| Parameters | Analysis condition | ||
|---|---|---|---|
| UPLC system | LC-30AD SHIMADZU | ||
| Detector | DAD, UV, 280 μm | ||
| Column | Kinetex XB.C18 100 Å (1.7 μm, 150×2.1 mm) | ||
| Mobile phase | A: 0.1% acetic acid in water (V/V, %) | ||
| B: 0.1% acetic acid in acetonitrile (V/V, %) | |||
| Gradient elution | Time (min) | A (%) | B (%) |
| 0.5 | 90 | 10 | |
| 2.5 | 60 | 40 | |
| 4.5 | 45 | 55 | |
| 6.0 | 40 | 60 | |
| 8.0 | 35 | 65 | |
| 9.5 | 35 | 65 | |
| 11.5 | 35 | 65 | |
| 13.0 | 30 | 70 | |
| 14.5 | 25 | 75 | |
| 16.0 | 20 | 80 | |
| 17.5 | 15 | 85 | |
| 20.0 | 10 | 90 | |
| 25.0 | 10 | 90 | |
| 26.0 | 90 | 10 | |
| 29.0 | 90 | 10 | |
| Injection volume (μL) | 2 | ||
| Flow rate (mL/min) | 0.3 | ||
| Column temp. (°C) | 30 | ||