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MB Sample ID: SA237963
Local Sample ID: | 0-3A-N |
Subject ID: | SU002473 |
Subject Type: | Cultured cells |
Subject Species: | Rattus norvegicus |
Taxonomy ID: | 10116 |
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Sample Preparation:
Sampleprep ID: | SP002479 |
Sampleprep Summary: | The MeOH/MTBE/H2O system was used for the extraction of lipid metabolites from cell samples. Cell culture dishes stored at -80 °C were first removed on ice and 1 mL of methanolic solution containing internal standards was added, consisting of 1 μg/mL FFA (C16:0)-d3 and FFA (C18:0)-d3, 0.87 μg/mL LPC (19:0) and SM (d18:1/12:0), 1.12 μg/mL PC (38: 0), 0.92 μg/mL PE (30:0), 0.67 μg/mL TG (45:0), and 0.47 μg/mL Cer (d18:1/17:0). The cells were transferred to 5 mL Eppendorf centrifuge tubes by scraping with a cell scraper, adding 2.5 mL of MTBE and vortexing for 30 s. The cells were further shaken for 30 min at 10 °C and 1000 rpm using a thermostatic mixer. 750 μL of ultrapure water was added, vortexed for 60 s and allowed to stand for 5 min, and then centrifuged at 12,000 g for 10 min at 6 °C. After separation of the two phases, 400 μL of the hydrophobic phase was collected and dried in a cryogenic vacuum centrifuge concentrator, and the resulting lyophilized sample was stored at -80 °C. |