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Updated: May 18, 2026

Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification
Published on: August 17, 2015
Quantitative mass spectrometry-based assay development and validation: from small molecules to proteins
Andrea Božović1, Vathany Kulasingam
1Laboratory Medicine Program, Toronto General Hospital, University Health Network, Toronto, Ontario, Canada M5G 2C4.
Mass spectrometry (MS) is a powerful tool for analyzing biomolecules in clinical diagnostics. This review covers method development and validation for small molecules and proteins using liquid chromatography tandem mass spectrometry (LC-MS/MS).
Area of Science:
- Analytical Chemistry
- Clinical Diagnostics
- Biochemistry
Background:
- Mass spectrometry (MS) is increasingly utilized in clinical laboratories for biomolecule analysis.
- Advancements in MS technology enhance its popularity for diagnostic assays.
- MS offers advantages like multi-analyte detection, low sample volume, and cost-effectiveness.
Purpose of the Study:
- To review method development and validation for clinical diagnostic assays using MS.
- To discuss small molecule analysis and protein quantification via LC-MS/MS.
- To highlight challenges in protein quantification and provide a future outlook for MS in laboratory medicine.
Main Methods:
- Focuses on liquid chromatography tandem mass spectrometry (LC-MS/MS).
- Discusses the critical phases of method development and validation.
- Covers both small molecule and protein quantification strategies.
Main Results:
- Explains the process of developing and validating MS-based assays for clinical use.
- Details the application of LC-MS/MS for small molecule and protein analysis.
- Identifies specific challenges encountered in protein quantification.
Conclusions:
- MS, particularly LC-MS/MS, is a valuable technique for clinical diagnostics.
- Method development and validation are crucial for reliable MS assay implementation.
- Future advancements will likely expand the role of MS in laboratory medicine.
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