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Precision and sensitivity of the measurement of 15 N enrichment in D-alanine from bacterial cell walls using positive/negative ion mass spectrometry

Author:
  • Anders Tunlid
  • Göran Odham
  • Robert H. Findlay
  • David C. White
Publishing year: 1985-01-01
Language: English
Pages: 237-245
Publication/Series: Journal of Microbiological Methods
Volume: 3
Issue: 3-4
Document type: Journal article
Publisher: Elsevier

Abstract english


Sensitive detection of cellular components from specific groups of microbes can be utilized as 'signatures' in the examination of microbial consortia from soils, sediments or biofilms. Utilizing capillary gas chromatography/mass spectrometry and stereospecific derivatizing agents, D-alanine, a component localized in the prokaryotic (bacteria) cell wall, can be detected reproducibly. Enrichments of D-
15
N]alanine determined in E. coli grown with [
15
N]ammonia can be determined with precision at 1.0 atom%. Chemical ionization with methane gas and the detection of negative ions (M-HF)
-
· and (M-F or M+H - HF)
-
formed from the heptafluorobutyryl D-2 butanol ester of D-alanine allowed as little as 8 pg (90 fmol) to be detected reproducibly. This method can be utilized to define the metabolic activity in terms of
15
N incorporation at the level of 10
3
-10
4
cells, as a function of the
15
N-
14
N ratio.

Keywords

  • Analytical Chemistry
  • Bacterial cell wall
  • Capillary gas chromatography/mass spectrometry
  • D-Alanine
  • Positive/negative ion chemical ionization
  • Selected ion monitoring
  • Stable isotope labeling

Other

Published
  • ISSN: 0167-7012
Anders Tunlid
E-mail: anders [dot] tunlid [at] biol [dot] lu [dot] se

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