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Capillary gas chromatography using electron capture or selected ion monitoring detection for the determination of muramic acid, diaminopimelic acid and the ratio of d/l-alanine in bacteria

Tunlid, Anders LU and Odham, Göran (1983) In Journal of Microbiological Methods 1(2). p.63-76
Abstract

Gas chromatographic analyses of muramic acid, diaminopimelic acid and D-alaline, which are specific components of the bacterial cell wall, have been performed using electron capture or selected ion monitoring detection. Intact cells or peptidogylycan preparations were hydrolyzed in HCl and DCl. After purification by cation exchange chromatography, followed by conversion to the N-heptafluobutyrliso-butyl esters, the components were separated on a 25 m fused silica column coated with SE-54 or on a chiral glass capillary column. The detection limits for muramic acid and diaminopimelic acid were about 10 pg using either detection method and the procedure has the potential sensitivity for detecting about 3 × 105 bacterial cells,... (More)

Gas chromatographic analyses of muramic acid, diaminopimelic acid and D-alaline, which are specific components of the bacterial cell wall, have been performed using electron capture or selected ion monitoring detection. Intact cells or peptidogylycan preparations were hydrolyzed in HCl and DCl. After purification by cation exchange chromatography, followed by conversion to the N-heptafluobutyrliso-butyl esters, the components were separated on a 25 m fused silica column coated with SE-54 or on a chiral glass capillary column. The detection limits for muramic acid and diaminopimelic acid were about 10 pg using either detection method and the procedure has the potential sensitivity for detecting about 3 × 105 bacterial cells, e.g., Escherichia coli. Mass spectrometric determination of the d/l ratio of alamine in intact cells of Group A streptococci, type M 15 and in peptidogylcan preparations thereof indicated the proportions 10.2% and 10.5% of D-alanine, respectively. The values uncorrected for racemization during acid hydrolysis were 10.3% and 10.7%, respectively.

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organization
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Contribution to journal
publication status
published
subject
keywords
Capillary GC, d/l-Alanine, Diaminopimelic acid, Electron capture detection, Muramic acid, Selected ion monitoring
in
Journal of Microbiological Methods
volume
1
issue
2
pages
63 - 76
publisher
Elsevier
external identifiers
  • scopus:0020959854
ISSN
0167-7012
DOI
10.1016/0167-7012(83)90016-7
language
English
LU publication?
yes
id
5eee6590-415f-4891-a319-1cf1d1c4e342
date added to LUP
2019-10-23 17:38:03
date last changed
2020-01-13 02:28:51
@article{5eee6590-415f-4891-a319-1cf1d1c4e342,
  abstract     = {<p>Gas chromatographic analyses of muramic acid, diaminopimelic acid and D-alaline, which are specific components of the bacterial cell wall, have been performed using electron capture or selected ion monitoring detection. Intact cells or peptidogylycan preparations were hydrolyzed in HCl and DCl. After purification by cation exchange chromatography, followed by conversion to the N-heptafluobutyrliso-butyl esters, the components were separated on a 25 m fused silica column coated with SE-54 or on a chiral glass capillary column. The detection limits for muramic acid and diaminopimelic acid were about 10 pg using either detection method and the procedure has the potential sensitivity for detecting about 3 × 10<sup>5</sup> bacterial cells, e.g., Escherichia coli. Mass spectrometric determination of the d/l ratio of alamine in intact cells of Group A streptococci, type M 15 and in peptidogylcan preparations thereof indicated the proportions 10.2% and 10.5% of D-alanine, respectively. The values uncorrected for racemization during acid hydrolysis were 10.3% and 10.7%, respectively.</p>},
  author       = {Tunlid, Anders and Odham, Göran},
  issn         = {0167-7012},
  language     = {eng},
  month        = {01},
  number       = {2},
  pages        = {63--76},
  publisher    = {Elsevier},
  series       = {Journal of Microbiological Methods},
  title        = {Capillary gas chromatography using electron capture or selected ion monitoring detection for the determination of muramic acid, diaminopimelic acid and the ratio of d/l-alanine in bacteria},
  url          = {http://dx.doi.org/10.1016/0167-7012(83)90016-7},
  doi          = {10.1016/0167-7012(83)90016-7},
  volume       = {1},
  year         = {1983},
}