{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000025","ANALYSIS_ID":"AN000045","VERSION":"1","CREATED_ON":"2016-09-17"},

"PROJECT":{"PROJECT_TITLE":"Metabolomics Analysis of Thermally Challenged Mayfly Larvae","PROJECT_TYPE":"MS/NMR","PROJECT_SUMMARY":"The purpose of this study was to examine the metabolic profiles of mayfly triangulifer) larvae subjected to a thermal challenge. This species is unusual terms of its ease of culture, and its suitability as a laboratory test Studies complementary to this metabolic study have focused on thermal effects metabolic rates (respiration) and gene expression patterns in this species and not support Prtners oxygen limitation paradigm. Our purpose here was to examine an environmentally realistic thermal challenge affects the physiology of this","INSTITUTE":"North Carolina State Unversity","DEPARTMENT":"Environmental and Molecular Toxicology","LABORATORY":"Buchwalter Laboratory","LAST_NAME":"Buchwalter","FIRST_NAME":"David","ADDRESS":"North Carolina State University Raleigh, NC 27695","EMAIL":"dbbuchwa@ncsu.edu","PHONE":"919-513-1129"},

"STUDY":{"STUDY_TITLE":"Metabolomics Analysis of Thermally Challenged Mayfly Larvae (GCMS analysis)","STUDY_TYPE":"Metabolomic analysis of mayflies","STUDY_SUMMARY":"The purpose of this study was to examine the metabolic profiles of mayfly triangulifer) larvae subjected to thermal challenge. This species is unusual in of its ease of culture, and its suitability as a laboratory test organism. Our here was to examine how an environmentally realistic thermal challenge affects physiology of this organism. In this study, we obtained several types of insect and we were able to show that GC-MS Metabolomics could be used to distinguish the different types of larvae.","INSTITUTE":"RTI International","DEPARTMENT":"Systems and Translational Sciences (STS)","LABORATORY":"NIH Eastern Regional Comprehensive Metabolomics Resource Core (RTI RCMRC)","LAST_NAME":"Sumner","FIRST_NAME":"Susan","ADDRESS":"3040 E. Cornwallis Road, Research Triangle Park, NC 27709, USA","EMAIL":"ssumner@rti.org","PHONE":"919-541-7479","NUM_GROUPS":"7","TOTAL_SUBJECTS":"13"},

