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MB Sample ID: SA055261
Local Sample ID: | 17 |
Subject ID: | SU000962 |
Subject Type: | Cell culture |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Cell Strain Details: | C57BL/6 |
Cell Primary Immortalized: | AT1 |
Cell Counts: | ~60% confluency 6 well plate |
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Sample Preparation:
Sampleprep ID: | SP000969 |
Sampleprep Summary: | The samples were “crash” deprotonized by methanol precipitation and spiked with D27-deuterated myristic acid (D27-C14:0) as an internal standard for retention-time locking and dried. The trimethylsilyl (TMS)-D27-C14:0 standard retention time (RT) was set at ~16.727 min. Reactive carbonyls were stabilized at 50 °C with methoxyamine hydrochloride in dry pyridine. Metabolites were made volatile with TMS groups using N-methyl-N-(trimethylsilyl) trifluoroacetamide or MSTFA with catalytic trimethylchlorosilane at 50 °C. GC/MS methods generally follow those of Roessner et al. (2000), Fiehn et al. (2008), and Kind et al. (2009), and used a 6890 N GC connected to a 5975 Inert single-quadrupole MS (Agilent Technologies, Santa Clara, CA). The two wall-coated, open-tubular (WCOT) GC columns connected in series are both from J&W/Agilent (part 122–5512), DB5-MS, 15 meters in length, 0.25 mm in diameter, with an 0.25-μm luminal film. Positive ions generated with conventional electron-ionization (EI) at 70 eV are scanned broadly from 600 to 50 m/z in the detector throughout the 45 min cycle time. |