A scan cycle comprised MS1 scan (m/z range from 350-1600, ion injection time of 20 ms, resolution 70000 and automatic gain control (AGC) 1x106) acquired in pro8ile mode, followed by 15 sequential dependent MS2 scans (resolution 17500) of the most intense ions ful8illing prede8ined selection criteria (AGC 2 x 105, maximum ion injection time 100 ms, isolation window of 1.4 m/z, 8ixed 8irst mass of 100 m/z, spectrum data type: centroid, intensity threshold 2 x 104, exclusion of unassigned, singly and >7 charged precursors, peptide match preferred, exclude isotopes on, dynamic exclusion time 45 s). The HCD collision energy was set to 27% of the normalized collision energy. Mass accuracy is checked before the start of samples analysis.