{
"METABOLOMICS WORKBENCH":{"VERSION":"1","CREATED_ON":"November 16, 2016, 2:50 pm"},

"PROJECT":{"PROJECT_TITLE":"Metabolic Dissection of Aberrant Glutamine Metabolism in Prostate Cancer","PROJECT_TYPE":"MS targeted analysis","PROJECT_SUMMARY":"In this project, we propose to define the specific role(s) of three crucial oncogenic signaling cascades (androgen receptor, Myc and AMPK) in regulating glutamine metabolism in prostate cancer using 13C-labeled glutamine with mass spectrometry-based metabolic flux analyses.","INSTITUTE":"Mayo Clinic","LABORATORY":"Mayo Clinic Metabolomics Resource Core","LAST_NAME":"Frigo","FIRST_NAME":"Daniel","ADDRESS":"Department of Biology and Biochemistry, University of Houston, Houston, Texas 77204-5001","EMAIL":"frigo@uh.edu","PHONE":"832-842-8824"},

"STUDY":{"STUDY_TITLE":"Timecourse of palmitate isotopomers in LNCaP Cells","STUDY_SUMMARY":"Timecourse of palmitate isotopomers in LNCaP Cell pellets","INSTITUTE":"Mayo Clinic","LABORATORY":"Mayo Clinic Metabolomics Resource Core","LAST_NAME":"Frigo","FIRST_NAME":"Daniel","ADDRESS":"Department of Biology and Biochemistry, University of Houston, Houston, Texas 77204-5001","EMAIL":"frigo@uh.edu","PHONE":"832-842-8824"},

"SUBJECT":{"SUBJECT_TYPE":"Human","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"DF-1",
"Sample ID":"ms5457-1",
"Factors":{"Treatment":"treated-vehicle (ethanol)","13C media incubation time":"24 hours"},
"Additional sample data":{"Amount":"3 million cells","1x PBS":"50","assay":"palmitate isotopomers"}
},
{
"Subject ID":"DF-2",
"Sample ID":"ms5457-2",
"Factors":{"Treatment":"treated-100 pM R1881","13C media incubation time":"24 hours"},
"Additional sample data":{"Amount":"3 million cells","1x PBS":"50","assay":"palmitate isotopomers"}
},
{
"Subject ID":"DF-3",
"Sample ID":"ms5457-3",
"Factors":{"Treatment":"treated-1 nM R1881","13C media incubation time":"24 hours"},
"Additional sample data":{"Amount":"3 million cells","1x PBS":"50","assay":"palmitate isotopomers"}
},
{
"Subject ID":"DF-4",
"Sample ID":"ms5457-4",
"Factors":{"Treatment":"treated-10 nM R1881","13C media incubation time":"24 hours"},
"Additional sample data":{"Amount":"3 million cells","1x PBS":"50","assay":"palmitate isotopomers"}
},
{
"Subject ID":"DF-8",
"Sample ID":"ms5457-5",
"Factors":{"Treatment":"treated-vehicle (ethanol)","13C media incubation time":"72 hours"},
"Additional sample data":{"Amount":"3 million cells","1x PBS":"50","assay":"palmitate isotopomers"}
},
{
"Subject ID":"DF-9",
"Sample ID":"ms5457-6",
"Factors":{"Treatment":"treated-100 pM R1881","13C media incubation time":"72 hours"},
"Additional sample data":{"Amount":"3 million cells","1x PBS":"50","assay":"palmitate isotopomers"}
},
{
"Subject ID":"DF-10",
"Sample ID":"ms5457-7",
"Factors":{"Treatment":"treated-1 nM R1881","13C media incubation time":"72 hours"},
"Additional sample data":{"Amount":"3 million cells","1x PBS":"50","assay":"palmitate isotopomers"}
},
{
"Subject ID":"DF-11",
"Sample ID":"ms5457-8",
"Factors":{"Treatment":"treated-10 nM R1881","13C media incubation time":"72 hours"},
"Additional sample data":{"Amount":"3 million cells","1x PBS":"50","assay":"palmitate isotopomers"}
},
{
"Subject ID":"DF-1",
"Sample ID":"ms5457-9",
"Factors":{"Treatment":"treated-vehicle (ethanol)","13C media incubation time":"24 hours"},
"Additional sample data":{"Amount":"10 mls spent media","1x PBS":"-","assay":"palmitate isotopomers"}
},
{
"Subject ID":"DF-2",
"Sample ID":"ms5457-10",
"Factors":{"Treatment":"treated-100 pM R1881","13C media incubation time":"24 hours"},
"Additional sample data":{"Amount":"10 mls spent media","1x PBS":"-","assay":"palmitate isotopomers"}
},
{
"Subject ID":"DF-3",
"Sample ID":"ms5457-11",
"Factors":{"Treatment":"treated-1 nM R1881","13C media incubation time":"24 hours"},
"Additional sample data":{"Amount":"10 mls spent media","1x PBS":"-","assay":"palmitate isotopomers"}
},
{
"Subject ID":"DF-4",
"Sample ID":"ms5457-12",
"Factors":{"Treatment":"treated-10 nM R1881","13C media incubation time":"24 hours"},
"Additional sample data":{"Amount":"10 mls spent media","1x PBS":"-","assay":"palmitate isotopomers"}
},
{
"Subject ID":"DF-8",
"Sample ID":"ms5457-13",
"Factors":{"Treatment":"treated-vehicle (ethanol)","13C media incubation time":"72 hours"},
"Additional sample data":{"Amount":"10 mls spent media","1x PBS":"-","assay":"palmitate isotopomers"}
},
{
"Subject ID":"DF-9",
"Sample ID":"ms5457-14",
"Factors":{"Treatment":"treated-100 pM R1881","13C media incubation time":"72 hours"},
"Additional sample data":{"Amount":"10 mls spent media","1x PBS":"-","assay":"palmitate isotopomers"}
},
{
"Subject ID":"DF-10",
"Sample ID":"ms5457-15",
"Factors":{"Treatment":"treated-1 nM R1881","13C media incubation time":"72 hours"},
"Additional sample data":{"Amount":"10 mls spent media","1x PBS":"-","assay":"palmitate isotopomers"}
},
{
"Subject ID":"DF-11",
"Sample ID":"ms5457-16",
"Factors":{"Treatment":"treated-10 nM R1881","13C media incubation time":"72 hours"},
"Additional sample data":{"Amount":"10 mls spent media","1x PBS":"-","assay":"palmitate isotopomers"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"LNCaP cell model"},

