| Instrument | Shimadzu HPLC system (HPLC-UV) | |
|---|---|---|
| Column | XBridge C18 (4.6×250 mm, 5 um) | |
| Mobile phase | Solution(A) : 0.1% TFA in D.W | |
| Solution(B) : Acetonitrile | ||
| Time (min) | B Conc. (%) | |
| 0.1 | 10 | |
| 5 | 10 | |
| 15 | 30 | |
| 20 | 47 | |
| 30 | 47 | |
| 33 | 10 | |
| 40 | − | |
| Oven temp. | 35°C | |
| Wavelength | 330 nm | |
| Injection volume | 10 μL | |
| Flow rate | 1 mL/min | |
| Run time | 40 min | |