"SUBJECT":{"SUBJECT_TYPE":"Animal","SUBJECT_SPECIES":"Centroptilum triangulifer;Atherix;Megaloptera","SPECIES_GROUP":"Insect"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"AD_01",
"Sample ID":"131221ajcsa17_1",
"Factors":{"species":"Athorix Diptoa","Treatment":"Athorix Diptoa control","wing pads":"no wing pads"}
},
{
"Subject ID":"AD_01",
"Sample ID":"131221ajcsa31_1",
"Factors":{"species":"Athorix Diptoa","Treatment":"Athorix Diptoa control","wing pads":"no wing pads"}
},
{
"Subject ID":"Pooled QC_01",
"Sample ID":"131221ajcsa09_1",
"Factors":{"species":"Mayfly, Megalopterans, Athorix Diptoa","Treatment":"Pooled QC sample","wing pads":"no wing pads"}
},
{
"Subject ID":"Pooled QC_02",
"Sample ID":"131221ajcsa15_1",
"Factors":{"species":"Mayfly, Megalopterans, Athorix Diptoa","Treatment":"Pooled QC sample","wing pads":"no wing pads"}
},
{
"Subject ID":"Pooled QC_03",
"Sample ID":"131221ajcsa21_1",
"Factors":{"species":"Mayfly, Megalopterans, Athorix Diptoa","Treatment":"Pooled QC sample","wing pads":"no wing pads"}
},
{
"Subject ID":"Pooled QC_01",
"Sample ID":"131221ajcsa23_1",
"Factors":{"species":"Mayfly, Megalopterans, Athorix Diptoa","Treatment":"Pooled QC sample","wing pads":"no wing pads"}
},
{
"Subject ID":"Pooled QC_02",
"Sample ID":"131221ajcsa29_1",
"Factors":{"species":"Mayfly, Megalopterans, Athorix Diptoa","Treatment":"Pooled QC sample","wing pads":"no wing pads"}
},
{
"Subject ID":"Pooled QC_03",
"Sample ID":"131221ajcsa35_1",
"Factors":{"species":"Mayfly, Megalopterans, Athorix Diptoa","Treatment":"Pooled QC sample","wing pads":"no wing pads"}
},
{
"Subject ID":"MF_01",
"Sample ID":"131221ajcsa11_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly control from different study","wing pads":"no wing pads"}
},
{
"Subject ID":"MF_01",
"Sample ID":"131221ajcsa24_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly control from different study","wing pads":"no wing pads"}
},
{
"Subject ID":"Control 01",
"Sample ID":"131221ajcsa12_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly control","wing pads":"no wing pads"}
},
{
"Subject ID":"Control 02",
"Sample ID":"131221ajcsa14_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly control","wing pads":"no wing pads"}
},
{
"Subject ID":"Control 03",
"Sample ID":"131221ajcsa20_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly control","wing pads":"no wing pads"}
},
{
"Subject ID":"Control 03",
"Sample ID":"131221ajcsa25_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly control","wing pads":"no wing pads"}
},
{
"Subject ID":"Control 01",
"Sample ID":"131221ajcsa26_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly control","wing pads":"no wing pads"}
},
{
"Subject ID":"Control 02",
"Sample ID":"131221ajcsa33_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly control","wing pads":"no wing pads"}
},
{
"Subject ID":"Control X",
"Sample ID":"131221ajcsa03_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly controlX","wing pads":"wing pads"}
},
{
"Subject ID":"Control X",
"Sample ID":"131221ajcsa04_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly controlX","wing pads":"wing pads"}
},
{
"Subject ID":"Control X",
"Sample ID":"131221ajcsa05_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly controlX","wing pads":"wing pads"}
},
{
"Subject ID":"Control X",
"Sample ID":"131221ajcsa06_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly controlX","wing pads":"wing pads"}
},
{
"Subject ID":"Control X",
"Sample ID":"131221ajcsa07_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly controlX","wing pads":"wing pads"}
},
{
"Subject ID":"Control X",
"Sample ID":"131221ajcsa19_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly controlX","wing pads":"wing pads"}
},
{
"Subject ID":"Control X",
"Sample ID":"131221ajcsa32_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly controlX","wing pads":"wing pads"}
},
{
"Subject ID":"Heat Treated_03",
"Sample ID":"131221ajcsa10_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly Heat Treated","wing pads":"no wing pads"}
},
{
"Subject ID":"Heat Treated_02",
"Sample ID":"131221ajcsa13_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly Heat Treated","wing pads":"no wing pads"}
},
{
"Subject ID":"Heat Treated_01",
"Sample ID":"131221ajcsa16_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly Heat Treated","wing pads":"no wing pads"}
},
{
"Subject ID":"Heat Treated_03",
"Sample ID":"131221ajcsa27_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly Heat Treated","wing pads":"no wing pads"}
},
{
"Subject ID":"Heat Treated_02",
"Sample ID":"131221ajcsa30_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly Heat Treated","wing pads":"no wing pads"}
},
{
"Subject ID":"Heat Treated_01",
"Sample ID":"131221ajcsa34_1",
"Factors":{"species":"Mayfly","Treatment":"Mayfly Heat Treated","wing pads":"no wing pads"}
},
{
"Subject ID":"MG_01",
"Sample ID":"131221ajcsa18_1",
"Factors":{"species":"Megalopterans","Treatment":"Megalopterans control","wing pads":"no wing pads"}
},
{
"Subject ID":"MG_01",
"Sample ID":"131221ajcsa28_1",
"Factors":{"species":"Megalopterans","Treatment":"Megalopterans control","wing pads":"no wing pads"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"whole insects were collected","COLLECTION_PROTOCOL_COMMENTS":"Original culture material was obtained from the Stroud Water Research Center PA). Larvae were reared at 22C and fed a periphytic diet. When larvae reached a size, they were separated from their food source for 12 hours to evacuate gut Larvae were then divided into two treatment groups a control group maintained the culturing temperature of 22C, and a thermal challenge group. The thermal group was subjected to a temperature increase at a rate of 1C per hour. This of temperature change is commonly observed in temperate streams. When the temperature reached 30C, larvae from both control and treatment groups were frozen in liquid nitrogen in groups of 12-13 larvae. Each grouping of 12-13 comprised a composite replicate for the metabolic profiling analysis. A total 3 replicates each for the control and thermal treatments were used in this Some larvae had developed dark wing pads during the experiment. This is a that the larvae were very close to emerging from the aquatic larval phase to a sub-adult phase. Enough of these larvae were present in the control treatment be removed from the control cohort and considered as an independent sample.","SAMPLE_TYPE":"whole insect","VOLUMEORAMOUNT_COLLECTED":"50 mg","STORAGE_CONDITIONS":"-80C"},

