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MB Sample ID: SA057827
| Local Sample ID: | BS_5 |
| Subject ID: | SU001006 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
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Sample Preparation:
| Sampleprep ID: | SP001013 |
| Sampleprep Summary: | Extraction, Solvent Removal, Resuspension, Solvent Removal, Two-step Derivatization and Addition of FAME Markers |
| Processing Method: | Aliquots of 30 µL serum were mixed with 1000 µL of degassed 3:3:2 Acetonitrile:Isopropanol:Water solution in a 2 mL snap cap tube. Samples were vortexed, shaken for 5 minutes then centrifuged at 4⁰C for 2 minutes at 14000rcf. The supernatant was recovered and split into two fractions. Fraction 1 (450 µL) was archived and fraction 2 (450 µL) was subjected to complete sample processing. Processing continued as follows. Samples were completely dried by vacuum centrifuge and reconstituted with 450 µL 1:1 acetonitrile:water solution. Samples were centrifuged for 2 minutes at 14000rcf and transferred to new snap cap tubes followed by complete evaporation at room temperature by vacuum centrifuge. |
| 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) |
