Ncs 3500rs ultimate 3000 binary rapid system

Liquid chromatography–high resolution mass spectrometry (LC-HRMS) was performed on a Thermo Scientific NCS-3500RS Ultimate 3000 Binary Rapid system coupled to a Thermo Scientific QExactive mass spectrometer (Thermo Scientific). The Ultimate 3000 RSLCnano system consisted of a degasser, a tertiary loading pump, a binary eluting pump, a column oven, and an RS Autosampler. The QExactive contained a heated electrospray ionization source (HESI-II) and was operated in positive ionization mode. The spray voltage was 3 kV, capillary temperature 350 °C, heater temperature 425 °C, S-lens RF level 50, sheath gas flow rate 50, auxiliary gas flow rate 13 and sweep gas 3 (manufacturer’s units). Nitrogen was used for spray stabilization, for collision-induced dissociation experiments in the HCD cell, and as the damping gas in the C-trap. The instrument was calibrated in the positive and negative modes every 25 h.