| Parameters | Conditions | ||
|---|---|---|---|
| HPLC model | ACQUITY H-class (Waters) | ||
| Column | Shiseido capcellpak C18 UG (5 μm, 4.6×250 mm) | ||
| Column Temperature | 35°C | ||
| Flow rate | 1.0 mL/min | ||
| Injection | 10 μL | ||
| Mobile phase | A : 0.1% Phosphoric acid in water | ||
| B : Methanol | |||
| Gradient step | Time (min) | A (%) | B (%) |
| 0 | 70 | 30 | |
| 8.5 | 65 | 35 | |
| 12.0 | 55 | 45 | |
| 15.0 | 45 | 55 | |
| 18.0 | 30 | 70 | |
| 25.0 | 70 | 30 | |
| 35.0 | 70 | 30 | |
| Detector | PDA detector (370 nm) | ||