Evaluation of extraction processes for intracellular metabolite profiling of mammalian cells: matching extraction approaches to cell type and metabolite targets

In this study we report on the optimisation of the technologies for generation of a global metabolomics profile for intracellular metabolites in Chinese hamster ovary (CHO) cells. We evaluated the effectiveness of a range of different extraction methods applied to CHO cells which had been quenched u...

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Bibliographic Details
Published in:Metabolomics 2010-09, Vol.6 (3), p.427-438
Main Authors: Sellick, Christopher A, Knight, David, Croxford, Alexandra S, Maqsood, Arfa R, Stephens, Gill M, Goodacre, Royston, Dickson, Alan J
Format: Article
Language:English
Subjects:
Antibody
Biochemistry
Biomedical and Life Sciences
Biomedicine
bioprocessing
Cell Biology
CHO
Developmental Biology
GC-MS
glutamate-ammonia ligase
Life Sciences
metabolomics
Molecular Medicine
Original Article
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Summary:In this study we report on the optimisation of the technologies for generation of a global metabolomics profile for intracellular metabolites in Chinese hamster ovary (CHO) cells. We evaluated the effectiveness of a range of different extraction methods applied to CHO cells which had been quenched using a previously optimised approach. The extraction methods tested included cold methanol, hot ethanol, acid, alkali and methanol/chloroform plus combinations of these. The extraction of metabolites using two 100% methanol extractions followed by a final water extraction recovered the largest range of metabolites. For the majority of metabolites, extracts generated in this manner exhibited the greatest recovery with high reproducibility. Therefore, this was the best extraction method for attaining a global metabolic profile from a single sample. However, another parallel extraction method (e.g. alkali) may also be required to maximise the range of metabolites recovered (e.g. non-polar metabolites).
ISSN:1573-3882
1573-3890
DOI:10.1007/s11306-010-0216-9