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

Adele Bourmaud, Sebastien Gallien, Bruno Domon*

*Corresponding author for this work

    Research output: Contribution to journalReview articlepeer-review

    183 Citations (Scopus)

    Abstract

    Targeted mass spectrometry-based approaches are nowadays widely used for quantitative proteomics studies and more recently have been implemented on high resolution/accurate mass (HRAM) instruments resulting in a considerable performance improvement. More specifically, the parallel reaction monitoring technique (PRM) performed on quadrupole-Orbitrap mass spectrometers, leveraging the high resolution and trapping capabilities of the instrument, offers a clear advantage over the conventional selected reaction monitoring (SRM) measurements executed on triple quadrupole instruments. Analyses performed in HRAM mode allow for an improved discrimination between signals derived from analytes and those resulting from matrix interferences translating in the reliable quantification of low abundance components. The purpose of the study defines various implementation schemes of PRM, namely: (i) exploratory experiments assessing the detectability of very large sets of peptides (100–1000), (ii) wide-screen analyses using (crude) internal standards to obtain statistically meaningful (relative) quantitative analyses, and (iii) precise/accurate quantification of a limited number of analytes using calibrated internal standards. Each of the three implementation schemes requires specific acquisition methods with defined parameters to appropriately control the acquisition during the actual peptide elution. This tutorial describes the different PRM approaches and discusses their benefits and limitations in terms of quantification performance and confidence in analyte identification.

    Original languageEnglish
    Pages (from-to)2146-2159
    Number of pages14
    JournalProteomics
    Volume16
    Issue number15-16
    DOIs
    Publication statusPublished - 1 Aug 2016

    Keywords

    • High resolution accurate mass
    • Parallel reaction monitoring
    • Precise quantification
    • Targeted proteomics
    • Technology

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