이전 사이트 이동

Grinded samples were prepared in 70% MeOH at a concentration of 25mg/mL and then sonicated (90 min, 2 times). After extract, these samples were centrifuged and their supernatant were collected. The collected supernatants were dilluted by water and filtered by 0.20 µm membrane filter (Ministart RC 15, Sartorius stedim Biotech, Germany).

LC separations were carried out on an Waters Acquity UPLC BEH C18 column (2.1 x 100 mm, 1.7 µm). Column temperature and flow rate were set to 40 °C and 0.3 mL/min, respectively. The mobile phases used were 0.1 % formic acid in water (A) and acetonitrile (B). The gradients were as follows: isocratic 20% B (0-9 min), 20-30% B (9-14 min), isocratic 30% B (14-17 min), 30-32% B (17-21 min), 32-42.5% B (21-26 min), 42.5-90% B (26-29 min), 90-20% B (29-30 min) followed by a 2 min re-equilibration phase at 20 % B, successively. The injection volume of the sample was set to 2 μl.

To obtain MS spectral data, LC-ESI-MS/MS analyses of samples were performed on a Waters Xevo G2 QTOF mass spectrometer (Manchester, UK) combined with a Waters Acquity UPLC system (Waters Co., Milford, MA, USA).