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MB Sample ID: SA176930
| Local Sample ID: | OB_12_A(raw) |
| Subject ID: | SU001989 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | wildtype |
| Age Or Age Range: | 4 weeks |
| Gender: | Male |
| Animal Animal Supplier: | Jackson Laboratory |
| Animal Housing: | UC Denver |
| Animal Feed: | High fat diet (HFD), 60 kcal% fat from Research diets Inc., # D12492 |
| Animal Water: | ad libitum |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001989 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | wildtype |
| Age Or Age Range: | 4 weeks |
| Gender: | Male |
| Animal Animal Supplier: | Jackson Laboratory |
| Animal Housing: | UC Denver |
| Animal Feed: | High fat diet (HFD), 60 kcal% fat from Research diets Inc., # D12492 |
| Animal Water: | ad libitum |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| OB_12_A(raw) | SA176930 | FL021938 | High Fat | Factor |
Collection:
| Collection ID: | CO001982 |
| Collection Summary: | Blood was collected and plasma/serum flash frozen in liquid nitrogen and stored in -80 degrees Celsius for biochemical/metabolic assessments. |
| Sample Type: | Blood (plasma) |
Treatment:
| Treatment ID: | TR002001 |
| Treatment Summary: | Sixteen-week-old diet-induced obese (DIO) C57BL/6J male mice [fed-high fat diet (HFD), 60 kcal% fat from Research diets Inc., # D12492) from 4 weeks of age] were purchased from Jackson labs (Bar Harbor, ME). Once received at the animal house facility in UC Denver, the mice were randomized into two groups (n=4/ group): a) Control-DIO group maintained on the 60 kcal% HFD for additional 40 days, and b) BMJ+DIO group maintained on 60 kcal% HFD fat for additional 40 days plus orally gavage with lyophilized bitter melon juice extract [(5mg per mouse; total volume of 100 μl, 200mg/kg body weight/ day), reconstituted in sterile deionized water immediately before use)] throughout the study. |
| Treatment: | Intervention |
| Treatment Compound: | Bitter melon juice |
| Treatment Route: | Oral gavage |
| Treatment Dose: | 200mg/kg body weight/ day |
| Animal Fasting: | On day 40 of study initiation, mice in control and treament groups were subjected to fasting for six hours prior to sacrifice. |
| Animal Endp Euthanasia: | On day 40, mice were sacrificed after CO2 asphyxiation and exsanguination. |
Sample Preparation:
| Sampleprep ID: | SP001995 |
| Sampleprep Summary: | Frozen plasma was stored at -80°C until sample preparation for untargeted liquid-chromatography mass spectrometry (LC-MS)-based metabolomics. Briefly, 100µL of sample underwent protein precipitation using methanol, followed by liquid-liquid extraction using methyl-tert butyl ether as previously described[1, 2] to obtain an aqueous fraction and a lipid fraction. The lipid fraction was dried down and resuspended in 100% methanol for LC-MS analysis. |
| Processing Storage Conditions: | Described in summary |
Combined analysis:
| Analysis ID | AN003108 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Agilent 1290 |
| Column | Agilent Zorbax RRHD SB-C18 (100 x 2.1mm,1.8um) |
| MS Type | ESI |
| MS instrument type | TOF |
| MS instrument name | Agilent 6210 TOF |
| Ion Mode | POSITIVE |
| Units | Abundance (mass spectral counts) |
Chromatography:
| Chromatography ID: | CH002295 |
| Chromatography Summary: | The samples from the lipid fraction were analyzed an Agilent 1290 series pump with an Agilent Zorbax Rapid Resolution HD (RRHD) SB-C18, 1.8 micron (2.1 x 100 mm) analytical column and an Agilent Zorbax SB-C18, 1.8 micron (2.1 x 5 mm) guard column. The autosampler tray temperature was set at 4°C, column temperature was set at 60°C, and the sample injection volume was 4µL. The flow rate was 0.7 mL/min with the following mobile phases: mobile phase A was water with 0.1% formic acid, and mobile phase B was 60:36:4 isopropyl alcohol:acetonitrile:water with 0.1% formic acid. Gradient elution was as follows: 0-1 minute 30-70% B, 1-7.92 minutes 70-100% B, 7.92-10.4 minutes 100% B, 10.4-10.5 minutes 100-30% B, followed by column re-equilibration with 30% B from 10.5-15.1 minutes. |
| Instrument Name: | Agilent 1290 |
| Column Name: | Agilent Zorbax RRHD SB-C18 (100 x 2.1mm,1.8um) |
| Column Temperature: | 60 |
| Flow Gradient: | 0-1 minute 30-70% B, 1-7.92 minutes 70-100% B, 7.92-10.4 minutes 100% B, 10.4-10.5 minutes 100-30% B, followed by column re-equilibration with 30% B from 10.5-15.1 minutes. |
| Flow Rate: | 0.7 mL/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 60% isopropanol/36% acetonitrile/4% water; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002890 |
| Analysis ID: | AN003108 |
| Instrument Name: | Agilent 6210 TOF |
| Instrument Type: | TOF |
| MS Type: | ESI |
| MS Comments: | The mass spectrometry conditions were as follows: Agilent 6210 Time-of-Flight mass spectrometer (TOF-MS) in positive ionization mode with dual electrospray (ESI) source, mass range 60-1600m/z, scan rate 2.03, gas temperature 300°C, gas flow 12.0L/min, nebulizer 30psi, skimmer 60V, capillary voltage 4000V, fragmentor 120V, reference masses 121.050873 and 922.009798 (Agilent reference mix). Proprietary analytical software for integration and peak picking |
| Ion Mode: | POSITIVE |
