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MB Sample ID: SA079580
Local Sample ID: | 7 |
Subject ID: | SU001210 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | C57BL6 |
Age Or Age Range: | 2-3 months |
Weight Or Weight Range: | 28-46 |
Gender: | Female |
Animal Animal Supplier: | Jackson Laboratory |
Animal Housing: | Single House |
Animal Light Cycle: | 12:12 |
Animal Feed: | CHOW diet (Envigo Teklad 6% fat 7002, Madison, WI) and Western (HFD; high-fat, high-sugar; 21% milk fat and 34% sucrose (Envigo TD.08811) |
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Subject:
Subject ID: | SU001210 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | C57BL6 |
Age Or Age Range: | 2-3 months |
Weight Or Weight Range: | 28-46 |
Gender: | Female |
Animal Animal Supplier: | Jackson Laboratory |
Animal Housing: | Single House |
Animal Light Cycle: | 12:12 |
Animal Feed: | CHOW diet (Envigo Teklad 6% fat 7002, Madison, WI) and Western (HFD; high-fat, high-sugar; 21% milk fat and 34% sucrose (Envigo TD.08811) |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
7 | SA079580 | FL012199 | HFD | Diet |
7 | SA079580 | FL012199 | 5week | Time |
Collection:
Collection ID: | CO001204 |
Collection Summary: | Approximately 20 mg of muscle tissue was homogenized in a glass homogenizer with 1.5 ml of 2:1 chloroform:methanol and then brought to 4 ml using the same ratio. The mixture was poured through a 2V grade qualitative 12.5 cm Whatman filter into a clean 10 ml glass tube. The volume in the tube was again brought up to 4 ml with the same 2:1 solution as above. One ml of water was added to the tube, vortexed for 20 seconds, and then centrifuged for 10 minutes at 2500 rpm. The top non-lipid portion was removed and the lower lipid-containing layer was dried under nitrogen. |
Sample Type: | Muscle |
Collection Method: | excision |
Collection Location: | femoral muscle |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR001225 |
Treatment Summary: | Male C57BL/6 mice, 3 months of age, (Jackson Laboratory, Bar Harbor, Maine) were allowed to acclimate for one week before experiment start. Mice were individually housed under controlled conditions (12:12 light-dark cycle, 50–60% humidity, and 25° C) and had ad libitum access to standard CHOW diet (Envigo Teklad 6% fat 7002, Madison, WI). Lipids in the CHOW diet consisted of an assortment of fatty acids where linoleic > oleic > palmitic > linolenic > stearic. Following a baseline glucose tolerance test (GTT), mice were grouped according to mean GTT and body mass into a standard 5 week CHOW (n = 10) or Western (HFD; high-fat, high-sugar; 21% milk fat and 34% sucrose (Envigo TD.08811); 5 (n = 5) and 13 week (n = 6)) diet group. The saturated fatty acids in HFD ranged from 4:0 to 18:0, however, palmitate (16:0) followed by steric (18:0) and myristic (14;0) where highest in quantity. |
Treatment: | Diet |
Treatment Compound: | Envigo TD.08811 |
Treatment Route: | oral |
Sample Preparation:
Sampleprep ID: | SP001218 |
Sampleprep Summary: | Approximately 20 mg of muscle tissue was homogenized in a glass homogenizer with 1.5 ml of 2:1 chloroform:methanol and then brought to 4 ml using the same ratio. The mixture was poured through a 2V grade qualitative 12.5 cm Whatman filter into a clean 10 ml glass tube. The volume in the tube was again brought up to 4 ml with the same 2:1 solution as above. One ml of water was added to the tube, vortexed for 20 seconds, and then centrifuged for 10 minutes at 2500 rpm. The top non-lipid portion was removed and the lower lipid-containing layer was dried under nitrogen.Lipid extracts were suspended in 100 uL of 2:1 Chloroform:Methanol. Injections were normalized such that equal amounts of lipid were analyzed for each sample, regardless of total lipid content of diet. 3 μL of extract was injected twice (n=2 replicates) onto a Waters Acquity UPLC system in discrete, randomized blocks. Next samples were separated using a Waters Acquity UPLC CSH Phenyl Hexyl column (1.7 µM, 1.0 x 100 mm), using a gradient from solvent A (water, 0.1% formic acid) to solvent B (Acetonitrile, 0.1% formic acid). Injections were made in 100% A, held at 100% A for 1 min, ramped to 98% B over 12 minutes, held at 98% B for 3 minutes, and then returned to starting conditions over 0.05 minutes and allowed to re-equilibrate for 3.95 minutes, with a 200 µL/min constant flow rate. The column and samples were held at 65 °C and 6 °C, respectively. The column eluent was infused into a Waters Xevo G2 TOF-MS with an electrospray source in positive mode, scanning 50-2000 m/z at 0.2 seconds per scan, alternating between MS (6 V collision energy) and MSE mode (15-30 V ramp). Calibration was performed using sodium iodide with 1 ppm mass accuracy. The capillary voltage was held at 2200 V, source temp at 150 °C, and nitrogen desolvation temp at 350 °C with a flow rate of 800 L/hr. |
Processing Storage Conditions: | -80℃ |
Extract Storage: | On ice |
Combined analysis:
Analysis ID | AN001890 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Waters Acquity UPLC |
Column | Acquity CSH PhenylHexyl |
MS Type | ESI |
MS instrument type | QTOF |
MS instrument name | Waters Synapt G2 XS |
Ion Mode | POSITIVE |
Units | Relative Abundance |
Chromatography:
Chromatography ID: | CH001367 |
Instrument Name: | Waters Acquity UPLC |
Column Name: | Acquity CSH PhenylHexyl |
Flow Gradient: | water + 0.1% Formic + 2 mM AmOH / Acetonitrile |
Flow Rate: | 200 uL/min |
Solvent A: | 100% water; 0.1% formic acid; 2 mM ammonium hydroxide |
Solvent B: | 100% acetonitrile |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS001746 |
Analysis ID: | AN001890 |
Instrument Name: | Waters Synapt G2 XS |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | Binary Data Format: .cdf |
Ion Mode: | POSITIVE |