{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001853","ANALYSIS_ID":"AN003002","VERSION":"1","CREATED_ON":"June 29, 2021, 1:55 pm"},

"PROJECT":{"PROJECT_TITLE":"Longitudinal Metabolomics of Human Plasma Reveals Robust Prognostic Markers of COVID-19 Disease Severity","PROJECT_SUMMARY":"There is an urgent need to identify which COVID-19 patients will develop life-threatening illness so that medical resources can be optimally allocated and rapid treatment can be administered early in the disease course, when clinical management is most effective. To aid in the prognostic classification of disease severity, we perform untargeted metabolomics on plasma from 339 patients, with samples collected at six longitudinal time points. Using the temporal metabolic profiles and machine learning, we build a predictive model of disease severity. We discover that a panel of metabolites measured at the time of study entry successfully determine disease severity. Through analysis of longitudinal samples, we confirm that the majority of these markers are directly related to disease progression and that their levels are restored to baseline upon disease recovery. Finally, we validate that these metabolites are also altered in a hamster model of COVID-19. Our results indicate that metabolic changes associated with COVID-19 severity can be effectively used to stratify patients and inform resource allocation during the pandemic.","INSTITUTE":"Washington University, St. Louis","DEPARTMENT":"Chemistry","LABORATORY":"Patti","LAST_NAME":"Patti","FIRST_NAME":"Gary","ADDRESS":"McMillen Chemistry Laboratory, Washington University 1 Brookings Dr @ Throop Drive, Rm 102, St. Louis, MO 63130-4899","EMAIL":"gjpattij@wustl.edu","PHONE":"314-935-3512"},

