This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We describe a prototype tandem mass spectrometer that is designed to increase the efficiency of linked-scan analyses by >100-fold over conventional linked-scan instruments. The key element of the mass spectrometer is a novel high ion capacity ion trap, combined in tandem configuration with a quadrupole collision cell and a quadrupole mass analyzer (i.e. a TrapqQ configuration). This ion trap can store >106 ions without significant degradation of its performance. The current mass resolution of the trap is 100?450 full width at half maximum for ions in the range 800?4000 m/z, yielding a 10?20 m/z selection window for ions ejected at any given time into the collision cell. The sensitivity of the mass spectrometer for detecting peptides is in the low femtomole range. We can envisage relatively straightforward modifications to the instrument that should improve both its resolution and sensitivity. We tested the tandem mass spectrometer for collecting precursor ion spectra of all the ions stored in the trap and demonstrated that we can selectively detect a phosphopeptide in a mixture of non-phosphorylated peptides. Based on this prototype instrument, we plan to construct a fully functional model of the mass spectrometer for detecting modification sites on proteins and profiling their abundances with high speed and sensitivity. Currently we are testing a second generation instrument coupled to an orthogonal time-of-flight analyzer for accurate mass readout of both the parent and fragment ions. Towards this goal we have: We have optimized the high capacity ion trap and the adjacent collision cell by incorporating a cylinder housing to use different gases. This will give us control over pressure in each region and allow us to use different gases in each region for better ion cooling. We changed our original quadrupole collision cell to a linearly accelerating (LINAC) collision cell. In the LINAC a small axial electric field is applied to the collision cell by inserting a T electrode with a DC gradient. This will minimize the residence time in LINAC and increase overall transmission efficiency. We shortened the entire instrument by removing several unnecessary quadrupoles. This will increase the overall transmission efficiency.
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