"TREATMENT":{"TREATMENT_SUMMARY":"LNCaP cells that were serum starved and incubated with 2mM [U-13C]glutamine and treated with dose response of a synthetic androgen (R1881) for 24 or 72 hours."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Add 50 uL 1x PBS solution to each vial containing cells. Cap, vortex, sonicate 2 x 20 sec bursts with vortexing inbetween and after Transfer entire contents of each vial into a labeled 1 dram vial. use 50 uL media for samples 9-16 use 50 uL plasma for QC Add 1.5 mL CHM solution to vial Add 20 uL 1:20 phenol red indicator Cap and vortex on multivortexer for 10 minutes Centrifuge @ 4C @ 3000 rpm for 10 minutes Aspirate and discard top phase hard to see the phase separation due to the light color (yellow). Dry bottom phase in speed vac Reconstitute samples and standards with 25 uL buffer A. (do triplicate injections)"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Agilent 1290 Infinity","COLUMN_NAME":"Waters Acquity BEH C18 (150 x 2.1mm, 1.7um)"},

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

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Agilent 6460 QQQ","INSTRUMENT_TYPE":"Triple quadrupole","MS_TYPE":"ESI","ION_MODE":"UNSPECIFIED"},

"MS_METABOLITE_DATA":{
"Units":"13C-palmitate % enrichment",

"Data":[{"Metabolite":"m0 palmitate isotopomers","ms5457-1":"88.99","ms5457-2":"87.46","ms5457-3":"87.81","ms5457-4":"83.06","ms5457-5":"83.13","ms5457-6":"77.56","ms5457-7":"78.33","ms5457-8":"76.18","ms5457-9":"99.97","ms5457-10":"99.83","ms5457-11":"99.82","ms5457-12":"99.91","ms5457-13":"99.69","ms5457-14":"99.67","ms5457-15":"99.86","ms5457-16":"99.99"},{"Metabolite":"m1 palmitate isotopomers","ms5457-1":"0.28","ms5457-2":"0.65","ms5457-3":"0.67","ms5457-4":"1.11","ms5457-5":"1.67","ms5457-6":"2.29","ms5457-7":"2.41","ms5457-8":"2.31","ms5457-9":"0.00","ms5457-10":"0.17","ms5457-11":"0.16","ms5457-12":"0.00","ms5457-13":"0.31","ms5457-14":"0.33","ms5457-15":"0.13","ms5457-16":"0.00"},{"Metabolite":"m2 palmitate isotopomers","ms5457-1":"5.91","ms5457-2":"5.95","ms5457-3":"6.14","ms5457-4":"8.03","ms5457-5":"9.81","ms5457-6":"13.05","ms5457-7":"12.92","ms5457-8":"14.00","ms5457-9":"0.03","ms5457-10":"0.00","ms5457-11":"0.00","ms5457-12":"0.08","ms5457-13":"0.00","ms5457-14":"0.00","ms5457-15":"0.00","ms5457-16":"0.00"},{"Metabolite":"m3 palmitate isotopomers","ms5457-1":"0.31","ms5457-2":"0.41","ms5457-3":"0.37","ms5457-4":"0.56","ms5457-5":"0.71","ms5457-6":"0.92","ms5457-7":"0.68","ms5457-8":"0.82","ms5457-9":"0.00","ms5457-10":"0.00","ms5457-11":"0.01","ms5457-12":"0.00","ms5457-13":"0.00","ms5457-14":"0.01","ms5457-15":"0.01","ms5457-16":"0.01"},{"Metabolite":"m4 