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MB Sample ID: SA084817

Local Sample ID:LB3
Subject ID:SU001274
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:C57BL6/J
Age Or Age Range:12 weeks old
Weight Or Weight Range:25-30 g
Gender:Male
Animal Animal Supplier:Jax Laboratories
Animal Housing:Conventional
Animal Light Cycle:regular 12h light /dark cycles
Animal Feed:ad libitum normal chow diet
Animal Water:ad libitum

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Subject:

Subject ID:SU001274
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:C57BL6/J
Age Or Age Range:12 weeks old
Weight Or Weight Range:25-30 g
Gender:Male
Animal Animal Supplier:Jax Laboratories
Animal Housing:Conventional
Animal Light Cycle:regular 12h light /dark cycles
Animal Feed:ad libitum normal chow diet
Animal Water:ad libitum

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
LB3SA084817FL0128245CTemperature
LB3SA084817FL012824LoxBlock-1Treatment

Collection:

Collection ID:CO001268
Collection Summary:Mice were anaesthetised with Avertine and blood was collected through cardiac puncture. Samples were let in room temperature for 30 minutes, and then centrifuged for 3 minutes at 14,000RPM. Serum fas collected and frozen at -20C for further lipidomics analysis.
Sample Type:Blood (serum)
Storage Conditions:-80℃

Treatment:

Treatment ID:TR001289
Treatment Summary:Mice were injected i.p. 15 minutes before the cold exposure, with 50ul of DMSO (vehicle) or LOXBlock-1. Then, the animas were placed in 5C or 22C temperature for 4 hours, until the samples were collected.

Sample Preparation:

Sampleprep ID:SP001282
Sampleprep Summary:Aliquots of 100 µL serum were taken and 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 AN002009
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:CH001453
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:MS001862
Analysis ID:AN002009
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). C18SPE cartridges were purchased from Biotage. All solvents were of HPLC or LC-MS/MS grade and were acquired from Sigma-Aldrich, Fisher Scientific, or VWR International.
Ion Mode:NEGATIVE
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