| HPLC | Jasco LC-4000 | ||
| Column | C18 (250 × 4.6 mm × 5 μm, YMC-Pack ODS-AQ) | ||
| Mobile phase | A: ACN (Acetonitrile), B: Water (D.W) | ||
| Detector | UV-vis detector (230 nm) | ||
| Run time | 28 min | ||
| Flow rate | 1 mL/min | ||
| Injection volume | 20 μL | ||
| Column temperature | 30°C | ||
| Method conditional (gradient) | Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 00.00 | 19 | 81 | |
| 05.00 | 19 | 81 | |
| 20.00 | 60 | 40 | |
| 20.01 | 100 | 0 | |
| 25.00 | 100 | 0 | |
| 25.01 | 19 | 81 | |
| 28.00 | 19 | 81 | |