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Higher energy collisional dissociation (HCD) fragmented (isolation width 1.2 m / z and 28% normalised collision energy) MS/MS scans in a cycle time of 1.1 s, the most abundant 25+ charged peaks in the MS scan were recorded in a data-dependent mode (Resolution 15,000, threshold 2e4, 15 s exclusion duration for the selected m / z 10 ppm)