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MB Sample ID: SA055390
| Local Sample ID: | 141001actsa46_1 |
| Subject ID: | SU000967 |
| Subject Type: | Animal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57/BL6N |
| Gender: | Male |
| Animal Housing: | SPF facility |
| Animal Light Cycle: | 12:12 h light-dark cycle |
| Animal Feed: | Ad libitum |
| Animal Water: | Ad libitum |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000967 |
| Subject Type: | Animal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57/BL6N |
| Gender: | Male |
| Animal Housing: | SPF facility |
| Animal Light Cycle: | 12:12 h light-dark cycle |
| Animal Feed: | Ad libitum |
| Animal Water: | Ad libitum |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 141001actsa46_1 | SA055390 | FL010149 | WT C57-Bl6 | Treatment* |
| 141001actsa46_1 | SA055390 | FL010149 | HFD | Genotype |
| 141001actsa46_1 | SA055390 | FL010149 | 24 weeks | TIME_POINT |
Collection:
| Collection ID: | CO000961 |
| Collection Summary: | Liver tissue from mice on the diets for 24 weeks for metabolomic analysis was collected, rinsed in cold PBS, excess fluid was blotted with a kim-wipe and tissue was immediately snap frozen in liquid nitrogen before storage at -80°C. Blood was collected by cardiac puncture and centrifuged at 9 rcf for 5 min at 4°C. Plasma was stored immediately at -20°C. |
| Sample Type: | Blood |
Treatment:
| Treatment ID: | TR000981 |
| Treatment Summary: | Male C57/BL6N mice weaned at 3weeks of age were randomly assigned to one of the four diets: 1) VIV chow: normal rodent chow, low in fat and high in fiber 2) HFD: 40 kcal% coconut oil 3) LA-HFD: 40 kcal% total fat soybean oil diet (21 kcal% from coconut oil and 19 kcal% from soybean oil) 4) PL-HFD: 40 kcal% total fat Plenish oil diet (21 kcal% from coconut oil and 19 kcal% from Plenish oil) |
Sample Preparation:
| Sampleprep ID: | SP000974 |
| Sampleprep Summary: | 1. Switch on bath to pre-cool at –20°C (±2°C validity temperature range) 2. Gently rotate or aspirate the blood samples for about 10s to obtain a homogenised sample. 3. Aliquot 30μl of plasma sample to a 1.0 mL extraction solution. The extraction solution has to be prechilled using the ThermoElectron Neslab RTE 740 cooling bath set to -20°C. 4. Vortex the sample for about 10s and shake for 5 min at 4°C using the Orbital Mixing Chilling/Heating Plate. If you are using more than one sample, keep the rest of the sample on ice (chilled at <0°C with sodium chloride). 5. Centrifuge samples for 2min at 14000 rcf using the centrifuge Eppendorf 5415 D. 6. Aliquot two 450μL portions of the supernatant. One for analysis and one for a backup sample. Store the backup aliquot in -20°C freezer. 7. Evaporate one 450μL aliquots of the sample in the Labconco Centrivap cold trap concentrator to complete dryness. 8. The dried aliquot is then re-suspended with 450 μL 50% acetonitrile (degassed as given above). 9. Centrifuged for 2 min at 14000 rcf using the centrifuge Eppendorf 5415. 10. Remove supernatant to a new Eppendorf tube. 11. Evaporate the supernatant to dryness in the Labconco Centrivap cold trap concentrator. 12. Submit to derivatization. |
| Sampleprep Protocol Filename: | SOP_Sample_preparation_of_blood_plasma_or_serum_samples_for_GCTOF_analysis.pdf |
Combined analysis:
| Analysis ID | AN001522 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Agilent |
| Column | Restek Rtx-5Sil (30m x 0.25mm,0.25um) |
| MS Type | EI |
| MS instrument type | GC Ion Trap |
| MS instrument name | Varian 210-MS GC Ion Trap |
| Ion Mode | POSITIVE |
| Units | Counts |
Chromatography:
| Chromatography ID: | CH001072 |
| Instrument Name: | Agilent |
| Column Name: | Restek Rtx-5Sil (30m x 0.25mm,0.25um) |
| Column Pressure: | 7.7 PSI (initial condition) |
| Column Temperature: | 50 - 330°C |
| Flow Rate: | 1 ml/min |
| Injection Temperature: | 50°C ramped to 250°C by 12°C/s |
| Sample Injection: | 0.5µl |
| Oven Temperature: | 50°C for 1 min, then ramped at 20°C/min to 330°C, held constant for 5 min |
| Transferline Temperature: | 230°C |
| 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: | MS001402 |
| Analysis ID: | AN001522 |
| Instrument Name: | Varian 210-MS GC Ion Trap |
| Instrument Type: | GC Ion Trap |
| MS Type: | EI |
| Ion Mode: | POSITIVE |
| Ion Source Temperature: | 250°C |
| Ionization Energy: | 70eV |
| Mass Accuracy: | Nominal |
| Source Temperature: | 250°C |
| Scan Range Moverz: | 85-500 |
| Scanning Cycle: | 17 Hz |
| Scanning Range: | 80-500 Da |
| Skimmer Voltage: | 1850 V |
