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MB Sample ID: SA052892
| Local Sample ID: | PM D4-18 e2 |
| Subject ID: | SU000938 |
| Subject Type: | Animal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Species Group: | Mammal |
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Subject:
| Subject ID: | SU000938 |
| Subject Type: | Animal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Species Group: | Mammal |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| PM D4-18 e2 | SA052892 | FL010009 | HFD | Diet |
Collection:
| Collection ID: | CO000932 |
| 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: | TR000952 |
| Treatment Summary: | Male C57/BL6N mice weaned at 3weeks of age were randomly assigned to one of the four diets-Viv chow, 40 kcal% coconut oil , 40 kcal% total fat soybean oil diet (21 kcal% from coconut oil and 19 kcal% from soybean oil) or 40 kcal% total fat Plenish oil diet (21 kcal% from coconut oil and 19 kcal% from Plenish oil) for 24 weeks. |
Sample Preparation:
| Sampleprep ID: | SP000945 |
| Sampleprep Summary: | Preparation of SPE on vacuum manifold: 1. Clean 60 mg Oasis HLB (Waters) spe column with 1 column volume ethyl acetate followed by 2 column volumes MeOH 2. Condition column with 2 column volume 0.1% acetic acid :5% MeOH solution 3. Close valves tightly when meniscus reaches frit over sorbent bed. Sample extraction by SPE on vacuum manifold: 4. If not in sample at collection, add 10 µL of 0.2 mg/mL of EDTA and BHT in MeOH: H2O (1:1) to sorbent bed of all columns 5. Vortex each sample a few seconds to homogenize just prior to loading. 6. Load 250µL plasma sample into SPE column. 7. Spike each column with 10µl 500nM surrogate solution 8. Load 1.5ml of 0.1% acetic acid: 5% MeOH solution to each column and let sit a few minutes. 9. Open valves to extract by gravity (use low vacuum only if necessary) 10. Wash SPE with 2 column volumes 0.1% acetic acid :5% MeOH solution 11. Pull aqueous plug from SPE with a few seconds high vacuum. 12. Further dry SPE with low vacuum. ~20min, -20psi. 13. For each sample, spike 2ml eppendorf tube with 6µl 30% glycerol in MeOH prior to eluate collection. 14. Elute spe into tube with 0.5 ml methanol followed by 1.5 mL of ethyl acetate 15. Evaporate extract until 2 µl glycerol plug is left, by Speed-vac. 16. Cap and freeze by liquid nitrogen (for shipping) or store at -80ºC until analysis. |
Combined analysis:
| Analysis ID | AN001467 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Agilent 1200SL |
| Column | Agilent Eclipse Plus C18 (150 x 2.1mm, 1.8um) |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | ABI Sciex 4000 QTrap |
| Ion Mode | NEGATIVE |
| Units | Concentration nmol/L |
Chromatography:
| Chromatography ID: | CH001031 |
| Instrument Name: | Agilent 1200SL |
| Column Name: | Agilent Eclipse Plus C18 (150 x 2.1mm, 1.8um) |
| Flow Rate: | 0.35 mL/min |
| Sample Injection: | 10 uL |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS001355 |
| Analysis ID: | AN001467 |
| Instrument Name: | ABI Sciex 4000 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| Ion Mode: | NEGATIVE |
| Ion Source Temperature: | 600 C |
