Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Mass Spectrometry: Carboxylic Acid, Ester, and Amide Fragmentation01:01

Mass Spectrometry: Carboxylic Acid, Ester, and Amide Fragmentation

984
The fragmentation patterns observed for compounds such as carboxylic acids, esters, and amides in the mass spectra include ⍺-cleavage and McLafferty rearrangement. Fragmentation by ⍺-cleavage preferentially occurs at the carbon-carbon bond at the ⍺-position next to the carboxylic group to generate a neutral radical and a cation. Long chain compounds with hydrogen at their γ-carbon undergo McLafferty rearrangement to give a radical cation and a neutral alkene.
For example,...
984
Mass Spectrometry: Overview01:19

Mass Spectrometry: Overview

3.6K
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 electrospray 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...
3.6K
Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

650
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...
650
Mass Spectrometry: Amine Fragmentation00:55

Mass Spectrometry: Amine Fragmentation

1.4K
Amines can be identified using mass spectroscopy based on their characteristic fragmentation patterns. The molecular ions of amines undergo fragmentation via ⍺-cleavage. The ⍺-cleavage of the carbon-carbon bonds in amines generates an alkyl radical and resonance-stabilized nitrogen-containing cation.
In amines, the number of nitrogen atoms affects the mass of the molecular ion, which is described by the nitrogen rule of mass spectrometry. This rule states that a compound containing...
1.4K
Mass Spectrometry of Amines01:19

Mass Spectrometry of Amines

4.1K
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 parent ion with an odd molecular weight. The remaining fragments have an even mass.
Amines undergo fragmentation through α cleavage, producing nitrogen-containing cations—iminium ions—and alkyl radicals. Mass spectra of aromatic and cyclic aliphatic amines exhibit...
4.1K
Mass Spectrometers01:16

Mass Spectrometers

4.8K
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:
4.8K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Single-Ion Imaging Native Mass Spectrometry: Unraveling the Structural Features and Dissociation Energetics of Macromolecular Assemblies.

Journal of the American Society for Mass Spectrometry·2026
Same author

Spatiotemporal Mapping of Drug Incorporation in Human Nails by MALDI-FTICR-MSI.

Journal of the American Society for Mass Spectrometry·2026
Same author

A Combined MS/MS and IMS Study Into the Fragmentation Pathway of Nifedipine.

Journal of mass spectrometry : JMS·2026
Same author

BMP-7 mRNA delivered by Fibrin-CaP scaffolds activates osteogenic programs in vivo as evidenced by transcriptomic and proteomic analyses.

Bioactive materials·2026
Same author

Isotope Decluttering Reduces Spectral Complexity while Maintaining Protein Structure.

Analytical chemistry·2026
Same author

The role of angiotensin II in cardiovascular disease-induced cancer growth.

Cardio-oncology (London, England)·2026
Same journal

Serine Octamer Substitution Reactions With α-Hydroxy Acids.

Rapid communications in mass spectrometry : RCM·2026
Same journal

Cryogen-Aided Chromatographic Purification of Millimole Quantities of Methane for Spectroscopic Clumped Isotope Analysis.

Rapid communications in mass spectrometry : RCM·2026
Same journal

Reaffirming the Consistency of the Original Stock of NBS 19 Limestone and Its Byproduct With Finer Grain Size for δ<sup>13</sup>C and δ<sup>18</sup>O Calibrations-Both Distributed Internationally.

Rapid communications in mass spectrometry : RCM·2026
Same journal

Three New Calcium Formate Reference Materials for δ<sup>13</sup>C Measurements and a Redetermination of the R(<sup>13</sup>C/<sup>12</sup>C) Ratio for VPDB Based on Proton Nuclear Magnetic Resonance Measurements.

Rapid communications in mass spectrometry : RCM·2026
Same journal

Correction to "Zinc/Silver-Based Hydrogen Sulfide Trapping as an Alternative to Cadmium for Sulfur Stable Isotope Analysis".

