Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA053344

Local Sample ID:	S29
Subject ID:	SU000947
Subject Type:	Sheep
Subject Species:	Ovis aries
Taxonomy ID:	9940
Gender:	Both

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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.

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.