| Parameter | Condition | ||
|---|---|---|---|
| | |||
| Instrument | Waters e2695 separation module | ||
| Detector | Waters 2996 UV/VIS detector | ||
| Wavelength | 238 nm | ||
| Column | X-Bridge C18 (4.6×250 mm, 3.5 µm, Waters) | ||
| Column temperature | 30°C | ||
| Sample temperature | 15°C | ||
| Flow rate | 0.6 mL/min | ||
| Run time | 40 min | ||
| Injection Volume | 20 µL | ||
| Mobile phase | Time (min) | A: Water (%) | B: Acetonitrile (%) |
| Initial | 95 | 5 | |
| 20 | 70 | 30 | |
| 22 | 5 | 95 | |
| 30 | 5 | 95 | |
| 32 | 95 | 5 | |
| 40 | 95 | 5 | |