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MB Sample ID: SA094661
Local Sample ID: | ncB5 |
Subject ID: | SU001385 |
Subject Type: | Mammal |
Subject Species: | Rattus norvegicus |
Taxonomy ID: | 10116 |
Genotype Strain: | Sprague Dawley |
Age Or Age Range: | 70 days old |
Gender: | Male |
Animal Animal Supplier: | Harlan |
Animal Housing: | polycarbonate-free caging |
Animal Light Cycle: | 14-hr light and 10-hr dark |
Animal Feed: | Phytoestrogen Reduced II 18-5, Ziegler Bros, Inc |
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Sample Preparation:
Sampleprep ID: | SP001393 |
Sampleprep Summary: | Lipids were extracted using a modified Bligh-Dyer method. Fifty µL of serum and 25 mg of crushed liver was used for the extraction. The extraction was carried out using 2:2:2 volume ratio of water/methanol/dichloromethane at room temperature after spiking internal standards 17:0 LPC, 17:0 PC, 17:0 PE, 17:0 PG, 17:0 ceramide, 17:0 SM, 17:0PS, 17:0PA, 17:0 TAG, 17:0 MAG, 16:0/18:1 DAG, 17:0 CE. The organic layer was collected and completely dried under nitrogen. Before MS analysis, the dried extract was resuspended in 100 μL of Buffer B (10:5:85 Acetonitrile/water/Isopropyl alcohol) containing 10 mM NH4OAc and subjected to LC/MS. The lipidome was separated using reverse-phase chromatography. |
Sampleprep Protocol Filename: | unbiased.liver.MS.method.pdf |