title: 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.
abstract: 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.
authors.affiliation: CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
authors.forename: Shili
authors.initials: S
authors.lastname: Chen
authors.forename: Miriam
authors.initials: M
authors.lastname: Hoene
authors.forename: Jia
authors.initials: J
authors.lastname: Li
authors.forename: Yanjie
authors.initials: Y
authors.lastname: Li
authors.forename: Xinjie
authors.initials: X
authors.lastname: Zhao
authors.forename: Hans-Ulrich
authors.initials: HU
authors.lastname: Häring
authors.forename: Erwin D
authors.initials: ED
authors.lastname: Schleicher
authors.forename: Cora
authors.initials: C
authors.lastname: Weigert
authors.forename: Guowang
authors.initials: G
authors.lastname: Xu
authors.forename: Rainer
authors.initials: R
authors.lastname: Lehmann
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008099:Liver / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry; D055432:Metabolomics / Q000379:methods*; D008738:Methyl Ethers / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009132:Muscles / Q000378:metabolism
chemical_list: D008738:Methyl Ethers; C043243:methyl tert-butyl ether