Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA076226

Local Sample ID:	C1
Subject ID:	SU001163
Subject Type:	Mammal
Subject Species:	Mesocricetus auratus
Taxonomy ID:	10036

Select appropriate tab below to view additional metadata details:

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:

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