"STUDY":{"STUDY_TITLE":"Longitudinal Metabolomics of Human Plasma Reveals Robust Prognostic Markers of COVID-19 Disease Severity","STUDY_SUMMARY":"There is an urgent need to identify which COVID-19 patients will develop life-threatening illness so that medical resources can be optimally allocated and rapid treatment can be administered early in the disease course, when clinical management is most effective. To aid in the prognostic classification of disease severity, we perform untargeted metabolomics on plasma from 339 patients, with samples collected at six longitudinal time points. Using the temporal metabolic profiles and machine learning, we build a predictive model of disease severity. We discover that a panel of metabolites measured at the time of study entry successfully determine disease severity. Through analysis of longitudinal samples, we confirm that the majority of these markers are directly related to disease progression and that their levels are restored to baseline upon disease recovery. Finally, we validate that these metabolites are also altered in a hamster model of COVID-19. Our results indicate that metabolic changes associated with COVID-19 severity can be effectively used to stratify patients and inform resource allocation during the pandemic.","INSTITUTE":"Washington University, St. Louis","DEPARTMENT":"Chemistry","LABORATORY":"Patti","LAST_NAME":"Patti","FIRST_NAME":"Gary","ADDRESS":"McMillen Chemistry Laboratory, Washington University 1 Brookings Dr @ Throop Drive, Rm 102, St. Louis, MO 63130-4899","EMAIL":"gjpattij@wustl.edu","NUM_GROUPS":"3","TOTAL_SUBJECTS":"56","NUM_FEMALES":"56","PHONE":"314-935-3512"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mesocricetus auratus"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"P H-433",
"Factors":{"Group":"Influenza","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-433_polar_pos.mzML,P H-433_polar_neg.mzML,P H-433_lipid_pos.mzML,P H-433_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-457",
"Factors":{"Group":"Influenza","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-457_polar_pos.mzML,P H-457_polar_neg.mzML,P H-457_lipid_pos.mzML,P H-457_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-458",
"Factors":{"Group":"Influenza","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-458_polar_pos.mzML,P H-458_polar_neg.mzML,P H-458_lipid_pos.mzML,P H-458_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-459",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-459_polar_pos.mzML,P H-459_polar_neg.mzML,P H-459_lipid_pos.mzML,P H-459_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-460",
"Factors":{"Group":"Influenza","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-460_polar_pos.mzML,P H-460_polar_neg.mzML,P H-460_lipid_pos.mzML,P H-460_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-461",
"Factors":{"Group":"Influenza","Day post infection":"6"},
"Additional sample data":{"RAW_FILE_NAME":"P H-461_polar_pos.mzML,P H-461_polar_neg.mzML,P H-461_lipid_pos.mzML,P H-461_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-462",
"Factors":{"Group":"Influenza","Day post infection":"6"},
"Additional sample data":{"RAW_FILE_NAME":"P H-462_polar_pos.mzML,P H-462_polar_neg.mzML,P H-462_lipid_pos.mzML,P H-462_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-463",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"6"},
"Additional sample data":{"RAW_FILE_NAME":"P H-463_polar_pos.mzML,P H-463_polar_neg.mzML,P H-463_lipid_pos.mzML,P H-463_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-464",
"Factors":{"Group":"Mock","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-464_polar_pos.mzML,P H-464_polar_neg.mzML,P H-464_lipid_pos.mzML,P H-464_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-466",
"Factors":{"Group":"Influenza","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-466_polar_pos.mzML,P H-466_polar_neg.mzML,P H-466_lipid_pos.mzML,P H-466_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-467",
"Factors":{"Group":"Influenza","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-467_polar_pos.mzML,P H-467_polar_neg.mzML,P H-467_lipid_pos.mzML,P H-467_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-468",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"6"},
"Additional sample data":{"RAW_FILE_NAME":"P H-468_polar_pos.mzML,P H-468_polar_neg.mzML,P H-468_lipid_pos.mzML,P H-468_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-469 -1",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"2"},
"Additional sample data":{"RAW_FILE_NAME":"P H-469 -1_polar_pos.mzML,P H-469 -1_polar_neg.mzML,P H-469 -1_lipid_pos.mzML,P H-469 -1_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-469 -2",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-469 -2 _polar_pos.mzML,P H-469 -2 _polar_neg.mzML,P H-469 -2 _lipid_pos.mzML,P H-469 -2 _lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-470",
"Factors":{"Group":"Mock","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-470_polar_pos.mzML,P H-470_polar_neg.mzML,P H-470_lipid_pos.mzML,P H-470_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-471",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-471_polar_pos.mzML,P H-471_polar_neg.mzML,P H-471_lipid_pos.mzML,P H-471_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-472",
"Factors":{"Group":"Mock","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-472_polar_pos.mzML,P H-472_polar_neg.mzML,P H-472_lipid_pos.mzML,P H-472_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-473",
"Factors":{"Group":"Influenza","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-473_polar_pos.mzML,P H-473_polar_neg.mzML,P H-473_lipid_pos.mzML,P H-473_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-474",
"Factors":{"Group":"Influenza","Day post infection":"2"},
"Additional sample data":{"RAW_FILE_NAME":"P H-474_polar_pos.mzML,P H-474_polar_neg.mzML,P H-474_lipid_pos.mzML,P H-474_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-475",
"Factors":{"Group":"Mock","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-475_polar_pos.mzML,P H-475_polar_neg.mzML,P H-475_lipid_pos.mzML,P H-475_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-476",
"Factors":{"Group":"Mock","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-476_polar_pos.mzML,P H-476_polar_neg.mzML,P H-476_lipid_pos.mzML,P H-476_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-477",
"Factors":{"Group":"Influenza","Day post infection":"6"},
"Additional sample data":{"RAW_FILE_NAME":"P H-477_polar_pos.mzML,P H-477_polar_neg.mzML,P H-477_lipid_pos.mzML,P H-477_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-478",
"Factors":{"Group":"Influenza","Day post infection":"2"},
"Additional sample data":{"RAW_FILE_NAME":"P H-478_polar_pos.mzML,P H-478_polar_neg.mzML,P H-478_lipid_pos.mzML,P H-478_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-479",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-479_polar_pos.mzML,P H-479_polar_neg.mzML,P H-479_lipid_pos.mzML,P H-479_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-480",
"Factors":{"Group":"Mock","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-480_polar_pos.mzML,P H-480_polar_neg.mzML,P H-480_lipid_pos.mzML,P H-480_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-481",
"Factors":{"Group":"Mock","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-481_polar_pos.mzML,P H-481_polar_neg.mzML,P H-481_lipid_pos.mzML,P H-481_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-482",
