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MB Sample ID: SA267007
| Local Sample ID: | ko3-2p |
| Subject ID: | SU002798 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | Wild-type and ATP10A knockouts |
| Age Or Age Range: | 16-20 weeks |
| Weight Or Weight Range: | 23-28 grams |
| Animal Housing: | temperature and humidity-controlled facilities |
| Animal Light Cycle: | 12 h light/dark cycles |
| Animal Feed: | standard chow or 60% HFD (D12492, Research Diets) ad libitum, |
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Sample Preparation:
| Sampleprep ID: | SP002804 |
| Sampleprep Summary: | 100 uL of plasma was collected from each mouse. An isotopically labeled lipid mixture (SPLASH LIPIDOMIX, Avanti) was added as an internal standard to each plasma sample. A liquid-liquid extraction was performed by adding 1 mL methyl tert-butyl ether (MTBE), vortexing, and subsequently centrifuging for 10 minutes at 10,000 rpm and 4˚C. The nonpolar, MTBE fraction (top liquid layer) containing the lipophilic components was removed and dried under vacuum centrifugation. Dried samples were stored at -80˚C until the day of MS analysis. For LC-IM-MS analysis, dried samples were resuspended in 100 µL IPA containing 40 µg/mL heptadecanoic acid and nonadecanoic acid, as well as 10 µg/mL glucosyl(β) sphingosine and N-heptadecanoyl-D-erythrosphingosine. |
| Processing Storage Conditions: | 4℃ |
| Extraction Method: | MTBE |
| Extract Enrichment: | vacuum centrifuge |
| Extract Storage: | -80℃ |
| Sample Resuspension: | 100 uL IPA |
| Sample Derivatization: | n/a |
| Sample Spiking: | 14 heavy-labeled lipids prior to extraction; 4 odd-chain lipids during reconstitution |
| Subcellular Location: | n/a |
