Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA326857

Local Sample ID:	BAT_HFD_neg_6
Subject ID:	SU003119
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	C57BL/6J
Age Or Age Range:	12 weeks
Weight Or Weight Range:	25-30g
Gender:	Male

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Subject:

Subject ID:	SU003119
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	C57BL/6J
Age Or Age Range:	12 weeks
Weight Or Weight Range:	25-30g
Gender:	Male

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
BAT_HFD_neg_6	SA326857	FL038831	HFD	Diet

Collection:

Collection ID:	CO003112
Collection Summary:	Animals were sacrificed immediately by cervical dislocation and tissues were rapidly extracted and immediately snap frozen in liquid nitrogen for metabolite profiling.
Sample Type:	Brown fat tissue

Treatment:

Treatment ID:	TR003128
Treatment Summary:	All mice were housed under a 12 h – 12 h light/dark cycle. Room-temperature mice were housed at 23˚C in ventilated cages with an ACH of 25. Mice were fed a standard diet (Lab Diet 5008) or high-fat diet (Research Diets; D12492; 60% Fat) and had free access to food and water.

Sample Preparation:

Sampleprep ID:	SP003125
Sampleprep Summary:	Animals were sacrificed immediately by cervical dislocation and tissues were rapidly extracted and immediately snap frozen in liquid nitrogen for metabolite profiling. For metabolite extraction, tissues were weighed and then homogenized with extraction buffer which consisted of 80% methanol containing D₈-Phenylalanine internal standards at a 40:1 volume to wet weight ratio. Samples were centrifuged at 16,000 x g at 4 °C for 15 min. Finally, 50 μL supernatant was transferred to the glass insert for LC-MS detection.

Sampleprep ID:

SP003125

Sampleprep Summary:

Animals were sacrificed immediately by cervical dislocation and tissues were rapidly extracted and immediately snap frozen in liquid nitrogen for metabolite profiling. For metabolite extraction, tissues were weighed and then homogenized with extraction buffer which consisted of 80% methanol containing D₈-Phenylalanine internal standards at a 40:1 volume to wet weight ratio. Samples were centrifuged at 16,000 x g at 4 °C for 15 min. Finally, 50 μL supernatant was transferred to the glass insert for LC-MS detection.

Combined analysis:

Analysis ID	AN004936
Analysis type	MS
Chromatography type	HILIC
Chromatography system	Thermo Vanquish
Column	Waters ACQUITY UPLC BEH Amide (100 x 2.1mm,1.7um)
MS Type	ESI
MS instrument type	Orbitrap
MS instrument name	Thermo orbitrap exploris 240
Ion Mode	NEGATIVE
Units	Peak area

Chromatography:

Chromatography ID:	CH003725
Instrument Name:	Thermo Vanquish
Column Name:	Waters ACQUITY UPLC BEH Amide (100 x 2.1mm,1.7um)
Column Temperature:	25 °C
Flow Gradient:	The linear gradient eluted from 95% B (0.0–1 min), 95% B to 65% B (1–7.0 min), 65% B to 40% B (7.0–8.0 min), 40% B (8.0–9.0 min), 40% B to 95% B (9.0–9.1 min), then stayed at 95% B for 5.9 min.
Flow Rate:	0.4 mL/min
Solvent A:	100% water; 25mM ammonium acetate; 25mM ammonium hydroxide
Solvent B:	100% acetonitrile
Chromatography Type:	HILIC

MS:

MS ID:	MS004679
Analysis ID:	AN004936
Instrument Name:	Thermo orbitrap exploris 240
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	ESI source parameters were set as follows: spray voltage, 3500 V or −2800 V, in positive or negative modes, respectively; vaporizer temperature, 350 °C; sheath gas, 50 arb; aux gas, 10 arb; ion transfer tube temperature, 325 °C. The full scan was set as: orbitrap resolution, 60,000; maximum injection time, 100 ms; scan range, 70–1050 Da. The ddMS2 scan was set as: orbitrap resolution, 30,000; maximum injection time, 60 ms; top N setting, 6; isolation width, 1.0 m/z; HCD collision energy (%), 30; Dynamic exclusion mode was set as auto. The data was analyzed by Compound Discoverer 3.3.
Ion Mode:	NEGATIVE