Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA237935

Local Sample ID:	C30-1C-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.

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.