Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA076223

Local Sample ID:	C5
Subject ID:	SU001163
Subject Type:	Mammal
Subject Species:	Mesocricetus auratus
Taxonomy ID:	10036

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Subject:

Subject ID:	SU001163
Subject Type:	Mammal
Subject Species:	Mesocricetus auratus
Taxonomy ID:	10036

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
C5	SA076223	FL011690	control	hibernation

Collection:

Collection ID:	CO001157
Collection Summary:	A total of 15 male 3-month-old Syrian hamsters had free access to food and water and were kept at 23C with an 8:16-h light/dark cycle for a four to six-week acclimatization period in our animal facility. All animals were euthanized by decapitation. Brains were then removed and immediately transferred to a N2(l)-containing recipient to freeze the tissues.
Sample Type:	Brain

Treatment:

Treatment ID:	TR001177
Treatment Summary:	the whole right hemisphere (300 mg approx.) was analyzed to decrease possible biological variability due to the brain region employed. Brain homogenate was prepared by adding cold (-20°C) methanol:water (1:1, v/v), (1:10 tissue:solvent). Tissue disruption was achieved with TissueLyser LT homogenizer (Qiagen, Germany) for metabolite extraction. Subsequently, 100 μL of brain tissue homogenate was vortex-mixed with 320 μL of methanol for 2 min, followed by the addition of 80 μL of MTBE for the extraction of non-polar compounds. Then, vials were rapidly capped and placed on a shaker for 1 h at room temperature. The extracted samples were centrifuged at 4000 g for 20 min at 20°C. For GC-MS analysis, 300 μL of supernatant was evaporated to dryness (SpeedVac Concentrator System, Thermo Fisher Scientific, Waltham, MA, USA). Methoxymation was then performed with 20 µL O-methoxyamine hydrochloride (15 mg/mL in pyridine) and vigorously vortex-mixed for 5 min. Vials were then incubated in darkness at room temperature for 16 hours. For silylation, 20 μL of BSTFA:TMCS (99:1) was added, vortex-mixed for 5 min, and capped vials were placed in the oven at 70°C for 1 h. Finally, 100 μL of heptane containing C18:0 methyl ester (10 ppm) as Internal Standard (IS) was added to each vial prior to injection. For LC-MS analysis, 90 μL of supernatant was transferred to an Ultra-High Performance Liquid Chromatography-Mass (UHPLC-MS) chromatography vial with insert and was directly injected into the system. For CE-MS analysis, 200 μL of initial brain homogenate was centrifuged separately at 16000 g for 30 min at 15°C. 150 μL of supernatant was evaporated to dryness using the SpeedVac, and re-suspended in 150 μL of 0.2 mM Methionine Sulfone (IS) in 0.1 M formic acid. Samples were vortex-mixed, sonicated, and then centrifuged at 16000 g for 20 min at 4°C. Finally, 100 μL of supernatant was transferred to a CE-MS vial for the analysis.

Treatment ID:

TR001177

Treatment Summary:

the whole right hemisphere (300 mg approx.) was analyzed to decrease possible biological variability due to the brain region employed. Brain homogenate was prepared by adding cold (-20°C) methanol:water (1:1, v/v), (1:10 tissue:solvent). Tissue disruption was achieved with TissueLyser LT homogenizer (Qiagen, Germany) for metabolite extraction. Subsequently, 100 μL of brain tissue homogenate was vortex-mixed with 320 μL of methanol for 2 min, followed by the addition of 80 μL of MTBE for the extraction of non-polar compounds. Then, vials were rapidly capped and placed on a shaker for 1 h at room temperature. The extracted samples were centrifuged at 4000 g for 20 min at 20°C. For GC-MS analysis, 300 μL of supernatant was evaporated to dryness (SpeedVac Concentrator System, Thermo Fisher Scientific, Waltham, MA, USA). Methoxymation was then performed with 20 µL O-methoxyamine hydrochloride (15 mg/mL in pyridine) and vigorously vortex-mixed for 5 min. Vials were then incubated in darkness at room temperature for 16 hours. For silylation, 20 μL of BSTFA:TMCS (99:1) was added, vortex-mixed for 5 min, and capped vials were placed in the oven at 70°C for 1 h. Finally, 100 μL of heptane containing C18:0 methyl ester (10 ppm) as Internal Standard (IS) was added to each vial prior to injection. For LC-MS analysis, 90 μL of supernatant was transferred to an Ultra-High Performance Liquid Chromatography-Mass (UHPLC-MS) chromatography vial with insert and was directly injected into the system. For CE-MS analysis, 200 μL of initial brain homogenate was centrifuged separately at 16000 g for 30 min at 15°C. 150 μL of supernatant was evaporated to dryness using the SpeedVac, and re-suspended in 150 μL of 0.2 mM Methionine Sulfone (IS) in 0.1 M formic acid. Samples were vortex-mixed, sonicated, and then centrifuged at 16000 g for 20 min at 4°C. Finally, 100 μL of supernatant was transferred to a CE-MS vial for the analysis.

