Return to study ST000095 main page
MB Sample ID: SA005397
| Local Sample ID: | T26-6 |
| 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 |
Select appropriate tab below to view additional metadata details:
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 ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| T26-6 | SA005397 | FL001362 | DDT | Treatment |
Collection:
| Collection ID: | CO000097 |
| Collection Summary: | - |
| 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: | 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 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 |
