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MB Sample ID: SA053336
| Local Sample ID: | S21 |
| Subject ID: | SU000947 |
| Subject Type: | Sheep |
| Subject Species: | Ovis aries |
| Taxonomy ID: | 9940 |
| Gender: | Both |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000947 |
| Subject Type: | Sheep |
| Subject Species: | Ovis aries |
| Taxonomy ID: | 9940 |
| Gender: | Both |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| S21 | SA053336 | FL010040 | Carotid | Tissue type |
| S21 | SA053336 | FL010040 | Hypoxic | Treatment group |
| S21 | SA053336 | FL010040 | Adult non-pregnant | Life stage |
Collection:
| Collection ID: | CO000941 |
| Collection Summary: | Part of the routinely flash frozen samples from the Loma Linda University School of Medicine, Center for Perinatal Biology. |
| Sample Type: | Tissue |
| Tissue Cell Identification: | Adipose tissue, Carotid, pulmonary arteries, uterine arteries |
Treatment:
| Treatment ID: | TR000961 |
| Treatment Summary: | To induce chronic hypoxia, pregnant and non-pregnant ewes were transported to the White Mountain Research Station that is owned and operated by the University of California. Animals were housed at the Barcroft Research Station (3800 m) for 100+ days prior to having the pregnant or non-pregnant ewes or 2 week old newborn lambs transported back to Loma Linda for study. |
Sample Preparation:
| Sampleprep ID: | SP000954 |
| Sampleprep Summary: | After sample randomization, weighed tissue (around 10mg for adipose and 50mg for arteries) in 2 mL polypropylene Eppendorf tube was enriched with deuterated surrogates in 20µL methanol (Tables S1 and S2 from Agrawal, K., L.A. Hassoun, N. Foolad, T.L. Pedersen, R.K. Sivamani, J.W. Newman. 2017. Sweat lipid mediator profiling: a non-invasive approach for cutaneous research. J. Lipid Res. 58:188–195 [EPub: Nov 7, 2016]. doi: 10.1194/jlr.M071738) and 5 μl of BHT/EDTA in 1:1 methanol/water (v/v). A total of 400 μl 1-cyclohexyl uredio, 3-dodecanoic acid / 1-phenyl ureido, 3-hexanoic acid (CUDA / PUHA) in 1:1 methanol/acetonitrile (v/v) was added, the sample ground with 3mm stainless steel beads (two for adipose tissue and 3 for blood vessel tissue) and agitation for 6 min at 1500RPM. Protein precipitate and debris were removed by centrifugation (10 min, 10,000 RCF, 6 °C). The supernatant was filtered by centrifugation through 1 µm PVDF membranes (Millipore, Billerica, MA) at 6 °C and 4,500 RCF for 3 min. The filtrate was stored in glass vials at -20 °C until UPLC-MS/MS analysis. |
Combined analysis:
| Analysis ID | AN001478 | AN001479 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Waters Acquity | Waters Acquity |
| Column | Aquity C18 BEH (100 x 2.1mm,1.7um) | Aquity C18 BEH (100 x 2.1mm,1.7um) |
| MS Type | ESI | ESI |
| MS instrument type | Triple quadrupole | Triple quadrupole |
| MS instrument name | ABI Sciex 6500 QTrap | ABI Sciex 6500 QTrap |
| Ion Mode | NEGATIVE | POSITIVE |
| Units | Concentration (nM) | Concentration (nM) |
Chromatography:
| Chromatography ID: | CH001037 |
| Instrument Name: | Waters Acquity |
| Column Name: | Aquity C18 BEH (100 x 2.1mm,1.7um) |
| Column Temperature: | 60 °C |
| Flow Gradient: | See protocol/methods file |
| Flow Rate: | 0.25 mL/min |
| Internal Standard: | See protocol/methods file |
| Retention Time: | See protocol/methods file |
| Sample Injection: | 5 µL |
| Solvent A: | 100% water; 0.1% acetic acid |
| Solvent B: | 90% acetonitrile/ 10% isopropanol |
| Analytical Time: | 20 min |
| Weak Wash Solvent Name: | 20% methanol, 10% isopropanol |
| Weak Wash Volume: | 600 µL |
| Strong Wash Solvent Name: | 50:50 Acetonitrile:Methanol |
| Strong Wash Volume: | 600 µL |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS001362 |
| Analysis ID: | AN001478 |
| Instrument Name: | ABI Sciex 6500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| Ion Mode: | NEGATIVE |
| MS ID: | MS001363 |
| Analysis ID: | AN001479 |
| Instrument Name: | ABI Sciex 6500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
