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MB Sample ID: SA034206
| Local Sample ID: | 231_CFS |
| Subject ID: | SU000640 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 53 ± 3.5 (23-69 yr) |
| Gender: | Male |
| Human Ethnicity: | White/non-Hispanic |
| Species Group: | Human |
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Sample Preparation:
| Sampleprep ID: | SP000647 |
| Sampleprep Summary: | Metabolites from plasma were extracted using pre-chilled methanol/acetonitrile (50:50) |
| Extraction Method: | Typically, 90 μl of plasma was thawed on ice and mixed with 5 μl of isotope standards and incubated for 10 min at 20°C to permit small molecules and vitamins in the internal standards to associate with plasma binding proteins. Macromolecules (protein, DNA, RNA, glycans, etc.) were precipitated by extraction with 4 volumes (400 μl) of cold (-20°C), acetonitrile:methanol (50:50) and incubated on crushed ice for 10 min, then removed by centrifugation at 16,000g x 10 min at 4°C. The supernatants containing the extracted metabolites and internal standards were transferred to labeled cryotubes and stored at -80°C for LCMS/MS analysis. |
| Extract Storage: | -80 |
| Sample Derivatization: | No |
| Sample Spiking: | 25-35 commercial stable isotope internal standards, and 57 stable isotope internal standards that were custom-synthesized in E. coli and S. cerevisiae by metabolic labeling with 13C-glucose and 13C bicarbonate |
