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MB Sample ID: SA343060
| Local Sample ID: | young_11 |
| Subject ID: | SU003287 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | Young donors (aged 25 years or younger) and older donors (aged 50 years or older). |
| Gender: | Male and female |
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Sample Preparation:
| Sampleprep ID: | SP003294 |
| Sampleprep Summary: | Lipids were extracted from ~20 mg heart tissue using a two-phase method with methyl-tert-butyl ether (MTBE) and water. Tissue was homogenised with steel beads in 250 µL methanol containing 0.01% (w/v) butylhydroxytoluene (BHT) and mass spectrometry internal standards: 2 nmoles PC(19:0/19:0); 1 nmole each of SM(d18:1/12:0), GluCer(d18:1/12:0), Cer(d18:1/17:0), PS(17:0/17:0), PE(17:0/17:0), PA(17:0/17:0), PI(d7-18:1/15:0), PG(17:0/17:0), CL(14:0/14:0/14:0/14:0), and TG(17:0/17:0/17:0); 0.5 nmoles each of DG(d7-18:1/15:0), CholE(17:0), LPC (17:0), LPE(17:1), and AcCa(d3-16:0); and 0.2 nmole each of Sph(d17:1), S1P(d17:1), LacCer(d18:1/12:0), and MG(d7-18:1). MTBE (850 µL) was added, and samples were sonicated in a 4°C water bath for 30 min. Mass spectrometry grade water (212 µL) was added to induce phase separation after vortexing and centrifugation at 2000g for 5 min. The upper organic phase was collected in 5 mL glass tubes and the aqueous phase was extracted twice more with 500 µL MTBE and 150 µL methanol followed by sonication for 15 min and phase separation with 125 µL water. Organic phases from the three extractions were combined and dried under vacuum in a Savant SC210 SpeedVac (ThermoFisher Scientific). Lipids were reconstituted in 400 µL 80% methanol/20% water/0.1% formic acid containing 0.01% (w/v) BHT and stored at -80 °C. |
| Sampleprep Protocol ID: | SP003225 |
