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MB Sample ID: SA123933
Local Sample ID: | 200 |
Subject ID: | SU001524 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001524 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
200 | SA123933 | FL014997 | Agilent 6545 centroid data | Data type |
Collection:
Collection ID: | CO001519 |
Collection Summary: | Human serum was frozen at -80 C. |
Sample Type: | Blood (serum) |
Treatment:
Treatment ID: | TR001539 |
Treatment Summary: | See sampleprep |
Sample Preparation:
Sampleprep ID: | SP001532 |
Sampleprep Summary: | 4 mL frozen commercial pooled human serum (Innovative Research, Novi, MI, USA) was thawed at 4 °C, vortexed and aliquoted into 50 µL portions in 2 mL Eppendorf vials. Every 50 µL portion was mixed with 250 µL cold methanol and vortexed to precipitate proteins.38 After 20 min incubation at −20 °C, these mixtures were centrifuged at 20,800 x g for 10 min at 4 °C. The supernatants were transferred into clean 2.0 mL Eppendorf vials and then dried in an Eppendorf Vacufuge (Brinkmann Instruments, Westbury, NY, USA). The residue in each Eppendorf vial was reconstituted in 50 µL H2O:ACN (2:3 v/v), vortexed and centrifuged at 20,800 x g for 10 min at 4 °C. The supernatant in all Eppendorf vials was pooled into a 5 mL Eppendorf vial, vortexed and centrifuged at xx,xxx x g (To Dan: I should have used 5000 rpm or a typical rpm in the big centrifuge in the biosample prep lab) for 10 min at 4 °C to further remove any solid residue. The resultant supernatant was aliquoted into 50 µL portions in 1.5 mL Eppendorf vials and stored at −80 °C. Prior to repeated injections to LC-MS in this work, 8 portions were diluted to 200 µL each with H2O:ACN (2:3 v/v), pooled into a 2 mL LC vial and vortexed. |
Combined analysis:
Analysis ID | AN002421 | AN002422 | AN002423 | AN002424 |
---|---|---|---|---|
Analysis type | MS | MS | MS | MS |
Chromatography type | HILIC | HILIC | HILIC | HILIC |
Chromatography system | Agilent 1260 Infinity | Agilent 1260 Infinity | Agilent 1290 Infinity | Agilent 1290 Infinity |
Column | Waters XBridge BEH Amide (15 cm x 2.1mm,2.5um) | Waters XBridge BEH Amide (15 cm x 2.1mm,2.5um) | Waters XBridge BEH Amide (15 cm x 2.1mm,2.5um) | Waters XBridge BEH Amide (15 cm x 2.1mm,2.5um) |
MS Type | ESI | ESI | ESI | ESI |
MS instrument type | QTOF | QTOF | QTOF | QTOF |
MS instrument name | Agilent 6520 QTOF | Agilent 6520 QTOF | Agilent 6545 QTOF | Agilent 6545 QTOF |
Ion Mode | POSITIVE | POSITIVE | POSITIVE | POSITIVE |
Units | Peak area | Peak area | Peak area | Peak area |
Chromatography:
Chromatography ID: | CH001778 |
Chromatography Summary: | The mobile phase consisted of (A) H2O:ACN (95:5, v/v) with 5 mM ammonium acetate and 0.1% acetic acid, and (B) H2O:ACN (5:95, v/v), 5 mM ammonium acetate and 0.1% acetic acid. Gradient elution was performed as follows: 100% mobile phase B for 1.5 min, 100 to 78% B from 1.5-6.0 min, 78 to 50% B from 6.0-9.0 min, 50% B from 9.0-15.0 min, restoration to 100% B from 15.0-17.0 min, and continued 100% B from 17.0-30.0 to equilibrate the LC column. The flow rate was 0.3 mL/min, the injection volume was 5 µL, followed by an H2O:ACN (5:95, v/v) needle wash for 10 s, and the column temperature was 35 oC. |
Instrument Name: | Agilent 1260 Infinity |
Column Name: | Waters XBridge BEH Amide (15 cm x 2.1mm,2.5um) |
Column Temperature: | 35 |
Flow Gradient: | 100% mobile phase B for 1.5 min, 100 to 78% B from 1.5-6.0 min, 78 to 50% B from 6.0-9.0 min, 50% B from 9.0-15.0 min, restoration to 100% B from 15.0-17.0 min, and continued 100% B from 17.0-30.0 to equilibrate the LC column. |
