Targeted Metabolomics
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- Abstract
- Table of Contents
- Materials
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- Literature Cited
Abstract
The metabolome is the terminal downstream product of the genome and consists of the total complement of all the low?molecular?weight molecules (metabolites) in a cell, tissue, or organism. Metabolomics aims to measure a wide breadth of small molecules in the context of physiological stimuli or disease states. Metabolomics methodologies fall into two distinct groups: untargeted metabolomics, an intended comprehensive analysis of all the measurable analytes in a sample including chemical unknowns, and targeted metabolomics, the measurement of defined groups of chemically characterized and biochemically annotated metabolites. The methodologies considered in this unit focus on the processes of conducting targeted metabolomics experiments, and the advantages of this general approach are highlighted herein. This unit outlines procedures for extracting nitrogenous metabolites (including amino acids), lipids, and intermediary metabolites (including TCA cycle oxoacids) from blood plasma. Specifically, protocols are described for analyzing these metabolites using targeted metabolomics experiments based on liquid chromatography?mass spectrometry. Curr. Protoc. Mol. Biol. 98:30.2.1?30.2.24. © 2012 by John Wiley & Sons, Inc.
Keywords: targeted metabolomics; liquid chromatography?mass spectrometry; multiple reaction monitoring
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PDF or HTML at Wiley Online LibraryTable of Contents
- Introduction
- Strategic Planning
- Basic Protocol 1: Targeted Metabolomic Analysis of Hydrophilic Metabolites and Amino Acids from Blood Plasma Using Hydrophilic‐Interaction Liquid Chromatography
- Support Protocol 1: Sample Preparation for Targeted Metabolomics: Extraction of Hydrophilic Metabolites and Amino Acids from Blood Plasma, Cerebrospinal Fluid, and Urine
- Basic Protocol 2: Targeted Metabolomic Analysis of Intermediary Metabolites from Blood Plasma Using Polar Reversed‐Phase Chromatography
- Support Protocol 2: Sample Preparation for Targeted Metabolomics: Extraction of Intermediary Metabolites from Blood Plasma
- Basic Protocol 3: Analysis of Targeted Metabolomic LC‐MS Data
- Commentary
- Literature Cited
- Figures
- Tables
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online LibraryMaterials
Basic Protocol 1: Targeted Metabolomic Analysis of Hydrophilic Metabolites and Amino Acids from Blood Plasma Using Hydrophilic‐Interaction Liquid Chromatography
Materials
Support Protocol 1: Sample Preparation for Targeted Metabolomics: Extraction of Hydrophilic Metabolites and Amino Acids from Blood Plasma, Cerebrospinal Fluid, and Urine
Materials
Basic Protocol 2: Targeted Metabolomic Analysis of Intermediary Metabolites from Blood Plasma Using Polar Reversed‐Phase Chromatography
Materials
Support Protocol 2: Sample Preparation for Targeted Metabolomics: Extraction of Intermediary Metabolites from Blood Plasma
Materials
Basic Protocol 3: Analysis of Targeted Metabolomic LC‐MS Data
Materials
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PDF or HTML at Wiley Online LibraryFigures
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Figure 30.2.1 Relationship between the human genome, proteome, and metabolome (from Gerszten and Wang, ). View Image -
Figure 30.2.2 (A ) Workflow for a LC triple‐quadrupole MS MRM targeted metabolomics experiment. (B ) The resolution of chromatography allows the separation of metabolites even in the case of a shared MRM transition. (C ) Specific precursor/product ion transitions in MRM experiments identify individual metabolites. View Image -
Figure 30.2.3 Example of an experiment to assess biological and technical variance (right, pre and post), variance in sample preparation (center), and the contribution of LC‐MS profiling to overall variance (left). View Image -
Figure 30.2.4 Graph of coefficient of variance (CV) versus integrated peak area for metabolites analyzed from pooled plasma using . CV decreases as integrated peak area increases. View Image
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Literature Cited
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