Nexera 1 lc 2040c 3d hplc system

HPLC-mass spectrometry analyses were carried out on a Nexera-I LC-2040C 3D HPLC system coupled to the LCMS-2020 system through an electrospray ionization (ESI) interface (Shimadzu Corp.). ESI-MS detection of the transformation products was performed out in the positive ion mode (+) in the full scan mode (mass-to-charge ratio, m/z 200–1000). HPLC-tandem mass spectrometry analyses in a product ion scan were performed with Agilent system 1290 Infinity II LC coupled to the 6470B system with a triple quadrupole in the positive ion mode (+), mass-to-charge ratio, m/z also 200–1000. HPLC-tandem mass spectrometry analyses were performed with an Agilent 6500 Series Accurate-Mass Quadrupole-Time of Flight (Q-TOF) mass spectrometer (Agilent Technologies, Santa Clara, CA, USA) controlled by Agilent MassHunter Workstation software. To ensure accurate mass during the experiment, the mass spectrometer was calibrated daily using calibration solution (ES-TOF reference mix, Agilent Technologies).

HPLC analyses of the described above the final supernatants obtained after electrochemical, enzymatic and cellular transformations were performed using a Nexera-I LC-2040C 3D HPLC system and the LabSolution software package (Shimadzu Corp., Kyoto, Japan). Samples were separated using a reversed-phase 5 μm Suplex pKb-100 analytical column (0.46 × 25 cm, C18) (Supelco, Bellefonte, PA, USA) warmed to 25 °C. The analyses were performed at a flow rate of 1 mL/min with the following mobile-phase system: a linear gradient from 15% to 80% methanol in ammonium formate buffer (0.05 M, pH 3.4) for 25 min, followed by linear gradient from 80% to 100% methanol in ammonium formate buffer for 3 min. The column was then re-equilibrated at initial conditions for 10 min between runs. The elution of each metabolite was monitored at 430 nm.