| Separation Variable |
Value |
| Column |
Zorbax Eclipse Plus C18 column
(250 × 4.6 mm, 5 μm) |
| Mobile phase |
0.1% formic acid (A) and Acetonitrile (B) |
| Diluent |
Acetonitrile and water (50:50) |
| Separation mode |
Gradient mode |
| |
Time |
0.1% formic acid (A) |
ACN (B) |
| 0 min |
60% |
40% |
| 1 min |
60% |
40% |
| 10 min |
10% |
90% |
| 18 min |
60% |
40% |
| 20 min |
60% |
40% |
| Flow rate |
1.0 mL min-1 |
| Temperature |
Ambient |
| Samples size |
20 μL |
| Detection wavelength |
252 nm |
| Retention time |
4.82 min |
|
