Improved Electron Transfer Dissociation (ETD) Duty Cycle and Spectral Signal to Noise Ratio in a Dual Cell Linear Ion Trap Christopher Mullen,1 Lee Earley,1 Jean-Jacques Dunyach,1 John E.P. Syka,1 Jeffrey Shabanowitz, 2 A. Michelle English, 2 Donald F. Hunt2 1 Thermo Fisher Scientific, San Jose, CA; 2 Department of Chemistry, University of Virginia, Charlottesville, VA

Methods: Thermo Scientific™ Orbitrap Fusion™ Tribrid™ mass spectrometer with the Thermo Scientific™ EASY-ETD™ ion source. Performing the ETD reaction in cation precursor population to the reagent anion species can independent trapping (CSIT) a number of cation precursor ch function of the space charge c Figure 2 shows the potentials injection events and the relativ Purpose: Improve ETD Signal to Noise (S/N) ratio and scan duty cycle. Results: Developed a methodology incorporating multiple fills of ETD products into a storage cell followed by a single m/z analysis leading to improved spectral...

Motivation for Employing Multiple Fills ETD ated to be a useful tool for the with labile PTM’s, peptides with he approach is that it leads to a al, compared to conventional action and the fact that the possibility to circumvent these z analysis in a 2D linear ion trap ss resolving quadrupole, OT-LT) architecture. linear ion trap is accomplished sel, and utilizing the low is cell. In this fashion, the e number of fills requested) a single m/z analysis in either priate transfer, in the OT mass p Fusion Tribrid MS. FIGURE 2. Cation sequestration and reagent injection conditions prior to the ETD...

jection conditions prior to the ual cell linear trap FIGURE 4. a) Angiotensin I b and y ion formation observed at high precursor initial targets resulting from harsh sequestration conditions. b) Fragment yield versus precursor target demonstrating the onset of b and y ion formation. AngioVsTarget_ETD_50msec #548 RT: 2.58 AV: 1 NL: 6.18E5 T: FTMS + p ESI Full ms2 [email protected] [150.0000-1500.0000] Relative Abundance ations Held In Back Section T is determined by the axial potential ured by a number of methods. We reaction of Angiotensin I as a ragment TIC plotted as a function of ressure trap...

FIGURE 6. Comparison of the Angiotensin I ETD c and z ion fragment TIC for an up to 8-fills ETD approach versus a single fill. The multiple fill data was taken at three individual precursor targets per fill corresponding to 1e4 (blue squares), 5e4 (red circles), and 1e5 (green triangles) charges/fill. erved at high precursor ditions. b) Fragment yield and y ion formation. ea vs. Precursor Target . Rxn Time = 70 msec rsor = Angio3+ zer = Orbitrap It is apparent (Figure 6) that the working range using the multiple fills approach is much greater than that for a single fill, while preserving soft...

nd z ion fragment TIC for an multiple fill data was taken at ding to 1e4 (blue squares), fill. The geometry of the Orbitrap Fusion Tribrid MS in conjunction with it’s ability to run scans in a parallel/pipelined fashion affords a significant decrease in scan acquisition time per spectrum for the multiple fills approach (Figure 8) as the ETD fills are run in parallel with OT m/z analysis. FIGURE 8. Scan acquisition time per spectrum for the ETD multiple fills experiment and the uScans approach taken at a variety of Orbitrap transient durations. Scan Time per Spectrum (sec) e multiple fills approach...

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