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MB Sample ID: SA155497
Local Sample ID: | HFD-PBS_3.4.3 |
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) |
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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_3.4.3 | SA155497 | FL018328 | Liver | Tissue |
HFD-PBS_3.4.3 | SA155497 | FL018328 | HFD | Diet |
HFD-PBS_3.4.3 | SA155497 | 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 |