Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA317516

Local Sample ID:	Con_7
Subject ID:	SU003036
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Age Or Age Range:	infant
Gender:	Not applicable
Human Race:	Pacific asian

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Sample Preparation:

Sampleprep ID:	SP003042
Sampleprep Summary:	~10 mg of the freeze-dried feces was homogenized well with an internal standard solution (200 μl of 1 μM C17 S1P solution). Then, 300 μl of chloroform, 200 μl of methanol, 50 μl of H2O, and 10 μl of 10N NaOH were added to each solution, and vortex well. The aqueous layer was collected after centrifugation, where S1P was separated from other lipids under alkaline condition. 50 uL of H2O was added to the remaining solution and the aqueous layer was collected again after centrifugation. Then, S1P was re-extracted into the chloroform phase under acidic conditions using 400 μl of chloroform and 20 μl of 10M HCl. This step was repeated three times. The chloroform was evaporated, and the dried samples were reconstituted with methanol prior to LC-MS/MS analysis.

Sampleprep ID:

SP003042

Sampleprep Summary:

~10 mg of the freeze-dried feces was homogenized well with an internal standard solution (200 μl of 1 μM C17 S1P solution). Then, 300 μl of chloroform, 200 μl of methanol, 50 μl of H2O, and 10 μl of 10N NaOH were added to each solution, and vortex well. The aqueous layer was collected after centrifugation, where S1P was separated from other lipids under alkaline condition. 50 uL of H2O was added to the remaining solution and the aqueous layer was collected again after centrifugation. Then, S1P was re-extracted into the chloroform phase under acidic conditions using 400 μl of chloroform and 20 μl of 10M HCl. This step was repeated three times. The chloroform was evaporated, and the dried samples were reconstituted with methanol prior to LC-MS/MS analysis.