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MB Sample ID: SA011564
| Local Sample ID: | PAW-6 |
| 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-6 | SA011564 | FL002879 | ON: 75% oxygen, normal litter | Treatment |
| PAW-6 | SA011564 | FL002879 | 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 |
