Table 22.
Mass Spectrometric Parameters (Data Dependent Acquisition)
| Ion Source Properties | |
| Ion Source Type | NSI |
| Spray Voltage | Static |
| Positive Ion (V) | 1800 |
| Gas Mode | Static |
| Sweep Gas (Arb) | 0 |
| Ion Transfer Tube Temp (°C) | 275 |
| FAIMS Mode | Not Installed |
| Tune Settings | |
| Application Mode | Peptide |
| Method Duration | 140 |
| Infusion Mode | Liquid Chromatography |
| Expected LC Peak Width (s) | 15 |
| Advanced Peak Determination | Checked |
| Default Charge State | 2 |
| Internal Mass Calibration | User-defined Lock Mass |
| Current Lock Mass for Positive Ion Mode | 445.12006 |
| Method | |
| Time Range (min) | 0–140 |
| Full Scan | 2 sec |
| MIPS | |
| Intensity | |
| Charge State | |
| Dynamic Exclusion | |
| ddMS2 | |
| Full Scan Properties | |
| Orbitrap Resolution | 120000 |
| Scan Range (m/z) | 375–1600 |
| RF Lens (%) | 40 |
| AGC Target | Custom |
| Normalized AGC Target (%) | 300 |
| Maximum Injection Time Mode | Custom |
| Maximum Injection Time (ms) | 100 |
| Microscans | 1 |
| Data Type | Profile |
| Polarity | Positive |
| Source Fragmentation | Unchecked |
| MIPS Properties | |
| Monoisotopic peak determination | Peptide |
| Relax restrictions when too few precursors are found | Checked |
| Intensity Properties | |
| Filter Type | Intensity Threshold |
| Intensity Threshold | 5.00E+03 |
| Charge State Properties | |
| Include charge state(s) | 2–6 |
| Include undetermined charge states | unchecked |
| Dynamic Exclusion Properties | |
| Dynamic Exclusion Mode | Custom |
| Exclude after n times | 1 |
| Exclusion duration (s) | 20 |
| Mass Tolerance | ppm |
| Low | 10 |
| High | 10 |
| Exclude isotopes | checked |
| Perform dependent scan on single charge state per precursor only | checked |
| Data Dependent MS2 Scan Properties | |
| Multiplex ions | unchecked |
| Isolation Window (m/z) | 1.4 |
| Isolation Offset | Off |
| Collision Energy Mode | Fixed |
| Collision Energy Type | Normalized |
| HCD Collision Energy (%) | 30 |
| Orbitrap Resolution | 7500 |
| TurboTMT | Off |
| Scan Range Mode | Define First Mass |
| First Mass (m/z) | 100 |
| AGC Target | Custom |
| Normalized AGC Target (%) | 100 |
| Maximum Injection Time Mode | Custom |
| Maximum Injection Time (ms) | 18 |
| Microscans | 1 |
| Data Type | Centroid |