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MB Sample ID: SA086081
| Local Sample ID: | Plasma_10 |
| Subject ID: | SU001284 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Male and female |
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Sample Preparation:
| Sampleprep ID: | SP001292 |
| Sampleprep Summary: | Plasma samples were thawed and 100 uL was prepared using methanol precipitation and liquid-liquid extraction as previously described (PMCID PMC4214365, PMCID PMC3734953) . In short, following the addition of standards, 400uL of ice cold MeOH was added to 100uL of plasma to precipitate proteins, then vortexed for 10 seconds, and centrifgued at 0oC for 15 min at 18,000 rpm to pellet precipitated protein. The supernatant was transferred to a glass culture tube and dried under N2. 3.1mL MTBE was added to the dried methanol residue, vortexed for 30 seconds, 750uL of water added to the tube, and vortex 10 seconds. The sample was then spun at 1000 rpm for 10 min at RT to form bilayer. 2.5 mL of MTBE layer was aliquoted and transferred to a new, clean glass culture tube. Then 3.0 mL MTBE was added to remaining water layer of sample, vortexed for 10 seconds, and centrifuged at 200 x g for 10 min at RT to form bilayer. 3mL of MTBE was aliquoted and combined with previous MTBE tube. This MTBE layer was dried under nitrogen at 35oC, and quickly re-suspend in 200uL Methanol, vortexed for 5 seconds and transfered to glass autosampler vial. The aqueous layer was dried under N2, 100uL of water quickly added to minimize oxidation. Then 400uL of ice cold MeOH was added, vortexed for 10 seconds, transfered to a 1.5 ml low retention Fisher microtube and frozen at -80oC for 25 min to crash out any remaining residual proteins. Then centrifuged at 0oC for 15 min at 18,000 xg. The supernatant was transferred to a clean microtube, dried in speed vac at 45oC. The dried supernatant was quickly resuspended in 100uL of 5% ACN in water, vortexed for 30 sec, then transferred to autosampler vials, and stored at -80C prior to MS analysis. |
| Processing Storage Conditions: | Described in summary |
| Extract Storage: | Described in summary |
