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MB Sample ID: SA051647
Local Sample ID: | S111 |
Subject ID: | SU000924 |
Subject Type: | Animal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Species Group: | Mammal |
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Sample Preparation:
Sampleprep ID: | SP000931 |
Sampleprep Summary: | Extraction, Solvent Removal, Resuspension, Solvent Removal, Two-step Derivatization and Addition of FAME Markers |
Processing Method: | 30 µL of serum were added to Eppendorf tube. Add 1000 µL of 3:3:2 ACN:ISP:H20 extraction solution to aliquots, shake and centrifuge for 2 minutes at 4C and 14000 rcf to precipitate proteins. Sample 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 and or 4⁰C cold room |
Extraction Method: | (step 1) 3:3:2 Acetonitrile:Isopropanol:H2O, (step 2) 1:1 Acetonitrile:H2O |
Extract Storage: | -80⁰C |
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) |