Simultaneous, Untargeted Metabolic Profiling of Polar and Nonpolar Metabolites by LC‐Q‐TOF Mass Spectrometry
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- Abstract
- Table of Contents
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Abstract
At its most ambitious, untargeted metabolomics aims to characterize and quantify all of the metabolites in a given system. Metabolites are often present at a broad range of concentrations and possess diverse physical properties complicating this task. Performing multiple sample extractions, concentrating sample extracts, and using several separation and detection methods are common strategies to overcome these challenges but require a great amount of resources. This protocol describes the untargeted, metabolic profiling of polar and nonpolar metabolites with a single extraction and using a single analytical platform. Curr. Protoc. Toxicol. 56:4.39.1?4.39.12. © 2013 by John Wiley & Sons, Inc.
Keywords: untargeted metabolomics; LC?MS/MS; hypothesis generation
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PDF or HTML at Wiley Online LibraryTable of Contents
- Introduction
- Basic Protocol 1: Metabolite Extraction
- Basic Protocol 2: LC‐MS/MS
- Basic Protocol 3: Metabolite Identification
- Commentary
- Literature Cited
- Figures
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online LibraryMaterials
Basic Protocol 1: Metabolite Extraction
Materials
Basic Protocol 2: LC‐MS/MS
Materials
Basic Protocol 3: Metabolite Identification
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Figure 4.39.1 (Above) Sample total ion chromatogram (TIC) from rat plasma; (Below) normalized extracted ion chromatograms for several polar and nonpolar metabolites. View Image -
Figure 4.39.2 Example of a PCA‐DA (principal component analysis‐discriminant analysis) scores plot. Legend denotes duration of exposure (skeletal muscle cells) to xanthohumol. View Image -
Figure 4.39.3 Example volcano plot of metabolomics dataset to investigate differences in metabolites for 15 versus 90‐minute treatment times. View Image -
Figure 4.39.4 Metabolite matching by MS and MS/MS. View Image -
Figure 4.39.5 Confirmation of experimental isotope ratio to theoretical isotope ratio. View Image -
Figure 4.39.6 Comparison of experimentally identified metabolite against chemical standard by (A ) LC‐MS and (B ) MS/MS. View Image
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Literature Cited
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