이전 사이트 이동

The branch and leaf of Acer pseudosieboldianum were extracted using 70% ethanol. After complete solvent removal by evaporation, the extracts were dissolved in DMSO at a concentration of 20 mg/mL. Each sample was assigned a Freshwater Biosources Culture Collection (FBCC) number and deposited at the Nakdonggang National Institute of Biological Resources (NNIBR). For metabolite analysis, extract samples (FBCC-EP1329 and FBCC-EP1331) were provided from NNIBR. For non-targeted analysis, peak identification was supported by quality control (QC) samples. These QC samples were prepared by mixing equal amounts of the Acer pseudosieboldianum branch extract (FBCC-EP1329) and leaf extract (FBCC-EP1331), and a seven-point QC set was generated within the concentration range of 700–1300 µg/mL. Both extracts were diluted to 1000 ppm for analysis. Prior to injection, all samples were freeze-dried to remove DMSO and reconstituted in 50% methanol containing 0.5 µg/mL of galangin as an internal standard. Subsequently, the samples were filtered through a 0.2 μm membrane filter and analyzed by UPLC-qTOF–MS.

Samples were injected into an ACQUITY UPLC I-Class system (Waters, Milford, MA, USA) equipped with a Phenomenex Luna Omega Polar C18 column (1.6 µm, 150 mm × 2.1 mm). The column oven temperature was maintained at 40 °C, the injection volume was 10 µL, and the flow rate was set to 0.3 mL/min. The mobile phases consisted of solvent A (0.5% formic acid in water, v/v) and solvent B (0.5% formic acid in acetonitrile, v/v). The gradient program was as follows: 0–5 min, 95% A; 5–20 min, 95–70% A; 20–28 min, 70–40% A; 28–30 min, 40–10% A; 30–32 min, 10% A; 32–35 min, 10–95% A; and 35–40 min, 95% A.

Non-targeted flavonoid profiling was performed on a Xevo G3-qTOF mass spectrometer (Waters, Milford, MA, USA) operated in MSe mode with a scan time of 0.5 sec and a mass range of m/z 50–1200. Ramp collision energy was set to 20–40 V. The source temperature was 100 °C, and both positive and negative ionization modes were used. In positive mode, the capillary voltage was 3.0 kV, sampling cone voltage was 40 V, desolvation temperature was 250 °C, and desolvation gas flow was 600 L/hr. In negative mode, the capillary voltage was 2.5 kV, sampling cone voltage was 40 V, desolvation temperature was 250 °C, and a desolvation gas flow of 800 L/hr.