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MB Sample ID: SA021102
Local Sample ID: | 150127cctsa36_1 |
Subject ID: | SU000440 |
Subject Type: | Animal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Gender: | Male |
Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU000440 |
Subject Type: | Animal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Gender: | Male |
Species Group: | Mammal |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
150127cctsa36_1 | SA021102 | FL005286 | Lung | Organ |
150127cctsa36_1 | SA021102 | FL005286 | High Fat Chow | Diet Assignment |
Collection:
Collection ID: | CO000434 |
Collection Summary: | C57BL/6 mice, 6-7 weeks of age upon diet assignment were fed for 150 days and weighed daily. Mice were euthanized with an overdose of pentobarbital IP and lungs were flash frozen. 6 mg of lung tissue was extracted for GC-TOFMS and HILIC-QTOFMS analysis. |
Sample Type: | Tissue |
Treatment:
Treatment ID: | TR000454 |
Treatment Summary: | Male C57BL/6N mice (6-7 weeks of age) were provided ad libitum access to one of three diets for 150 days: low fat (10% kcals) control (CTRL) chow, high fat (45% kcals) with sugar (HFS) chow or very high (60% kcals) fat (VHF) chow. Body weight and food intake were measured daily. |
Treatment Doseduration: | 150 Days |
Sample Preparation:
Sampleprep ID: | SP000447 |
Sampleprep Summary: | 1. Weigh 50 mg tissue sample in to a 25 ml conical polypropylene centrifuge tube. 2. Add 2.5mL extraction solvent to the tissue sample and homogenize for 45 seconds ensuring that sample resembles a powder. In between samples, clean the homogenizer in solutions of methanol, acetone, water, and the extraction solvent. 3. Centrifuge the samples at 2500 rpm. for 5 minutes. Aliquot 2 X 500μl supernatant, one for analysis and one for a backup sample. Store backup aliquot in the -20°C freezer. 4. Evaporate one 500μl aliquot of the sample in the Labconco Centrivap cold trap concentrator to complete dryness 5. The dried aliquot is then re-suspended with 500l 50% acetonitrile (degassed as given) 6. Centrifuge for 2 min at 14000 rcf using the centrifuge Eppendorf 5415. 7. Remove supernatant to a new Eppendorff tube. 8. Evaporate the supernatant to dryness in the the Labconco Centrivap cold trap concentrator. 9. Submit to derivatization. |
Sampleprep Protocol Filename: | SOP Extraction of Mammalian Tissue Samples.pdf |
Combined analysis:
Analysis ID | AN000661 |
---|---|
Analysis type | MS |
Chromatography type | GC |
Chromatography system | Leco Pegasus 4D GC |
Column | Restek Corporation Rtx-5Sil MS |
MS Type | EI |
MS instrument type | GC x GC-TOF |
MS instrument name | Leco Pegasus 4D GCxGC TOF |
Ion Mode | POSITIVE |
Units | counts |
Chromatography:
Chromatography ID: | CH000478 |
Methods Filename: | Data_Dictionary_Fiehn_laboratory_GCTOF_MS_primary_metabolism_10-15-2013_general.pdf |
Instrument Name: | Leco Pegasus 4D GC |
Column Name: | Restek Corporation Rtx-5Sil MS |
Column Pressure: | 7.7 PSI |
Column Temperature: | 50-330C |
Flow Rate: | 1 ml/min |
Injection Temperature: | 50 C ramped to 250 C by 12 C/s |
Sample Injection: | 0.5 uL |
Oven Temperature: | 50°C for 1 min, then ramped at 20°C/min to 330°C, held constant for 5 min |
Transferline Temperature: | 230C |
Washing Buffer: | Ethyl Acetate |
Sample Loop Size: | 30 m length x 0.25 mm internal diameter |
Randomization Order: | Excel generated |
Chromatography Type: | GC |
MS:
MS ID: | MS000587 |
Analysis ID: | AN000661 |
Instrument Name: | Leco Pegasus 4D GCxGC TOF |
Instrument Type: | GC x GC-TOF |
MS Type: | EI |
Ion Mode: | POSITIVE |
Ion Source Temperature: | 250 C |
Ionization Energy: | 70 eV |
Mass Accuracy: | Nominal |
Source Temperature: | 250 C |
Scan Range Moverz: | 85-500 Da |
Scanning Cycle: | 17 Hz |
Scanning Range: | 85-500 Da |
Skimmer Voltage: | 1850 V |