Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA084780

Local Sample ID:	Thermoneutral 5 male
Subject ID:	SU001273
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

Select appropriate tab below to view additional metadata details:

Subject:

Subject ID:	SU001273
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
Thermoneutral 5 male	SA084780	FL012819	30C	Temperature
Thermoneutral 5 male	SA084780	FL012819	7 days	Time of exposure

Collection:

Collection ID:	CO001267
Collection Summary:	Mice were anaesthethized with isoflurane, the blood collected through cardiac puncture, and left under room temperature for 30 minutes. After that, the blood was centrifuged for 3 minutes at 14,000 rpm. Then, serum was transferred for a clean 1.5ml microbe and frost.
Sample Type:	Blood (serum)
Storage Conditions:	-20℃

Treatment:

Treatment ID:	TR001288
Treatment Summary:	We divided the C57BL6/J mice in 6 groups: females housed in 30C degree for 1 week and females housed in 5C degree for 1 week; Males housed in 30C degree for 1 week and females housed in 5C degree for 1 week; males housed in 22C for 1 hour and males housed in 5C for 1 hour.

Sample Preparation:

Sampleprep ID:	SP001281
Sampleprep Summary:	Aliquots of 100 µL serum or were taken. A mixture of deuterium-labeled internal standards was added to each aliquot, followed by 3x volume of sample of cold methanol (MeOH). Samples were vortexed for 5 minutes and stored at −20 °C overnight. Cold samples were centrifuged at 14,000g for 10 minutes, and the supernatant was then transferred to a new tube and 3 mL of acidified H2O (pH 3.5) was added to each sample prior to C18 SPE. The methyl formate fractions were collected, dried under nitrogen, and reconstituted in 50 µL MeOH:H2O (1:1, by vol). Samples were transferred to 0.5 mL tubes and centrifuged at 20,000g at 4 °C for 10 minutes. (35ul) of supernatant was transferred to LC–MS/MS vials for analysis using the BERG LC–MS/MS mediator lipidomics platform.

Sampleprep ID:

SP001281

Sampleprep Summary:

Aliquots of 100 µL serum or were taken. A mixture of deuterium-labeled internal standards was added to each aliquot, followed by 3x volume of sample of cold methanol (MeOH). Samples were vortexed for 5 minutes and stored at −20 °C overnight. Cold samples were centrifuged at 14,000g for 10 minutes, and the supernatant was then transferred to a new tube and 3 mL of acidified H2O (pH 3.5) was added to each sample prior to C18 SPE. The methyl formate fractions were collected, dried under nitrogen, and reconstituted in 50 µL MeOH:H2O (1:1, by vol). Samples were transferred to 0.5 mL tubes and centrifuged at 20,000g at 4 °C for 10 minutes. (35ul) of supernatant was transferred to LC–MS/MS vials for analysis using the BERG LC–MS/MS mediator lipidomics platform.

Combined analysis:

Analysis ID	AN002008
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	Ekspert MicroLC 200 system
Column	Phenomenex Synergi Fusion-RP capillary C18 (150 × 0.5 mm,4um)
MS Type	ESI
MS instrument type	QTOF
MS instrument name	ABI Sciex 5600+ TripleTOF
Ion Mode	NEGATIVE
Units	Peak Area

Chromatography:

Chromatography ID:	CH001452
Instrument Name:	Ekspert MicroLC 200 system
Column Name:	Phenomenex Synergi Fusion-RP capillary C18 (150 × 0.5 mm,4um)
Chromatography Type:	Reversed phase

MS:

MS ID:	MS001861
Analysis ID:	AN002008
Instrument Name:	ABI Sciex 5600+ TripleTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	MS spectra were acquired in high-resolution mode (>30,000) using a 100-ms accumulation time per spectrum. Full-scan MS/MS was acquired in high sensitivity mode, with an accumulation time optimized per cycle. Collision energy was set using rolling collision energy with a spread of 15V. The identity of a component was confirmed using PeakView® software (SCIEX), and quantification was performed using MultiQuant™ software (SCIEX).
Ion Mode:	NEGATIVE