Absorption Mode Broadband 2D MS for Proteomics and Metabolomics

Area of Science:

• Analytical Chemistry
• Spectrometry

Background:

• Two-dimensional mass spectrometry (2D MS) correlates precursor and fragment ions without isolation.
• Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) data processing requires phase correction for optimal results.
• Previous work showed phase-corrected absorption mode improved signal-to-noise ratio (SNR) and resolving power by a factor of 2 on limited data sets.

Purpose of the Study:

• Expand phase-corrected absorption mode data processing to all 2D mass spectra.
• Apply this method to complex biological and chemical samples.
• Demonstrate significant improvements in SNR and resolving power.

Main Methods:

• Developed and applied phase-corrected absorption mode data processing to 2D mass spectra.
• Utilized Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS).
• Analyzed top-down proteomics of oxidized ubiquitin proteoforms and ergot alkaloid extracts.

Main Results:

• Phase-corrected absorption mode processing was successfully applied to 2D MS data of varying sizes and frequency ranges.
• Significant improvements in SNR and resolving power were observed compared to standard magnitude mode.
• Enhanced sequence coverage in top-down proteomics and precursor-fragment correlation accuracy in metabolomics were achieved.

Conclusions:

• Phase-corrected absorption mode data processing is a powerful technique for enhancing 2D MS data.
• This method offers substantial benefits for both top-down proteomics and metabolomics.
• The improved SNR and resolving power facilitate more accurate and comprehensive analyses.