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MB Sample ID: SA206355
| Local Sample ID: | WT_1.raw |
| Subject ID: | SU002238 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002238 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| WT_1.raw | SA206355 | FL025283 | Wild type | Sample Source |
Collection:
| Collection ID: | CO002231 |
| Collection Summary: | Wild-type (WT) and Sirt5-/- (SIRT5 KO) mice on 129 background were maintained on a standard chow diet (Teklad Global 18% Protein Rodent Diet, ENVIGO, Cat.#2018) until they were euthanized. Metabolites extracted from eight WT and eight Sirt5-/- mouse livers were analyzed by LC/MS. Intracellular metabolites and acyl-CoA from cells were harvested as described previously using 80% methanol/water as solvent (PMID: 24410464, PMID: 24894601, PMID: 25795660) and dry pellets were stored in -80 °C freezer until ready for LC-MS analysis. For acyl-CoA and 5-methyltetrahydrofolate analysis, dry pellets were reconstituted into 30 μL of sample solvent (water containing 50 mM ammonium acetate), and 8 μL was injected into the LC-MS. For non-acyl-CoA polar metabolite analysis, pellets were reconstituted into 30 μL of sample solvent (water:methanol:acetonitrile, 2:1:1, v/v), and 3 μL was injected into the LC-MS. |
| Sample Type: | Liver |
Treatment:
| Treatment ID: | TR002250 |
| Treatment Summary: | Eight Wild-type (WT) and eight Sirt5-/- (SIRT5 KO) mice on 129 background were maintained on a standard chow diet (Teklad Global 18% Protein Rodent Diet, ENVIGO, Cat.#2018) until they were euthanized. Mice had free access to food and water. Mouse body weight and food intake were monitored weekly. |
Sample Preparation:
| Sampleprep ID: | SP002244 |
| Sampleprep Summary: | Intracellular metabolites and acyl-CoA from mouse liver tissues were harvested as described previously using 80% methanol/water as solvent (PMID: 24410464, PMID: 24894601, PMID: 25795660) and dry pellets were stored in -80 °C freezer until ready for LC-MS analysis. For acyl-CoA and 5-methyltetrahydrofolate analysis, dry pellets were reconstituted into 30 μL of sample solvent (water containing 50 mM ammonium acetate), and 8 μL was injected into the LC-MS. For non-acyl-CoA polar metabolite analysis, pellets were reconstituted into 30 μL of sample solvent (water:methanol:acetonitrile, 2:1:1, v/v), and 3 μL was injected into the LC-MS. |
Combined analysis:
| Analysis ID | AN003523 | AN003524 | AN003525 |
|---|---|---|---|
| Analysis type | MS | MS | MS |
| Chromatography type | HILIC | HILIC | Reversed phase |
| Chromatography system | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 |
| Column | Waters Xbridge amide (100 x 2.1mm,3.5um) | Waters Xbridge amide (100 x 2.1mm,3.5um) | Phenomenex Luna C18 (100 x 2.0mm,3um) |
| MS Type | ESI | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE | POSITIVE |
| Units | ion counts | ion counts | ion counts |
Chromatography:
| Chromatography ID: | CH002602 |
| Chromatography Summary: | HILIC method is for general metabolomics analysis. |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Waters Xbridge amide (100 x 2.1mm,3.5um) |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH002603 |
| Chromatography Summary: | RPLC is for acyl-CoA and folate analysis. |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Phenomenex Luna C18 (100 x 2.0mm,3um) |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003281 |
| Analysis ID: | AN003523 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | It was operated in the full-scan mode with a scan range of 60-900 and the resolution set at 70 000 (at m/z 200).LC-MS data were analyzed using Sieve 2.0 (Thermo Scientific), and the integrated area under metabolite peak was used to compare the relative abundance of each metabolite in different samples in the same batch. |
| Ion Mode: | POSITIVE |
| MS ID: | MS003282 |
| Analysis ID: | AN003524 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | It was operated in the full-scan mode with a scan range of 60-900 and the resolution set at 70 000 (at m/z 200).LC-MS data were analyzed using Sieve 2.0 (Thermo Scientific), and the integrated area under metabolite peak was used to compare the relative abundance of each metabolite in different samples in the same batch. |
| Ion Mode: | NEGATIVE |
| MS ID: | MS003283 |
| Analysis ID: | AN003525 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | It was operated in the full-scan mode with a scan range of 150-1000 and the resolution set at 70 000 (at m/z 200).LC-MS data were analyzed using Sieve 2.0 (Thermo Scientific), and the integrated area under metabolite peak was used to compare the relative abundance of each metabolite in different samples in the same batch. |
| Ion Mode: | POSITIVE |
