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MB Sample ID: SA011548
Local Sample ID: | PAW-4 |
Subject ID: | SU000273 |
Subject Type: | Animal |
Subject Species: | Rattus norvegicus |
Taxonomy ID: | 10116 |
Age Or Age Range: | 2 weeks |
Animal Housing: | Litters of 10 (N group) or 17 (R group) |
Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU000273 |
Subject Type: | Animal |
Subject Species: | Rattus norvegicus |
Taxonomy ID: | 10116 |
Age Or Age Range: | 2 weeks |
Animal Housing: | Litters of 10 (N group) or 17 (R group) |
Species Group: | Mammal |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
PAW-4 | SA011548 | FL002875 | AN: Room Air, normal litter (no restricted nutritional intake) | Treatment |
PAW-4 | SA011548 | FL002875 | Plasma | Tissue |
Collection:
Collection ID: | CO000265 |
Collection Summary: | - |
Collection Protocol Filename: | NewmanLab-F&BOxys-Protocol-PT.pdf |
Sample Type: | Lung and plasma |
Collection Method: | Dissection |
Storage Conditions: | - 80°C |
Storage Vials: | Eppendorf tubes |
Treatment:
Treatment ID: | TR000285 |
Treatment Summary: | Litters of 10 pups (N, normal intake) or litters of 17 pups (R, restricted intake). Normal and restriced intake pups were exposed to 75% oxygen (O, oxygen) in a Plexiglass chamber or to 21% oxygen (A, room air) continuously for 14 days, |
Treatment Protocol Filename: | NewmanLab-F&BOxys-Protocol-PT.pdf |
Treatment: | Intervention |
Treatment Doseduration: | 1 week |
Treatment Vehicle: | Food availability, oxygen exposure |
Animal Endp Euthanasia: | 2 weeks |
Animal Endp Tissue Coll List: | Lung and plasma |
Animal Endp Tissue Proc Method: | Dissected, placed on dry ice |
Sample Preparation:
Sampleprep ID: | SP000279 |
Sampleprep Summary: | Tissues were pulverized on dry ice, enriched with deuterated surrogates, antioxidants (BHT and EDTA), and the total lipid extract was isolated using 10:8:11 cyclohexane/2-propanol/ammonium acetate (v/v/v). Subsequently, esterified oxylipins were liberated via sodium methoxide. Next, the remaining extract was eluted through a Waters Oasis-HLB cartridge. Plasma was added directly to a Waters Ostro plate, mixed with the surrogates, antioxidants, and acetonitrile and the extract eluted through. Both tissue and plasma extracts were dried, reconstituted in 100?L 50:50 methanol:acetonitrile with internal standards 1?phenyl?3?hexanoic acid urea and 1?cyclohexyl?3-dodecanoic acid urea (Sigma-Aldrich, St. Louis, MO) and filtered at 0.1 µm. |
Sampleprep Protocol Filename: | NewmanLab-F&BOxys-Protocol-PT.pdf |
Extraction Method: | Liquid:liquid with HLB solid phase extraction (Tissue); Ostro plate (Plasma) |
Extract Cleanup: | HLB solid phase extraction (Tissue); Ostro plate (Plasma) |
Extract Storage: | - 20 °C |
Sample Resuspension: | 100 µL |
Sample Spiking: | See sample prep protocol file |
Organ: | Lung and plasma |
Combined analysis:
Analysis ID | AN000400 | AN000401 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | ||
Chromatography system | Waters Acquity | Waters Acquity |
Column | ||
MS Type | ESI | ESI |
MS instrument type | Triple quadrupole | Triple quadrupole |
MS instrument name | ABI Sciex API 4000 QTrap | ABI Sciex API 4000 QTrap |
Ion Mode | NEGATIVE | NEGATIVE |
Units | % Area Ratio | nM |
Chromatography:
Chromatography ID: | CH000283 |
Chromatography Summary: | Oxylipin analysis |
Methods Filename: | NewmanLab-F&BOxys-Protocol-PT.pdf |
Instrument Name: | Waters Acquity |
Column Temperature: | 60 C |
Flow Gradient: | See protocol/methods file |
Flow Rate: | 0.25 |
Internal Standard: | See protocol/methods file |
Retention Time: | See protocol/methods file |
Sample Injection: | 5 L |
Solvent A: | 100% water; 0.1% acetic acid |
Solvent B: | 90% acetonitrile/ 10% isopropanol |
Analytical Time: | 16 min |
Weak Wash Solvent Name: | 20% methanol, 10% isopropanol |
Weak Wash Volume: | 600 L |
Strong Wash Solvent Name: | 50:50 Acetonitrile:Methanol |
Strong Wash Volume: | 600 L |
Sample Loop Size: | 17 L |
MS:
MS ID: | MS000344 |
Analysis ID: | AN000400 |
Instrument Name: | ABI Sciex API 4000 QTrap |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
MS Comments: | Plasma results |
Ion Mode: | NEGATIVE |
MS ID: | MS000345 |
Analysis ID: | AN000401 |
Instrument Name: | ABI Sciex API 4000 QTrap |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
MS Comments: | Lung results |
Ion Mode: | NEGATIVE |