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MB Sample ID: SA098930
| Local Sample ID: | Preeclampsia_9064 |
| Subject ID: | SU001434 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
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Sample Preparation:
| Sampleprep ID: | SP001442 |
| Sampleprep Summary: | Plasma samples were stored at − 80 °C until the time of analysis, which was conducted by the Michigan Regional Comprehensive Metabolomics Resource Core. Lipids were extracted using a modified BlighDyer Method. The extraction was performed using water/methanol/dichloromethane (2:2:2 v/v/v) at room temperature after the addition of internal standards. The organic layer was then collected and dried under a stream of nitrogen before being re-suspended in 100 μL of Buffer B (5%H2O:10%ACN:85%IPA containing 10 mM NH4OAc) and analyzed using a liquid chromatography tandem mass spectrometry (LC/MS/MS) lipidomics assay. The lipid extract was injected onto a 1.8 μm particle 50 × 2.1 mm internal diameter Waters Acquity HSS T3 column (Waters, Milford, MA) that was heated to 55 °C. For chromatographic elution, we used a linear gradient over a 20 min total run time. A 60% Solvent A and 40% Solvent B gradient was used for the first 10 min. Then the gradient was ramped in a linear fashion to 100% Solvent B which was maintained for 7 min. Thereafter, the system was switched back to 60% Solvent B and 40% Solvent A for 3 min. The flow rate used for these experiments was 0.4 mL/min and the injection volume was 5 μL. The column was equilibrated for 3 min before the next injection and run at a flow rate of 0.400uL/min for a total run time of 20 min. Data were acquired in positive and negative modes using data-dependent MS/MS with dynamic mass exclusion. Pooled human plasma samples and pooled experimental samples (prepared by combining small aliquots of each experimental sample) were used to control for the quality of sample preparation and analysis. A randomization scheme was used to distribute pooled samples within the set and a mixture of pure authentic standards was used to monitor instrument performance on a regular basis. |
| Processing Storage Conditions: | -80℃ |
| Extract Storage: | Described in summary |
