Magic c18 aq 200a reverse phase media

LC-MS/MS analysis was performed using a LTQ Orbitrap XL mass spectrometer (Thermo Scientific). The LC system configured in a vented format consisted of a fused-silica nanospray needle packed in-house with Magic C18 AQ 100A reverse-phase media (Michrom Bioresources Inc, 25 cm) and a trap (2 cm) containing Magic C18 AQ 200A reverse-phase media. The inner diameters of the analytical and trap columns were 75 μm and 100 μm, respectively. The peptide samples were loaded onto the column and chromatographic separation was performed using a two-mobile-phase solvent system consisting of 0.1% formic acid in water (A) and 0.1% acetic acid in acetonitrile (B) over 60 min from 5% B to 40% B. The mass spectrometer operated in a data-dependent MS/MS mode over the m/z range of 400-1800. For each cycle, the five most abundant ions from each MS scan were selected for MS/MS analysis using 35% normalized collision energy. Selected ions were dynamically excluded for 45 seconds.