期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1298, 期 -, 页码 9-16出版社
ELSEVIER
DOI: 10.1016/j.chroma.2013.05.019
关键词
Liquid chromatography/mass spectrometry; Metabolic profiling; Sample pretreatment; Methyl tert-butyl ether (MTBE); Biopsy; Metabolomics
资金
- Sino-German Center for Research Promotion (DFG) [GZ 753, LE 1391/1-1]
- Sino-German Center for Research Promotion (NSFC) [GZ 753]
- State Key Science & Technology Project for Infectious Diseases of China [2012ZX100002011, 2012ZX10002009]
- Stiftung far Pathobiochemie und Molekulare Diagnostik of the German Society of Clinical Chemistry and Laboratory Medicine
- Deutsche Forschungsgemeinschaft [GRK 1302]
- German Federal Ministry of Education and Research (BMBF)
- Kompetenznetz Diabetes mellitus (Competence Network for Diabetes mellitus)
- Federal Ministry of Education and Research [FkZ 01 GI 1104A]
- foundation and creative research group project (NSFC) [21175132, 21021004]
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. (C) 2013 Elsevier B.V. All rights reserved.
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