Flow Rate: | 0.3 mL/min |
Solvent A: | 95% water/5% acetonitrile; 0.1% acetic acid; 5 mM ammonium acetate |
Solvent B: | 5% water/95% acetonitrile; 0.1% acetic acid; 5 mM ammonium acetate |
Chromatography Type: | HILIC |
Chromatography ID: | CH001779 |
Chromatography Summary: | The mobile phase consisted of (A) H2O:ACN (95:5, v/v) with 5 mM ammonium acetate and 0.1% acetic acid, and (B) H2O:ACN (5:95, v/v), 5 mM ammonium acetate and 0.1% acetic acid. Gradient elution was performed as follows: 100% mobile phase B for 1.5 min, 100 to 78% B from 1.5-6.0 min, 78 to 50% B from 6.0-9.0 min, 50% B from 9.0-15.0 min, restoration to 100% B from 15.0-17.0 min, and continued 100% B from 17.0-30.0 to equilibrate the LC column. The flow rate was 0.3 mL/min, the injection volume was 5 µL, followed by an H2O:ACN (5:95, v/v) needle wash for 10 s, and the column temperature was 35 oC. |
Instrument Name: | Agilent 1260 Infinity |
Column Name: | Waters XBridge BEH Amide (15 cm x 2.1mm,2.5um) |
Column Temperature: | 35 |
Flow Gradient: | 100% mobile phase B for 1.5 min, 100 to 78% B from 1.5-6.0 min, 78 to 50% B from 6.0-9.0 min, 50% B from 9.0-15.0 min, restoration to 100% B from 15.0-17.0 min, and continued 100% B from 17.0-30.0 to equilibrate the LC column. |
Flow Rate: | 0.3 mL/min |
Solvent A: | 95% water/5% acetonitrile; 0.1% acetic acid; 5 mM ammonium acetate |
Solvent B: | 5% water/95% acetonitrile; 0.1% acetic acid; 5 mM ammonium acetate |
Chromatography Type: | HILIC |
Chromatography ID: | CH001780 |
Chromatography Summary: | The mobile phase consisted of (A) H2O:ACN (95:5, v/v) with 5 mM ammonium acetate and 0.1% acetic acid, and (B) H2O:ACN (5:95, v/v), 5 mM ammonium acetate and 0.1% acetic acid. Gradient elution was performed as follows: 100% mobile phase B for 1.5 min, 100 to 78% B from 1.5-6.0 min, 78 to 50% B from 6.0-9.0 min, 50% B from 9.0-15.0 min, restoration to 100% B from 15.0-17.0 min, and continued 100% B from 17.0-30.0 to equilibrate the LC column. The flow rate was 0.3 mL/min, the injection volume was 5 µL, followed by an H2O:ACN (5:95, v/v) needle wash for 10 s, and the column temperature was 35 oC. |
Instrument Name: | Agilent 1290 Infinity |
Column Name: | Waters XBridge BEH Amide (15 cm x 2.1mm,2.5um) |
Column Temperature: | 35 |
Flow Gradient: | 100% mobile phase B for 1.5 min, 100 to 78% B from 1.5-6.0 min, 78 to 50% B from 6.0-9.0 min, 50% B from 9.0-15.0 min, restoration to 100% B from 15.0-17.0 min, and continued 100% B from 17.0-30.0 to equilibrate the LC column. |
Flow Rate: | 0.3 mL/min |
Solvent A: | 95% water/5% acetonitrile; 0.1% acetic acid; 5 mM ammonium acetate |
Solvent B: | 5% water/95% acetonitrile; 0.1% acetic acid; 5 mM ammonium acetate |
Chromatography Type: | HILIC |
Chromatography ID: | CH001781 |
Chromatography Summary: | The mobile phase consisted of (A) H2O:ACN (95:5, v/v) with 5 mM ammonium acetate and 0.1% acetic acid, and (B) H2O:ACN (5:95, v/v), 5 mM ammonium acetate and 0.1% acetic acid. Gradient elution was performed as follows: 100% mobile phase B for 1.5 min, 100 to 78% B from 1.5-6.0 min, 78 to 50% B from 6.0-9.0 min, 50% B from 9.0-15.0 min, restoration to 100% B from 15.0-17.0 min, and continued 100% B from 17.0-30.0 to equilibrate the LC column. The flow rate was 0.3 mL/min, the injection volume was 5 µL, followed by an H2O:ACN (5:95, v/v) needle wash for 10 s, and the column temperature was 35 oC. |
Instrument Name: | Agilent 1290 Infinity |
