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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

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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 IDMB Sample IDFactor Level IDLevel ValueFactor Name
HFD_2SA167188FL019901HFDTreatment

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
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