| Description |
Nexera ultra-high-performance liquid chromatograph (SHIMADZU, Kyoto, JAPAN) hyphenated with Q-Exactive high-resolution tandem mass spectrometer equipped with heated electrospray ionization (ESI) source (Thermo Fisher Scientific, Waltham, MA, USA) was used to analyze the metabolic profiling in both ESI positive and ESI negative ion modes. An ACQUITY UPLC HSS T3 columnwas employed in positive and negative modes. The binary gradient elution system consisted of (A) water (containing 0.1% formic acid, v/v) and (B) ACN (containing 0.1 % formic acid, v/v) and separation was achieved using the following gradient: 0.01 min, 5% B; 2 min, 5% B; 4 min, 30% B; 8 min, 50% B; 10 min, 80% B; 14 min, 100% B; 15 min, 100% B; 15.1 min, 5% and 16 min, 5% B. The flow rate was 0.35 mL/min, and the column temperature was 45.All the samples were kept at 10 during the analysis. The injection volume was 2 μL.The mass range was from m/z 100 to 1200. The resolution was set at 70,000 for the full MS scans and 17,500 for HCD MS/MS scans. The collision energy was set at 10, 20, and 40 eV. The mass spectrometer operated as follows: spray voltage, 3500 V (+) and 3000 V -; sheath gas flow rate, 35 arbitrary units; auxiliary gas flow rate, 8 arbitrary units; capillary temperature, 320; Aux gas heater temperature, 350; S-lens RF level, 50. |
