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MB Sample ID: SA193792
Local Sample ID: | CarnsTG3 |
Subject ID: | SU002138 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Age Or Age Range: | 8-12 weeks |
Gender: | Male |
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Sample Preparation:
Sampleprep ID: | SP002144 |
Sampleprep Summary: | To extract polar metabolites, 800 µL of methanol was added to 200 µL of homogenized heart sample. Mixture was vortexed for 2 min and then placed on ice for 10 min, followed by another 2 min of vortex mixing, and centrifuged at 15,000 rpm for 20 min at 4 °C. Supernatant was transferred into a glass vial and dried in a SpeedVac evaporator to remove methanol, followed by lyophilization to remove water. The dried metabolite extract was dissolved with 30 µL of 20 mg/mL methoxyamine hydrochloride pyridine solution followed by vigorous vortex mixing for 1 min. Methoxylation was performed by sonicating the sample for 20 min and incubation at 60 °C for 1 h. Derivatization was performed by adding 20 µL of N-trimethylsilyl-N-methyl trifluoroacetamide (MSTFA) or N-Methyl-N-tert-butyldimethysilyltrifuoroacetamide (MTBSTFA) with 1% trimethylchlorosilane to the glass vial. Samples were incubated for 1 h at 60 °C and the mixture was transferred to a GC vial for analysis. Pooled samples were prepared by mixing 30 µL of derivatized metabolite extract from each sample to monitor the instrumental variations during the course of GC×GC-MS analysis. |