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Related Concept Videos

¹H NMR of Labile Protons: Deuterium (²H) Substitution00:48

¹H NMR of Labile Protons: Deuterium (²H) Substitution

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This lesson illustrates the role of deuterium substitution in simplifying the NMR spectrum of compounds comprising labile protons. One method employed is the use of deuterium. Amongst the three isotopes of hydrogen, deuterium (2H) has a nucleus composed of one proton and one neutron. When the D2O solvent is added to a pure dry ethanol solution, its labile proton is substituted with deuterium.
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High-Resolution Mass Spectrometry (HRMS)01:15

High-Resolution Mass Spectrometry (HRMS)

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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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¹³C NMR: Distortionless Enhancement by Polarization Transfer (DEPT)01:20

¹³C NMR: Distortionless Enhancement by Polarization Transfer (DEPT)

1.8K
When proton-coupled carbon-13 spectra are simplified by a broadband proton decoupling technique, structural information about the coupled protons is lost. Distortionless enhancement by polarization transfer (DEPT) is a technique that provides information on the number of hydrogens attached to each carbon in a molecule. While the DEPT experiment utilizes complex pulse sequences, the pulse delay and flip angle are specifically manipulated. The resulting signals have different phases depending on...
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Mass Spectrometry: Isotope Effect01:13

Mass Spectrometry: Isotope Effect

5.0K
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: 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.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
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Tandem Mass Spectrometry01:21

Tandem Mass Spectrometry

2.9K
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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Related Experiment Video

Updated: Mar 26, 2026

A Hydrogen-Deuterium Exchange Mass Spectrometry HDX-MS Platform for Investigating Peptide Biosynthetic Enzymes
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Hydrogen Exchange Mass Spectrometry.

Leland Mayne1

  • 1Johnson Research Foundation, Department of Biochemistry and Biophysics, Perelman School of Medicine, The University of Pennsylvania, Philadelphia, Pennsylvania, USA.

Methods in Enzymology
|January 22, 2016
PubMed
Summary
This summary is machine-generated.

Hydrogen exchange (HX) methods coupled with mass spectrometry (MS) now enable high-resolution structural studies of larger proteins. This advancement provides unprecedented insights into protein structure, energetics, and dynamics.

Keywords:
Epitope mappingHDX-MSHX-MSHydrogen exchangeHydrogen–deuterium exchangeMass spectrometryProtein dynamicsProtein foldingProtein stabilityProtein structure

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Area of Science:

  • Biochemistry
  • Structural Biology
  • Analytical Chemistry

Background:

  • Hydrogen exchange (HX) techniques are crucial for understanding protein structure, energetics, and dynamics.
  • Traditional HX methods have limitations in studying larger proteins and achieving high structural resolution.

Purpose of the Study:

  • To introduce the integration of mass spectrometry (MS) with HX analysis.
  • To extend the capabilities of HX studies to larger protein systems.
  • To achieve high structural resolution in protein dynamics analysis.

Main Methods:

  • Utilizing hydrogen exchange (HX) labeling experiments.
  • Employing mass spectrometry (MS) for analyzing HX data.
  • Integrating fragmentation-separation techniques with MS-based HX analysis.

Main Results:

  • The combined HX-MS approach allows for the study of larger proteins.
  • High structural resolution is achievable for protein dynamics.
  • New information regarding protein structure and energetics can be obtained.

Conclusions:

  • The synergy of HX and MS significantly advances protein structural studies.
  • This methodology offers enhanced capabilities for investigating protein behavior.
  • The discussed experimental aspects are key for successful HX-MS data acquisition and analysis.