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Mass Spectrometry: Overview01:19

Mass Spectrometry: Overview

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Mass spectrometry is an analytical technique used to determine the molecular mass and molecular formula of a compound. The basic principle of mass spectrometry is to generate ions from the analyte molecule and measure these ion abundances against their molecular mass. One common type of ionization, known as electron ionization or EI, bombards the analyte molecules in the gas phase with high-energy electron beams. The electron beams displace an electron from the molecule and leave behind a...
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Tandem Mass Spectrometry01:21

Tandem Mass Spectrometry

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Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and reduce chemical noise during analyte detection. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called...
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Mass Spectrometry: Isotope Effect01:13

Mass Spectrometry: Isotope Effect

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Most elements exist in nature as a mixture of isotopes. The isotopes differ in weight due to their respective number of neutrons. The molecular weight of a molecule is different depending on the specific isotope of its elements involved. As a result, the mass spectrum of the molecule exhibits peaks from the same fragment at multiple positions. The positions of these mass signals depend on the mass differences between isotopes. Furthermore, the intensity of these signals is dependent on the...
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Mass Spectrometry of Amines01:15

Mass Spectrometry of Amines

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In mass spectroscopy, amines undergo fragmentation to give parent ions with odd molecule weights. This observed mass spectrum follows the nitrogen rule; a molecule with an odd number of nitrogen atoms produces a molecular ion with an odd molecular weight. Amines undergo fragmentation through α cleavage, producing nitrogen-containing cations—iminium ions—and alkyl radicals. Mass spectra of aromatic and cyclic aliphatic amines exhibit strong molecular ion peaks, but acyclic...
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Chemical Ionization (CI) Mass Spectrometry01:21

Chemical Ionization (CI) Mass Spectrometry

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The molecular ion peak of a molecule in the mass spectrum provides vital information for molecular identification. However, conventional electron impact ionization can lead to the rapid dissociation of some molecular ions before they reach the detector. A milder ionization method is required to increase the lifetime of such ionized analyte molecules. Chemical ionization (CI) is a gas-phase protonation reaction useful for mass-analyzing analyte molecules that are easily protonated to yield the...
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Mass Spectrometry: Alkene Fragmentation00:59

Mass Spectrometry: Alkene Fragmentation

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Alkenes lose one electron from the unsaturated π bond upon ionization and form stable molecular ions. Further fragmentation of alkenes occurs through three different reaction pathways. The most prominent fragmentation is the cleavage at the allylic position. The resultant allylic carbocation is resonance stabilized. In the mass spectra of terminal alkenes, this fragment appears at a mass-to-charge ratio of 41. In the internal alkenes, where there are two choices of allylic cleavage, the...
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Quantitative Metabolomics of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry
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Mass Spectrometry-Based Metabolomics.

Young-Mo Kim1, Heino M Heyman2

  • 1Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, WA, USA. young-mo.kim@pnnl.gov.

Methods in Molecular Biology (Clifton, N.J.)
|June 8, 2018
PubMed
Summary
This summary is machine-generated.

Mass spectrometry-based metabolomics offers detailed insights into biological systems. This study outlines essential workflows for both global and targeted metabolomic analyses using chromatography-mass spectrometry techniques.

Keywords:
Data processingDerivatizationGas chromatographyLibrary constructionLiquid chromatographyMass spectrometryMetabolite extraction

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Systems Biology

Background:

  • Metabolomics, utilizing mass spectrometry, quantifies and qualifies the metabolome within biological systems.
  • A standard metabolomics workflow involves sample preparation, potential derivatization, instrumental analysis, data processing, and statistical interpretation.

Purpose of the Study:

  • To present general metabolomics workflows.
  • To cover both global and targeted metabolomic analyses.
  • To detail methods using chromatography coupled with mass spectrometry.

Main Methods:

  • Sample preparation with rapid metabolism quenching.
  • Optional chemical derivatization for enhanced analysis.
  • Instrumental measurement using Gas Chromatography-Mass Spectrometry (GC-MS) or Liquid Chromatography-Mass Spectrometry (LC-MS).
  • Data processing and statistical analysis for interpretation.

Main Results:

  • Established robust workflows for metabolomic studies.
  • Demonstrated applicability for both broad (global) and specific (targeted) metabolite profiling.
  • Highlighted the versatility of GC-MS and LC-MS platforms.

Conclusions:

  • The presented workflows provide a foundation for comprehensive metabolomic investigations.
  • These methods enable quantitative and qualitative analysis of biological samples.
  • Effective metabolomics relies on meticulous sample handling, precise instrumentation, and rigorous data analysis.