"Factors":{"Group":"Mock","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-482_polar_pos.mzML,P H-482_polar_neg.mzML,P H-482_lipid_pos.mzML,P H-482_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-483",
"Factors":{"Group":"Influenza","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-483_polar_pos.mzML,P H-483_polar_neg.mzML,P H-483_lipid_pos.mzML,P H-483_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-484",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-484_polar_pos.mzML,P H-484_polar_neg.mzML,P H-484_lipid_pos.mzML,P H-484_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-485",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-485_polar_pos.mzML,P H-485_polar_neg.mzML,P H-485_lipid_pos.mzML,P H-485_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-486",
"Factors":{"Group":"Influenza","Day post infection":"6"},
"Additional sample data":{"RAW_FILE_NAME":"P H-486_polar_pos.mzML,P H-486_polar_neg.mzML,P H-486_lipid_pos.mzML,P H-486_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-487",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-487_polar_pos.mzML,P H-487_polar_neg.mzML,P H-487_lipid_pos.mzML,P H-487_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-488",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-488_polar_pos.mzML,P H-488_polar_neg.mzML,P H-488_lipid_pos.mzML,P H-488_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-489",
"Factors":{"Group":"Mock","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-489_polar_pos.mzML,P H-489_polar_neg.mzML,P H-489_lipid_pos.mzML,P H-489_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-490",
"Factors":{"Group":"Mock","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-490_polar_pos.mzML,P H-490_polar_neg.mzML,P H-490_lipid_pos.mzML,P H-490_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-491",
"Factors":{"Group":"Influenza","Day post infection":"6"},
"Additional sample data":{"RAW_FILE_NAME":"P H-491_polar_pos.mzML,P H-491_polar_neg.mzML,P H-491_lipid_pos.mzML,P H-491_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-492",
"Factors":{"Group":"Influenza","Day post infection":"2"},
"Additional sample data":{"RAW_FILE_NAME":"P H-492_polar_pos.mzML,P H-492_polar_neg.mzML,P H-492_lipid_pos.mzML,P H-492_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-495",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"6"},
"Additional sample data":{"RAW_FILE_NAME":"P H-495_polar_pos.mzML,P H-495_polar_neg.mzML,P H-495_lipid_pos.mzML,P H-495_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-496",
"Factors":{"Group":"Influenza","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-496_polar_pos.mzML,P H-496_polar_neg.mzML,P H-496_lipid_pos.mzML,P H-496_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-497",
"Factors":{"Group":"Influenza","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-497_polar_pos.mzML,P H-497_polar_neg.mzML,P H-497_lipid_pos.mzML,P H-497_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-498",
"Factors":{"Group":"Influenza","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-498_polar_pos.mzML,P H-498_polar_neg.mzML,P H-498_lipid_pos.mzML,P H-498_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-499",
"Factors":{"Group":"Influenza","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-499_polar_pos.mzML,P H-499_polar_neg.mzML,P H-499_lipid_pos.mzML,P H-499_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-501",
"Factors":{"Group":"Mock","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-501_polar_pos.mzML,P H-501_polar_neg.mzML,P H-501_lipid_pos.mzML,P H-501_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-502",
"Factors":{"Group":"Influenza","Day post infection":"2"},
"Additional sample data":{"RAW_FILE_NAME":"P H-502_polar_pos.mzML,P H-502_polar_neg.mzML,P H-502_lipid_pos.mzML,P H-502_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-504",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"6"},
"Additional sample data":{"RAW_FILE_NAME":"P H-504_polar_pos.mzML,P H-504_polar_neg.mzML,P H-504_lipid_pos.mzML,P H-504_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-505",
"Factors":{"Group":"Influenza","Day post infection":"2"},
"Additional sample data":{"RAW_FILE_NAME":"P H-505_polar_pos.mzML,P H-505_polar_neg.mzML,P H-505_lipid_pos.mzML,P H-505_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-506",
"Factors":{"Group":"Influenza","Day post infection":"2"},
"Additional sample data":{"RAW_FILE_NAME":"P H-506_polar_pos.mzML,P H-506_polar_neg.mzML,P H-506_lipid_pos.mzML,P H-506_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-507",
"Factors":{"Group":"Mock","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-507_polar_pos.mzML,P H-507_polar_neg.mzML,P H-507_lipid_pos.mzML,P H-507_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-509",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"6"},
"Additional sample data":{"RAW_FILE_NAME":"P H-509_polar_pos.mzML,P H-509_polar_neg.mzML,P H-509_lipid_pos.mzML,P H-509_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-510",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-510_polar_pos.mzML,P H-510_polar_neg.mzML,P H-510_lipid_pos.mzML,P H-510_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-511",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"14"},
"Additional sample data":{"RAW_FILE_NAME":"P H-511_polar_pos.mzML,P H-511_polar_neg.mzML,P H-511_lipid_pos.mzML,P H-511_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-512",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"6"},
"Additional sample data":{"RAW_FILE_NAME":"P H-512_polar_pos.mzML,P H-512_polar_neg.mzML,P H-512_lipid_pos.mzML,P H-512_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-513",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"2"},
"Additional sample data":{"RAW_FILE_NAME":"P H-513_polar_pos.mzML,P H-513_polar_neg.mzML,P H-513_lipid_pos.mzML,P H-513_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-514",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"4"},
"Additional sample data":{"RAW_FILE_NAME":"P H-514_polar_pos.mzML,P H-514_polar_neg.mzML,P H-514_lipid_pos.mzML,P H-514_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-515",
"Factors":{"Group":"Influenza","Day post infection":"6"},
"Additional sample data":{"RAW_FILE_NAME":"P H-515_polar_pos.mzML,P H-515_polar_neg.mzML,P H-515_lipid_pos.mzML,P H-515_lipid_neg.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"P H-516",
"Factors":{"Group":"SARS-CoV-2","Day post infection":"2"},
"Additional sample data":{"RAW_FILE_NAME":"P H-516_polar_pos.mzML,P H-516_polar_neg.mzML,P H-516_lipid_pos.mzML,P H-516_lipid_neg.mzML"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Outbred female LVG golden Syrian hamsters (6-8 weeks of age) were obtained from Charles River Laboratories (Kingston, NY). The hamsters were anesthetized by intraperitoneal injection of a mixture of ketamine and xylazine prior to intranasal inoculation with 0.1 mL of 1e5 plaque-forming units (PFU) of SARS-CoV-2 (WA-1) or H1N1 influenza A virus (A/California/04/2009). On day 2, 4, 6, and 14 after infection, 3-6 anesthetized hamsters per infection group were euthanized by exsanguination followed by intracardiac injection of veterinary euthanasia solution (SleepAway; Fort Dodge). Plasma samples were treated by exposure to germicidal UV-C light.","SAMPLE_TYPE":"Blood (plasma)"},

