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MB Sample ID: SA005403

Local Sample ID:T18-3
Subject ID:SU000114
Subject Type:Animal
Subject Species:Mus musculus
Taxonomy ID:10090
Age Or Age Range:9 months
Gender:Female
Animal Housing:Singly housed
Animal Feed:High fat diet (HFD, 4.73 kcal/g, 20% protein, 35% carbohydrate, and 45% fat per kcal; D12451, Research Diets, New Brunswick, NJ)
Species Group:Mammal

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Subject:

Subject ID:SU000114
Subject Type:Animal
Subject Species:Mus musculus
Taxonomy ID:10090
Age Or Age Range:9 months
Gender:Female
Animal Housing:Singly housed
Animal Feed:High fat diet (HFD, 4.73 kcal/g, 20% protein, 35% carbohydrate, and 45% fat per kcal; D12451, Research Diets, New Brunswick, NJ)
Species Group:Mammal

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
T18-3SA005403FL001362DDTTreatment

Collection:

Collection ID:CO000097
Collection Protocol Filename:Newman_Lab_Bile_Acids_Extraction_&_Analysis_Protocol_La_Merrill_WCMC_P&F.pdf
Sample Type:Liver tissue
Collection Method:Dissection
Storage Conditions:- 80°C
Storage Vials:Eppendorf tubes

Treatment:

Treatment ID:TR000115
Treatment Summary:1.7 mg DDT/kg body weight was administered to dams daily on gestational day 11 through postnatal day 5 and litters were culled to 6 mice at postnatal day 6. Mice were metabolically screened from 2-6 months when they consumed a high fat diet. Mice were sacrificed at 9 months of age.
Treatment Protocol Filename:Newman_Lab_Bile_Acid_Protocols_La_Merrill_WCMC_P&F.pdf
Treatment:Intervention
Treatment Compound:p,p`-DDT, o,p`-DDT
Treatment Route:Oral
Treatment Dose:77.2% p,p`-DDT (98.5% purity neat, AccuStandard, New Haven, CT) and 22.8% o,p`-DDT (100% purity neat, AccuStandard) were dissolved at a final concentration of 0.17 g DDT mixture/L organic olive oil
Treatment Doseduration:From 11.5 days post coitus to postnatal day 5
Treatment Vehicle:Olive oil
Animal Endp Euthanasia:9 months
Animal Endp Tissue Coll List:Liver
Animal Endp Tissue Proc Method:Dissected, placed on dry ice

Sample Preparation:

Sampleprep ID:SP000110
Sampleprep Summary:A 15 mg liver sample (n=1 female/litter in 7 VEH and 8 DDT litters) was pulverized on dry ice, enriched with deuterated bile acid surrogates, butylated hydroxytoluene and ethylinediaminetetraacetic acid, and extracted twice with 500 uL methanol. The combined extract was dried, reconstituted in 100uL 50:50 methanol:acetonitrile with internal standards 1-phenyl-3-hexanoic acid urea and 1-cyclohexyl-3-dodecanoic acid urea (Sigma-Aldrich, St. Louis, MO) and filtered at 0.1 µm.
Sampleprep Protocol Filename:Newman_Lab_Bile_Acid_Protocols_La_Merrill_WCMC_P&F.pdf
Processing Method:Pulverization on dry ice
Extraction Method:Methanol
Extract Concentration Dilution:Dried with Rotoevaporator
Extract Storage:Placed in 10 °C autosampler and immediately analyzed
Sample Spiking:Spiked with deuterated bile acid surrogates.
Organ:Liver

Combined analysis:

Analysis ID AN000151
Analysis type MS
Chromatography type Reversed phase
Chromatography system Waters Acquity UPLC
Column Waters Acquity BEH C18 (100 x 2mm, 1.7um)
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name ABI Sciex API 6500 QTrap
Ion Mode NEGATIVE
Units % abundance

Chromatography:

Chromatography ID:CH000107
Chromatography Summary:Targeted UPLC-MS/MS method
Methods Filename:Newman_Lab_Bile_Acids_Extraction_&_Analysis_Protocol_La_Merrill_WCMC_P&F.pdf
Instrument Name:Waters Acquity UPLC
Column Name:Waters Acquity BEH C18 (100 x 2mm, 1.7um)
Column Temperature:60 C
Flow Gradient:Yes
Flow Rate:0.4mL/min
Internal Standard:See protocol/methods file
Sample Injection:5 L
Solvent A:0.1% formic acid
Solvent B:0.1% formic acid in acetonitrile
Analytical Time:16 min
Weak Wash Solvent Name:20% methanol, 10% isopropanol
Weak Wash Volume:600 L
Strong Wash Solvent Name:50:50 Acetonitrile:Methanol
Strong Wash Volume:600 L
Sample Loop Size:17 L
Chromatography Type:Reversed phase

MS:

MS ID:MS000127
Analysis ID:AN000151
Instrument Name:ABI Sciex API 6500 QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
Ion Mode:NEGATIVE
Ion Source Temperature:See protocol/methods file
Ion Spray Voltage:See protocol/methods file
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