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Crosslinking of guanine with tyrosine and glutathione is studied by on-line electrochemistry electrospray ionization mass spectrometry (EC ESI MS) in the positive ion mode with a high resolution FT ICR mass spectrometer without the use of an external oxidizer in the sample.
The instrument with nanospray ion source was operated as following: external calibration weekly, top 5 MS (in the ion trap), 1 microscan, resolution (FT full scan): 30,000 m/Δm50%, MS threshold: 8,000 (~S/N 2 1), MS target: 1e, MS target: 5e, exclusion mass tolerance: 10 ppm, charge state selection: 2+, 3+.
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High resolution FT-MS analysis was performed on a Thermo LTQ-Orbitrap XL.
These were consistent with the molecular formula obtained from the ultra-high-mass resolution FT-ICR-MS (see below).
When we sought to determine the relative abundance of human-derived and murine-derived proteins, we used extracted ion chromatograms (XIC's) for each peptide precursor ion in the intervening high resolution FT-MS scans of the LC-MS/MS runs.
The LTQ Orbitrap XL was configured with a TOP 5 methodology comprising a 60 K resolution FT-MS full scan followed by IT-MS/MS scans for the 5 most intense peptide ions.
The QCL data at 2 containstains ∼550 bands and is presented undersampled by a factor of 2 such that it matches the 4 cm–1 resolution FT-IR systems.
MS and MS scans were acquired using data-dependent acquisition based on a full 30,000 resolution FT-MS scan to sequence the top 6 most intense ions FT-MS using CID fragmentation and 7,500 resolution FT-MS Orbitrap scans with a single repeat count (5 s repeat duration) followed by a 10 s dynamic exclusion with a 10 ppm mass window based on a maximal exclusion list of 500 entries.
Only peptides having a spectral probability of less than 1×10−10 and a length of at least six amino acids were retained for matching to peptide feature data generated using high resolution FT-MS instrumentation (LTQ-Orbitrap; Thermo Scientific, San Jose CA) as described previously [ 68].
MS spectra were acquired in the high-resolution FT mass analyzer and MS spectra were acquired in the linear ion trap mass analyzer.
In this work, we show the capabilities of high-resolution FT alone and combined with MS/MS for the characterization of modern coating components.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com