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MB Sample ID: SA046823
Local Sample ID: | AZ 37 |
Subject ID: | SU000870 |
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: | SU000870 |
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 |
---|---|---|---|---|
AZ 37 | SA046823 | FL009245 | Pravastatin | Treatment |
AZ 37 | SA046823 | FL009245 | Left Lung | Lung |
AZ 37 | SA046823 | FL009245 | 8 | Time point (day) |
Collection:
Collection ID: | CO000864 |
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. Lung wash fluid was collected at day 0, 8 and 12 of each treatment. |
Sample Type: | Bronchoalveolar lavage |
Treatment:
Treatment ID: | TR000884 |
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. Lung wash fluid was collected at day 0, 8 and 12 of each treatment. |
Sample Preparation:
Sampleprep ID: | SP000877 |
Sampleprep Summary: | Oxylipins, endocannabinoids, and fatty acids were isolated by solid phase extraction on 60 mg Waters Oasis-HLB cartridges (Milford, MA), as previously described by Luria et al (1). Prior to extraction, cartridges were washed with 1 column volume ethyl acetate followed by 2 column volumes methanol and conditioned with 2 mL of 95:5 v/v water/methanol (MeOH) with 0.1% acetic acid. The column reservoir was spiked with 5 µL anti-oxidant solution, (0.2 mg/ml solution BHT/EDTA in 1:1 MeOH:water), and 10 μL 1000nM analytical surrogates (See Table 2 below for specific compounds). Sample aliquots (500 µL media) were then introduced to the column reservoir and diluted with 1 column volume wash solution (5% MeOH, 0.1% acetic acid) and allowed to gravity extract into tubes containing 10% bleach solution to decontaminate sample waste. SPE cartridges were dried by vacuum @ -7.5in Hg for 20 min. Analytes were then eluted by gravity with 0.5 mL MeOH, followed by 1.25 mL Ethyl Acetate, into 2 mL autosampler vials containing 10 µL 20% glycerol solution in MeOH. Eluent was dried by vacuum evaporation for 35 min, and residues were re-constituted with 100uL of 100 nM internal standard solution containing 1-cyclohexyl ureido,3-dodecanoic acid (CUDA) and 1-Phenyl 3-Hexadecanoic Acid Urea (PHAU), in 50:50 MeOH:ACN and let sit for 10 min at ambient temperature. Vials were then chilled 15 min on wet ice, and extracts were transferred to a centrifugal filter (0.1 µm Durapore, Millipore, Billerica, MA), centrifuged for 3 min at 6ºC at <4500g (rcf) and transferred to 150 uL glass inserts within 2 mL amber vials, and capped. Extracts were stored at -20ºC until analysis by UPLC-MS/MS. The internal standard was used to quantify the recovery of the deuterated extraction surrogates by ratio response. |
Combined analysis:
Analysis ID | AN001364 | AN001365 |
---|---|---|
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 | NEGATIVE | POSITIVE |
Units | Concentration (nM) | Concentration (nM) |
Chromatography:
Chromatography ID: | CH000952 |
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 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: | MS001256 |
Analysis ID: | AN001364 |
Instrument Name: | ABI Sciex 6500 QTrap |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
Ion Mode: | NEGATIVE |
MS ID: | MS001257 |
Analysis ID: | AN001365 |
Instrument Name: | ABI Sciex 6500 QTrap |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
Ion Mode: | POSITIVE |