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MB Sample ID: SA167151
Local Sample ID: | CFD_4 |
Subject ID: | SU001872 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | C57BL6/J |
Age Or Age Range: | 6-7 weeks |
Gender: | Male |
Animal Housing: | temperature-controlled room under specific-pathogen free (SPF) or standard animal housing conditions with free access to food and water |
Animal Feed: | semi-purified normal chow diet (control fat diet (CFD), 7% w/w fat and 16% total energy from fat; AIN93G, Specialty Feeds Pty Ltd, Australia) or high fat diet (HFD, 36% w/w fat and 59% total energy from fat; SF03-002, Specialty Feeds Pty Ltd, Australia) |
Animal Water: | ad libitum |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001872 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | C57BL6/J |
Age Or Age Range: | 6-7 weeks |
Gender: | Male |
Animal Housing: | temperature-controlled room under specific-pathogen free (SPF) or standard animal housing conditions with free access to food and water |
Animal Feed: | semi-purified normal chow diet (control fat diet (CFD), 7% w/w fat and 16% total energy from fat; AIN93G, Specialty Feeds Pty Ltd, Australia) or high fat diet (HFD, 36% w/w fat and 59% total energy from fat; SF03-002, Specialty Feeds Pty Ltd, Australia) |
Animal Water: | ad libitum |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
CFD_4 | SA167151 | FL019896 | CFD | Treatment |
Collection:
Collection ID: | CO001865 |
Collection Summary: | The efferent mesenteric lymphatic duct was cannulated in isoflurane anaesthetised non-fasted mice or rats and lymph fluid was collected for up to 4 h. For the lymphatic drug transport study, lymph was collected in mice that were fasted for 3-4 h. The mesenteric lymph duct cannulation was performed as described previously in rats4 and mice5 and mesenteric lymph fluid was collected continuously for up to 6 h. |
Sample Type: | Mesenteric lymph |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR001885 |
Treatment Summary: | Celecoxib was incorporated into HFD feed at a dose equivalent to ~29 mg/kg/day (based on average food intake). Mice were fed for 15 weeks before the analysis. |
Sample Preparation:
Sampleprep ID: | SP001878 |
Sampleprep Summary: | For lipidomics analysis, lipid was extracted from mesenteric lymph by chloroform:methanol (1:3). Samples were vortexed for 1 h at 4°C and then centrifuged at 16,000 g for 10 min. Supernatant was carefully transferred to another tube and stored at -80°C until analysis. Before LC-MS analysis, the extract was dried with nitrogen and reconstituted in 20 µl water and 180 µl butanol-methanol (1:1 v/v). The reconstituted extract was vortexed (Vortex mixer, Ratek) for 200 sec with 20 cycles of 5 sec spin and 20 sec vortex. The extract was sonicated in a water bath for 1 hour which was maintained at <20°C by sonicating the samples on ice. The samples were then centrifuged for 10 min at 16,000 g and the supernatant was transferred to LC-MS vials and stored at 4°C prior to analysis. |
Processing Storage Conditions: | 4℃ |
Extract Storage: | -80℃ |
Combined analysis:
Analysis ID | AN002915 | AN002916 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | Reversed phase | Reversed phase |
Chromatography system | Thermo Dionex Ultimate 3000 RS | Thermo Dionex Ultimate 3000 RS |
Column | Sigma Aldrich,Supleco,C8 (100 x 2.1mm,2.7um) | Sigma Aldrich,Supleco,C8 (100 x 2.1mm,2.7um) |
MS Type | ESI | ESI |
MS instrument type | Orbitrap | Orbitrap |
MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
Ion Mode | POSITIVE | NEGATIVE |
Units | height | height |
Chromatography:
Chromatography ID: | CH002159 |
Chromatography Summary: | Lipidomics analysis was performed using reversed phase liquid chromatography and high-resolution mass spectrometry. Samples (10 µl) were injected onto a Dionex Ultimate 3000 UHPLC system (Thermo Scientific, Australia) fitted with an analytical C8 column (100 x 2.1 mm; 2.7 µm, Sigma Aldrich, Australia). Chromatography was performed using solvent A (2 mM formic acid, 8 mM ammonium formate, 40% v/v isopropanol) and solvent B (2 mM formic acid, 8 mM ammonium formate, 98% v/v isopropanol) as mobile phases with a 30 min gradient starting at 0% B and increasing to 35% B from 0 to 8 min, then to 50% B from 8-16 min, then to 80% B from 16-19 min, then finally to 100% B by 23 min. 100% B was maintained for a further 3 min before equilibrating to 0% by 28 min and washing for a further 2 min. |
Instrument Name: | Thermo Dionex Ultimate 3000 RS |
Column Name: | Sigma Aldrich,Supleco,C8 (100 x 2.1mm,2.7um) |
Column Temperature: | 40C |
Flow Gradient: | 0 min - 0%B, 8 min - 35%B, 16 min - 50%B, 19 min - 80%B, 23 min 100%B, 26 min - 100%B, 28 min - 0%B |
Flow Rate: | 0.2 ml/min |
Injection Temperature: | 4 |
Solvent A: | 40% isopropanol/60% water; 2 mM formic acid; 8 mM ammonium formate, |
Solvent B: | 98% isopropanol/2% water; 2 mM formic acid; 8 mM ammonium formate |
Analytical Time: | 30 min |
Oven Temperature: | 40C |
Sample Loop Size: | 25 uL |
Sample Syringe Size: | 25 uL |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS002707 |
Analysis ID: | AN002915 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Pos and neg data were acquired during the same chromatographic run using polarity switching. Full scan acquisition mode, resolution 140k, AGC target 3e6, max IT 200 ms, mass range 140-2000 m/z, number of scans 1 |
Ion Mode: | POSITIVE |
Mass Accuracy: | 3 ppm |
Automatic Gain Control: | 3e6 |
Desolvation Gas Flow: | 34 |
Interface Voltage: | 3.5 kV |
MS ID: | MS002708 |
Analysis ID: | AN002916 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Pos and neg data were acquired during the same chromatographic run using polarity switching. Full scan acquisition mode, resolution 140k, AGC target 3e6, max IT 200 ms, mass range 140-2000 m/z, number of scans 1 |
Ion Mode: | NEGATIVE |
Mass Accuracy: | 3 ppm |
Automatic Gain Control: | 3e6 |
Desolvation Gas Flow: | 34 |
Interface Voltage: | 3.5 kV |