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MB Sample ID: SA037925
| Local Sample ID: | P1 B08 |
| Subject ID: | SU000682 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Male |
| Subject Comments: | Individuals with peripheral arterial disease, as defined by ankle brachial indices ≥0.4 and ≤0.8 and the presence of classic claudication symptoms |
| Species Group: | Human |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000682 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Male |
| Subject Comments: | Individuals with peripheral arterial disease, as defined by ankle brachial indices ≥0.4 and ≤0.8 and the presence of classic claudication symptoms |
| Species Group: | Human |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| P1 B08 | SA037925 | FL008010 | v3 | Visit |
| P1 B08 | SA037925 | FL008010 | A | Blood Draw |
Collection:
| Collection ID: | CO000676 |
| Collection Summary: | Blood was drawn form the subject at the baseline (Visit 2 (V2)) before (B) and after (A) the treadmill exercise test. Further, blood was collected after 12 weeks of supervised walking therapy (Visit 3 (V3)), before (B) and after (A) the treadmill exercise test. |
| Sample Type: | Plasma |
Treatment:
| Treatment ID: | TR000696 |
| Treatment Summary: | Subjects with peripheral arterial disease, as defined by ankle brachial indices ≥0.4 and ≤0.8 and the presence of classic claudication symptom were subjected to the treadmill exercise test. Further, they underwent 12 weeks of supervised walking therapy and subjected to the treadmill exercise test again. |
Sample Preparation:
| Sampleprep ID: | SP000689 |
| Sampleprep Summary: | Lipid mediator extraction and quantification: Oxylipins, endocannabinoids, and fatty acids were isolated from plasma using Ostro Sample Preparation Plates (Waters; Milford, MA) in the presence of BHT/EDTA and quantified by UPLC-MS/MS using internal standard methods. Briefly, in the Ostro Plate well, 100µl plasma aliquots were mixed with 5µl of BHT/EDTA (1:1 v/v MeOH/water) and 5µl methanol containing a suite of deuterated surrogates. The samples were then mixed with 300µL of acetonitrile with 1% formic acid and eluted with 15 mmHg vacuum for 10 min into 200µL glass inserts containing 10µl of 20% glycerol in methanol. Solvent was removed by vacuum centrifugation and the glycerol plug was stored at -80Co until analysis. Prior acquisition, samples were reconstituted in 100µl of 1:1 methanol:acetonitrile containing 100nM of 1-cyclohexyl ureido, 3-dodecanoic acid (CUDA) and 1-phenyl ureido, 3-hexadecanoic acid (PUHA). Residues within extracts were separated on a 2.1 x 150mm 0.17µm BEH column (Waters) and detected by electrospray ionization with multi reaction monitoring on a API 6500 QTRAP (Sciex; Redwood City, CA) and quantified against 7-9 point calibration curves of authentic standards using modifications of previously reported methods (Grapov, D., et al. 2012. PLoS One 7: e48852.; doi: 10.1371/journal.pone.0048852). |
Combined analysis:
| Analysis ID | AN001005 | AN001006 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Waters Acquity | Waters Acquity |
| Column | Waters Acquity BEH C18 (100 x 2mm,1.7um) | Waters Acquity BEH C18 (100 x 2mm,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 | POSITIVE | NEGATIVE |
| Units | Concentration pmol/g | Concentration pmol/g |
Chromatography:
| Chromatography ID: | CH000722 |
| Methods Filename: | 2017_6500_OxyEndo_Analysis_protocol.pdf |
| Instrument Name: | Waters Acquity |
| Column Name: | Waters Acquity BEH C18 (100 x 2mm,1.7um) |
| Column Temperature: | 60 °C |
| Flow Gradient: | See protocol/methods file |
| Flow Rate: | 0.25 uL |
| 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: | MS000900 |
| Analysis ID: | AN001005 |
| Instrument Name: | ABI Sciex 6500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Endocannabinoid Analysis |
| Ion Mode: | POSITIVE |
| MS ID: | MS000901 |
| Analysis ID: | AN001006 |
| Instrument Name: | ABI Sciex 6500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Oxylipin Analysis |
| Ion Mode: | NEGATIVE |