"TREATMENT":{"TREATMENT_SUMMARY":"The hamsters were anesthetized by intraperitoneal injection of a mixture of ketamine and xylazine prior to intranasal inoculation with 0.1 mL of 1e5 plaque-forming units (PFU) of SARS-CoV-2 (WA-1) or H1N1 influenza A virus (A/California/04/2009)."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Hamster plasma samples were diluted 1:4 with methanol (v/v), vortexed for 30 seconds, and incubated at -20 C for 2 hours. Samples were centrifuged for 10 minutes at 13,500 x g at 4°C and supernatant was transferred to a new centrifuge tube, concentrated, and stored at -80 C until reconstitution. Hamster plasma was thawed on ice. A 50 µL aliquot was transferred onto the solid-phase-extraction (SPE)-system CAPTIVA-EMR Lipid 96-wellplate (Agilent Technologies) before addition of 250 µL of acetonitrile containing 1% formic acid (v/v) and 10 µM internal standard (consisting of uniformly 13C and 15N labeled amino acids from Cambridge Isotope Laboratories, Inc). The samples were mixed for 1 min at 360 rpm on an orbital shaker at room temperature prior to a 10 min incubation period at 4 C. Afterwards, 200 µL 80% acetonitrile in water (v/v) were added to the samples. The samples were mixed on an orbital shaker (360 rpm) for an additional 10 min at room temperature. The samples were then eluted into a 96-deepwell collection plate by centrifugation (10 min, 57 x g, 4 C followed by 2 min, 1000 x g, 4 C). Polar eluates were stored at -80 C until the day of LC/MS analysis. The SPE-plates were then washed twice with 500 µL 80% acetonitrile in water (v/v). Lipids still bound to the SPE-material were then released into a second elution plate, in two elution steps applying 2x 500 µL 1:1 methyl tert-butyl ether:methanol (v/v) onto the SPE cartridge and centrifuging for 2 min at 1000 g and 4 C. The combined eluates were dried under a stream of nitrogen (Biotage SPE Dry Evaporation System) at room temperature and reconstituted with 100 µL 1:1 2-propanol:methanol (v/v) prior to LC/MS analysis."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"An aliquot of 2 µL of polar metabolite extract was subjected to LC/MS analysis by using an Agilent 1290 Infinity II liquid-chromatography (LC) system coupled to an Agilent 6540 Quadrupole-Time-of-Flight (Q-TOF) mass spectrometer with a dual Agilent Jet Stream electrospray ionization source. Polar metabolites were separated on a SeQuant® ZIC®-pHILIC column (100 x 2.1 mm, 5 µm, polymer, Merck-Millipore) including a ZIC®-pHILIC guard column (2.1 mm x 20 mm, 5 µm). The column compartment temperature was maintained at 40 C and the flow rate was set to 250 µLmin-1. The mobile phases consisted of A: 95% water, 5% acetonitrile, 20 mM ammonium bicarbonate, 0.1% ammonium hydroxide solution (25% ammonia in water), 2.5 µM medronic acid, and B: 95% acetonitrile, 5% water, 2.5 µM medronic acid. The following linear gradient was applied: 0 to 1 min, 90% B; 12 min, 35% B; 12.5 to 14.5 min, 25% B; 15 min, 90% B followed by a re-equilibration phase of 4 min at 400 µLmin-1 and 2 min at 250 µLmin-1.","CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Agilent 1290 Infinity II","COLUMN_NAME":"SeQuant® ZIC®-pHILIC column (100 x 2.1 mm, 5 µm, polymer, Merck-Millipore) including a ZIC®-pHILIC guard column (2.1 mm x 20 mm, 5 µm)"},

