{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001328","ANALYSIS_ID":"AN002214","VERSION":"1","CREATED_ON":"March 18, 2020, 10:34 am"},

"PROJECT":{"PROJECT_TITLE":"Pulmonary Tuberculosis Mice Project","PROJECT_TYPE":"Natural disease progression assessment","PROJECT_SUMMARY":"The goal of this multiplatform, non-targeted metabolomics study was to explore the metabolic alterations occurring during the natural progression of pulmonary tuberculosis in a murine model of disease (C57BL/6 genotype). For this purpose, we used gas chromatography, capillary electrophoresis, and reversed-phase liquid chromatography coupled to high-resolution mass analyzers (GC-EI-QTOF/MS, CE-ESI(+)-QTOF/MS, LC-ESI(+)-QTOF/MS and LC-ESI(-)-QTOF/MS to analyze lung extracts of age and sex-matched uninfected mice (UW, n=4), Mycobacterium tuberculosis-infected mice at 4 weeks post-infection (4W, n=4) and Mycobacterium tuberculosis-infected mice at 9 weeks post-infection. All data were acquired in MS1 mode, following a canonical non-targeted workflow. The potential benefit for the general research community arising from the combination of these analyses is to generate mass spectrometry data which has not been previously described for this model of the disease. This data may serve as a foundation for the validation of observations, data modeling, and biochemical interpretation of MS-based metabolic changes found in the comparisons between the different sample groups","INSTITUTE":"Universidad San Pablo-CEU, CEU Universities","DEPARTMENT":"Departamento de Quimica y Bioquimica","LABORATORY":"Centro de Metabolomica y Bioanalisis (CEMBIO)","LAST_NAME":"Fernandez Garcia","FIRST_NAME":"Miguel","ADDRESS":"Facultad de Farmacia, Universidad San Pablo-CEU, CEU Universities, Urbanización Montepríncipe, 28660 Boadilla del Monte, Spain","EMAIL":"miguel.fernandezgarcia@ceu.es","PHONE":"+34690090778"},

"STUDY":{"STUDY_TITLE":"Multiplatform, non-targeted analysis of lung extracts of uninfected and Mtb-infected C57BL/6 mice at 4 and 9 weeks p.i.","STUDY_SUMMARY":"The goal of this multiplatform, non-targeted metabolomics study was to explore the metabolic alterations occurring during the natural progression of pulmonary tuberculosis in a murine model of disease (C57BL/6 genotype). For this purpose, we used gas chromatography, capillary electrophoresis, and reversed-phase liquid chromatography coupled to high-resolution mass analyzers (GC-EI-QTOF/MS, CE-ESI(+)-QTOF/MS, LC-ESI(+)-QTOF/MS and LC-ESI(-)-QTOF/MS to analyze lung extracts of age and sex-matched uninfected mice (UW, n=4), Mycobacterium tuberculosis-infected mice at 4 weeks post-infection (4W, n=4) and Mycobacterium tuberculosis-infected mice at 9 weeks post-infection. All data were acquired in MS1 mode, following a canonical non-targeted workflow.","INSTITUTE":"Universidad San Pablo-CEU, CEU Universities","DEPARTMENT":"Departamento de Quimica y Bioquimica","LABORATORY":"Centro de Metabolomica y Bioanalisis (CEMBIO)","LAST_NAME":"Fernandez Garcia","FIRST_NAME":"Miguel","ADDRESS":"Facultad de Farmacia, Universidad San Pablo-CEU, CEU Universities, Urbanización Montepríncipe, 28660 Boadilla del Monte, Spain","EMAIL":"miguel.fernandezgarcia@ceu.es","PHONE":"+34690090778","NUM_GROUPS":"3","TOTAL_SUBJECTS":"13","NUM_MALES":"6","NUM_FEMALES":"7"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","GENOTYPE_STRAIN":"C57BL/6","AGE_OR_AGE_RANGE":"8-10 weeks old","GENDER":"Male and female","ANIMAL_FEED":"ad libitum","ANIMAL_WATER":"ad libitum"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"Mouse #1",
