Orbitrap q exactive high field mass spectrometer

Liquid chromatography was performed on a Vanquish HPLC system controlled by Chromeleon software (ThermoFisher Scientific) and coupled to an Orbitrap Q-Exactive High-Field mass spectrometer controlled by Xcalibur software (ThermoFisher Scientific). Methanolic extracts prepared as described above were separated on a Thermo Hypersil Gold C18 column (150 mm × 2.1 mm, particle size 1.9 μM, part no. 25002-152130) maintained at 40 °C with a flow rate of 0.5 mL/min. Solvent A is 0.1% formic acid (Fisher Chemical Optima LC/MS grade; A11750) in water (Fisher Chemical Optima LC/MS grade; W6-4); solvent B is 0.1% formic acid in acetonitrile (Fisher Chemical Optima LC/MS grade; A955-4). A/B gradient started at 1% B for 3 min after injection and increased linearly to 98% B at 20 min, followed by 5 min at 98% B, then back to 1% B over 0.1 min, and finally held at 1% B for the remaining 2.9 min to reequilibrate the column (28 min total method time). Mass spectrometer parameters were spray voltage, −3.0 kV/+3.5 kV; capillary temperature, 380 °C; probe heater temperature, 400 °C; sheath, auxiliary, and sweep gas, 60, 20, and 2 AU, respectively; S-Lens RF level, 50; resolution, 120,000 at m/z 200; and AGC target, 3E6. Each sample was analyzed in negative (ESI−) and positive (ESI+) electrospray ionization modes with m/z range 100 to 1,000.