Return to study ST001795 main page

MB Sample ID: SA167155

Local Sample ID:CFD_6
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 IDMB Sample IDFactor Level IDLevel ValueFactor Name
CFD_6SA167155FL019896CFDTreatment

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
  logo