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MB Sample ID: SA254079
Local Sample ID: | BOT11 |
Subject ID: | SU002621 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
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Sample Preparation:
Sampleprep ID: | SP002627 |
Sampleprep Summary: | Serum samples were thawed on ice, followed by metabolite extraction of the non-polar (lipids) metabolome. Extraction solvent was prepared by adding 725 μL of the isotopically labeled lipid standard mixture to 43.5 mL 2-propanol (1:60 ratio) and was kept on ice. This cold extraction solvent was added to serum samples in a 3:1 ratio (solvent: serum) for protein precipitation followed by vortex mixing for 15 seconds. Samples were centrifuged at 13,000 rpm for 7 minutes and the resulting supernatant was transferred to LC vials. The supernatant was stored at -80 °C until UHPLC-MS analysis, which was performed within one week of preparation. A blank sample, prepared with LC-MS grade water, underwent the same sample preparation process as the serum samples and was analyzed with the rest of the samples. Pooled quality control (QC) samples were prepared by combining 5-10 μL aliquots of the supernatant of each serum sample. This pooled QC sample was analyzed after every 10 LC-MS runs to monitor instrument stability through the course of the experiment. Samples were randomized for sample preparation and were run in a randomized order on consecutive days. |