palmitate isotopomers","ms5457-1":"2.92","ms5457-2":"3.44","ms5457-3":"3.19","ms5457-4":"4.55","ms5457-5":"3.34","ms5457-6":"4.42","ms5457-7":"4.16","ms5457-8":"4.76","ms5457-9":"0.00","ms5457-10":"0.00","ms5457-11":"0.00","ms5457-12":"0.00","ms5457-13":"0.00","ms5457-14":"0.00","ms5457-15":"0.00","ms5457-16":"0.00"},{"Metabolite":"m5 palmitate isotopomers","ms5457-1":"0.08","ms5457-2":"0.13","ms5457-3":"0.09","ms5457-4":"0.15","ms5457-5":"0.12","ms5457-6":"0.17","ms5457-7":"0.10","ms5457-8":"0.18","ms5457-9":"0.00","ms5457-10":"0.00","ms5457-11":"0.00","ms5457-12":"0.00","ms5457-13":"0.00","ms5457-14":"0.00","ms5457-15":"0.00","ms5457-16":"0.00"},{"Metabolite":"m6 palmitate isotopomers","ms5457-1":"1.06","ms5457-2":"1.36","ms5457-3":"1.18","ms5457-4":"1.78","ms5457-5":"0.90","ms5457-6":"1.21","ms5457-7":"1.05","ms5457-8":"1.33","ms5457-9":"0.00","ms5457-10":"0.00","ms5457-11":"0.00","ms5457-12":"0.00","ms5457-13":"0.00","ms5457-14":"0.00","ms5457-15":"0.00","ms5457-16":"0.00"},{"Metabolite":"m7 palmitate isotopomers","ms5457-1":"0.00","ms5457-2":"0.00","ms5457-3":"0.00","ms5457-4":"0.00","ms5457-5":"0.00","ms5457-6":"0.01","ms5457-7":"0.02","ms5457-8":"0.01","ms5457-9":"0.00","ms5457-10":"0.00","ms5457-11":"0.00","ms5457-12":"0.00","ms5457-13":"0.00","ms5457-14":"0.00","ms5457-15":"0.00","ms5457-16":"0.00"},{"Metabolite":"m8 palmitate isotopomers","ms5457-1":"0.35","ms5457-2":"0.45","ms5457-3":"0.44","ms5457-4":"0.58","ms5457-5":"0.24","ms5457-6":"0.30","ms5457-7":"0.26","ms5457-8":"0.33","ms5457-9":"0.00","ms5457-10":"0.00","ms5457-11":"0.00","ms5457-12":"0.00","ms5457-13":"0.00","ms5457-14":"0.00","ms5457-15":"0.00","ms5457-16":"0.00"},{"Metabolite":"m9 palmitate isotopomers","ms5457-1":"0.00","ms5457-2":"0.00","ms5457-3":"0.00","ms5457-4":"0.00","ms5457-5":"0.00","ms5457-6":"0.00","ms5457-7":"0.00","ms5457-8":"0.00","ms5457-9":"0.00","ms5457-10":"0.00","ms5457-11":"0.00","ms5457-12":"0.00","ms5457-13":"0.00","ms5457-14":"0.00","ms5457-15":"0.00","ms5457-16":"0.00"},{"Metabolite":"m10 palmitate isotopomers","ms5457-1":"0.11","ms5457-2":"0.15","ms5457-3":"0.13","ms5457-4":"0.17","ms5457-5":"0.07","ms5457-6":"0.08","ms5457-7":"0.07","ms5457-8":"0.09","ms5457-9":"0.00","ms5457-10":"0.00","ms5457-11":"0.00","ms5457-12":"0.00","ms5457-13":"0.00","ms5457-14":"0.00","ms5457-15":"0.00","ms5457-16":"0.00"},{"Metabolite":"m11 palmitate isotopomers","ms5457-1":"0.00","ms5457-2":"0.00","ms5457-3":"0.00","ms5457-4":"0.00","ms5457-5":"0.00","ms5457-6":"0.00","ms5457-7":"0.00","ms5457-8":"0.00","ms5457-9":"0.00","ms5457-10":"0.00","ms5457-11":"0.00","ms5457-12":"0.00","ms5457-13":"0.00","ms5457-14":"0.00","ms5457-15":"0.00","ms5457-16":"0.00"},{"Metabolite":"m12 