Rapid communications in mass spectrometry : RCM·2026
Same journal

The Possibility of X-Type Ions With Linear Saturated Non-Reducing Termini Dissociated From Potasiated Saikosaponins.

Rapid communications in mass spectrometry : RCM·2026
See all related articles

Related Experiment Video

Updated: May 10, 2025

A Mass Spectrometry-Based Proteomics Approach for Global and High-Confidence Protein R-Methylation Analysis
09:40

A Mass Spectrometry-Based Proteomics Approach for Global and High-Confidence Protein R-Methylation Analysis

Published on: April 28, 2022

2.2K

Buffer 4-Ethylmorpholinium/Acetate: Exploring a New Alternative Buffer for Native Mass Spectrometry.

Darya Hadavi1, Che Yee Ng1, Yuandi Zhao1

  • 1Maastricht Multi Modal Molecular Imaging (M4i) Institute, Division of Imaging Mass Spectrometry (IMS), Maastricht University, Maastricht, The Netherlands.

Rapid Communications in Mass Spectrometry : RCM
|April 21, 2025
PubMed
Summary
This summary is machine-generated.

A new 4-ethylmorpholinium/acetate (4EM/A) buffer preserves protein structure and bioactivity in native mass spectrometry (MS) studies. This buffer offers enhanced conformational stability and enables analysis of challenging proteins like cardiac troponin T.

Keywords:
charge state distributionethylmorpholineion mobility spectrometrynative mass spectrometryprotein complexes

More Related Videos

Using a Cyclic Ion Mobility Spectrometer for Tandem Ion Mobility Experiments
08:40

Using a Cyclic Ion Mobility Spectrometer for Tandem Ion Mobility Experiments

Published on: January 20, 2022

4.1K
Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies
10:01

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies

Published on: November 28, 2017

19.5K

Related Experiment Videos

Last Updated: May 10, 2025

A Mass Spectrometry-Based Proteomics Approach for Global and High-Confidence Protein R-Methylation Analysis
09:40

A Mass Spectrometry-Based Proteomics Approach for Global and High-Confidence Protein R-Methylation Analysis

Published on: April 28, 2022

2.2K
Using a Cyclic Ion Mobility Spectrometer for Tandem Ion Mobility Experiments
08:40

Using a Cyclic Ion Mobility Spectrometer for Tandem Ion Mobility Experiments

Published on: January 20, 2022

4.1K
Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies
10:01

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies

Published on: November 28, 2017

19.5K

Area of Science:

  • Biophysical Chemistry
  • Analytical Chemistry
  • Structural Biology

Background:

  • Native mass spectrometry (MS) requires volatile, electrospray ionization (ESI)-compatible buffers.
  • Traditional buffers like ammonium acetate (AA) can induce protein unfolding and acidification during ESI-MS.
  • A buffer is needed to maintain protein native state and simulate physiological conditions.

Purpose of the Study:

  • To evaluate the 4-ethylmorpholinium/acetate (4EM/A) buffer for native MS analysis.
  • To compare 4EM/A with ammonium acetate (AA) for protein and protein complex analysis.
  • To assess the impact of 4EM/A on protein bioactivity and conformational integrity.

Main Methods:

  • Compared 4EM/A buffer to AA buffer for proteins (5–103 kDa) and protein complexes.
  • Analyzed native-MS profiles, collision cross-section (CCS) values, and arrival time distributions (ATDs).
  • Assessed protein bioactivity and analyzed challenging proteins like the human cardiac troponin complex (cTn complex).

Main Results:

  • 4EM/A resulted in lower protein charge states, preserving native folding during nano-ESI and MS.
  • Ion mobility showed reduced conformational variation in 4EM/A, indicating enhanced stability and compactness.
  • Lysozyme exhibited increased bioactivity in 4EM/A; 4EM/A enabled first-time native-MS analysis of cardiac troponin T (cTnT).

Conclusions:

  • Introduced 4EM/A (pKa 7.72/4.76) as a superior buffer for native MS.
  • 4EM/A maintains protein and protein complex bioactivity and conformational integrity.
  • 4EM/A is a promising tool for advanced native MS applications.