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Development and optimization of a metabolomic method for analysis of adherent cell cultures

Danielsson, Anders LU ; Moritz, Thomas; Mulder, Hindrik LU and Spégel, Peter LU (2010) In Analytical Biochemistry 404(1). p.30-39
Abstract
In this investigation, a gas chromatography/mass spectrometry (GC/MS)-based metabolomic protocol for adherent cell cultures was developed using statistical design of experiments. Cell disruption, metabolite extraction, and the GC/MS settings were optimized aiming at a gentle, unbiased, sensitive, and high-throughput metabolomic protocol. Due to the heterogeneity of the metabolome and the inherent selectivity of all analytical techniques, development of unbiased protocols is highly complex. Changing one parameter of the protocol may change the response of many groups of metabolites. In this investigation, statistical design of experiments and multivariate analysis also allowed such interaction effects to be taken into account. The protocol... (More)
In this investigation, a gas chromatography/mass spectrometry (GC/MS)-based metabolomic protocol for adherent cell cultures was developed using statistical design of experiments. Cell disruption, metabolite extraction, and the GC/MS settings were optimized aiming at a gentle, unbiased, sensitive, and high-throughput metabolomic protocol. Due to the heterogeneity of the metabolome and the inherent selectivity of all analytical techniques, development of unbiased protocols is highly complex. Changing one parameter of the protocol may change the response of many groups of metabolites. In this investigation, statistical design of experiments and multivariate analysis also allowed such interaction effects to be taken into account. The protocol was validated with respect to linear range, precision, and limit of detection in a clonal rat insulinoma cell line (INS-1 832/13). The protocol allowed high-throughput profiling of metabolites covering the major metabolic pathways. The majority of metabolites displayed a linear range from a single well in a 96-well plate up to a 10 cm culture dish. The method allowed a total of 47 analyses to be performed in 24 h. (C) 2010 Elsevier Inc. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Mass spectrometry, Gas chromatography, Metabolomics, Statistical design of experiments, Cell culture
in
Analytical Biochemistry
volume
404
issue
1
pages
30 - 39
publisher
Elsevier
external identifiers
  • wos:000279803400005
  • scopus:77954086132
  • pmid:20417172
ISSN
1096-0309
DOI
10.1016/j.ab.2010.04.013
language
English
LU publication?
yes
id
c1924d26-1506-4f7b-8c46-d0e78239487c (old id 1657721)
date added to LUP
2010-08-24 12:55:11
date last changed
2018-05-29 10:33:47
@article{c1924d26-1506-4f7b-8c46-d0e78239487c,
  abstract     = {In this investigation, a gas chromatography/mass spectrometry (GC/MS)-based metabolomic protocol for adherent cell cultures was developed using statistical design of experiments. Cell disruption, metabolite extraction, and the GC/MS settings were optimized aiming at a gentle, unbiased, sensitive, and high-throughput metabolomic protocol. Due to the heterogeneity of the metabolome and the inherent selectivity of all analytical techniques, development of unbiased protocols is highly complex. Changing one parameter of the protocol may change the response of many groups of metabolites. In this investigation, statistical design of experiments and multivariate analysis also allowed such interaction effects to be taken into account. The protocol was validated with respect to linear range, precision, and limit of detection in a clonal rat insulinoma cell line (INS-1 832/13). The protocol allowed high-throughput profiling of metabolites covering the major metabolic pathways. The majority of metabolites displayed a linear range from a single well in a 96-well plate up to a 10 cm culture dish. The method allowed a total of 47 analyses to be performed in 24 h. (C) 2010 Elsevier Inc. All rights reserved.},
  author       = {Danielsson, Anders and Moritz, Thomas and Mulder, Hindrik and Spégel, Peter},
  issn         = {1096-0309},
  keyword      = {Mass spectrometry,Gas chromatography,Metabolomics,Statistical design of experiments,Cell culture},
  language     = {eng},
  number       = {1},
  pages        = {30--39},
  publisher    = {Elsevier},
  series       = {Analytical Biochemistry},
  title        = {Development and optimization of a metabolomic method for analysis of adherent cell cultures},
  url          = {http://dx.doi.org/10.1016/j.ab.2010.04.013},
  volume       = {404},
  year         = {2010},
}