"Sample ID":"CEMS_UW1",
"Factors":{"Pulmonary TB status":"Control - uninfected"},
"Additional sample data":{"Sex":"Female","Analytical platform":"CE-ESI(+)-TOF/MS","RAW_FILE_NAME":"CEMS_UW1.d"}
},
{
"Subject ID":"Mouse #2",
"Sample ID":"CEMS_UW2",
"Factors":{"Pulmonary TB status":"Control - uninfected"},
"Additional sample data":{"Sex":"Female","Analytical platform":"CE-ESI(+)-TOF/MS","RAW_FILE_NAME":"CEMS_UW2.d"}
},
{
"Subject ID":"Mouse #3",
"Sample ID":"CEMS_UW3",
"Factors":{"Pulmonary TB status":"Control - uninfected"},
"Additional sample data":{"Sex":"Male","Analytical platform":"CE-ESI(+)-TOF/MS","RAW_FILE_NAME":"CEMS_UW3.d"}
},
{
"Subject ID":"Mouse #4",
"Sample ID":"CEMS_UW4",
"Factors":{"Pulmonary TB status":"Control - uninfected"},
"Additional sample data":{"Sex":"Male","Analytical platform":"CE-ESI(+)-TOF/MS","RAW_FILE_NAME":"CEMS_UW4.d"}
},
{
"Subject ID":"Mouse #5",
"Sample ID":"CEMS_4W1",
"Factors":{"Pulmonary TB status":"4 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"CE-ESI(+)-TOF/MS","RAW_FILE_NAME":"CEMS_4W1.d"}
},
{
"Subject ID":"Mouse #6",
"Sample ID":"CEMS_4W2",
"Factors":{"Pulmonary TB status":"4 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"CE-ESI(+)-TOF/MS","RAW_FILE_NAME":"CEMS_4W2.d"}
},
{
"Subject ID":"Mouse #7",
"Sample ID":"CEMS_4W3",
"Factors":{"Pulmonary TB status":"4 weeks post-infection"},
"Additional sample data":{"Sex":"Male","Analytical platform":"CE-ESI(+)-TOF/MS","RAW_FILE_NAME":"CEMS_4W3.d"}
},
{
"Subject ID":"Mouse #8",
"Sample ID":"CEMS_4W4",
"Factors":{"Pulmonary TB status":"4 weeks post-infection"},
"Additional sample data":{"Sex":"Male","Analytical platform":"CE-ESI(+)-TOF/MS","RAW_FILE_NAME":"CEMS_4W4.d"}
},
{
"Subject ID":"Mouse #9",
"Sample ID":"CEMS_9W1",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"CE-ESI(+)-TOF/MS","RAW_FILE_NAME":"CEMS_9W1.d"}
},
{
"Subject ID":"Mouse #10",
"Sample ID":"CEMS_9W2",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"CE-ESI(+)-TOF/MS","RAW_FILE_NAME":"CEMS_9W2.d"}
},
{
"Subject ID":"Mouse #11",
"Sample ID":"CEMS_9W3",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"CE-ESI(+)-TOF/MS","RAW_FILE_NAME":"CEMS_9W3.d"}
},
{
"Subject ID":"Mouse #12",
"Sample ID":"CEMS_9W4",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Male","Analytical platform":"CE-ESI(+)-TOF/MS","RAW_FILE_NAME":"CEMS_9W4.d"}
},
{
"Subject ID":"Mouse #13",
"Sample ID":"CEMS_9W5",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Male","Analytical platform":"CE-ESI(+)-TOF/MS","RAW_FILE_NAME":"CEMS_9W5.d"}
},
{
"Subject ID":"Mouse #1",
"Sample ID":"GCMS_UW1",
"Factors":{"Pulmonary TB status":"Control - uninfected"},
"Additional sample data":{"Sex":"Female","Analytical platform":"GC-EI-QTOF/MS","RAW_FILE_NAME":"GCMS_UW1.d"}
},
{
"Subject ID":"Mouse #2",
"Sample ID":"GCMS_UW2",
