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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1657721
- author
- Danielsson, Anders
LU
; Moritz, Thomas
; Mulder, Hindrik
LU
and Spégel, Peter
LU
- organization
- publishing date
- 2010
- 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
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Molecular Metabolism (013212001), Analytical Chemistry (S/LTH) (011001004)
- id
- c1924d26-1506-4f7b-8c46-d0e78239487c (old id 1657721)
- date added to LUP
- 2016-04-01 10:33:23
- date last changed
- 2022-03-04 20:43:09
@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}},
keywords = {{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}},
doi = {{10.1016/j.ab.2010.04.013}},
volume = {{404}},
year = {{2010}},
}