Return to study ST001687 main page
MB Sample ID: SA155498
| Local Sample ID: | HFD-PBS_3.5.1 |
| Subject ID: | SU001764 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57Bl/6N |
| Age Or Age Range: | 35 weeks |
| Gender: | Male |
| Animal Animal Supplier: | KU Leuven Animal Laboratory |
| Animal Feed: | control diet (E15742-33 ssniff Spezialdiäten GmbH) or 60% high fat diet (S8655-E220 sniff Spezialdiäten GmbH) |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001764 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57Bl/6N |
| Age Or Age Range: | 35 weeks |
| Gender: | Male |
| Animal Animal Supplier: | KU Leuven Animal Laboratory |
| Animal Feed: | control diet (E15742-33 ssniff Spezialdiäten GmbH) or 60% high fat diet (S8655-E220 sniff Spezialdiäten GmbH) |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| HFD-PBS_3.5.1 | SA155498 | FL018328 | Liver | Tissue |
| HFD-PBS_3.5.1 | SA155498 | FL018328 | HFD | Diet |
| HFD-PBS_3.5.1 | SA155498 | FL018328 | No | DEN |
Collection:
| Collection ID: | CO001757 |
| Collection Summary: | At endpoint, mice were sacrificed by injecting approximately 50 µl of a 60 mg/ml Nembutal solution (Vetoquinol). Tissues were immediately excised, washed in ice-cold saline, placed into pre-labelled bags and frozen using a liquid nitrogen-cooled biosqueezer. The bags were then placed in liquid nitrogen until all collections were finished and finally stored at -80°C until further processing. Where necessary tumors were rapidly separated from normal tissue prior to freezing with tumor and normal tissue being stored separately. |
| Sample Type: | Liver |
| Collection Method: | Surgical removal, tissue snap-freezing |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR001777 |
| Treatment Summary: | 2-week old mice were injected i.p. diethylnitrosamine (DEN, 25 mg/Kg) in phosphate buffered saline (3.17 mg/ml). At 6-weeks of age, mice were randomized into two groups: control diet (CD, E15742-33 ssniff Spezialdiäten GmbH) or high fat diet (HFD, S8655-E220 sniff Spezialdiäten GmbH). |
| Treatment Compound: | dietylnitrosamine (DEN) |
| Treatment Route: | i.v. |
| Treatment Dose: | 25mg/kg |
| Treatment Doseduration: | single dose at 2-weeks age |
| Treatment Vehicle: | phosphate buffered saline (PBS) |
| Animal Endp Euthanasia: | 50 µl of 60 mg/ml Nembutal solution (Vetoquinol) |
| Animal Endp Clinical Signs: | 20% increase in abdomen diameter and hard mass detection via palpation, loss of ambulation, loss of skin elasticity, labored respiration, or weight loss over 20 % of initial body weight |
Sample Preparation:
| Sampleprep ID: | SP001770 |
| Sampleprep Summary: | Lipids were extracted from liver samples with 700 μl of sample (100 μl of plasma diluted in PBS, or 700 μl of homogenized cells) was mixed with 800 μl 1 N HCl:CH3OH 1:8 (v/v), 900 μl CHCl3, 200 μg/ml of the antioxidant 2,6-di-tert-butyl-4-methylphenol (BHT; Sigma Aldrich) and 3 μl of SPLASH® LIPIDOMIX® Mass Spec Standard (#330707, Avanti Polar Lipids). After vortexing and centrifugation, the lower organic fraction was collected and evaporated using a Savant Speedvac spd111v (Thermo Fisher Scientific) at room temperature and the remaining lipid pellet was stored at - 20°C under argon. Just before mass spectrometry analysis, lipid pellets were reconstituted in 100% ethanol. |
Combined analysis:
| Analysis ID | AN002754 | AN002755 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | Shimadzu Nexera X2 | Shimadzu Nexera X2 |
| Column | Waters Xbridge Amide (150 x 4.6mm,3.5um) | Waters Xbridge Amide (150 x 4.6mm,3.5um) |
| MS Type | ESI | ESI |
| MS instrument type | Triple quadrupole | Triple quadrupole |
| MS instrument name | ABI Sciex 6500+ QTRAP | ABI Sciex 6500+ QTRAP |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | nmol lipid/mg DNA | nmol lipid/mg DNA |
Chromatography:
| Chromatography ID: | CH002035 |
| Instrument Name: | Shimadzu Nexera X2 |
| Column Name: | Waters Xbridge Amide (150 x 4.6mm,3.5um) |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS002551 |
| Analysis ID: | AN002754 |
| Instrument Name: | ABI Sciex 6500+ QTRAP |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | - |
| Ion Mode: | POSITIVE |
| MS ID: | MS002552 |
| Analysis ID: | AN002755 |
| Instrument Name: | ABI Sciex 6500+ QTRAP |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | - |
| Ion Mode: | NEGATIVE |
