Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA024363

Local Sample ID:	S_7
Subject ID:	SU000497
Subject Type:	Murine
Subject Species:	Mus musculus
Taxonomy ID:	10090
Species Group:	Mammal

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Subject:

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

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
S_7	SA024363	FL006166	Veh	Treatment

Collection:

Collection ID:	CO000491
Collection Summary:	Trunk blood was collected into centrifuge tubes and centrifuged for 15 min at ~1,000 x g at 4o C. Supernatant (serum) was extracted, frozen on dry ice, and stored at -80o C.
Sample Type:	Blood

Treatment:

Treatment ID:	TR000511
Treatment Summary:	BALB/cByJ male mice (4-5 weeks in age; Jackson Laboratory, Bar Harbor, ME) were given four treatments of either vehicle or OT (1.0 mg/kg; IP) across 8 days, with at least 48 hours between each injection. Mice were evaluated in a 3-chamber choice test for social preference 24 hr after the final injection.

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.

Combined analysis:

Analysis ID	AN000741
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	Waters Acquity
Column	Waters Acquity CSH C18 (100 x 2.1mm,1.7um)
MS Type	ESI
MS instrument type	Orbitrap
MS instrument name	Orbitrap
Ion Mode	POSITIVE
Units	MS

Chromatography:

Chromatography ID:	CH000533
Instrument Name:	Waters Acquity
Column Name:	Waters Acquity CSH C18 (100 x 2.1mm,1.7um)
Chromatography Type:	Reversed phase

MS:

MS ID:	MS000658
Analysis ID:	AN000741
Instrument Name:	Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
Ion Mode:	POSITIVE