{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001985","ANALYSIS_ID":"AN003236","VERSION":"1","CREATED_ON":"October 28, 2021, 9:04 am"},

"PROJECT":{"PROJECT_TITLE":"Profiling Plasmodium falciparum parasites and human red blood cells after treatment with MMV693183","PROJECT_SUMMARY":"This project includes data for two distinct biological replicates of 2.5-hour treatment experiments on either magnetically purified Plasmodium falciparum parasites or uninfected human red blood cells. The study also includes an equal number of untreated replicates as a control and treatment with compound MMV693183 at 240 nM, 24 nM, and 2.4 nM.","INSTITUTE":"Pennsylvania State University","LAST_NAME":"Munro","FIRST_NAME":"Justin","ADDRESS":"Millenium Science Complex, University Park, Pennsylvania, 16802, USA","EMAIL":"jtm5518@psu.edu","PHONE":"484-792-1415","FUNDING_SOURCE":"T32 DK120509"},

"STUDY":{"STUDY_TITLE":"Profiling Plasmodium falciparum parasites and human red blood cells after treatment with MMV693183","STUDY_SUMMARY":"Compound MMV693183 was added to either Plasmodium falciparum or uninfected red blood cells. The concentrations used were 240 nM, 24 nM, and 2.4 nM. Untreated parasites and red blood cells were also used as controls.","INSTITUTE":"Pennsylvania State University","LAST_NAME":"Llinás","FIRST_NAME":"Manuel","ADDRESS":"W126 Millennium Science Complex, University Park, PENNSYLVANIA, 16802, USA","EMAIL":"manuel@psu.edu","PHONE":"8148673527"},

"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Plasmodium falciparum","TAXONOMY_ID":"5833","GENOTYPE_STRAIN":"3D7","CELL_COUNTS":"1*10^8"},
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],
"COLLECTION":{"COLLECTION_SUMMARY":"Parasites were magnetically purified at the early trophozoite stage from cultures of 5-10% parasitemia. Samples were generated with 5 mL of media containing 1*10^8 cells (either parasites or human red blood cells) for the treatment condition. Excess media was removed and the cells were centrifuged and washed once with PBS. Metabolism was quenched with 90% methanol containing 0.25 micromolar of universally labeled C-13, N-15 aspartate. The samples were centrifuged and the metabolites in the methanol were collected and transferred to a new tube to be dried with nitrogen.","COLLECTION_PROTOCOL_FILENAME":"Metabolite_Extraction_for_LCMS_2017.pdf","SAMPLE_TYPE":"Cultured cells","COLLECTION_LOCATION":"Millenium Science Complex, University Park, Pennsylvania","STORAGE_CONDITIONS":"-80℃","COLLECTION_VIALS":"1.5 mL eppendorf","STORAGE_VIALS":"1.5 mL eppendorf","COLLECTION_TUBE_TEMP":"On ice","TISSUE_CELL_QUANTITY_TAKEN":"1x10^8 cells per sample in 1 mL total volume"},

"TREATMENT":{"TREATMENT_SUMMARY":"In all conditions 1*10^8 cells were cultured in 5 mL of RPMI 1640 media for 2.5 hours. At the start of the incubation period, cells were either not exposed to any drug or exposed to 240 nM, 24 nM, or 2.4 nM MMV693183. Dates reflect date of running an individual sample on the analytical platform.","TREATMENT_COMPOUND":"MMV693183","TREATMENT_ROUTE":"Transfer to media by pipette","TREATMENT_DOSE":"5 microliters of drug to a final concentration of 240 nM, 24 nM, or 2.4 nM.","TREATMENT_VEHICLE":"DMSO","CELL_GROWTH_CONTAINER":"6-well plate","CELL_GROWTH_CONFIG":"5 mL in each sample well, 1x10^8 infected red blood cells per well, conditions performed in triplicate","CELL_MEDIA":"RPMI 1640 containing Albumax, Gentamycin, Hypoxanthine, HEPES, Sodium Bicarbonate"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Once samples were obtained, metabolism was quenched with 90% methanol containing 0.25 uM labeled aspartate, cells were centrifuged, and the supernatant was removed by nitrogen drying. Sample was reconstituted in 3% HPLC-grade methanol and run on the instrument.","PROCESSING_METHOD":"Wash, spin, quench, spin, dry, store at -80 C until run on the instrument, resuspend","PROCESSING_STORAGE_CONDITIONS":"On ice","EXTRACTION_METHOD":"90% methanol extraction","EXTRACT_CLEANUP":"Centrifugation and nitrogen drying","SAMPLE_RESUSPENSION":"100 uL 3% methanol with 1 uM chlorpropamide","SAMPLE_SPIKING":"0.25 uM Labelled Aspartate, 1 uM chlorpropamide"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Ion-pairing method using reverse-phase chromatography setup.","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Dionex Ultimate 3000","COLUMN_NAME":"Waters XSelect HSS (2.1 x 100 mm, 2.5 um)","FLOW_RATE":"0.200 mL/minute","COLUMN_TEMPERATURE":"30","SOLVENT_A":"3% methanol, 15 mM acetic acid, 10 mM tributylamine","SOLVENT_B":"100% methanol"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","OPERATOR_NAME":"Justin Munro"},

"MS":{"INSTRUMENT_NAME":"Thermo Exactive Plus Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"Data was centroided using MSConvert and converted to .mzXML for utilization in MzMine and El-Maven software."},

"MS_METABOLITE_DATA":{
"Units":"Peak Abundance (normalized, blank subtracted, and corrected for baseline noise)",

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}