{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001167","ANALYSIS_ID":"AN001930","VERSION":"1","CREATED_ON":"April 9, 2019, 6:10 pm"},

"PROJECT":{"PROJECT_TITLE":"Comprehensive Profiling by Non-targeted Stable Isotope Tracing Capillary Electrophoresis-Mass Spectrometry","PROJECT_SUMMARY":"Prostate cell RWPE1 and prostate cancer cell VCaP were labeled with 13C6-Glucose for 6 time points 0h, 0.25h, 4h, 12h, 24h and 6 days. Then CE-MS based metabolomics analysis were peformed.","INSTITUTE":"Dalian Institute Of Chemical Physics, Chinese Academy Of Sciences","DEPARTMENT":"Group, 1808","LAST_NAME":"Wang","FIRST_NAME":"Zhichao","ADDRESS":"457, Zhongshan Road, Dalian, Shenyang","EMAIL":"wangzc05@dicp.ac.cn","PHONE":"+86-15998625250","PROJECT_COMMENTS":"A Novel Tool Complementing Metabolomics Analyses of Polar Metabolites"},

"STUDY":{"STUDY_TITLE":"Comprehensive Profiling by Non-targeted Stable Isotope Tracing Capillary Electrophoresis-Mass Spectrometry","STUDY_SUMMARY":"We developed a stable-isotope tracing capillary electrophoresis (CE)-MS metabolomics approach to cover polar metabolites as well as isotopologues in a non-targeted way. An in-house developed software enables high throughput processing of complex multi-dimensional data. The practicability is demonstrated analysing 13C-U-glucose exposed prostate cancer and non-cancer cells.","INSTITUTE":"Dalian Institute Of Chemical Physics, Chinese Academy Of Sciences","LAST_NAME":"Wang","FIRST_NAME":"Zhichao","ADDRESS":"457, Zhongshan Road","EMAIL":"wangzc05@dicp.ac.cn","PHONE":"+86-15998625250","STUDY_COMMENTS":"Comprehensive Profiling by Non-targeted Stable Isotope Tracing Capillary Electrophoresis-Mass Spectrometry - A Novel Tool Complementing Metabolomics Analyses of Polar Metabolites"},

"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"12RWPE1_0-1",
"Factors":{"Cell lines":"RWPE1","Time points":"0h"}
},
{
"Subject ID":"-",
"Sample ID":"12RWPE1_0-2",
"Factors":{"Cell lines":"RWPE1","Time points":"0h"}
},
{
"Subject ID":"-",
"Sample ID":"12RWPE1_0-3",
"Factors":{"Cell lines":"RWPE1","Time points":"0h"}
},
{
"Subject ID":"-",
"Sample ID":"12RWPE1_0-4",
"Factors":{"Cell lines":"RWPE1","Time points":"0h"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_0.25-2",
"Factors":{"Cell lines":"RWPE1","Time points":"0.25h"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_0.25-3",
"Factors":{"Cell lines":"RWPE1","Time points":"0.25h"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_0.25-4",
"Factors":{"Cell lines":"RWPE1","Time points":"0.25h"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_4-2",
"Factors":{"Cell lines":"RWPE1","Time points":"4h"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_4-3",
"Factors":{"Cell lines":"RWPE1","Time points":"4h"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_4-4",
"Factors":{"Cell lines":"RWPE1","Time points":"4h"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_12-1",
"Factors":{"Cell lines":"RWPE1","Time points":"12h"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_12-2",
"Factors":{"Cell lines":"RWPE1","Time points":"12h"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_12-3",
"Factors":{"Cell lines":"RWPE1","Time points":"12h"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_12-4",
"Factors":{"Cell lines":"RWPE1","Time points":"12h"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_24-1",
"Factors":{"Cell lines":"RWPE1","Time points":"24h"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_24-2",
"Factors":{"Cell lines":"RWPE1","Time points":"24h"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_24-3",
"Factors":{"Cell lines":"RWPE1","Time points":"24h"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_24-4",
"Factors":{"Cell lines":"RWPE1","Time points":"24h"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_P3-1",
"Factors":{"Cell lines":"RWPE1","Time points":"6days"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_P3-2",
"Factors":{"Cell lines":"RWPE1","Time points":"6days"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_P3-3",
"Factors":{"Cell lines":"RWPE1","Time points":"6days"}
},
{
"Subject ID":"-",
"Sample ID":"13RWPE1_P3-4",
"Factors":{"Cell lines":"RWPE1","Time points":"6days"}
},
{
"Subject ID":"-",
"Sample ID":"12VCaP_0-1",
"Factors":{"Cell lines":"VCaP","Time points":"0h"}
},
{
"Subject ID":"-",
"Sample ID":"12VCaP_0-2",
"Factors":{"Cell lines":"VCaP","Time points":"0h"}
},
{
"Subject ID":"-",
"Sample ID":"12VCaP_0-3",
"Factors":{"Cell lines":"VCaP","Time points":"0h"}
},
{
"Subject ID":"-",
"Sample ID":"12VCaP_0-4",
"Factors":{"Cell lines":"VCaP","Time points":"0h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_0.25-1",
"Factors":{"Cell lines":"VCaP","Time points":"0.25h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_0.25-2",
"Factors":{"Cell lines":"VCaP","Time points":"0.25h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_0.25-3",
"Factors":{"Cell lines":"VCaP","Time points":"0.25h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_0.25-4",
"Factors":{"Cell lines":"VCaP","Time points":"0.25h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_4-1",
"Factors":{"Cell lines":"VCaP","Time points":"4h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_4-2",
"Factors":{"Cell lines":"VCaP","Time points":"4h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_4-3",
"Factors":{"Cell lines":"VCaP","Time points":"4h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_4-4",
"Factors":{"Cell lines":"VCaP","Time points":"4h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_12-1",
"Factors":{"Cell lines":"VCaP","Time points":"12h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_12-2",
"Factors":{"Cell lines":"VCaP","Time points":"12h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_12-3",
"Factors":{"Cell lines":"VCaP","Time points":"12h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_12-4",
"Factors":{"Cell lines":"VCaP","Time points":"12h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_24-1",
"Factors":{"Cell lines":"VCaP","Time points":"24h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_24-2",
"Factors":{"Cell lines":"VCaP","Time points":"24h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_24-3",
"Factors":{"Cell lines":"VCaP","Time points":"24h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_24-4",
"Factors":{"Cell lines":"VCaP","Time points":"24h"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_P3-1",
"Factors":{"Cell lines":"VCaP","Time points":"6days"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_P3-2",
"Factors":{"Cell lines":"VCaP","Time points":"6days"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_P3-3",
"Factors":{"Cell lines":"VCaP","Time points":"6days"}
},
{
"Subject ID":"-",
"Sample ID":"13VCaP_P3-4",
"Factors":{"Cell lines":"VCaP","Time points":"6days"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"cells were grown in RPMI 1640 medium containing 2 g/L glucose with 10% dFBS, and were harvested at 80% confluence. One hour before the isotope switch, the medium was aspirated and replaced with fresh unlabeled medium[1]. Subsequently, VCaP and RWPE-1 were cultured in RPMI 1640 medium with 2 g/L [U−13C]-glucose and 10% dFBS for 0 h, 0.25 h, 4 h, 12 h, 24 h and long-term for 6 days performing thereby 3 passages. 10 cm petri dish with 10 mL of media was used for each sample, and 4 independent biological replicates were prepared for the labeling experiments. Unlabeled samples were treated in the same way with medium containing unlabeled glucose. After the cultures have been grown for the targeted amount of time, the medium was aspirated completely, then cells were washed thrice with 5% D-mannitol solution, followed by inactivation with liquid nitrogen immediately.","SAMPLE_TYPE":"Cultured cells"},

