Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA024364

Local Sample ID:	S_18
Subject ID:	SU000497
Subject Type:	Murine
Subject Species:	Mus musculus
Taxonomy ID:	10090
Species Group:	Mammal

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Sample Preparation:

Sampleprep ID:	SP000504
Sampleprep Summary:	30 µL of each serum sample was transferred to a new, pre-labeled 1.5 mL LoBind eppendorf tube to make the individual study samples, and 12 µL was transferred to a new, pre-labeled 1.5 mL LoBind Eppendorf to make the total pool. The total pool was aliquoted into 3 Total Pool QC Samples and 4 Column Equilibrium samples with volumes of 30 µL in 1.5 mL LoBind Eppendorf tubes. 600 uL of Lipid Extraction Solvent (PC(17:0/17:0) 50 µg/mL in 2:1 dichloromethane:methanol) was added to all samples, and samples were vortexed at 4,000 rpm for 2mins on multiple-tube vortexer. 120 µL of H2O was added to each tube and the samples were vortexed at 4,000 rpm for 1min. Samples sat at room temperature for 10 minutes, and were centrifuged at 16,000 rcf for 10 min at 10°C. 240 µL of the lower lipid-rich DCM layer was transferred into new pre-labeled 1.5 mL LoBind Eppendorf tubes, and rubber stoppers were inserted. Samples were placed at -80˚C for 60 mins, and then lyophilized for 19.5 hrs. 240 µL of ACN/IPA/H2O (65:30:5 v/v/v) was added to each sample, and the samples were vortexed at 4,000 rpm for 2mins and centrifuged at 16,000 rcf for 4 min. 225 µL of the supernatant was transferred to pre-labeled autosampler vials and 10 µL was injected into OrbiTrap Velos.

Sampleprep ID:

SP000504

Sampleprep Summary:

30 µL of each serum sample was transferred to a new, pre-labeled 1.5 mL LoBind eppendorf tube to make the individual study samples, and 12 µL was transferred to a new, pre-labeled 1.5 mL LoBind Eppendorf to make the total pool. The total pool was aliquoted into 3 Total Pool QC Samples and 4 Column Equilibrium samples with volumes of 30 µL in 1.5 mL LoBind Eppendorf tubes. 600 uL of Lipid Extraction Solvent (PC(17:0/17:0) 50 µg/mL in 2:1 dichloromethane:methanol) was added to all samples, and samples were vortexed at 4,000 rpm for 2mins on multiple-tube vortexer. 120 µL of H2O was added to each tube and the samples were vortexed at 4,000 rpm for 1min. Samples sat at room temperature for 10 minutes, and were centrifuged at 16,000 rcf for 10 min at 10°C. 240 µL of the lower lipid-rich DCM layer was transferred into new pre-labeled 1.5 mL LoBind Eppendorf tubes, and rubber stoppers were inserted. Samples were placed at -80˚C for 60 mins, and then lyophilized for 19.5 hrs. 240 µL of ACN/IPA/H2O (65:30:5 v/v/v) was added to each sample, and the samples were vortexed at 4,000 rpm for 2mins and centrifuged at 16,000 rcf for 4 min. 225 µL of the supernatant was transferred to pre-labeled autosampler vials and 10 µL was injected into OrbiTrap Velos.