Column Name: | Waters XBridge BEH Amide (15 cm x 2.1mm,2.5um) |
Column Temperature: | 35 |
Flow Gradient: | 100% mobile phase B for 1.5 min, 100 to 78% B from 1.5-6.0 min, 78 to 50% B from 6.0-9.0 min, 50% B from 9.0-15.0 min, restoration to 100% B from 15.0-17.0 min, and continued 100% B from 17.0-30.0 to equilibrate the LC column. |
Flow Rate: | 0.3 mL/min |
Solvent A: | 95% water/5% acetonitrile; 0.1% acetic acid; 5 mM ammonium acetate |
Solvent B: | 5% water/95% acetonitrile; 0.1% acetic acid; 5 mM ammonium acetate |
Chromatography Type: | HILIC |
MS:
MS ID: | MS002258 |
Analysis ID: | AN002421 |
Instrument Name: | Agilent 6520 QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | The ESI conditions were as follows: Electrospray ion source ESI Agilent Jet Stream Technology in positive ionization mode; voltage 3.8 kV; desolvation temperature 325 °C; cone flow 20 L/h; desolvation gas flow 600 L/h; nebulizer pressure 45 psi, N2 drying gas; MS scan rate of 1.03 spectra/s across the range m/z 60-1000. The software, Progenesis Qi, was used to process raw data. In the results, every compound (feature) had at least two ions, like an ion and its isotope ion or adduct ion. Compounds with single ions were filtered out. If a compound had m/z information only, the m/z was directly used in the compound name. If a compound had neutral mass information, the neutral mass was converted to m/z. |
Ion Mode: | POSITIVE |
MS ID: | MS002259 |
Analysis ID: | AN002422 |
Instrument Name: | Agilent 6520 QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | The ESI conditions were as follows: Electrospray ion source ESI Agilent Jet Stream Technology in positive ionization mode; voltage 3.8 kV; desolvation temperature 325 °C; cone flow 20 L/h; desolvation gas flow 600 L/h; nebulizer pressure 45 psi, N2 drying gas; MS scan rate of 1.03 spectra/s across the range m/z 60-1000. The software, Progenesis Qi, was used to process raw data. In the results, every compound (feature) had at least two ions, like an ion and its isotope ion or adduct ion. Compounds with single ions were filtered out. If a compound had m/z information only, the m/z was directly used in the compound name. If a compound had neutral mass information, the neutral mass was converted to m/z. |
Ion Mode: | POSITIVE |
MS ID: | MS002260 |
Analysis ID: | AN002423 |
Instrument Name: | Agilent 6545 QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | (Agilent 6545) MS scan rate of 1.03 spectra/s across the range m/z 60-1000. The software, Progenesis Qi, was used to process raw data. In the results, every compound (feature) had at least two ions, like an ion and its isotope ion or adduct ion. Compounds with single ions were filtered out. If a compound had m/z information only, the m/z was directly used in the compound name. If a compound had neutral mass information, the neutral mass was converted to m/z. |
Ion Mode: | POSITIVE |
MS ID: | MS002261 |
Analysis ID: | AN002424 |
Instrument Name: | Agilent 6545 QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | (Agilent 6545) MS scan rate of 1.03 spectra/s across the range m/z 60-1000. The software, Progenesis Qi, was used to process raw data. In the results, every compound (feature) had at least two ions, like an ion and its isotope ion or adduct ion. Compounds with single ions were filtered out. If a compound had m/z information only, the m/z was directly used in the compound name. If a compound had neutral mass information, the neutral mass was converted to m/z. |
Ion Mode: | POSITIVE |