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Peptide Identification Using Tandem Mass Spectrometry

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...

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Updated: May 22, 2025

Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification

Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification

Published on: August 17, 2015

MRM-based LC-MS method for accurate C-peptide quantitation.

Will Grothoff¹, Ivan Khodakivskyi¹, Aleks Shin¹

¹Pathology & Anatomical Sciences, University of Missouri, 1 Hospital Drive, Columbia, MO 65211, USA.

Journal of Mass Spectrometry and Advances in the Clinical Lab

|March 17, 2025

Summary

This summary is machine-generated.

We developed a mass spectrometry method for accurate C-peptide quantitation, improving diabetes diagnosis. This method shows high accuracy and precision, supporting clinical decision-making for diabetes mellitus.

Keywords:

C-peptide Diabetes mellites LC-MS Standardization

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Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry

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Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry

Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry

Published on: July 31, 2011

Area of Science:

Biochemistry
Clinical Chemistry
Analytical Chemistry

Background:

C-peptide is a biomarker for beta-cell function and diabetes mellitus.
Current C-peptide assays lack standardization, limiting clinical utility.
Accurate C-peptide measurement is crucial for diabetes type differentiation and management.

Purpose of the Study:

To develop and validate a mass spectrometry-based method for precise C-peptide quantitation.
To establish a reliable assay for C-peptide measurement in clinical settings.
To address the need for standardized C-peptide measurements.

Main Methods:

Developed a liquid chromatography-mass spectrometry (LC-MS) method using multiple reaction monitoring (MRM).
Employed isotope-labeled C-peptide as an internal standard.
Utilized solid-phase extraction and anion exchange for sample enrichment, followed by Glu-C digestion.

Main Results:

The assay demonstrated excellent linearity and a low limit of quantitation (0.058 nmol/L).
High precision was achieved with inter-day CV < 9.6% and intra-day CV < 8.9%.
The method showed strong correlation (r² = 0.95) with the reference method and minimal interference.

Conclusions:

The developed LC-MS/MRM method provides accurate C-peptide quantitation.
This method can serve as a valuable supplemental tool to existing reference methods.
It contributes to refining the C-peptide reference system and improving diabetes diagnostics.