"TREATMENT":{"TREATMENT_SUMMARY":"One hour before the isotope switch, the medium was aspirated and replaced with fresh unlabeled medium[1]. Subsequently, VCaP and RWPE-1 were cultured in RPMI 1640 medium with 2 g/L [U−13C]-glucose and 10% dFBS for 0 h, 0.25 h, 4 h, 12 h, 24 h and long-term for 6 days performing thereby 3 passages. 10 cm petri dish with 10 mL of media was used for each sample, and 4 independent biological replicates were prepared for the labeling experiments. Unlabeled samples were treated in the same way with medium containing unlabeled glucose."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"After the cultures have been grown for the targeted amount of time, the medium was aspirated completely, then cells were washed thrice with 5% D-mannitol solution, followed by inactivation with liquid nitrogen immediately. Cells were handled on ice during the preparation. 1 mL of precooled methanol, containing 13.3 μM of MetS and CSA, was added to each cell culture dish. Cells were harvested using a cell scraper, and transferred into a 5 mL tube. Then, 1 mL of chloroform was added and vortexed for 5 min, followed by 400 μL of Milli-Q water and 10 min of vortexing. After 5 min of standing, the mixture was centrifuged to form a two-phase system (10,000 g, 4 °C, 15 min). The upper layer was transferred and centrifugally filtered through a Millipore 5-kDa cutoff filter (USA) (13,000 g, 3h at 4°C). The filtrate was dried and stored at -80 °C. Finally, the upper extract was dissolved in Milli-Q water containing 50 μM of 3-AP, DPA, DHD and TMA for capillary electrophoresis-time of flight mass spectrometry (CE-TOF/MS) analysis."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"CE","INSTRUMENT_NAME":"Agilent 1290 Infinity II","COLUMN_NAME":"a fused silica capillary (50 µm i.d. × 80 cm)","FLOW_RATE":"internal pressure of 15 mbar and positive voltage of 30 kV","COLUMN_TEMPERATURE":"20","SOLVENT_A":"solution of 25 mM ammonium acetate and 75 mM diammonium hydrogen phosphate (pH 8.5)","INJECTION_TEMPERATURE":"20"},

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

"MS":{"INSTRUMENT_NAME":"Agilent 6224 TOF","INSTRUMENT_TYPE":"Triple quadrupole","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"capillary voltage of 3.5 kV, fragmentor of 125 V and mass range of 50-1,000 were set in the TOF/MS analysis. During sample analysis, reference masses were applied to ensure the real-time calibration of exact mass measurement."},

"MS_METABOLITE_DATA":{
"Units":"Relative intensity after normalization",

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}

}