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Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

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Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
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Mass spectrometry is a powerful characterization technique that can identify and separate a wide variety of compounds ranging from chemical to biological entities, based on their mass-to-charge ratio (m/z). The instruments that allow this detection, known as mass spectrometers, have three components: an ion source, a mass analyzer, and a detector. These spectrometers differ based on the nature of their ion source and analyzers.Matrix-assisted laser desorption ionization (MALDI) is a commonly...
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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 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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Mass Spectrometers

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This lesson details the instrumentation of a mass spectrometer—a physical instrument to perform mass spectrometry on analyte molecules and record the characteristic mass spectra. This is achieved via three chief functions:
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High-Resolution Mass Spectrometry (HRMS)01:15

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The resolution of a mass spectrometer depends on the efficiency of separating ions with different ion masses. The mass of an atom is approximated to the sum of the masses of protons and neutrons inside, considering the masses of protons and neutrons as equal. However, the masses of the proton (1.6726 × 10−24 g) and neutron (1.6749 × 10−24 g) are not truly equal. There is a minor error in the expression of atomic masses relative to the simplest atom of hydrogen. For...
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A Strategy for Sensitive, Large Scale Quantitative Metabolomics
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LC-Mass Spectrometry for Metabolomics.

Allyson L Dailey1

  • 1Department of Chemistry and Biochemistry, George Mason University, 10920 George Mason Circle, MS1A9, Manassas, VA, 20110, USA. adailey3@gmu.edu.

Methods in Molecular Biology (Clifton, N.J.)
|May 15, 2017
PubMed
Summary

New liquid chromatography-mass spectrometry (LC-MS) methods enhance metabolomics research by enabling the detection of previously uncaptured metabolites in tissues and feces. This protocol details sample preparation and data acquisition for comprehensive analysis.

Keywords:
FecesGlobal metabolomicsLC-MSMetabolomicsReversed phaseSample processingTissues

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Area of Science:

  • Metabolomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Metabolomics research is advancing with new technologies.
  • Gas chromatography-mass spectrometry (GC-MS) has limitations in metabolite detection.
  • Liquid chromatography-mass spectrometry (LC-MS) offers expanded metabolite profiling capabilities.

Purpose of the Study:

  • To describe a protocol for global metabolomic analysis.
  • To detail sample preparation and data acquisition procedures.
  • To facilitate the study of metabolites in biological samples like tissues and feces.

Main Methods:

  • Utilizing liquid chromatography-mass spectrometry (LC-MS).
  • Customizing LC columns and mass spectrometer parameters.
  • Developing a protocol for sample preparation.
  • Establishing a data acquisition strategy.

Main Results:

  • LC-MS enables the detection of a wider range of metabolites compared to GC-MS.
  • The described protocol allows for tailored instrument configuration.
  • Successful application to global metabolomic analysis of tissues and feces.

Conclusions:

  • The protocol refines metabolomic analysis through advanced LC-MS technology.
  • Customizable LC-MS enhances the ability to capture diverse metabolites.
  • This method provides a robust approach for comprehensive metabolomic studies.