Uplc sunfire c18 column

The Prep-HPLC analysis was performed as described in our recent studies [9 (link),10 (link)]. The extract sample (10 mg/mL) was dissolved in ultrapure water (analytical grade, Merck KGaA, Darmstadt, Germany), centrifugated at 8000× g for 20 min, and then passed through 0.22 μm membrane (Shanghai Sangon Biological Engineeing Technology and Service Co., Ltd., Shanghai, China). The filtration was subjected to Prep-HPLC using a Waters 2707 (Waters, Milford, MA, USA) linked with a UPLC Sunfire C18 column (5 μm, 10 × 250 mm) (Waters, Milford, MA, USA) with the parameters described recently by Liu et al. [9 (link)]. After the Prep-HPLC, the isolated fragments were individually collected, concentrated, and evaporated using the rotary evaporator (IKA, Staufen, Germany), and then redissolved in ultrapure water for further analyses.

Aliquots (10 mg/mL) of freeze-dried samples resolved in H₂O (Analytical grade, Merck KGaA, Darmstadt, Germany) were centrifuged at 12,000 rpm for 20 min. The supernatant was filtered through 0.22 µm membrane (Sangon, Shanghai, China), and the filtration was collected for further analysis. Prep-HPLC was run using Waters 2707 (Waters, Milford, Massachusetts, USA) linked with UPLC Sunfire C18 column (5 μm, 10 × 250 mm) (Waters, Massachusetts, USA) at the following parameters: column temperature, 40 °C; injection volume, 100 μL; and mobile phase of methanol (eluent A) and water (eluent B) at a flow rate of 4 mL/min (isocratic elution: 0–15 min, 20% eluent A and 80% eluent B). Photo-diode array (PDA) spectra were measured in the wavelength ranging from 200 to 600 nm.

Aliquots of the extracted samples (10 mg/mL) were resolved, centrifuged, filtered, and subjected for the Prep-HPLC Analysis, using Waters 2707 (Waters, Milford, MA, USA) linked with UPLC Sunfire C18 column (5 µm, 10 × 250 mm) (Waters, Milford, MA, USA) with the same parameters and elution conditions described in our recent reports [15 (link),16 (link)].