| Parameter | Conditions | ||
|---|---|---|---|
| Instrument | Waters 1525 binary HPLC pump system (Miami, FL, USA) | ||
| Detector | Waters 2489 UV/VIS detector 205 nm (Miami, FL, USA) | ||
| Column | YMC Pack-Pro C18 column (250×4.6 mm, 5 ¼m, YMC Co., Ltd., Kyoto, Japan) | ||
| Column temperature | 30°C | ||
| Flow rate | 0.9 mL/min | ||
| Elent | A: 0.1% trifluoroacetic acid/water B: acetonitrile | ||
| Injection volume | 10 | ||
| Gradient condition | |||
| 0 | 80 | 20 | |
| 15 | 80 | 20 | |
| 55 | 40 | 60 | |
| 57 | 5 | 95 | |
| 62 | 5 | 95 | |
| 65 | 80 | 20 | |
| 75 | 80 | 20 | |