Parallel reaction monitoring using quadrupole-Orbitrap mass spectrometer: Principle and applications

Area of Science:

• Proteomics
• Analytical Chemistry
• Mass Spectrometry

Background:

• Targeted mass spectrometry is crucial for quantitative proteomics.
• High-resolution/accurate mass (HRAM) instruments significantly improve performance.
• Parallel Reaction Monitoring (PRM) on quadrupole-Orbitrap instruments surpasses conventional Selected Reaction Monitoring (SRM) on triple quadrupoles.

Purpose of the Study:

• To describe various implementation schemes of PRM for quantitative proteomics.
• To assess PRM's utility in exploratory, wide-screen, and precise quantification.
• To guide the selection of appropriate acquisition methods for PRM analyses.

Main Methods:

• Implementation of PRM on quadrupole-Orbitrap mass spectrometers.
• Development of specific acquisition methods tailored to peptide elution.
• Comparison of PRM with conventional SRM techniques.

Main Results:

• HRAM-based PRM enables better discrimination of analyte signals from matrix interferences.
• PRM allows for reliable quantification of low-abundance peptides.
• Different PRM schemes (exploratory, wide-screen, precise) offer distinct advantages.

Conclusions:

• PRM on HRAM instruments provides enhanced performance for quantitative proteomics.
• Specific acquisition strategies are vital for optimizing PRM experiments.
• PRM offers improved quantification performance and confidence in analyte identification.