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MB Sample ID: SA079575
| Local Sample ID: | 20 |
| 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 |
|---|---|---|---|---|
| 20 | SA079575 | FL012199 | HFD | Diet |
| 20 | SA079575 | 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 QTOF |
| 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 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Binary Data Format: .cdf |
| Ion Mode: | POSITIVE |
