Orbitrap velos pro system

A UHPLC system equipped with a Vanquish binary pump H system (Thermo Fisher Scientific, Waltham, MA, USA) was united with an auto-sampler and column compartment. A Phenomenex KINETEX^® C18 column (100 mm × 2.1 mm, 1.7 µm particle size; Torrance, CA, USA) was used to chromatographic separation. The injection volume was 5 µL. The column temperature was set to 40 °C, and the flow rate was 0.3 mL/min. The mobile phase consisted of 0.1% v/v formic acid in water (A) and acetonitrile (B). The gradient parameters were configured as follows: Five percent solvent B was maintained initially for 1 min, followed by a linear increase to 100% solvent B over 9 min and then sustained at 100% solvent B for 1 min, with a gradual decrease to 5% solvent B over 3 min. The total run time was 14 min. The MS data were collected in a range of 100–1500 m/z (in negative- and positive-ion mode) using an Orbitrap Velos Pro system (Thermo Fisher Scientific, Waltham, MA, USA) coupled with an ion-trap mass spectrometer coupled with a heated electrospray ionization (HESI-II) probe. The probe heater and capillary temperatures were set to 300 °C and 350 °C, respectively. The capillary voltage was set to 2.5 kV in negative mode (positive mode 3.7 kV).

The dried extracts were re-dissolved in 80% MeOH for UHPLC-LTQ-Orbitrap-MS/MS analysis. All analytes were set to 20,000 ppm by using daidzein (0.25 mg/ml) as IS.
A UHPLC system was equipped with a Vanquish binary pump H system (Thermo Fisher Scientific, Waltham, MA, USA) coupled with an autosampler and column compartment. Chromatographic separation was performed with a Phenomenex KINETEX^® C18 column (100 mm × 2.1 mm, 1.7 μm particle size; Torrance, CA, USA); and the injection volume was 5 μl. The column temperature was set at 40°C, and the flow rate was 0.3 ml/min. The MS data were collected in the range of 100–1,500 m/z (under both negative- and positive-ion modes) using an Orbitrap Velos Pro™ system, which was combined with an ion-trap mass spectrometer (Thermo Fisher Scientific) coupled with a HESI-II probe. All samples were analyzed based on the analytical methods described by Kwon et al. (20 (link)).