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(B ) Schematic of Multiplexed OH (hydroxyl radical) Cleavage Analysis (MOHCA -seq workflow, showing randoMOHCA -seqation of radical sources, fragmentation, adapter ligations, and analysis by sequencing.
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In this work, detailed fragmentation pathways were elucidated by further dissociation of each fragment ion in the ion spectrum, essentially, by incorporating fragmentor voltage induced dissociation (in-source fragmentation) and isolation of fragments in a quadrupole cell Q1.
The in-source fragmentation of MeHgCys showed an important fragment with a mass loss of 17 amu with abundance as high as the MH+ molecular ion.
This is an indication that the group forming this fragment has been lost during in-source fragmentation.
However, at 40 V and higher, the overall intensity of both Gln and pGlu dramatically declined with subsequent appearance of a fragment at 84.01 m/ z, which corresponded to in-source fragmentation of the molecules.
Similar to the conventional in-source fragmentation technique, the in-source desolvation is more gentle, which is designed to fragment clusters and droplets other than ions.
The fragmentor voltage was optimized to produce adequate in-source fragmentation; values of 100, 120,140,160,180 and 200 V were tested to obtain the fragments of each compound in the scan spectra.
Unintended in-source fragmentation (IS-CID) could be ruled out in these particular cases since no fragments were observed in the scans without applying HCD fragmentation.
c) In-source fragmentation of compound 3c.
c) In-source fragmentation of compound 3a.
A special case of proactive fragmentation is the source fragmentation.
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