Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA332472

Local Sample ID:	Heart_504
Subject ID:	SU003191
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	C57BL/6
Gender:	Male

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Sample Preparation:

Sampleprep ID:	SP003197
Sampleprep Summary:	All tissue sample was first homogenized in liquid nitrogen and then 10-20 mg tissues were weighed into a new 1.7 ml Eppendorf tube. Ice cold extraction solvent (400 l 80% methanol/water) and 10 l metformin-d6 (50 ng/l in water) was added to each sample. Geno/Grinder homogenizer was used (1500 rpm, 1 to 2 min) to further break down the tissue chunk and form an even suspension. 200 ul supernatant containing polar metabolites was removed after centrifugation at 20,000 g at 4 °C for 10 min and transferred to LC vial for polar metabolite analysis using LC-HRMS. To extract metabolites and lipids from mouse plasma, 10 l plasma was mixed with 10 l water containing internal standards (50 ng/l metformin-d6, 5 mM [U-13C]-glucose and [U-13C]-lactate, and 80 l ice cold methanol was added. After vortex for 1 min, the mixture was centrifuged with a speed of 20,000 g at 4 °C for 10 min, and 20 l was transferred directly to LC vial without solvent evaporation, followed by LC-HRMS analysis (injection volume, 3 l) of polar metabolites such as glucose and metformin.

Sampleprep ID:

SP003197

Sampleprep Summary:

All tissue sample was first homogenized in liquid nitrogen and then 10-20 mg tissues were weighed into a new 1.7 ml Eppendorf tube. Ice cold extraction solvent (400 l 80% methanol/water) and 10 l metformin-d6 (50 ng/l in water) was added to each sample. Geno/Grinder homogenizer was used (1500 rpm, 1 to 2 min) to further break down the tissue chunk and form an even suspension. 200 ul supernatant containing polar metabolites was removed after centrifugation at 20,000 g at 4 °C for 10 min and transferred to LC vial for polar metabolite analysis using LC-HRMS. To extract metabolites and lipids from mouse plasma, 10 l plasma was mixed with 10 l water containing internal standards (50 ng/l metformin-d6, 5 mM [U-13C]-glucose and [U-13C]-lactate, and 80 l ice cold methanol was added. After vortex for 1 min, the mixture was centrifuged with a speed of 20,000 g at 4 °C for 10 min, and 20 l was transferred directly to LC vial without solvent evaporation, followed by LC-HRMS analysis (injection volume, 3 l) of polar metabolites such as glucose and metformin.