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Related Concept Videos

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: Overview01:19

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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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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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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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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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Semi-Quantitative Analysis of Peptidoglycan by Liquid Chromatography Mass Spectrometry and Bioinformatics
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Quantitative Extraction and Mass Spectrometry Analysis at a Single-Cell Level.

Ruichuan Yin1, Venkateshkumar Prabhakaran2, Julia Laskin1

  • 1Department of Chemistry , Purdue University , West Lafayette , Indiana 47907 , United States.

Analytical Chemistry
|June 7, 2018
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Summary

Localized electroosmotic extraction enables quantitative mass spectrometry of sub-cellular biomolecules. This method precisely extracts picoliter volumes from live cells for sensitive metabolite analysis.

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Area of Science:

  • Analytical Chemistry
  • Cell Biology
  • Biochemistry

Background:

  • Quantitative live cell mass spectrometry at the subcellular level is challenging.
  • Precise extraction of sub-picoliter volumes is required for sensitive analysis and quantification without prior separation.

Purpose of the Study:

  • To demonstrate localized electroosmotic extraction for quantitative mass spectrometry of biomolecules from live cells.
  • To enable analysis of picoliter volumes extracted from subcellular locations.

Main Methods:

  • Localized electroosmotic extraction using a nanopipette and two electrodes.
  • Direct analysis of extracted analytes via nanoelectrospray ionization (nanoESI) mass spectrometry.
  • Quantification using a stable isotope-labeled standard (glucose-d2).

Main Results:

  • Detection of over 50 metabolites, including sugars and flavonoids, in 2-5 pL of Allium cepa cytoplasm.
  • Linear relationship observed between glucose concentration and signal ratio with glucose-d2 standard, indicating quantitative capability.
  • Efficient separation of hydrophilic and hydrophobic analytes based on electrolyte partitioning.

Conclusions:

  • Localized electroosmotic extraction is a viable method for quantitative mass spectrometry of small cellular volumes.
  • The technique provides insights into metabolite properties and enables sensitive subcellular analysis.
  • This approach advances the field of live cell metabolomics.