1290 infinity lc instrument

UPLC analysis was performed on an Agilent 1290 Infinity LC instrument (Agilent, Waldbronn, Germany) consisting of a quaternary pump, an auto-sampler, a column compartment and a diode-array detector. The samples were separated on a Zorbax SB-C₁₈ column (3.5 μm, 100 mm × 2.1 mm i.d.). The mobile phase was a stepwise gradient of 0.1% formic acid aqueous solution (A) and methanol (B) as follows: 10% B at 0–3 min; 10–65% B at 3–10 min; 65–80% B at 10–10.5 min and maintained at 80% B from 10.5 min to 15 min. The UV wavelength was set at 254 nm. The column temperature and the flow rate were set at 20 °C and 0.2 mL/min, respectively.
The UPLC system was connected to an Agilent 6530 QTOF mass spectrometer (Santa Clara, CA, USA) equipped with a dual electrospray ionization (ESI) interface using the following operation parameters: capillary voltage, 3.5 kV in (−)ESI mode or 4.0 kV in (+)ESI mode; nebuliser, 40 psig; drying gas (nitrogen) flow rate, 10.0 L/min; gas temperature, 325 °C; fragmentor, 175 V; skimmer voltage, 65 V; OCT 1 RF Vpp, 750 V. Mass spectra were recorded across the range m/z 100–1500. The MS/MS collision energy was set at 20–30 V.

The metabolite profiles of GAL were determined using an LC-QTOF-MS instrument (Agilent 1290 Infinity LC instrument coupled to an Agilent 6540 series QTOF-MS equipped with an ESI source and a diode-array detector (DAD)) in positive ion mode. Chromatographic separation was conducted on an Agilent Poroshell 120 EC-C18 column (150 mm length x 4.6 mm inner diameter, particle size 2.7 μm). The gradient elution was performed using 0.1% formic acid water (mobile phase A) and acetonitrile (mobile phase B) at a flow rate of 200 μL/min. The injection volume was 1.0 μL, and the column temperature was maintained at 35°C. Data acquisition was controlled using the Mass Hunter Workstation Software Qualitative Analysis (version B.08.00, Agilent Technologies, California, United States of America). The phytochemical compounds present in the extract samples were identified by comparing retention times, mass data, and fragmentation patterns with a compound database in the library search of the Agilent MassHunter Personal Compound Database and Library (Agilent Technologies). Peaks with similarity scores of 80% compared to the database were selected to confirm peak identification (Sun et al., 2015 (link); Zhu et al., 2022 (link)).