Simplified absolute metabolite quantification by gas
chromatography-isotope dilution mass spectrometry on the basis of
commercially available source material
In the field of metabolomics, GC-MS has rather established itself as a
tool for semi-quantitative strategies like metabolic fingerprinting or
metabolic profiling. Absolute quantification of intra- or extracellular
metabolites is nowadays mostly accomplished by application of diverse
LC-MS techniques. Only few groups have so far adopted GC-MS technology
for this exceptionally challenging task. Besides numerous and deeply
investigated problems related to sample generation, the pronounced
matrix effects in biological samples have led to the almost mandatory
application of isotope dilution mass spectrometry (IDMS) for the
accurate determination of absolute metabolite concentrations.
Nevertheless, access to stable isotope labeled internal standards
(ILIS), which are in many cases commercially unavailable, is quite
laborious and very expensive. Here we present an improved and simplified
gas chromatography-isotope dilution mass spectrometry (GC-IDMS) protocol
for the absolute determination of intra- and extracellular metabolite
levels. Commercially available C-13-labeled algal cells were used as a
convenient source for the preparation of internal standards. Advantages
as well as limitations of the described method are discussed. (C) 2011
Elsevier B.V. All rights reserved.
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
number
32
pages
3859-3870
publisher
ELSEVIER SCIENCE BV
volume
879
type
Article
issn
1570-0232
funding-acknowledgement
transnational research initiative ``Systems Biology of Microorganisms
(SysMO)'' within network MOSES: ``MicroOrganisnn Systems Biology:
Energy and Saccharomyces cerevisiae''
research-areas
Biochemistry & Molecular Biology; Chemistry
doc-delivery-number
868NO
usage-count-since-2013
37
web-of-science-categories
Biochemical Research Methods; Chemistry, Analytical
The authors would like to thank Lara Bogner, Andreas Freund, Achim
Hauck, Mira Lenfers-Lucker and Alexander Muller (Institute of
Biochemical Engineering (IBVT), University of Stuttgart, Germany) for
the experimental support. The research was funded by the transnational
research initiative ``Systems Biology of Microorganisms (SysMO)''
within network MOSES: ``MicroOrganisnn Systems Biology: Energy and
Saccharomyces cerevisiae'' http://www.sysmo.net/.
%0 Journal Article
%1 ISI:000298530200013
%A Vielhauer, Oliver
%A Zakhartsev, Maksim
%A Horn, Thomas
%A Takors, Ralf
%A Reuss, Matthias
%C PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS
%D 2011
%I ELSEVIER SCIENCE BV
%J JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
%K myown
%N 32
%P 3859-3870
%R 10.1016/j.jchromb.2011.10.036
%T Simplified absolute metabolite quantification by gas
chromatography-isotope dilution mass spectrometry on the basis of
commercially available source material
%U https://doi.org/10.1016/j.jchromb.2011.10.036
%V 879
%X In the field of metabolomics, GC-MS has rather established itself as a
tool for semi-quantitative strategies like metabolic fingerprinting or
metabolic profiling. Absolute quantification of intra- or extracellular
metabolites is nowadays mostly accomplished by application of diverse
LC-MS techniques. Only few groups have so far adopted GC-MS technology
for this exceptionally challenging task. Besides numerous and deeply
investigated problems related to sample generation, the pronounced
matrix effects in biological samples have led to the almost mandatory
application of isotope dilution mass spectrometry (IDMS) for the
accurate determination of absolute metabolite concentrations.
Nevertheless, access to stable isotope labeled internal standards
(ILIS), which are in many cases commercially unavailable, is quite
laborious and very expensive. Here we present an improved and simplified
gas chromatography-isotope dilution mass spectrometry (GC-IDMS) protocol
for the absolute determination of intra- and extracellular metabolite
levels. Commercially available C-13-labeled algal cells were used as a
convenient source for the preparation of internal standards. Advantages
as well as limitations of the described method are discussed. (C) 2011
Elsevier B.V. All rights reserved.
@article{ISI:000298530200013,
abstract = {{In the field of metabolomics, GC-MS has rather established itself as a
tool for semi-quantitative strategies like metabolic fingerprinting or
metabolic profiling. Absolute quantification of intra- or extracellular
metabolites is nowadays mostly accomplished by application of diverse
LC-MS techniques. Only few groups have so far adopted GC-MS technology
for this exceptionally challenging task. Besides numerous and deeply
investigated problems related to sample generation, the pronounced
matrix effects in biological samples have led to the almost mandatory
application of isotope dilution mass spectrometry (IDMS) for the
accurate determination of absolute metabolite concentrations.
