| Column | ACQUITY UPLC BEH C18 1.7 um, 2.1 100 mm |
| Mobile phase | A solution : 25mM Ammonium acetate (pH 5.0) in 20% MeOH |
| B solution : Acetonitrile |
| Gradient condition | Time (min) | A (%) | B (%) | Curve |
| Initial | 75 | 25 | - |
| 3.0 | 65 | 35 | 6 |
| 3.5 | 60 | 40 | 6 |
| 4.0 | 75 | 25 | 1 |
| 5.0 | 75 | 25 | 1 |
| Flow rate | 0.35 mL/min |
| Injection volume | 10 uL |
| MS condition |
| Ionization mode : ESI negative Desolvation temperature : 500°C |
| Capillary voltage : 3.0 V | Desolvation gas flow: 150 L/hr |
| Source temperature : 150°C | Collision gas flow: 0.14mL/min |
| Cone gas flow :500°C | |