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MB Sample ID: SA094682

Local Sample ID:HFB2
Subject ID:SU001387
Subject Type:Mammal
Subject Species:Rattus norvegicus
Taxonomy ID:10116
Genotype Strain:Sprague Dawley
Age Or Age Range:240 days
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) or D09100301 (Research Diets, Inc)

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Sample Preparation:

Sampleprep ID:SP001395
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.serum.MS.method.HFD.pdf
unbiased.serum.MS.method.pdf
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