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MB Sample ID: SA052451
| Local Sample ID: | 4629 BM2-PHZ |
| Subject ID: | SU000927 |
| Subject Type: | Animals |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57/B6 |
| Age Or Age Range: | 8-16 weeks |
| Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000927 |
| Subject Type: | Animals |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57/B6 |
| Age Or Age Range: | 8-16 weeks |
| Species Group: | Mammal |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 4629 BM2-PHZ | SA052451 | FL009974 | control chow | Diet of subject mice |
| 4629 BM2-PHZ | SA052451 | FL009974 | 12 | Feeding period (wk) |
| 4629 BM2-PHZ | SA052451 | FL009974 | 1 | PHZ dose (mg) |
Collection:
| Collection ID: | CO000921 |
| Collection Summary: | Mouse plasma was collected by obtaining peripheral blood via peri-orbital bleeding. Approximately 100 ul blood is placed in a microvette vial with pre-loaded KEDTA and then centrifuged at 14,000 rpm for 10 minutes at 4 degrees. Plasma is removed and stored at -80 C until analysis. |
| Sample Type: | Blood |
Treatment:
| Treatment ID: | TR000941 |
| Treatment Summary: | Phenylhydrazine was administered at 1mg once on day 1 and plasma was collected on day 4. Vitamin A deficient diet: Teklad, TD.86143. Control diet: Teklad, TD.91280. Fatty acid deficient diet: Teklad, TD.03314. Control diet: Teklad, TD.130321. |
Sample Preparation:
| Sampleprep ID: | SP000934 |
| Sampleprep Summary: | For free fatty acids analysis, 100 μL of plasma extraction sample or 400 μL pull-down samples was used for DMAPA (dimethylaminopropylamine) derivatization for improving MS sensitivity of free fatty acids. Prior to the derivatization, 50 ng of AA-d8 (deuterated arachidonic acid –d8) as the internal standard was added to each sample. The solvents in the sample were dried under a stream of nitrogen. To derivatize the sample, 50 μL of 50 mM EDC (N-ethyl-N-(3-dimethylaminopropyl)-carbodiimide hydrochloride) and 50 mM DMAPA and 50 mM DMAP (4-dimethylaminopyridine) were added to the dried fatty acid samples and heated at 50 ◦C for 30 minutes. The samples were dried under nitrogen and then dissolved in 1 mL of ethanol : water for MS. |
Combined analysis:
| Analysis ID | AN001452 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Shimadzu 20AD |
| Column | Agilent Eclipse XDB-C18 (100 x 3.0mm,3.5um) |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | ABI Sciex API 4000 QTrap |
| Ion Mode | POSITIVE |
| Units | peak area ratio |
Chromatography:
| Chromatography ID: | CH001021 |
| Chromatography Summary: | For DMAPA derivatized Fatty acid analysis. Shimadzu 20AD two pump system was used. Solvent A: 0.1% formic acid in water. Solvent B: 0.1% in Acetonitrile. The initial LC condition was 30% B. The solvent gradient was programmed from 30% B to 99% B in 4 min and was hold for 1 min at the flow rate of 1 mL/min. Then, it was immediately brought back to 30 % B and was hold for 2 min. An Agilent Eclipse XDB-C18 column (3.0 x100 mm x 3.5 µm) was used for this analysis. |
| Instrument Name: | Shimadzu 20AD |
| Column Name: | Agilent Eclipse XDB-C18 (100 x 3.0mm,3.5um) |
| Column Temperature: | 60 C |
| Flow Gradient: | 30% B to 99% B in 4 min |
| Flow Rate: | 1 mL/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% acetonitrile; 0.1% formic acid |
| Analytical Time: | 7 min |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS001342 |
| Analysis ID: | AN001452 |
| Instrument Name: | ABI Sciex API 4000 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Units of measurement:peak area ratio: analyte peak area/peak area of internal standard |
| Ion Mode: | POSITIVE |
