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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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Axoplasm Isolation from Rat Sciatic Nerve
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Mapping Lipid Distribution in Rat Sciatic Nerve Using Imaging Mass Spectrometry.

Roberto Fernández1, Jone Garate1, Beatriz Abad2

  • 1Faculty of Science and Technology, Department of Physical Chemistry, University of the Basque Country (UPV/EHU), Leioa, Spain.

Methods in Molecular Biology (Clifton, N.J.)
|July 15, 2018
PubMed
Summary

This study presents a new protocol for analyzing lipid expression and distribution in rat sciatic nerves. The method combines MALDI-IMS and UHPLC-MS/MS to map lipid species across nerve areas.

Keywords:
Imaging mass spectrometryLipidomicsMALDISciatic nerveUHPLC

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Area of Science:

  • Neuroscience
  • Biochemistry
  • Analytical Chemistry

Background:

  • Lipids are crucial cellular components involved in signaling and biological functions.
  • Lipid dysregulation often indicates cellular dysfunction.
  • Understanding lipid profiles in neural tissues is vital for neuroscience research.

Purpose of the Study:

  • To develop and describe a protocol for analyzing lipid expression in rat sciatic nerves.
  • To investigate the spatial distribution of lipids within different anatomical regions of the sciatic nerve.
  • To provide a comprehensive method for lipidomic analysis in neural tissues.

Main Methods:

  • The protocol integrates Matrix-Assisted Laser Desorption/Ionization - Imaging Mass Spectrometry (MALDI-IMS) for spatial lipid analysis.
  • Ultra-High-Performance Liquid Chromatography coupled with tandem Mass Spectrometry (UHPLC-MS/MS) is employed for lipid identification and quantification.
  • These techniques are combined to achieve high-throughput lipidomic profiling and tissue mapping.

Main Results:

  • The protocol successfully identifies and maps a wide range of lipid species within the rat sciatic nerve.
  • It allows for the detailed analysis of lipid expression patterns across distinct anatomical areas of the nerve.
  • The study demonstrates the feasibility of combining MALDI-IMS and UHPLC-MS/MS for comprehensive neural lipidomics.

Conclusions:

  • The described protocol offers a powerful approach for detailed lipidomic analysis of nerve tissues.
  • This method enables the cartography of lipid distribution, aiding in the understanding of nerve function and disease.
  • The combined techniques provide unprecedented insights into the complex lipid landscape of the sciatic nerve.