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MB Sample ID: SA049109

Local Sample ID:C10
Subject ID:SU000899
Subject Type:Sheep
Subject Species:Ovis aries
Taxonomy ID:9940
Gender:Both
Species Group:Mammal

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Subject:

Subject ID:SU000899
Subject Type:Sheep
Subject Species:Ovis aries
Taxonomy ID:9940
Gender:Both
Species Group:Mammal

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
C10SA049109FL009550AdultLife stage
C10SA049109FL009550PregnantPregnancy status
C10SA049109FL009550NormoxicTreatment group
C10SA049109FL009550VenousSample origin

Collection:

Collection ID:CO000893
Collection Summary:Whole blood was immediately separated by centrifugation and the plasma frozen in liquid nitrogen and stored at -80C for later analysis.
Sample Type:Blood
Blood Serum Or Plasma:Venous and arterial plasma

Treatment:

Treatment ID:TR000913
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:SP000906
Sampleprep Summary:After randomization, a 50 μl plasma aliquot in a 1.5 mL polypropylene Eppendorf tube was enriched with deuterated surrogates in 10 µL 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 200 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 mixed by vortexing and protein precipitate was 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 AN001404 AN001405
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:CH000984
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:MS001296
Analysis ID:AN001404
Instrument Name:ABI Sciex 6500 QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
Ion Mode:NEGATIVE
  
MS ID:MS001297
Analysis ID:AN001405
Instrument Name:ABI Sciex 6500 QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
Ion Mode:POSITIVE
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