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Related Concept Videos

Peptide Identification Using Tandem Mass Spectrometry01:33

Peptide Identification Using Tandem Mass Spectrometry

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...
Tandem Mass Spectrometry01:21

Tandem Mass Spectrometry

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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Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization
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Semi-automated alignment and quantification of peaks using parallel factor analysis for comprehensive two-dimensional

Robert C Allen1, Sarah C Rutan

  • 1Virginia Commonwealth University, 1001 West Main Street, P.O. Box 842006, Richmond, VA 23284-2006, USA.

Analytica Chimica Acta
|March 27, 2012
PubMed
Summary

A new semi-automated method aligns peaks in comprehensive two-dimensional liquid chromatography data without reference injections. This technique improves quantification accuracy for complex samples, enhancing analytical reliability.

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Quantification of Site-specific Protein Lysine Acetylation and Succinylation Stoichiometry Using Data-independent Acquisition Mass Spectrometry

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Area of Science:

  • Analytical Chemistry
  • Chromatography
  • Spectroscopy

Background:

  • Comprehensive two-dimensional liquid chromatography (LCxLC) generates complex data.
  • Accurate peak alignment is crucial for quantitative analysis in LCxLC.
  • Existing alignment methods often require reference injections, which can be impractical.

Purpose of the Study:

  • To develop a semi-automated peak alignment method for four-way LCxLC-diode array detector (DAD) data.
  • To enable accurate quantification without the need for reference injections.
  • To improve the reliability of LCxLC data analysis.

Main Methods:

  • Utilized parallel factor analysis (PARAFAC) for data decomposition.
  • Developed a spectral signature-based method to align peaks across injections.
  • Shifted chromatographic component profiles based on spectral matching and peak position.
  • Calculated peak shifts using median and maximum data points in chromatographic dimensions.

Main Results:

  • Achieved target analyte recoveries within 2% of 100% for simulated data.
  • Demonstrated quantification precision of 4% or better for spectrally similar, coeluting peaks in urine.
  • Validated the method against LC-LC MS/MS for phenytoin in wastewater, showing agreement within method precision.

Conclusions:

  • The developed semi-automated alignment method is effective for four-way LCxLC-DAD data.
  • The method provides accurate and precise quantification, even for challenging samples.
  • This approach enhances the utility of LCxLC for complex mixture analysis.