| LC | Column | CAPCELL CORE C18 (2.1 mm×150 mm×2.7 μm) |
| Column oven | 40°C |
| Injection volume | 2 μL |
| Flow rate | 0.3 mL/min |
| Mobile phase | A: 0.1% formic acid, 5 mM ammonium formate in water B: 0.1% formic acid, 5 mM ammonium formate in methanol |
| Gradient program | Time (min) | A (%) | B (%) |
| 0 | 95 | 5 |
| 1.0 | 95 | 5 |
| 1.5 | 70 | 30 |
| 12.0 | 2 | 98 |
| 16.0 | 2 | 98 |
| 16.1 | 95 | 5 |
| 20.0 | 95 | 5 |
| MS/MS | Ionization mode | Electrospray ionization (ESI) |
| Ion source | HESI |
| Spray voltage | 3.20 kV |
| Capillary temperature | 320°C |
| Aux gas heater temperature | 350°C |
| Sheath gas flow (N2) | 40 L/min |
| Aux gas flow (N2) | 15 L/min |
| Sweep gas flow (N2) | 1 L/min |