title: Development of high throughput 96-blade solid phase microextraction-liquid chromatrography-mass spectrometry protocol for metabolomics.
abstract: In metabolomics, the workflow for quantitative and comprehensive metabolic mapping of cellular metabolites can be a very challenging undertaking. Sampling and sample preparation play a significant role in untargeted analysis, as they may affect the composition of the analyzed metabolome. In the current work, different solid phase microextraction (SPME) coating chemistries were developed and applied to provide simultaneous extraction of a wide range of both hydrophobic and hydrophilic cellular metabolites produced by a model organism, Escherichia coli. Three different LC-MS methods were also evaluated for analysis of extracted metabolites. Finally, over 200 cellular metabolites were separated and detected with widely varying hydrophobicities ranging within -7 < log P < 15, including amino acids, peptides, nucleotides, carbohydrates, polycarboxylic acids, vitamins, phosphorylated compounds, and lipids such as hydrophobic phospholipids, prenol lipids, and fatty acids at the stationary phase of the E. coli life cycle using the developed 96-blade SPME-LC-MS method.
authors.affiliation: Department of Chemistry, University of Waterloo, 200 University Avenue West, Waterloo, Ontario N2L 3G1, Canada.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Mousavi
authors.affiliation: Department of Chemistry, University of Waterloo, 200 University Avenue West, Waterloo, Ontario N2L 3G1, Canada.
authors.forename: Barbara
authors.initials: B
authors.lastname: Bojko
authors.affiliation: Department of Chemistry, University of Waterloo, 200 University Avenue West, Waterloo, Ontario N2L 3G1, Canada. Electronic address: janusz@uwaterloo.ca.
authors.forename: Janusz
authors.initials: J
authors.lastname: Pawliszyn
mesh_terms: D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000378:metabolism*; D055432:Metabolomics; D025341:Principal Component Analysis; D052617:Solid Phase Microextraction; D053719:Tandem Mass Spectrometry
keywords: 96 blade system; Escherichia coli; High throughput; In-vivo solid phase micro extraction; Metabolomics; Optimization