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MB Sample ID: SA332515
Local Sample ID: | Heart_492 |
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. |