UPLC-HRMS Analysis of H. atra Extracts

UPLC-HRMS analysis of the H. atra extracts was carried out according to Rivera-Mondragon et al. (2019), with modifications [54 (link)], using a XEVO-G2-XS QTOF mass spectrometer (Waters, Milford, MA, USA) coupled with an Acquity UPLC system. The system was operated with MassLynx 4.1 software (Waters, Milford, MA, USA). An HSS T3 RP18 column (1.8 µm, 2.1 × 100 mm) (Waters, Milford, MA, USA) was used to obtain separation. The following samples were analyzed using a UPLC Acquity system coupled with a Xevo G2-XS Q-Tof mass spectrometer (Waters, Milford, MA, USA), including subfractions MB, MC, ME, MF, and MG, and the Me fraction and Bu fraction. A total of two isolated compounds were also analyzed by UPLC-HRMS. The mobile phases used were (A) H₂O + 0.1% FA and (B) ACN + 0.1% FA, and the gradient was set as follows: 3% of B (0–1 min), 100% of B (17–19 min), and 3% of B (21–25 min). The flow rate was 0.4 mL/min. The following settings were used for the mass spectrometer: a cone gas flow of 50 L/h; a desolvation gas flow of 1000 L/h; a source temperature of 120 °C; and desolvation at 550 °C. The samples were analyzed in MSe mode, thus obtaining information from the molecular ions and mass fragmentation data simultaneously. The MS data were recorded in ESI+ and ESI- mode with an MS scan range from m/z 50 to 1500.