguson A. R. The regulation of glutamine metabolism in Candida utilis: studies with 15NH3 to measure in vivo rates of glutamine synthesis. J Gen Microbiol. 1974 Jan;80(1):143–158. doi: 10.1099/00221287-80-1-143. [DOI] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES