Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA301913

Local Sample ID:	posi-7
Subject ID:	SU002921
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	C57BL/6
Age Or Age Range:	14-15 weeks old
Gender:	Male

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Sample Preparation:

Sampleprep ID:	SP002927
Sampleprep Summary:	For extraction of water-soluble metabolites from plasma samples, 200 µl of methanol was added to 5 µl plasma. This mixture was incubated on ice for 10 minutes, then centrifuged at 17,000g for 10 minutes at 4°C. The supernatant was transferred to an MS vial to run on LCMS. For extraction of hydrophobic metabolites (lipids) from plasma samples, 120 µl of isopropanol kept on dry ice was added to 3 µl plasma. This mixture was vortexed for 10 seconds and kept on dry ice for 10 minutes, then centrifuged at 17,000g for 10 minutes at 4°C. The supernatant was loaded into glass inserts in an MS vial to run on LCMS. For extraction of water-soluble metabolites from tissue samples, frozen tissues were first ground to a powder using a cryomill (Restch, Newtown, PA). Extraction solution kept on ice (40:40:20 methanol:acetonitrile:water) was added to ~10 mg of ground tissue (40 µl solution per mg tissue), vortexed, and kept on ice for 10 minutes, then centrifuged at 17,000g for 10 minutes at 4°C. The supernatant was collected and centrifuged again at the same speed for 10 minutes, and this second supernatant was transferred to an MS vial to run on LCMS.

Sampleprep ID:

SP002927

Sampleprep Summary:

For extraction of water-soluble metabolites from plasma samples, 200 µl of methanol was added to 5 µl plasma. This mixture was incubated on ice for 10 minutes, then centrifuged at 17,000g for 10 minutes at 4°C. The supernatant was transferred to an MS vial to run on LCMS. For extraction of hydrophobic metabolites (lipids) from plasma samples, 120 µl of isopropanol kept on dry ice was added to 3 µl plasma. This mixture was vortexed for 10 seconds and kept on dry ice for 10 minutes, then centrifuged at 17,000g for 10 minutes at 4°C. The supernatant was loaded into glass inserts in an MS vial to run on LCMS. For extraction of water-soluble metabolites from tissue samples, frozen tissues were first ground to a powder using a cryomill (Restch, Newtown, PA). Extraction solution kept on ice (40:40:20 methanol:acetonitrile:water) was added to ~10 mg of ground tissue (40 µl solution per mg tissue), vortexed, and kept on ice for 10 minutes, then centrifuged at 17,000g for 10 minutes at 4°C. The supernatant was collected and centrifuged again at the same speed for 10 minutes, and this second supernatant was transferred to an MS vial to run on LCMS.