"Factors":{"Pulmonary TB status":"Control - uninfected"},
"Additional sample data":{"Sex":"Female","Analytical platform":"GC-EI-QTOF/MS","RAW_FILE_NAME":"GCMS_UW2.d"}
},
{
"Subject ID":"Mouse #3",
"Sample ID":"GCMS_UW3",
"Factors":{"Pulmonary TB status":"Control - uninfected"},
"Additional sample data":{"Sex":"Male","Analytical platform":"GC-EI-QTOF/MS","RAW_FILE_NAME":"GCMS_UW3.d"}
},
{
"Subject ID":"Mouse #4",
"Sample ID":"GCMS_UW4",
"Factors":{"Pulmonary TB status":"Control - uninfected"},
"Additional sample data":{"Sex":"Male","Analytical platform":"GC-EI-QTOF/MS","RAW_FILE_NAME":"GCMS_UW4.d"}
},
{
"Subject ID":"Mouse #5",
"Sample ID":"GCMS_4W1",
"Factors":{"Pulmonary TB status":"4 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"GC-EI-QTOF/MS","RAW_FILE_NAME":"GCMS_4W1.d"}
},
{
"Subject ID":"Mouse #6",
"Sample ID":"GCMS_4W2",
"Factors":{"Pulmonary TB status":"4 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"GC-EI-QTOF/MS","RAW_FILE_NAME":"GCMS_4W2.d"}
},
{
"Subject ID":"Mouse #7",
"Sample ID":"GCMS_4W3",
"Factors":{"Pulmonary TB status":"4 weeks post-infection"},
"Additional sample data":{"Sex":"Male","Analytical platform":"GC-EI-QTOF/MS","RAW_FILE_NAME":"GCMS_4W3.d"}
},
{
"Subject ID":"Mouse #8",
"Sample ID":"GCMS_4W4",
"Factors":{"Pulmonary TB status":"4 weeks post-infection"},
"Additional sample data":{"Sex":"Male","Analytical platform":"GC-EI-QTOF/MS","RAW_FILE_NAME":"GCMS_4W4.d"}
},
{
"Subject ID":"Mouse #9",
"Sample ID":"GCMS_9W1",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"GC-EI-QTOF/MS","RAW_FILE_NAME":"GCMS_9W1.d"}
},
{
"Subject ID":"Mouse #10",
"Sample ID":"GCMS_9W2",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"GC-EI-QTOF/MS","RAW_FILE_NAME":"GCMS_9W2.d"}
},
{
"Subject ID":"Mouse #11",
"Sample ID":"GCMS_9W3",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"GC-EI-QTOF/MS","RAW_FILE_NAME":"GCMS_9W3.d"}
},
{
"Subject ID":"Mouse #12",
"Sample ID":"GCMS_9W4",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Male","Analytical platform":"GC-EI-QTOF/MS","RAW_FILE_NAME":"GCMS_9W4.d"}
},
{
"Subject ID":"Mouse #13",
"Sample ID":"GCMS_9W5",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Male","Analytical platform":"GC-EI-QTOF/MS","RAW_FILE_NAME":"GCMS_9W5.d"}
},
{
"Subject ID":"Mouse #1",
"Sample ID":"LCMSpos_UW1",
"Factors":{"Pulmonary TB status":"Control - uninfected"},
"Additional sample data":{"Sex":"Female","Analytical platform":"LC-ESI(+)-QTOF/MS","RAW_FILE_NAME":"LCMSpos_UW1.d"}
},
{
"Subject ID":"Mouse #2",
"Sample ID":"LCMSpos_UW2",
"Factors":{"Pulmonary TB status":"Control - uninfected"},
"Additional sample data":{"Sex":"Female","Analytical platform":"LC-ESI(+)-QTOF/MS","RAW_FILE_NAME":"LCMSpos_UW2.d"}
},
{
"Subject ID":"Mouse #3",
"Sample ID":"LCMSpos_UW3",
"Factors":{"Pulmonary TB status":"Control - uninfected"},
"Additional sample data":{"Sex":"Male","Analytical platform":"LC-ESI(+)-QTOF/MS","RAW_FILE_NAME":"LCMSpos_UW3.d"}
},
{
"Subject ID":"Mouse #4",
"Sample ID":"LCMSpos_UW4",
"Factors":{"Pulmonary TB status":"Control - uninfected"},
"Additional sample data":{"Sex":"Male","Analytical platform":"LC-ESI(+)-QTOF/MS","RAW_FILE_NAME":"LCMSpos_UW4.d"}
},
{