Sample Preparation:

Sampleprep ID:	SP001170
Sampleprep Summary:	the whole right hemisphere (300 mg approx.) was analyzed to decrease possible biological variability due to the brain region employed. Brain homogenate was prepared by adding cold (-20°C) methanol:water (1:1, v/v), (1:10 tissue:solvent). Tissue disruption was achieved with TissueLyser LT homogenizer (Qiagen, Germany) for metabolite extraction. Subsequently, 100 μL of brain tissue homogenate was vortex-mixed with 320 μL of methanol for 2 min, followed by the addition of 80 μL of MTBE for the extraction of non-polar compounds. Then, vials were rapidly capped and placed on a shaker for 1 h at room temperature. The extracted samples were centrifuged at 4000 g for 20 min at 20°C. For GC-MS analysis, 300 μL of supernatant was evaporated to dryness (SpeedVac Concentrator System, Thermo Fisher Scientific, Waltham, MA, USA). Methoxymation was then performed with 20 µL O-methoxyamine hydrochloride (15 mg/mL in pyridine) and vigorously vortex-mixed for 5 min. Vials were then incubated in darkness at room temperature for 16 hours. For silylation, 20 μL of BSTFA:TMCS (99:1) was added, vortex-mixed for 5 min, and capped vials were placed in the oven at 70°C for 1 h. Finally, 100 μL of heptane containing C18:0 methyl ester (10 ppm) as Internal Standard (IS) was added to each vial prior to injection. For LC-MS analysis, 90 μL of supernatant was transferred to an Ultra-High Performance Liquid Chromatography-Mass (UHPLC-MS) chromatography vial with insert and was directly injected into the system. For CE-MS analysis, 200 μL of initial brain homogenate was centrifuged separately at 16000 g for 30 min at 15°C. 150 μL of supernatant was evaporated to dryness using the SpeedVac, and re-suspended in 150 μL of 0.2 mM Methionine Sulfone (IS) in 0.1 M formic acid. Samples were vortex-mixed, sonicated, and then centrifuged at 16000 g for 20 min at 4°C. Finally, 100 μL of supernatant was transferred to a CE-MS vial for the analysis

Sampleprep ID:

SP001170

Sampleprep Summary:

Combined analysis:

Analysis ID	AN001813	AN001814	AN001815	AN001816
Analysis type	MS	MS	MS	MS
Chromatography type	GC	Reversed phase	CE	Reversed phase
Chromatography system	Agilent 7890A	Agilent 1290 Infinity	Agilent 7100 CE	Agilent 1290 Infinity
Column	Agilent 122-5532G	Agilent Zorbax Eclipse Plus C8 (150 x 2.1mm, 1.8 um)	Fused-silica capillary from Agilent Technologies	Agilent Zorbax Eclipse Plus C8 (150 x 2.1mm, 1.8 um)
MS Type	EI	ESI	ESI	ESI
MS instrument type	Single quadrupole	QTOF	TOF	QTOF
MS instrument name	Agilent 5975C	Agilent 6550 QTOF	Agilent 6224 TOF	Agilent 6550 QTOF
Ion Mode	POSITIVE	POSITIVE	POSITIVE	NEGATIVE
Units	peak area	peak area	peak area	peak area

Chromatography:

Chromatography ID:	CH001279
Instrument Name:	Agilent 7890A
Column Name:	Agilent 122-5532G
Chromatography Type:	GC

Chromatography ID:	CH001280
Instrument Name:	Agilent 1290 Infinity
Column Name:	Agilent Zorbax Eclipse Plus C8 (150 x 2.1mm, 1.8 um)
Chromatography Type:	Reversed phase

Chromatography ID:	CH001281
Instrument Name:	Agilent 7100 CE
Column Name:	Fused-silica capillary from Agilent Technologies
Chromatography Type:	CE

Chromatography ID:	CH001282
Instrument Name:	Agilent 1290 Infinity
Column Name:	Agilent Zorbax Eclipse Plus C8 (150 x 2.1mm, 1.8 um)
Chromatography Type:	Reversed phase

MS:

MS ID:	MS001672
Analysis ID:	AN001813
Instrument Name:	Agilent 5975C
Instrument Type:	Single quadrupole
MS Type:	EI
Ion Mode:	POSITIVE

MS ID:	MS001673
Analysis ID:	AN001814
Instrument Name:	Agilent 6550 QTOF
Instrument Type:	QTOF
MS Type:	ESI
Ion Mode:	POSITIVE

MS ID:	MS001674
Analysis ID:	AN001815
Instrument Name:	Agilent 6224 TOF
Instrument Type:	TOF
MS Type:	ESI
Ion Mode:	POSITIVE

MS ID:	MS001675
Analysis ID:	AN001816
Instrument Name:	Agilent 6550 QTOF
Instrument Type:	QTOF
MS Type:	ESI
Ion Mode:	NEGATIVE