"TREATMENT":{"TREATMENT_SUMMARY":"Metabolomics profile of mayfly larvae exposed to heat","TREATMENT_PROTOCOL_COMMENTS":"Heat Treated |  Control |  ControlX","TREATMENT":"Abiotic","ANIMAL_ACCLIMATION_DURATION":"exposed to heat (22C to 30C ramp); fasted for 8 hours | control (fasted for 8 | control (fasted for 6 hours)"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Insect samples homogenized in 50% acetonitrile at a concentration of 50 mg/mL. dried and aliquot taken for NMR. Sample returned and extract processed for analysis. Samples were derivatized using 2 step derivatization method with methoxy amine followed by TMS derivatization with MSTFA).","SAMPLEPREP_PROTOCOL_FILENAME":"RTI_DB_INSECT_Metabolomics_Procedure_March12_2014.docx","PROCESSING_METHOD":"Homogenization and Solvent Removal with SpeedVac","PROCESSING_STORAGE_CONDITIONS":"4C","EXTRACTION_METHOD":"50% Acetonitrile","EXTRACT_STORAGE":"-80C","SAMPLE_RESUSPENSION":"resuspended in D2O + Chenomix ISTD solution","SAMPLE_DERIVATIZATION":"Methoximation with methoxy amine at 40 mg/ml in pyridine followed by TMS with MSTFA","SAMPLE_SPIKING":"FAMES (C8-C30) added to MSTFA as retention time markers"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"GC/MS analysis is based on Oliver Fiehn methods","CHROMATOGRAPHY_TYPE":"GC","INSTRUMENT_NAME":"Leco Pegasus IV GC","COLUMN_NAME":"Restek Rtx-5Sil MS (30 x 0.25mm, 0.25um)","COLUMN_PRESSURE":"10.4 psi","COLUMN_TEMPERATURE":"50C(0.5 min)-330C(@20C/min)-330C(5 min)","FLOW_GRADIENT":"constant flow","FLOW_RATE":"1.0 mL/min","INJECTION_TEMPERATURE":"250C","RETENTION_INDEX":"FAMEs markers (C8-C30)","RETENTION_TIME":"FAMEs markers (C8-C30)","ANALYTICAL_TIME":"20 minutes","OVEN_TEMPERATURE":"50C(0.5 min)-330C(@20C/min)-330C(5 min)","TRANSFERLINE_TEMPERATURE":"280C","SAMPLE_SYRINGE_SIZE":"10 L"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","LABORATORY_NAME":"Eastern RCMRC (RTI)","ACQUISITION_DATE":"41629","SOFTWARE_VERSION":"Chromatof 4.50.8.0","OPERATOR_NAME":"James Carlson","DETECTOR_TYPE":"TOF (+ ion mode)","DATA_FORMAT":"netcdf"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Leco Pegasus IV TOF","INSTRUMENT_TYPE":"GC x GC-TOF","MS_TYPE":"EI","ION_MODE":"POSITIVE","CAPILLARY_TEMPERATURE":"280C (transfer line)","HELIUM_FLOW":"1.0 ml/min","ION_SOURCE_TEMPERATURE":"250C","IONIZATION_ENERGY":"70 eV","SOURCE_TEMPERATURE":"250C","DATAFORMAT":"netcdf","RESOLUTION_SETTING":"1200","SCAN_RANGE_MOVERZ":"50-800","SCANNING":"20 spectra/sec"},

"MS_METABOLITE_DATA":{
"Units":"peak area",

"Data":[{"Metabolite":"1-monostearin","131221ajcsa17_1":"24875.0000","131221ajcsa31_1":"25027.0000","131221ajcsa09_1":"25911.0000","131221ajcsa15_1":"39491.0000","131221ajcsa21_1":"47425.0000","131221ajcsa23_1":"38390.0000","131221ajcsa29_1":"51679.0000","131221ajcsa35_1":"33464.0000","131221ajcsa11_1":"18674.0000","131221ajcsa24_1":"35774.0000","131221ajcsa12_1":"42528.0000","131221ajcsa14_1":"35605.0000","131221ajcsa20_1":"49061.0000","131221ajcsa25_1":"48047.0000","131221ajcsa26_1":"45112.0000","131221ajcsa33_1":"48701.0000","131221ajcsa03_1":"500102.0000","131221ajcsa04_1":"546187.0000","131221ajcsa05_1":"573895.0000","131221ajcsa06_1":"539082.0000","131221ajcsa07_1":"532260.0000","131221ajcsa19_1":"546499.0000","131221ajcsa32_1":"607085.0000","131221ajcsa10_1":"39327.0000","131221ajcsa13_1":"31364.0000","131221ajcsa16_1":"44129.0000","131221ajcsa27_1":"49550.0000","131221ajcsa30_1":"49499.0000","131221ajcsa34_1":"53750.0000","131221ajcsa18_1":"85510.0000","131221ajcsa28_1":"81723.0000"},{"Metabolite":"2_5-dihydroxypyrazine 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