"Subject ID":"Mouse #5",
"Sample ID":"LCMSpos_4W1",
"Factors":{"Pulmonary TB status":"4 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"LC-ESI(+)-QTOF/MS","RAW_FILE_NAME":"LCMSpos_4W1.d"}
},
{
"Subject ID":"Mouse #6",
"Sample ID":"LCMSpos_4W2",
"Factors":{"Pulmonary TB status":"4 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"LC-ESI(+)-QTOF/MS","RAW_FILE_NAME":"LCMSpos_4W2.d"}
},
{
"Subject ID":"Mouse #7",
"Sample ID":"LCMSpos_4W3",
"Factors":{"Pulmonary TB status":"4 weeks post-infection"},
"Additional sample data":{"Sex":"Male","Analytical platform":"LC-ESI(+)-QTOF/MS","RAW_FILE_NAME":"LCMSpos_4W3.d"}
},
{
"Subject ID":"Mouse #8",
"Sample ID":"LCMSpos_4W4",
"Factors":{"Pulmonary TB status":"4 weeks post-infection"},
"Additional sample data":{"Sex":"Male","Analytical platform":"LC-ESI(+)-QTOF/MS","RAW_FILE_NAME":"LCMSpos_4W4.d"}
},
{
"Subject ID":"Mouse #9",
"Sample ID":"LCMSpos_9W1",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"LC-ESI(+)-QTOF/MS","RAW_FILE_NAME":"LCMSpos_9W1.d"}
},
{
"Subject ID":"Mouse #10",
"Sample ID":"LCMSpos_9W2",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"LC-ESI(+)-QTOF/MS","RAW_FILE_NAME":"LCMSpos_9W2.d"}
},
{
"Subject ID":"Mouse #11",
"Sample ID":"LCMSpos_9W3",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"LC-ESI(+)-QTOF/MS","RAW_FILE_NAME":"LCMSpos_9W3.d"}
},
{
"Subject ID":"Mouse #12",
"Sample ID":"LCMSpos_9W4",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Male","Analytical platform":"LC-ESI(+)-QTOF/MS","RAW_FILE_NAME":"LCMSpos_9W4.d"}
},
{
"Subject ID":"Mouse #13",
"Sample ID":"LCMSpos_9W5",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Male","Analytical platform":"LC-ESI(+)-QTOF/MS","RAW_FILE_NAME":"LCMSpos_9W5.d"}
},
{
"Subject ID":"Mouse #1",
"Sample ID":"LCMSneg_UW1",
"Factors":{"Pulmonary TB status":"Control - uninfected"},
"Additional sample data":{"Sex":"Female","Analytical platform":"LC-ESI(-)-QTOF/MS","RAW_FILE_NAME":"LCMSneg_UW1.d"}
},
{
"Subject ID":"Mouse #2",
"Sample ID":"LCMSneg_UW2",
"Factors":{"Pulmonary TB status":"Control - uninfected"},
"Additional sample data":{"Sex":"Female","Analytical platform":"LC-ESI(-)-QTOF/MS","RAW_FILE_NAME":"LCMSneg_UW2.d"}
},
{
"Subject ID":"Mouse #3",
"Sample ID":"LCMSneg_UW3",
"Factors":{"Pulmonary TB status":"Control - uninfected"},
"Additional sample data":{"Sex":"Male","Analytical platform":"LC-ESI(-)-QTOF/MS","RAW_FILE_NAME":"LCMSneg_UW3.d"}
},
{
"Subject ID":"Mouse #4",
"Sample ID":"LCMSneg_UW4",
"Factors":{"Pulmonary TB status":"Control - uninfected"},
"Additional sample data":{"Sex":"Male","Analytical platform":"LC-ESI(-)-QTOF/MS","RAW_FILE_NAME":"LCMSneg_UW4.d"}
},
{
"Subject ID":"Mouse #5",
"Sample ID":"LCMSneg_4W1",
"Factors":{"Pulmonary TB status":"4 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"LC-ESI(-)-QTOF/MS","RAW_FILE_NAME":"LCMSneg_4W1.d"}
},
{
"Subject ID":"Mouse #6",
"Sample ID":"LCMSneg_4W2",
"Factors":{"Pulmonary TB status":"4 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"LC-ESI(-)-QTOF/MS","RAW_FILE_NAME":"LCMSneg_4W2.d"}
},
{
"Subject ID":"Mouse #7",
"Sample ID":"LCMSneg_4W3",
"Factors":{"Pulmonary TB status":"4 weeks post-infection"},
