LC-MS/MS parameters
| LC system . | Ultimate 3000 . |
|---|---|
| Gradient | 5 min (4% buffer B), 80 min (26%), 5 min (32%), 5 min (95%), 5 min (95%), 1 min (4%), 9 min (4%) |
| Mass spectrometer | Fusion lumos |
| Scan cycle | 3 s |
| MS1 resolution | 120,000 |
| MS1 scan range | 400–1,600 daltons |
| Injection time | 50 s |
| AGC for MS1 | 106 |
| APD | On |
| Included charge states | 2–6 |
| Exclusion duration | 60 s |
| Isolation windows | 1.6 dalton |
| NCE | 35% |
| MS2 resolution | 60,000 |
| Injection time | Dynamic |
| AGC for MS2 | 125,000 |
| LC system . | Ultimate 3000 . |
|---|---|
| Gradient | 5 min (4% buffer B), 80 min (26%), 5 min (32%), 5 min (95%), 5 min (95%), 1 min (4%), 9 min (4%) |
| Mass spectrometer | Fusion lumos |
| Scan cycle | 3 s |
| MS1 resolution | 120,000 |
| MS1 scan range | 400–1,600 daltons |
| Injection time | 50 s |
| AGC for MS1 | 106 |
| APD | On |
| Included charge states | 2–6 |
| Exclusion duration | 60 s |
| Isolation windows | 1.6 dalton |
| NCE | 35% |
| MS2 resolution | 60,000 |
| Injection time | Dynamic |
| AGC for MS2 | 125,000 |
LC-MS/MS, liquid chromatography–tandem mass spectrometry; APD, advanced peak determination; NCE, normalized collision energy; AGC, automatic gain control.