Nevertheless, access to stable isotope labeled internal standards
(ILIS), which are in many cases commercially unavailable, is quite
laborious and very expensive. Here we present an improved and simplified
gas chromatography-isotope dilution mass spectrometry (GC-IDMS) protocol
for the absolute determination of intra- and extracellular metabolite
levels. Commercially available C-13-labeled algal cells were used as a
convenient source for the preparation of internal standards. Advantages
as well as limitations of the described method are discussed. (C) 2011
Elsevier B.V. All rights reserved.}},
added-at = {2018-06-08T11:28:10.000+0200},
address = {{PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS}},
affiliation = {{Vielhauer, O (Reprint Author), Univ Stuttgart, Inst Biochem Engn, Allmandring 31, D-70569 Stuttgart, Germany.
Vielhauer, Oliver; Zakhartsev, Maksim; Horn, Thomas; Takors, Ralf; Reuss, Matthias, Univ Stuttgart, Inst Biochem Engn, D-70569 Stuttgart, Germany.}},
author = {Vielhauer, Oliver and Zakhartsev, Maksim and Horn, Thomas and Takors, Ralf and Reuss, Matthias},
author-email = {{vielhauer@ibvt.uni-stuttgart.de}},
biburl = {https://puma.ub.uni-stuttgart.de/bibtex/274f468aebbe99c337e7578fffc317563/ralftakors},
da = {{2018-01-26}},
doc-delivery-number = {{868NO}},
doi = {{10.1016/j.jchromb.2011.10.036}},
funding-acknowledgement = {{transnational research initiative ``Systems Biology of Microorganisms
(SysMO){''} within network MOSES: ``MicroOrganisnn Systems Biology:
Energy and Saccharomyces cerevisiae{''}}},
funding-text = {{The authors would like to thank Lara Bogner, Andreas Freund, Achim
Hauck, Mira Lenfers-Lucker and Alexander Muller (Institute of
Biochemical Engineering (IBVT), University of Stuttgart, Germany) for
the experimental support. The research was funded by the transnational
research initiative ``Systems Biology of Microorganisms (SysMO){''}
within network MOSES: ``MicroOrganisnn Systems Biology: Energy and
Saccharomyces cerevisiae{''} {[}http://www.sysmo.net/].}},
interhash = {e0f4af3a025146fac0de7f6e30a95bd9},
intrahash = {74f468aebbe99c337e7578fffc317563},
issn = {{1570-0232}},
journal = {{JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES}},
journal-iso = {{J. Chromatogr. B}},
keywords = {myown},
keywords-plus = {{NUCLEAR-MAGNETIC-RESONANCE; CENTRAL CARBON METABOLISM; IN-VIVO ANALYSIS;
ESCHERICHIA-COLI; INTRACELLULAR METABOLITES; SACCHAROMYCES-CEREVISIAE;
LIQUID-CHROMATOGRAPHY; CAPILLARY-ELECTROPHORESIS;
ELECTROSPRAY-IONIZATION; FUNCTIONAL GENOMICS}},
language = {{English}},
month = {{DEC 15}},
number = {{32}},
number-of-cited-references = {{90}},
orcid-numbers = {{Zakhartsev, Maksim/0000-0002-7973-9902}},
pages = {{3859-3870}},
publisher = {{ELSEVIER SCIENCE BV}},
research-areas = {{Biochemistry \& Molecular Biology; Chemistry}},
times-cited = {{22}},
timestamp = {2018-06-08T09:28:10.000+0200},
title = {{Simplified absolute metabolite quantification by gas
chromatography-isotope dilution mass spectrometry on the basis of
commercially available source material}},
type = {{Article}},
unique-id = {{ISI:000298530200013}},
url = {https://doi.org/10.1016/j.jchromb.2011.10.036},
usage-count-last-180-days = {{2}},
usage-count-since-2013 = {{37}},
volume = {{879}},
web-of-science-categories = {{Biochemical Research Methods; Chemistry, Analytical}},
year = {{2011}}
}