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MB Sample ID: SA011605

Local Sample ID:P25
Subject ID:SU000274
Subject Type:Animal
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:C57BL/6
Species Group:Mammal

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Subject:

Subject ID:SU000274
Subject Type:Animal
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:C57BL/6
Species Group:Mammal

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
P25SA011605FL002898Chronic Alcohol ModelGroups
P25SA011605FL002898No IPATreatment
P25SA011605FL002898WTStrain

Collection:

Collection ID:CO000266
Collection Summary:-
Sample Type:Blood

Treatment:

Treatment ID:TR000286
Treatment Summary:Control vs IPA Supplemented Wild Type and SL Mice
Treatment:Indole-3-propionic Acid (IPA) Supplemention
Treatment Compound:Indole-3-propionic Acid (IPA)
Treatment Route:Drinking Water
Treatment Dose:0, 1, and 10 mM IPA

Sample Preparation:

Sampleprep ID:SP000280
Sampleprep Summary:Aliquots of plasma samples (20 µL) were transferred into pre-labeled 2.0 mL low-bind Eppendorf tubes for experimental samples. For all samples 100 µL Protein Precipitation Buffer with internal standard [2.5 ng/ml of 2-methyl-4-chlorophenyl-acetic acid (MCPA) in methanol] was added to each tube and were vortexed for 2 min at 5000 rpm. The samples were centrifuged at 16,000 rcf for 4 min. A 90 µL of the supernatant wastransferred into pre-labeled auto-sampler vials and diluted by adding 90 µL of water with 10% Formic Acid. Samples were then mixed on a multiple tube vortexer for 5 min at 5000 rpm. 10 µL of samples were injected onto API-4000 Q-trap.
Processing Storage Conditions:4°C
Extract Storage:-80C
Sample Resuspension:Methanol, H2O, formic acid
Sample Spiking:2-methyl-4-chlorophenyl-acetic acid (MCPA)

Combined analysis:

Analysis ID AN000402
Analysis type MS
Chromatography type Reversed phase
Chromatography system Waters Acquity
Column Waters Acquity BEH C18 (50 x 2.1mm,1.7um)
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name ABI Sciex API 4000 QTrap
Ion Mode NEGATIVE
Units

Chromatography:

Chromatography ID:CH000284
Chromatography Summary:Reversed-Phase Gradient Seperation
Methods ID:RTI-RCMRC-IPA-Analysis
Methods Filename:RTI-RCMRC-IPA-Analysis
Instrument Name:Waters Acquity
Column Name:Waters Acquity BEH C18 (50 x 2.1mm,1.7um)
Column Pressure:6000-10000
Column Temperature:50 C
Flow Gradient:Time(min) Flow Rate %A %B Curve ;Initial 0.600 99.0 1.0 6;0.75 0.600 99.0 1.0 6 ;2.75 0.600 1.0 99.0 6 ;3.00 0.600 1.0 99.0 6 ;3.10 0.600 99.0 1.0 6 ;3.50 0.600 99.0 1.0 6;
Flow Rate:0.6 ml/min
Injection Temperature:8 C
Internal Standard:2-methyl-4-chlorophenyl-acetic acid (MCPA)
Solvent A:95% water/5% acetonitrile; 0.15% acetic acid
Solvent B:95% acetonitrile/5% water; 0.15% acetic acid
Analytical Time:3.5 mins
Weak Wash Solvent Name:95:5 Water:ACN with 0.15% Acetic Acid
Weak Wash Volume:600 uL
Strong Wash Solvent Name:95:5 ACN:Water with 0.15% Acetic Acid
Strong Wash Volume:600 uL
Target Sample Temperature:8 C
Sample Loop Size:10 uL
Sample Syringe Size:100 uL
Randomization Order:Yes
Chromatography Type:Reversed phase

MS:

MS ID:MS000346
Analysis ID:AN000402
Instrument Name:ABI Sciex API 4000 QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
Ion Mode:NEGATIVE
Collision Energy:-20
Collision Gas:6
Ion Source Temperature:450 C
Ion Spray Voltage:-4500 volts
Acquisition Parameters File:RTI-RCMRC-IPA-Analysis
Processing Parameters File:RTI-RCMRC-IPA-Analysis
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