Related Experiment Video
Updated: Jul 26, 2025
09:06
Enhanced Sample Multiplexing of Tissues Using Combined Precursor Isotopic Labeling and Isobaric Tagging cPILOT
Published on: May 1, 2017
7.0K
DiffN Selection of Tandem Mass Spectrometry Precursors
Tyler S Larson1, Cameron D Worthington1, Matthew D Verber2
1Department of Chemistry, University of North Carolina at Chapel Hill, Kenan and Caudill Laboratories, Chapel Hill, North Carolina 27599-3290, United States.
Analytical Chemistry
|June 13, 2023
Summary
A new DiffN data-dependent acquisition approach improves biomarker discovery by selecting ions based on differential intensity between samples, outperforming the traditional TopN method for lipidomic analysis.
Area of Science:
- Mass spectrometry
- Lipidomics
- Biomarker discovery
Background:
- Current data-dependent acquisition (DDA) methods like TopN rely on absolute ion intensity, potentially missing low-abundance biomarkers.
- Identifying differentially abundant molecules is crucial for understanding biological processes and disease states.
Purpose of the Study:
- To introduce and validate a novel DDA approach, DiffN, for enhanced biomarker discovery.
- To compare the efficacy of DiffN against the conventional TopN approach in lipidomic analyses.
Main Methods:
- Development of the DiffN approach utilizing a dual nano-electrospray ionization source for parallel sample analysis.
- Application of DiffN and TopN DDA methods to compare lipid profiles of colorectal cancer cell lines (SW480 and SW620).
Main Results:
- The DiffN approach successfully targets ions with the largest fold changes between samples.
- DiffN demonstrated a higher probability of identifying significant lipid biomarkers compared to TopN in colorectal cancer cell lines.
- Analysis of SW480 and SW620 cell lines revealed differences in lipid abundance, highlighting DiffN's utility.
Conclusions:
- The DiffN DDA approach offers a more efficient strategy for selecting precursor ions of interest in lipidomics.
- DiffN enhances the likelihood of biomarker discovery by focusing on relative intensity differences.
- This method holds potential for broader applications in shotgun analyses of various molecule classes.
Related Concept Videos
01:21Tandem Mass Spectrometry
1.1K
Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and signal-to-noise ratio for the analyte. 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 collision-induced...
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 collision-induced...
1.1K
01:33Peptide Identification Using Tandem Mass Spectrometry
6.6K
Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
6.6K
01:21Mass Spectrometry: Complex Analysis
830
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...
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
830
01:16Mass Spectrometers
5.7K
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:
5.7K
01:19Mass Spectrometry: Overview
5.5K
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...
5.5K
01:20Mass Spectrometry: Molecular Fragmentation Overview
3.3K
The ionization of a molecule into a molecular ion inside the mass spectrometer causes instability in the molecule's structure due to the loss of an electron. This eventually leads to the fragmentation or breaking of some bonds in the molecule. The fragmentation occurs predominantly at specific bonds to yield relatively stable fragments.
One type of fragmentation pattern is the cleavage of a single bond in the molecular ion. The cleavage leads to a radical and a cation. The cleavage can...
One type of fragmentation pattern is the cleavage of a single bond in the molecular ion. The cleavage leads to a radical and a cation. The cleavage can...
3.3K