{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001911","ANALYSIS_ID":"AN003108","VERSION":"1","CREATED_ON":"August 20, 2021, 10:13 am"},

"PROJECT":{"PROJECT_TITLE":"Metabolome changes induced by bitter melon (Momordica charantia)- intake in a high-fat diet induced obesity model","PROJECT_TYPE":"LCMS quanitative analysis","PROJECT_SUMMARY":"Quanitative analysis of metabolomics on samples from murine samples (KEGG Pathway MAPS, LIPIDMAPS, Human Metabolome Database)","INSTITUTE":"University of Colorado Anschutz Medical Campus","DEPARTMENT":"Pharmaceutical Sciences","LABORATORY":"Dr. Rajesh Agarwal's Laboratory and","LAST_NAME":"Agarwal","FIRST_NAME":"Rajesh","ADDRESS":"12850 E. Montview Blvd, C238, Room V20-2118, Aurora, CO 80045, USA","EMAIL":"Rajesh.Agarwal@cuanschutz.edu","PHONE":"303-724-4055","FUNDING_SOURCE":"This work was supported by the National Institutes of Health/ National Cancer Institute grant R01CA195708, the National Institutes of Health Office of Dietary Supplements supplemental funds R01CA195708-02S1, and the pilot funding from the office of the Associate Dean for Research and Graduate Education (ADR), School of Pharmacy, UC AMC (to RA), and the National Cancer Institute diversity supplement grant R01CA195708-04S1 (to DR and RA).","CONTRIBUTORS":"Dominique Reed, Dileep Kumar, Sushil Kumar, Komal Raina, Reenu Punia, Charmion Cruickshank-Quinn, Boris Tabakoff, Nichole Reisdorph, Michael Wempe, Chapla Agarwal"},

"STUDY":{"STUDY_TITLE":"A high-fat diet induced obesity murine model treated with bitter melon (Momordica charantia)","STUDY_TYPE":"A 40-day Intervention experiment","STUDY_SUMMARY":"Metabolic effects of lyophilized bitter melon juice (BMJ) extract (oral gavage 200mg/kg/body weight-daily for 40 days) intake were evaluated in diet-induced obese C57BL/6J male mice [fed-high fat diet (HFD), 60 kcal% fat]. Changes in metabolite abundance levels in lipid-phase plasma [liquid chromatography mass spectrometry (LC-MS)-based metabolomics] after BMJ intervention were assessed.","INSTITUTE":"University of Colorado Anschutz Medical Campus","DEPARTMENT":"Pharmaceutical Sciences","LABORATORY":"Dr. Rajesh Agarwal's Laboratory","LAST_NAME":"Agarwal","FIRST_NAME":"Rajesh","ADDRESS":"12850 E. Montview Blvd, C238, Room V20-2118, Aurora, CO 80045, USA","EMAIL":"Rajesh.Agarwal@cuanschutz.edu","PHONE":"303-724-4055","NUM_GROUPS":"2","TOTAL_SUBJECTS":"8","NUM_MALES":"8"},

"SUBJECT":{"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"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"OB_07(raw)",
"Factors":{"Factor":"High Fat"},
"Additional sample data":{"RAW_FILE_NAME":"OB_07"}
},
{
"Subject ID":"-",
"Sample ID":"OB_11(raw)",
"Factors":{"Factor":"High Fat"},
"Additional sample data":{"RAW_FILE_NAME":"OB_11"}
},
{
"Subject ID":"-",
"Sample ID":"OB_12_A(raw)",
"Factors":{"Factor":"High Fat"},
"Additional sample data":{"RAW_FILE_NAME":"OB_12_A"}
},
{
"Subject ID":"-",
"Sample ID":"OB+BMJ_03(raw)",
"Factors":{"Factor":"Treated"},
"Additional sample data":{"RAW_FILE_NAME":"OB+BMJ_03"}
},
{
"Subject ID":"-",
"Sample ID":"OB+BMJ_04(raw)",
"Factors":{"Factor":"Treated"},
"Additional sample data":{"RAW_FILE_NAME":"OB+BMJ_04"}
},
{
"Subject ID":"-",
"Sample ID":"OB+BMJ_05(raw)",
"Factors":{"Factor":"Treated"},
"Additional sample data":{"RAW_FILE_NAME":"OB+BMJ_05"}
}
],
"COLLECTION":{"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_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."},

"SAMPLEPREP":{"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"},

"CHROMATOGRAPHY":{"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.","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Agilent 1290","COLUMN_NAME":"Agilent Zorbax RRHD SB-C18 (100 x 2.1mm, 1.8um)"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Agilent 6210 TOF","INSTRUMENT_TYPE":"TOF","MS_TYPE":"ESI","ION_MODE":"POSITIVE","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","MS_RESULTS_FILE":"ST001911_AN003108_Results.txt UNITS:Abundance (mass spectral counts) Has m/z:Neutral masses Has RT:Yes RT units:Minutes"}

}