"Additional sample data":{"Sex":"Male","Analytical platform":"LC-ESI(-)-QTOF/MS","RAW_FILE_NAME":"LCMSneg_4W3.d"}
},
{
"Subject ID":"Mouse #8",
"Sample ID":"LCMSneg_4W4",
"Factors":{"Pulmonary TB status":"4 weeks post-infection"},
"Additional sample data":{"Sex":"Male","Analytical platform":"LC-ESI(-)-QTOF/MS","RAW_FILE_NAME":"LCMSneg_4W4.d"}
},
{
"Subject ID":"Mouse #9",
"Sample ID":"LCMSneg_9W1",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"LC-ESI(-)-QTOF/MS","RAW_FILE_NAME":"LCMSneg_9W1.d"}
},
{
"Subject ID":"Mouse #10",
"Sample ID":"LCMSneg_9W2",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"LC-ESI(-)-QTOF/MS","RAW_FILE_NAME":"LCMSneg_9W2.d"}
},
{
"Subject ID":"Mouse #11",
"Sample ID":"LCMSneg_9W3",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Female","Analytical platform":"LC-ESI(-)-QTOF/MS","RAW_FILE_NAME":"LCMSneg_9W3.d"}
},
{
"Subject ID":"Mouse #12",
"Sample ID":"LCMSneg_9W4",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Male","Analytical platform":"LC-ESI(-)-QTOF/MS","RAW_FILE_NAME":"LCMSneg_9W4.d"}
},
{
"Subject ID":"Mouse #13",
"Sample ID":"LCMSneg_9W5",
"Factors":{"Pulmonary TB status":"9 weeks post-infection"},
"Additional sample data":{"Sex":"Male","Analytical platform":"LC-ESI(-)-QTOF/MS","RAW_FILE_NAME":"LCMSneg_9W5.d"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Both male and female age-matched C57BL/6 mice (8-10 weeks old) were infected with Mtb H37Rv in animal BSL-3 laboratory and monitored with food and water ad libitum. Mice were sacrificed by anesthesia with isoflurane followed by gentle cervical dislocation as approved by institutional Animal Protocol Number (APN): 08591. Mice experimental procedures were approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Alabama at Birmingham, USA. For mice studies, we adhered as per national/international regulation of “Public Health Service Policy on Humane Care and Use of Laboratory Animals” (NIH), and “Animal Welfare Act and Animal Welfare Regulations” (USDA). Mouse genotype was confirmed by PCR and Western blotting. Mtb H37Rv was grown at 37 °C with shaking in BD Difco Middlebrook 7H9 media supplemented with 0.2 % glycerol, ADS (Albumin, Dextrose, NaCl) with 0.02 % tyloxapol. Mice were infected with 5 x 104 Mtb H37Rv via the intratracheal route. Lungs were collected from uninfected (n = 4), and Mtb-infected mice at 4- and 9-weeks post-infection (n = 4 and n = 5, respectively), and stored immediately at -80 °C for further processing and metabolite extraction.","SAMPLE_TYPE":"Lung","STORAGE_CONDITIONS":"-80℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"Mice were infected with 5 x 104 Mtb H37Rv via the intratracheal route and zero timepoint and monitored for disease progression until lung collection at 4 or 9 weeks post-infection","TREATMENT":"Infection","TREATMENT_ROUTE":"Intratracheal","TREATMENT_DOSE":"5 x 10^4 Mtb H37Rv"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"After lung collection, 1 mL of 50 % methanol was added to 100 mg of Mtb-infected or uninfected lung tissue and homogenized in a dounce homogenizer to prepare a uniform suspension. For CE-TOF/MS, 200 uL of homogenate were mixed with 200 L of 0.2 M formic acid and vortexed for 2 