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. 2011 Jun;22(6):1105-8.
doi: 10.1007/s13361-011-0119-7. Epub 2011 Apr 9.

Implementing photodissociation in an Orbitrap mass spectrometer

Lisa A Vasicek  1 , Aaron R Ledvina, Jared Shaw, Jens Griep-Raming, Michael S Westphall, Joshua J Coon, Jennifer S Brodbelt
Affiliations

Implementing photodissociation in an Orbitrap mass spectrometer

Lisa A Vasicek et al. J Am Soc Mass Spectrom. 2011 Jun.

Abstract

We modified a dual pressure linear ion trap Orbitrap to permit infrared multiphoton dissociation (IRMPD) in the higher energy collisional dissociation (HCD) cell for high resolution analysis. A number of parameters, including the pressures of the C-trap and HCD cell, the radio frequency (rf) amplitude applied to the C-trap, and the HCD DC offset, were evaluated to optimize IRMPD efficiency and maintain a high signal-to-noise ratio. IRMPD was utilized for characterization of phosphopeptides, supercharged peptides, and N-terminal modified peptides, as well as for top-down protein analysis. The high resolution and high mass accuracy capabilities of the Orbitrap analyzer facilitated confident assignment of product ions arising from IRMPD.

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Figures

Figure 1
Figure 1
Schematic showing implementation of IRMPD in the HCD cell of a dual-cell linear ion trap-Orbitrap
Figure 2
Figure 2
Effect of pressure on photodissociation efficiency and signal intensity. IRMPD of doubly charged phosphorylated peptide TSTEPQpYQPGENL, [M + 2H]2+, (10 μM in 49.5/49.5/1 methanol/water/acetic acid), following 1 ms of 48 W irradiation at P1 (A), P2 (B), and P3 (C). rf Amplitude=1180 Vp-p (corresponding to first m/z=100), collision energy = −11 V
Figure 3
Figure 3
IRMPD of the 12+ charged state of ubiquitin (10 μM in 49.5/49.5/1 methanol/water/acetic acid) following 25 ms of 48 W irradiation, C-trap rf amplitude=2300 Vp-p (corresponding to first m/z=200), collision energy = −11 V. Inset shows baseline resolution of the y5810+ ion with a mass accuracy of 3.5 ppm relative to the theoretical m/z of 653.7562
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References

    1. Brodbelt JS, Wilson JJ. Infrared Multiphoton Dissociation in Quadrupole Ion Traps. Mass Spectrom Rev. 2009;28:390–424. - PubMed
    1. Wiesner J, Premsler T, Sickmann A. Application of Electron Transfer Dissociation (ETD) for the Analysis of Post-Translational Modifications. Proteomics. 2008;8:4466–4483. - PubMed
    1. Zubarev R. Electron capture dissociation and other ion-electron fragmentation reactions. In: Lifshitz C, Laskin J, editors. Principles of Mass Spectrometry Applied to Biomolecules. 2006. pp. 475–517.
    1. Ly T, Julian RR. Ultraviolet Photodissociation: Developments Towards Applications for Mass-Spectrometry-Based Proteomics. Angew Chem Int Ed. 2009;48:7130–7137. - PubMed
    1. Coon JJ. Collisions or Electrons? Protein Sequence Analysis in the 21st Century. Anal Chem. 2009;81:3208–3215. - PMC - PubMed

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