Sieve v2.2 sp2

For tandem mass spectrometry analysis, a Thermo ESI-Q-Orbitrap MS mass spectrometer was connected to the UltiMate 3000 UHPLC instrument via ESI interface. Ultra-high purity nitrogen (N₂) was used as the collision gas and the sheath/auxiliary gas. The ESI source parameters were set as follows: ion spray voltage 3.2 kV, capillary temperature 350 °C, ion source heater temperature 300 °C, sheath gas (N₂) 40 L/h, auxiliary gas (N₂) 10 L/min, and a normalized collision energy (NCE) of −35 V was used. The Orbitrap analyzer scanned the mass range from m/z 150 to 1500 in negative ion mode. Monitoring time was 0–17 min. Detection was obtained by full mass-dd mass mode. The MS data were recorded in both profile and centroid formats. Data recording and processing were performed using the Xcalibur 4.0 software (Thermo Fisher Scientific, Inc., Waltham, MA, USA). The accuracy error threshold was fixed at 5 ppm.
Software-aided, automatic background subtraction and components extraction technique was used to generate a peak list containing all the components profiled from the aerial part of A.mongolicum. Sieve v2.2 SP2 (Thermo Fisher Scientific) was used for the automatic components extraction: time range, 1–17 min; BP minimum count, 10,000; BP minimum scans, 5; Background SN, 3; MZ Step, 10; and Frame, >1.

Spirostanol saponins identification was carried out by using a Thermo ESI-Q-Exactive-Orbitrap MS (in the positive ESI mode (capillary voltage: 3.2 kV). Ultra-high purity nitrogen (N₂) and high purity nitrogen (N₂) were used as the collision gas and the sheath/auxiliary gas, respectively. The ESI source parameters were 350 °C for capillary temperature, 300 °C for ion source heater temperature, 40 arbitrary units for sheath gas (N₂), 10 arbitrary units for auxiliary gas (N₂), and the collision energy of the quadrupole ranged between 15 and 45 V were used. The mass range of the Orbitrap analyzer scanner was m/z 150 to 1500. Selected precursors analyzed more than two times were actively excluded from analysis for 60 s. Monitoring time was 0–65 min. Data recording and processing were performed using the Xcalibur 4.0 software (Thermo Fisher Scientific, Inc., Waltham, MA, USA). The accuracy error threshold was fixed at 5 ppm. Automatic components extraction was accomplished by a Sieve v2.2 SP2 (Thermo Fisher Scientific) with the time range from 0 to 65 min and the BP minimum count at 10,000. Mass resolution is 70,000 and 17,500 for MS and MS², respectively.