Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA250400

Local Sample ID:	pos-Pre-TBI-28
Subject ID:	SU002597
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Gender:	Male and female

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Sample Preparation:

Sampleprep ID:	SP002603
Sampleprep Summary:	GC-MS： 200μL of methanol containing 0.5μg/ml tridecanoic acid (IS) was added into 50μL plasma. After vortexmixing, the solution was centrifuged at 13000 rpm for 15 min at 4 ℃. Then the supernatant was transferred into a new eppendorf tube and dried by vacuum (LABCONCO, USA). The dried sample was re-dissolved in 50μL methoxyamine solution (20 mg/mL) for the oximation reaction (37℃,1.5h), which was followed by a silylation reaction with 50μL MSTFA (37℃,1.5h). Finally, the derivatized sample was centrifuged at 13 000 rpm for 15 min and the supernatant was used for the GC-MS analysis. LC-MS： The internal standards (ISs) were dissolved in acetonitrile including d4-choline (d4-CA, 8μg/mL), C19:0 lysoPC (4μg/mL), C17:0-C17:0 PC (8μg/mL), C17:0-C17:0 PE (8μg/mL), d3-C16:0 free fatty acids (d3-C16:0 FFA, 8μg/mL), d4-cholic acid (CA, 4μg/mL) and d4-chenodeoxycholic acid (d4-CDCA, 4μg/mL). For LC-MS analysis, 200 μL ISs was added into 50 μL plasma sample for the protein precipitation. After vortexing, the sample was centrifuged for 10 min (14,000 rpm, 4 ℃), the supernatant (180 μL ) was redissolved with 100μ L 50% methanol before LC-MS analysis.

Sampleprep ID:

SP002603

Sampleprep Summary:

GC-MS： 200μL of methanol containing 0.5μg/ml tridecanoic acid (IS) was added into 50μL plasma. After vortexmixing, the solution was centrifuged at 13000 rpm for 15 min at 4 ℃. Then the supernatant was transferred into a new eppendorf tube and dried by vacuum (LABCONCO, USA). The dried sample was re-dissolved in 50μL methoxyamine solution (20 mg/mL) for the oximation reaction (37℃,1.5h), which was followed by a silylation reaction with 50μL MSTFA (37℃,1.5h). Finally, the derivatized sample was centrifuged at 13 000 rpm for 15 min and the supernatant was used for the GC-MS analysis. LC-MS： The internal standards (ISs) were dissolved in acetonitrile including d4-choline (d4-CA, 8μg/mL), C19:0 lysoPC (4μg/mL), C17:0-C17:0 PC (8μg/mL), C17:0-C17:0 PE (8μg/mL), d3-C16:0 free fatty acids (d3-C16:0 FFA, 8μg/mL), d4-cholic acid (CA, 4μg/mL) and d4-chenodeoxycholic acid (d4-CDCA, 4μg/mL). For LC-MS analysis, 200 μL ISs was added into 50 μL plasma sample for the protein precipitation. After vortexing, the sample was centrifuged for 10 min (14,000 rpm, 4 ℃), the supernatant (180 μL ) was redissolved with 100μ L 50% methanol before LC-MS analysis.