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MB Sample ID: SA057627

Local Sample ID:BL_84
Subject ID:SU001001
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090

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Sample Preparation:

Sampleprep ID:SP001008
Sampleprep Summary:Extraction, Solvent Removal, Resuspension, Solvent Removal, Two-step Derivatization and Addition of FAME Markers
Processing Method:Liver tissue were frozen on dry ice and weighed at approximate 100 mg to a MagNa Lyser tube with 10 to 15 ceramic beads inside. The ice-cold homogenization buffer (50% acetonitrile) was added with the mass/volume ratio at 1 mg tissue per 5 µL buffer. The homogenization was carried on at 4,500 rpm for two 30 sec pulses by the MagNA Lyser. The homogenized mixture was centrifuged at 14,000 rpm for 5 min at -4 °C for the supernatant. Aliquots of 100 µL homogenized supernatant of the study sample (equal to 20 mg tissue) were mixed with 1000 µL of cold degassed 3:3:2 ACN:ISP:H20 solution in a 2 mL snap cap tube. Samples are vortex and shaken for 5 min, and then centrifuged at 4 °C for 2 min at 14000 rcf. The supernatant was split into (2)-450 µL fractions, one fraction was archived in -80°C the second fraction was subject t complete sample processing. Transfer supernatant to a new vial. Evaporate supernatant to complete dryness at room temp.
Processing Storage Conditions:On ice
Extraction Method:(step 1) 3:3:2 Acetonitrile:Isopropanol:H2O, (step 2) 1:1 Acetonitrile:H2O
Extract Storage:-80℃
Sample Resuspension:Samples resuspended in 200 µL of 1:1 ACN:H2O. Evaporated to dryness w/speed vac.
Sample Derivatization:Formation of methoximes by adding 10µL of 40mg/mL MeOx in pyridine to each sample. Placed onto Thermomixer for 90 min at 30⁰C. Then FAME retention index markers and MSTFA added to each sample for derivatization step. Samples returned to Thermomixer for 45 min at 70⁰C.
Sample Spiking:Fatty acid methyl esters (FAME)
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