The UHPLC analysis was conducted utilizing an LC30 system (Shimadzu Co., Tokyo, Japan) equipped with an HSS T3 C18 (100 mm × 2.1 mm × 1.8 μm) column sourced from Waters Co. (Milford, MA, USA). The mobile phases consisted of solvent A (0.1 % formic acid in water) and solvent B (acetonitrile). The elution program was optimized based on a previous protocol (Chen et al., 2021 (link)): 0–2 min, 5.0 % B; 2–5 min, 5 %-10 % B; 5–14 min, 10 %-40 % B; 14–24 min, 40 %-100 % B; 24–26.5 min, 100 %-100 % B; 26.5–27 min, 100 %-5% B. A re-equilibration step was carried out for three minutes at 5 % B. The system operated at a flow rate of 0.3 mL/min, with an injection volume of 5 µL and a column temperature of 40 °C.
Mass spectrometry (SCIEX Co., Framingham, MA, USA) was performed in positive ion mode, with a mass range of 50–1000 m/z. The following settings were used for data collection: declustering potential at 50 V, collision energy at 10 V, and a temperature of 450 °C, curtain gas 35 psi, nebulizer gas (nitrogen) pressure at 2 bar, capillary voltage at 4,200 V (L. Zhang et al., 2023b). The MS/MS data collection was carried out in IDA mode, where the top 10 ions of each spectrum were fragmented using collision-induced dissociative energy of 35 ± 10 eV (Li, Zhou, et al., 2023).
Mass spectrometry (SCIEX Co., Framingham, MA, USA) was performed in positive ion mode, with a mass range of 50–1000 m/z. The following settings were used for data collection: declustering potential at 50 V, collision energy at 10 V, and a temperature of 450 °C, curtain gas 35 psi, nebulizer gas (nitrogen) pressure at 2 bar, capillary voltage at 4,200 V (L. Zhang et al., 2023b). The MS/MS data collection was carried out in IDA mode, where the top 10 ions of each spectrum were fragmented using collision-induced dissociative energy of 35 ± 10 eV (Li, Zhou, et al., 2023).