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Chen, S.L.* ; Hoene, M. ; Li, J.* ; Li, Y.J.* ; Zhao, X.J.* ; Häring, H.-U. ; Schleicher, E.D. ; Weigert, C. ; Xu, G.W.* ; Lehmann, R.

Simultaneous extraction of metabolome and lipidome with methyl tert-butyl ether from a single small tissue sample for ultra-high performance liquid chromatography/mass spectrometry.

J. Chromatogr. A 1298, 9-16 (2013)
DOI PMC
Open Access Green as soon as Postprint is submitted to ZB.
A common challenge for scientists working with animal tissue or human biopsy samples is the limitation of material and consequently, the difficulty to perform comprehensive metabolic profiling within one experiment. Here, we present a novel approach to simultaneously perform targeted and non-targeted metabolomics as well as lipidomics from one small piece of liver or muscle tissue by ultra-high performance liquid chromatography/mass spectrometry (UHPLC/MS) following a methyl tert-butyl ether (MTBE)-based extraction. Equal relative amounts of the resulting polar and non-polar fractions were pooled, evaporated and reconstituted in the appropriate solvent for UHPLC/MS analysis. This mix was comparable or superior in yield and reproducibility to a standard 80% methanol extraction for the profiling of polar and lipophilic metabolites (free carnitine, acylcarnitines and FFA). The mix was also suitable for non-targeted metabolomics, an easy measure to increase the metabolite coverage by 30% relative to using the polar fraction alone. Lipidomics was performed from an aliquot of the non-polar fraction. This novel strategy could successfully be applied to one mouse soleus muscle with a dry weight of merely 2.5 mg. By enabling a simultaneous profiling of lipids and metabolites with mixed polarity while saving material for molecular, biochemical or histological analyses, our approach may open up new perspectives toward a comprehensive investigation of small, valuable tissue samples.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Liquid Chromatography/mass Spectrometry ; Metabolic Profiling ; Sample Pretreatment ; Methyl Tert-butyl Ether (mtbe) ; Biopsy ; Metabolomics; Fatty-acid-composition ; Mass-spectrometry ; Muscle ; Resistance ; Disease ; Plasma ; Brain ; Liver ; Serum ; Model
ISSN (print) / ISBN 0021-9673
e-ISSN 1873-3778
Journal Journal of Chromatography A
Quellenangaben Volume: 1298, Issue: , Pages: 9-16 Article Number: , Supplement: ,
Publisher Elsevier
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) Institute of Diabetes Research and Metabolic Diseases (IDM)