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MB Sample ID: SA167188
Local Sample ID: | HFD_2 |
Subject ID: | SU001873 |
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: | SU001873 |
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 |
---|---|---|---|---|
HFD_2 | SA167188 | FL019901 | HFD | Treatment |
Collection:
Collection ID: | CO001866 |
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: | TR001886 |
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 CFD diet (one group) and HFD diet (three groups). After 15 weeks two of the HFD groups were switched on HFD diet supplemented with Celecoxib and Celecoxib prodrug while other two groups continued unchanged CFD and HFD diets. Analysis at 22-34 weeks. |
Sample Preparation:
Sampleprep ID: | SP001879 |
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 | AN002917 | AN002918 |
---|---|---|
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: | CH002160 |
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: | MS002709 |
Analysis ID: | AN002917 |
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: | MS002710 |
Analysis ID: | AN002918 |
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 |