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MB Sample ID: SA052488
Local Sample ID: | NFC-PHZ-23 |
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 |
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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 |
---|---|---|---|---|
NFC-PHZ-23 | SA052488 | FL009976 | control chow | Diet of subject mice |
NFC-PHZ-23 | SA052488 | FL009976 | 4 | Feeding period (wk) |
NFC-PHZ-23 | SA052488 | FL009976 | 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 |