Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA274734

Local Sample ID:	49
Subject ID:	SU002835
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

Select appropriate tab below to view additional metadata details:

Sample Preparation:

Sampleprep ID:	SP002841
Sampleprep Summary:	Metabolites were extracted from frozen and crushed skeletal muscle (~10 mg) or serum (20 uL) with 1 mL of solvent mixture (methanol:chloroform:water = 10:4:4, v/v/v) containing piperazine-1,4-bis (2-ethanesulfonic acid) (PIPES) (0.63 umol/L), 2-bromohypoxanthine (0.13 umol/L), free fatty acid (FA) 16:0 (13C16) (0.13 umol/L), diacylglycerol (DG) 15:0-18:1 (d7) (0.15 umol/L), and triacylglycerol (TG) 15:0-18:1 (d7) -15:0 (0.35 umol/L) as internal standards. The samples were vigorously mixed for 1 min and sonicated for 5 min. The extracts were then centrifuged at 16,000 × g for 5 min at 4°C, and the resultant supernatant was collected. Protein concentrations in the pellets were determined using a Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, Massachusetts, USA). The collected supernatant (700 uL) was mixed with 235 uL of chloroform, and 155 uL of water, and then centrifuged at 16,000 × g for 5 min at 4°C. The aqueous (upper) layer (400 µL) was evaporated under vacuum, dried extracts were stored at −80°C until targeted polar metablome analysis. Prior to analysis, the dried aqueous layer was reconstituted in 50 uL of water. The organic (lower) layer (200 µL) was dried under a nitrogen stream, dried extracts were stored at −80°C until targeted lipidome analysis. Prior to analysis, the organic aqueous layer was reconstituted in 80 µL of methanol/chloroform (1/1, v/v)

Sampleprep ID:

SP002841

Sampleprep Summary:

Metabolites were extracted from frozen and crushed skeletal muscle (~10 mg) or serum (20 uL) with 1 mL of solvent mixture (methanol:chloroform:water = 10:4:4, v/v/v) containing piperazine-1,4-bis (2-ethanesulfonic acid) (PIPES) (0.63 umol/L), 2-bromohypoxanthine (0.13 umol/L), free fatty acid (FA) 16:0 (13C16) (0.13 umol/L), diacylglycerol (DG) 15:0-18:1 (d7) (0.15 umol/L), and triacylglycerol (TG) 15:0-18:1 (d7) -15:0 (0.35 umol/L) as internal standards. The samples were vigorously mixed for 1 min and sonicated for 5 min. The extracts were then centrifuged at 16,000 × g for 5 min at 4°C, and the resultant supernatant was collected. Protein concentrations in the pellets were determined using a Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, Massachusetts, USA). The collected supernatant (700 uL) was mixed with 235 uL of chloroform, and 155 uL of water, and then centrifuged at 16,000 × g for 5 min at 4°C. The aqueous (upper) layer (400 µL) was evaporated under vacuum, dried extracts were stored at −80°C until targeted polar metablome analysis. Prior to analysis, the dried aqueous layer was reconstituted in 50 uL of water. The organic (lower) layer (200 µL) was dried under a nitrogen stream, dried extracts were stored at −80°C until targeted lipidome analysis. Prior to analysis, the organic aqueous layer was reconstituted in 80 µL of methanol/chloroform (1/1, v/v)