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MB Sample ID: SA236564
Local Sample ID: | TFA-T4 |
Subject ID: | SU002443 |
Subject Type: | Mammal |
Subject Species: | Rattus norvegicus |
Taxonomy ID: | 10116 |
Age Or Age Range: | 26 weeks of age |
Weight Or Weight Range: | 320-460 grams |
Gender: | Male |
Animal Animal Supplier: | RRRC (University of Missouri) |
Animal Housing: | Center for Comparative Medicine UConn |
Animal Feed: | Modified AIN-93G diet from Research Diets |
Animal Water: | ab libitum |
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Sample Preparation:
Sampleprep ID: | SP002449 |
Sampleprep Summary: | Samples (0.85 ml) were spiked with 5 ng each (in 150 μl methanol) of 15(S)-HETE-d8,14(15)-EpETrE-d11, Resolvin D2-d5, Leukotriene B4-d4, and Prostaglandin E1-d4 as internal standards for recovery and quantitation and mixed thoroughly. The samples were then extracted for PUFA metabolites using C18 extraction columns as described earlier [1-4]. Briefly, the internal standard spiked samples were applied to conditioned C18 cartridges, washed with 15% methanol in water followed by hexane and dried under vacuum. The cartridges were eluted with 0.5 ml methanol. The eluate was dried under a gentle stream of nitrogen. The residue was redissolved in 50 μl methanol-25 mM aqueous ammonium acetate (1:1) and subjected to LC-MS analysis. |
Sampleprep Protocol Filename: | Methods of Eicosanoid Detection.pdf |