Nexera lcms 9030

A chemical profile was obtained using ultra-high-performance liquid chromatography coupled with photodiode array detection (UPLC-PAD) on an ACQUITY UPLC system (Waters, Milford, MA, USA), which consisted of a photodiode array detector, quaternary pump, degasser, column oven, and autosampler. The separation of metabolites was performed using Phenomenex^® Kinetex C18 100 Å (75 mm × 2.1 mm; 2.6 µm). The mobile phase was formic acid 0.1% (component A) and acetonitrile (component B) as follows: 0–8 min, 16% B; 8–10 min, 16–20% B; 10–12 min, 20% B; 12–15 min, 20–22% B; 15–18 min, 22–27% B; 18–20 min, 27–30% B; 20–23 min, 30–35% B; 23–28 min, 35% B; 28–34 min, 35–90% B; and then back to 16% B in 1 min [52 (link)]. The samples were injected automatically at 3 µL. The column temperature was maintained at 30 °C. The flow rate was set at 0.4 mL/min. The photo diode array (PDA) detection was set at 340 nm. Ultra-high-performance liquid chromatography–electrospray ionization–quadrupole time-of-flight–mass spectrometry (UHPLC-ESI-QToF-MS) analysis was performed on Nexera LCMS 9030 Shimadzu Scientific-Instruments (Columbia, MD, USA) equipment. The chromatographic conditions were the same as those described previously: mass spectrum detection for negative ion mode. The capillary potentials were set at +3 kV, drying gas temperature 250 °C, and the flow rate of drying gas 350 L/min.

NMR spectroscopic data were recorded with a 400 MHz (400.13 MHz for ¹H and 100.61 MHz for ¹³C{1H}, and 376 MHz for ¹⁹F{1H}) or 500 MHz (500.03 MHz for ¹H{1H} and 125.73 MHz for ¹³C{1H}) spectrometer (Bruker Avance III, Billerica, MA, USA) in CDCl₃ and DMSO-d₆ and were referenced to residual solvent proton signals (δ_H = 7.26, and δ_H = 2.50, respectively) and solvent carbon signals (δ_C = 77.0, δC = 39.5, respectively). Mass spectra were recorded with a HRMS-ESI-qTOF spectrometer Nexera LCMS-9030 (electrospray ionization mode) (Shimadzu Europa, Duisburg, Germany). Chlorobenzene (PhCl) was distilled from P₂O₅ and dried over molecular sieves 4Å (>24 h). Column chromatography was carried out on silica gel grade 60 (0.040−0.063 mm) 230−400 mesh. Kieselgel on aluminum was used for TLC (UV 254). All imines were prepared in our laboratory according to common protocol for condensation of equimolar amounts of amine with aldehyde in dichloromethane (DCM) (overnight, r.t.) in the presence of anhydrous MgSO₄.

A chemical profile was obtained using ultra-high-performance liquid chromatography coupled with photodiode array detection (UPLC-PAD) on an ACQUITY UPLC system (Waters, Milford, MA, USA), which consisted of a photodiode array detector, quaternary pump, degasser, column oven, and autosampler. The separation of metabolites was performed using Phenomenex^® Kinetex C18 100 Å (75 mm × 2.1 mm; 2.6 µm). The mobile phase was formic acid 0.1% (component A) and acetonitrile (component B) as follows: 0–8 min, 16% B; 8–10 min, 16–20% B; 10–12 min, 20% B; 12–15 min, 20–22% B; 15–18 min, 22–27% B; 18–20 min, 27–30% B; 20–23 min, 30–35% B; 23–28 min, 35% B; 28–34 min, 35–90% B; and then back to 16% B in 1 min [52 (link)]. The samples were injected automatically at 3 µL. The column temperature was maintained at 30 °C. The flow rate was set at 0.4 mL/min. The photo diode array (PDA) detection was set at 340 nm. Ultra-high-performance liquid chromatography–electrospray ionization–quadrupole time-of-flight–mass spectrometry (UHPLC-ESI-QToF-MS) analysis was performed on Nexera LCMS 9030 Shimadzu Scientific-Instruments (Columbia, MD, USA) equipment. The chromatographic conditions were the same as those described previously: mass spectrum detection for negative ion mode. The capillary potentials were set at +3 kV, drying gas temperature 250 °C, and the flow rate of drying gas 350 L/min.