Lund University is celebrating 350 years. Read more on lunduniversity.lu.se

Menu

Javascript is not activated in your browser. This website needs javascript activated to work properly.

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

Author:
  • Anders Danielsson
  • Thomas Moritz
  • Hindrik Mulder
  • Peter Spégel
Publishing year: 2010
Language: English
Pages: 30-39
Publication/Series: Analytical Biochemistry
Volume: 404
Issue: 1
Document type: Journal article
Publisher: Elsevier

Abstract english

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.

Keywords

  • Endocrinology and Diabetes
  • Mass spectrometry
  • Gas chromatography
  • Metabolomics
  • Statistical design of experiments
  • Cell culture

Other

Published
  • ISSN: 1096-0309
Peter Spegel
E-mail: peter.spegel [at] chem.lu.se

Researcher

Centre for Analysis and Synthesis

1

Lund University Diabetes Centre, CRC, SUS Malmö, Entrance 72, House 91:12. SE-205 02 Malmö. Telephone: +46 40 39 10 00