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MB Sample ID: SA227242
Local Sample ID: | 16 |
Subject ID: | SU002402 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002402 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
16 | SA227242 | FL027592 | ndufa9f/f | Genotype |
Collection:
Collection ID: | CO002395 |
Collection Summary: | Liver was frozen on liqN2. Liver was crushed on liqN2, and metabolites extracted with 80% meOH. MeOH was dried, and samples resuspended in MPA and injected onto a qTOF. |
Sample Type: | Liver |
Treatment:
Treatment ID: | TR002414 |
Treatment Summary: | 8 weeks post AAV injection the mouse liver was pulverized, metabolites extracted |
Sample Preparation:
Sampleprep ID: | SP002408 |
Sampleprep Summary: | 80% methanol containing metabolites was dried overnight in a speedvac. Dried pellets were derivatized using methoxyamine and MTBSTFA. Samples were immediately injected. |
Combined analysis:
Analysis ID | AN003783 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Agilent 1290 |
Column | Waters Acquity UPLC HSS T3 (150 x 2.1mm,1.8um) |
MS Type | ESI |
MS instrument type | QTOF |
MS instrument name | Agilent 6550 QTOF |
Ion Mode | UNSPECIFIED |
Units | area |
Chromatography:
Chromatography ID: | CH002797 |
Chromatography Summary: | Data acquisition was performed by reverse-phase chromatography on a 1290 UHPLC liquid chromatography (LC) system interfaced to a high-resolution mass spectrometry (HRMS) 6550 iFunnel Q-TOF mass spectrometer (MS) (Agilent Technologies, CA). The MS was operated in both positive and negative (ESI+ and ESI-) modes. Analytes were separated on an Acquity UPLC® HSS T3 column (1.8 μm, 2.1 x 150 mm, Waters, MA). The column was kept at room temperature. Mobile phase A composition was 0.1% formic acid in water and mobile phase B composition was 0.1% formic acid in 100% ACN. The LC gradient was 0 min: 1% B; 5 min: 5% B; 15 min: 99% B; 23 min: 99% B; 24 min: 1% B; 25 min: 1% B. The flow rate was 250 μL min-1. The sample injection volume was 5 μL. ESI source conditions were set as follows: dry gas temperature 225 °C and flow 18 L min-1, fragmentor voltage 175 V, sheath gas temperature 350 °C and flow 12 L min-1, nozzle voltage 500 V, and capillary voltage +3500 V in positive mode and −3500 V in negative. The instrument was set to acquire over the full m/z range of 40–1700 in both modes, with the MS acquisition rate of 1 spectrum s-1 in profile format. Raw data files were processed using Profinder B.08.00 SP3 software (Agilent Technologies, CA) with an in-house database containing retention time and accurate mass information on 600 standards from Mass Spectrometry Metabolite Library (IROA Technologies, MA) which was created under the same analysis conditions. The in-house database matching parameters were mass tolerance 10 ppm; retention time tolerance 0.5 min. Peak integration result was manually curated in Profinder for improved consistency |
Chromatography Comments: | The LC gradient was 0 min: 1% B; 5 min: 5% B; 15 min: 99% B; 23 min: 99% B; 24 min: 1% B; 25 min: 1% B. |
Instrument Name: | Agilent 1290 |
Column Name: | Waters Acquity UPLC HSS T3 (150 x 2.1mm,1.8um) |
Flow Gradient: | 0 min: 1% B; 5 min: 5% B; 15 min: 99% B; 23 min: 99% B; 24 min: 1% B; 25 min: 1% B |
Flow Rate: | 250 uL/min |
Injection Temperature: | 37 |
Sample Injection: | 5 uL |
Solvent A: | 100% water; 0.1% formic acid |
Solvent B: | 100% acetonitrile; 0.1% formic acid |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS003526 |
Analysis ID: | AN003783 |
Instrument Name: | Agilent 6550 QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | The MS was operated in both positive and negative (ESI+ and ESI-) modes. Raw data files were processed using Profinder B.08.00 SP3 software (Agilent Technologies, CA) with an in-house database containing retention time and accurate mass information on 600 standards from Mass Spectrometry Metabolite Library (IROA Technologies, MA) which was created under the same analysis conditions. The in-house database matching parameters were mass tolerance 10 ppm; retention time tolerance 0.5 min. Peak integration result was manually curated in Profinder for improved consistency |
Ion Mode: | UNSPECIFIED |