| Instrument | SCL-40 controller unit that includes SPD-M40 UV, LC-40D xs, SIL-40C xs, and CTO-40C (Shimadzu Scientific Company, Tokyo, Japan) |
|---|---|
| Column | Agilent Eclipse Plus C18 (100 × 2.1 mm, 1.8 µm) |
| Detector | UV detector (204nm) |
| Flow rate | 0.5 ml/min |
| Injection volume | 10 µL |
| Gradient elution | Water (A), acetonitrile (B) |
| 0–7 min, 81% A, 19% B | |
| 7–11 min, 80% A, 20% B | |
| 11-14 min, 74% A, 26% B | |
| 14–25 min, 67% A, 33% B | |
| 25-35 min, 50% A, 50% B | |
| 35-40 min, 40% A, 60% B | |
| 40-42 min, 81% A, 19% B | |
| 42-45 min, 81% A, 19% B |