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MB Sample ID: SA057718
Local Sample ID: | 1_9_2018_islet_ugluc_timecourse_30mins_8 |
Subject ID: | SU001004 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | C57BL/6N |
Age Or Age Range: | 4 weeks old (young) or 10-15 months old (old) |
Gender: | Male and female |
Animal Animal Supplier: | Charles River Labs |
Animal Feed: | standard chow diet (PicoLab Rodent Diet 20 pellets, #5053 |
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Sample Preparation:
Sampleprep ID: | SP001011 |
Sampleprep Summary: | Metabolites were extracted from pellets using a bligh and dyer-based methanol/chloroform/water extraction with inclusion of norvaline as a polar internal standard. Briefly, 250 μl MeOH, 250 μl CHCL3, 100 μl water containing norvaline were added to pellets. This was vortexed for 10 minutes followed by centrifugation at 10,000 g for 5 minutes at 4°C. The upper phase was separated and dried under vacuum at 4°C until dry. Polar metabolites were derivatized in 2% (w/v) methoxyamine hydrochloride in pyridine and incubated at 45°C for 60 minutes. Samples were then silylated with N-tertbutyldimethylsilyl-N-methyltrifluoroacetamide (MtBSTFA) with 1% tert-butyldimethylchlorosilane (tBDMCS) at 45°C for 30 minutes. Polar derivatives were analyzed by GC-MS using a DB-35MS column (30m x 0.25 mm i.d. x 0.25 μm) installed in an Agilent 7890B gas chromatograph (GC) interfaced with an Agilent 5977A mass spectrometer (MS) with an XTR EI source using the following temperature program: 100 °C initial, increase by 3.5 °C/min to 255 °C, increase by 15 °C/min to 320 °C and hold for 3 min. |