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The isolated precursors were fragmented by collision induced dissociation (CID) at 30% collision energy with the maximum injection time of 35 ms for 3 s (top speed setting) and detected in the ion trap, followed by multinotch (simultaneous) isolation of the top 10 MS2 fragment ions within the m/z range 4001,400, fragmentation (MS3) by higher-energy collision dissociation (HCD) at 55% collision energy and detection in the Orbitrap at 50,000 resolution m/z range 100500 and maximum injection time 105 ms