Metabolite Profiling of Plant Extracts

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)).