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MB Sample ID: SA242309
Local Sample ID: | 3_pre |
Subject ID: | SU002509 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Male and female |
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Combined analysis:
Analysis ID | AN003941 |
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Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Waters NanoAcquity |
Column | Waters ACQUITY UPLC BEH C18 (150 mm x 75 μm, 1.7 μm, 150 Å) |
MS Type | ESI |
MS instrument type | Orbitrap |
MS instrument name | Thermo Q Exactive Orbitrap |
Ion Mode | POSITIVE |
Units | 1 |
Chromatography:
Chromatography ID: | CH002918 |
Chromatography Summary: | Four-plex pooled samples were reconstituted in 0.1% formic acid before injection. The HPLC-MS/MS analysis was conducted using a Waters nanoACQUITY UPLC coupled with Thermo Q Exactive Orbitrap MS. The separation column was in-house made with an emitter tip and dimensions of 75 μm inner diameter × 15 cm length. The column was packed with 1.7 μm, 150 Å, ethylene-bridged-hybrid (BEH) C18 material (Waters, Milford, MA). Mobile phase A was water containing 0.1% formic acid, and mobile phase B was acetonitrile containing 0.1% formic acid. The flow rate was set as 0.3 μL/min, and the LC gradient was 55 min and set as follows: 0−10 min, 3%-30% solvent B; 10−30 min, 30−80% B; 30-30.5 min, 80%-95% B; 30.5− 40.5 min, 95% B; 40.5−41 min, 95%-3% B; 41-55 min, 3% B. Positive ionization mode was used and full MS scans were acquired from m/z 180 to 800 at a resolution of 60 k, automatic gain control (AGC) was set as 5x 10^5, and a maximum injection time as set as 30 ms. The top 20 precursors were selected for normalized collision energy (NCE) dissociation (NCE = 30) with an isolation window of m/z 1, fixed first m/z 110, dynamic exclusion of 5 seconds, charge exclusion of >2, and a resolution of 35 k. |
Instrument Name: | Waters NanoAcquity |
Column Name: | Waters ACQUITY UPLC BEH C18 (150 mm x 75 μm, 1.7 μm, 150 Å) |
Column Temperature: | Room temperature |
Flow Gradient: | LC gradient was 55 min and set as follows: 0−10 min, 3%-30% solvent B; 10−30 min, 30−80% B; 30-30.5 min, 80%-95% B; 30.5− 40.5 min, 95% B; 40.5−41 min, 95%-3% B; 41-55 min, 3% B. |
Flow Rate: | 0.3 μL/min |
Solvent A: | 100% water; 0.01% formic acid |
Solvent B: | 100% acetonitrile; 0.01% formic acid |
Chromatography Type: | Reversed phase |