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

"MS":{"INSTRUMENT_NAME":"Agilent 6540 QTOF","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"gas temperature 200 C, drying gas flow 10 Lmin-1, nebulizer pressure 44 psi, sheath gas temperature 300 C, sheath gas flow 12 Lmin-1, VCap 3000 V, nozzle voltage 2000 V, Fragmentor 100 V, Skimmer 65 V, Oct 1 RF Vpp 750 V, and m/z range 50-1700. Data were acquired under continuous reference mass correction at m/z 121.0509 and 922.0890 for positive ion mode and m/z 119.0363 and 966.0007 for negative ion mode. Polar metabolite identifications were supported by matching the retention time, accurate mass, and MS/MS fragmentation data to our in-house retention time and MS/MS library created from authentic reference standards (Mass Spectrometry Metabolite Library supplied by IROA Technologies, Millipore Sigma, St. Louis, MO, USA) and online MS/MS libraries (Human Metabolome Database (HMDB, https://hmdb.ca), Mass Bank of North America (MoNA, https://mona.fiehnlab.ucdavis.edu/), and mzCloud (https://mzcloud.org). Lipid iterative MS/MS data were annotated with the Agilent Lipid Annotator software. All data files were then analyzed in Skyline (Version 20.1.0.155) to obtain peak areas."},

"MS_METABOLITE_DATA":{
"Units":"Relative Intensity",

"Data":[{"Metabolite":"Kynurenate","P H-433":"7847","P H-457":"5644","P H-458":"6149","P H-459":"4111","P H-460":"5202","P H-461":"4387","P H-462":"3507","P H-463":"6240","P H-464":"2883","P H-466":"3373","P H-467":"1876","P H-468":"4151","P H-469 -1":"4577","P H-469 -2":"3635","P H-470":"3042","P H-471":"4603","P H-472":"1813","P H-473":"4381","P H-474":"2359","P H-475":"1042","P H-476":"5736","P H-477":"3883","P H-478":"5379","P H-479":"2719","P H-480":"5121","P H-481":"5350","P H-482":"2853","P H-483":"3048","P H-484":"6147","P H-485":"3681","P H-486":"1822","P H-487":"2938","P H-488":"3763","P H-489":"4948","P H-490":"5686","P H-491":"4683","P H-492":"13170","P H-495":"3583","P H-496":"4966","P H-497":"3873","P H-498":"3683","P H-499":"5790","P H-501":"4059","P H-502":"9966","P H-504":"4923","P H-505":"18032","P H-506":"4135","P H-507":"3354","P H-509":"985","P H-510":"6914","P H-511":"8827","P H-512":"3368","P H-513":"6169","P H-514":"2727","P H-515":"4381","P H-516":"3838"},{"Metabolite":"L-Serine","P H-433":"10919","P H-457":"10529","P H-458":"11324","P H-459":"14306","P H-460":"10556","P H-461":"15640","P H-462":"127829","P H-463":"16647","P H-464":"11556","P H-466":"34426","P H-467":"9625","P H-468":"221841","P H-469 -1":"134268","P H-469 -2":"16787","P H-470":"8318","P H-471":"10903","P H-472":"16656","P H-473":"8934","P H-474":"201378","P H-475":"19059","P H-476":"39802","P H-477":"108634","P H-478":"178561","P H-479":"23273","P H-480":"79789","P H-481":"13978","P H-482":"11925","P H-483":"14747","P H-484":"11827","P H-485":"10075","P H-486":"96741","P H-487":"14515","P H-488":"15896","P H-489":"18540","P H-490":"14618","P H-491":"9056","P H-492":"141421","P H-495":"10253","P H-496":"12899","P H-497":"111522","P H-498":"11832","P H-499":"197382","P H-501":"11730","P H-502":"153336","P H-504":"13164","P H-505":"101536","P H-506":"125491","P H-507":"12190","P H-509":"184130","P H-510":"12234","P H-511":"13808","P H-512":"219381","P H-513":"188114","P H-514":"14925","P H-515":"10053","P H-516":"209251"}],

"Metabolites":[{"metabolite_name":"Kynurenate"},{"metabolite_name":"L-Serine"}]
}

}