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MB Sample ID: SA046949
Local Sample ID: | Zeki R 95 |
Subject ID: | SU000874 |
Subject Type: | Animal |
Subject Species: | Macaca mulatta |
Taxonomy ID: | 9544 |
Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU000874 |
Subject Type: | Animal |
Subject Species: | Macaca mulatta |
Taxonomy ID: | 9544 |
Species Group: | Mammal |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
Zeki R 95 | SA046949 | FL009282 | Right Lung | Tissue |
Zeki R 95 | SA046949 | FL009282 | Simvastatin | Treatment |
Collection:
Collection ID: | CO000868 |
Collection Summary: | Monkeys were treated with placebo or Provastatin for 12 days. Further, after the wash out period animals were treated with Simvastatin for 12 days. Plasma was collected at day 0, 8 and 12 of each treatment. |
Sample Type: | Tissue |
Tissue Cell Identification: | Trachea and lung tissue |
Treatment:
Treatment ID: | TR000888 |
Treatment Summary: | Monkeys were treated (by inhalation) with placebo or Provastatin for 12 days. Further, after the wash out period animals were treated with Simvastatin for 12 days. Plasma was collected at day 0, 8 and 12 of each treatment. |
Sample Preparation:
Sampleprep ID: | SP000881 |
Sampleprep Summary: | Oxylipins, endocannabinoids, bile acids and fatty acids were isolated using a Waters Ostro Sample Preparation Plate (Milford, MA). Lung and trachea samples were aliquoted (~40-60mg) into 2mL polypropylene tubes and spiked with a 5 µL anti-oxidant solution (0.2 mg/ml solution BHT/EDTA in 1:1 MeOH:water) and 10 μL 1000nM analytical deuterated surrogates. A total of 50 µL of methanol, 550µL isopropanol w/ 10mM ammonium formate & 1% formic acid and 100 uL water were added and the tube was placed in a Geno/Grinder for 30 sec before being centrifuged at 10,000g for 5 min at room temp. The supernate was then transferred into the plate wells and samples were eluted into glass inserts containing 10 μL 20% glycerol by applying a vacuum at 15 Hg for 10 min. Eluent was dried by speed vacuum for 35 min at the medium BP setting, before switching to an aqueous setting for an additional 35 min. Once dry, samples were re-constituted with the internal standard 1-cyclohexyl ureido, 3-dodecanoic acid (CUDA) and 1-Phenyl 3-Hexadecanoic Acid Urea (PHAU) at 100 nM (50:50 MeOH:CAN), vortexed 1 min, transferred to a spin filter (0.1 µm, Millipore, Billerica, MA), centrifuged for 3 min at 6ºC at <4500g (rcf), before being transferred to 2 mL LC-MS amber vials. Extracts were stored at -20ºC until analysis by UPLC-MS/MS. The internal standard was used to quantify the recovery of surrogate standards. |
Combined analysis:
Analysis ID | AN001371 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Waters Acquity |
Column | Aquity C18 BEH (100 x 2.1mm,1.7um) |
MS Type | ESI |
MS instrument type | Triple quadrupole |
MS instrument name | ABI Sciex 6500 QTrap |
Ion Mode | NEGATIVE |
Units | Concentration (nM) |
Chromatography:
Chromatography ID: | CH000956 |
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: | 0.4 mL/min |
Internal Standard: | See protocol/methods file |
Retention Time: | See protocol/methods file |
Sample Injection: | 5 µL |
Solvent A: | 100% water; 0.1% formic acid |
Solvent B: | 100% acetonitrile; 0.1% formic acid |
Analytical Time: | 16 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: | MS001263 |
Analysis ID: | AN001371 |
Instrument Name: | ABI Sciex 6500 QTrap |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
Ion Mode: | NEGATIVE |