palmitate isotopomers","ms5457-1":"0.00","ms5457-2":"0.00","ms5457-3":"0.00","ms5457-4":"0.00","ms5457-5":"0.00","ms5457-6":"0.00","ms5457-7":"0.00","ms5457-8":"0.00","ms5457-9":"0.00","ms5457-10":"0.00","ms5457-11":"0.00","ms5457-12":"0.00","ms5457-13":"0.00","ms5457-14":"0.00","ms5457-15":"0.00","ms5457-16":"0.00"},{"Metabolite":"m13 palmitate isotopomers","ms5457-1":"0.00","ms5457-2":"0.00","ms5457-3":"0.00","ms5457-4":"0.00","ms5457-5":"0.00","ms5457-6":"0.00","ms5457-7":"0.00","ms5457-8":"0.00","ms5457-9":"0.00","ms5457-10":"0.00","ms5457-11":"0.00","ms5457-12":"0.00","ms5457-13":"0.00","ms5457-14":"0.00","ms5457-15":"0.00","ms5457-16":"0.00"},{"Metabolite":"m14 palmitate isotopomers","ms5457-1":"0.00","ms5457-2":"0.00","ms5457-3":"0.00","ms5457-4":"0.00","ms5457-5":"0.00","ms5457-6":"0.00","ms5457-7":"0.00","ms5457-8":"0.00","ms5457-9":"0.00","ms5457-10":"0.00","ms5457-11":"0.00","ms5457-12":"0.00","ms5457-13":"0.00","ms5457-14":"0.00","ms5457-15":"0.00","ms5457-16":"0.00"},{"Metabolite":"m15 palmitate isotopomers","ms5457-1":"0.00","ms5457-2":"0.00","ms5457-3":"0.00","ms5457-4":"0.00","ms5457-5":"0.00","ms5457-6":"0.00","ms5457-7":"0.00","ms5457-8":"0.00","ms5457-9":"0.00","ms5457-10":"0.00","ms5457-11":"0.00","ms5457-12":"0.00","ms5457-13":"0.00","ms5457-14":"0.00","ms5457-15":"0.00","ms5457-16":"0.00"},{"Metabolite":"m16 palmitate isotopomers","ms5457-1":"0.00","ms5457-2":"0.00","ms5457-3":"0.00","ms5457-4":"0.00","ms5457-5":"0.00","ms5457-6":"0.00","ms5457-7":"0.00","ms5457-8":"0.00","ms5457-9":"0.00","ms5457-10":"0.00","ms5457-11":"0.00","ms5457-12":"0.00","ms5457-13":"0.00","ms5457-14":"0.00","ms5457-15":"0.00","ms5457-16":"0.00"}],

"Metabolites":[{"metabolite_name":"m0 palmitate isotopomers"},{"metabolite_name":"m1 palmitate isotopomers"},{"metabolite_name":"m2 palmitate isotopomers"},{"metabolite_name":"m3 palmitate isotopomers"},{"metabolite_name":"m4 palmitate isotopomers"},{"metabolite_name":"m5 palmitate isotopomers"},{"metabolite_name":"m6 palmitate isotopomers"},{"metabolite_name":"m7 palmitate isotopomers"},{"metabolite_name":"m8 palmitate isotopomers"},{"metabolite_name":"m9 palmitate isotopomers"},{"metabolite_name":"m10 palmitate isotopomers"},{"metabolite_name":"m11 palmitate isotopomers"},{"metabolite_name":"m12 palmitate isotopomers"},{"metabolite_name":"m13 palmitate isotopomers"},{"metabolite_name":"m14 palmitate isotopomers"},{"metabolite_name":"m15 palmitate isotopomers"},{"metabolite_name":"m16 palmitate isotopomers"}]
}

}