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MB Sample ID: SA235376
Local Sample ID: | Naive_249 |
Subject ID: | SU002434 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Age Or Age Range: | 12-24 weeks |
Gender: | Male |
Animal Animal Supplier: | Jackson |
Animal Light Cycle: | 12L/12D |
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Sample Preparation:
Sampleprep ID: | SP002440 |
Sampleprep Summary: | Whole blood was extracted from animals at the time of euthanization from the heart chamber. Blood was collected into blood collection tubes (Fisher Scientific; #02-675-185) and spun for 10 min at 11,000g. Serum was collected and frozen in liquid nitrogen. 25uL of plasma was extracted with 500uL of acetonitrile by vortexing and centrifugation at 10min at 14,000rpm. 450uL of supernatant was transferred to new tube and dried via SpeedVac. Dried samples were reconstituted with 25uL of 50% methanol and transferred to autosampler vials. Injection volume =10uL in PRM mode for detection and quantification of 10 different analytes. Metabolite mixture was analyzed on Thermo Orbitrap IDX MS system coupled to a Vanquish UPLC system. Samples were transported via the autosampler (10uL injection volume) onto a Waters BEH C18 column. Runtime was 15min in PRM mode. Buffer A: 0.1% formic acid in water. Buffer B: 0.1% formic acid in methanol. LC Gradient: 0min: 0% B, 8min: 50% B, 9 min: 98% B, 13min: 98% B. Recalibration of system up to 15 min at 0% B for next injection. |
Processing Storage Conditions: | Described in summary |
Extract Storage: | Described in summary |