min. The samples were cleared by centrifugation at 16000 x g for 10 min at 4 °C and the supernatant was filter-sterilized using 0.22 um spin-X columns (Sigma). For GC-QTOF/MS and LC-QTOF/MS, 200 uL of each sample homogenate were mixed with 800 L of 80:20 methanol:MTBE (Methyl tert-butyl ether) and vortexed for 2 min. Metabolites were then extracted for 1 h with shaking at room temperature and then centrifuged at 4000 x g at 20 °C for 20 min. Supernatants were sterile filtered using 0.22 um Spin-X columns. All samples were passed through a Millipore filter (30-kDa cutoff) to remove large proteins. Samples were dried under high vacuum and stored at -80 °C until further platform-specific processing and analysis. For CE-ESI(+)-TOF/MS analysis, The dried samples were resuspended in Milli-Q water containing 0.1 mM formic acid and 0.2 mM methionine sulfone (internal standard) (Sigma-Aldrich, Germany) by vortexing for 1 min. After subsequent centrifugation (12600 x g, 15 min), the resulting clear solution was analyzed. For GC-QTOF/MS analysis, The above described dried samples were re-suspended in 450 µL of MeOH:H2O:MTBE (74:10:16) and after centrifugation at 12600 x g, 15 min at 4 °C, the supernatant was transferred to a vial with insert and evaporated to dryness under high vacuum. The obtained dried extracts were derivatized by a MPS autosampler for GC/MS analysis as previously described (Fiehn O., 2006). For LC-QTOF/MS analysis, The above described dried samples were resuspended in 200 µL of methanol:water:MTBE (7.4:1:1.6), vortexed for 1.5 h and centrifuged (4000 x g, 10 min, 4 °C). Clear solutions were analyzed.","SAMPLE_DERIVATIZATION":"In GC-EI-QTOF/MS analyses, Briefly, aldehyde and keto groups were first converted to O-methyloximes by reaction with 10 µL pyridine containing 15 mg/mL O-methoxyamine (Sigma-Aldrich, Germany) for 60 min at 70 °C. In a second step, acid hydrogen-containing metabolites were trimethylsilylated by reaction with 10 µL N,O-Bis(trimethylsilyl)trifluoroacetamide (BSTFA) (Sigma-Aldrich, Germany) to enhance the GC/MS metabolite coverage"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"The separation occurred in a fused-silica capillary (Agilent Technologies) (total length, 96 cm; i.d., 50 μm) under normal polarity with a background electrolyte containing 1.0 M formic acid in 10 % (v/v) methanol at 20 °C. Sheath liquid (6 µL·min-1) was methanol/water (1/1, v/v) containing 1.0 mM formic acid with two reference masses to allow correction and high mass resolution in the MS. Samples were hydrodynamically injected at 50 mbar for 35 s and stacked by injecting background electrolyte at 100 mbar for 10 s.","CHROMATOGRAPHY_TYPE":"CE","INSTRUMENT_NAME":"Agilent 7100 CE","COLUMN_NAME":"Agilent bare-fused silica capillary (96 cm x 50 um i.d.)"},

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

"MS":{"INSTRUMENT_NAME":"Agilent 6224 TOF","INSTRUMENT_TYPE":"TOF","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"The optimized MS parameters were: fragmentor 125 V, Skimmer 65 V, octopole 750 V, nebulizer pressure 10 psi, drying gas temperature at 200 °C and flow rate 10.0 L·min-1. The capillary voltage was 3500 V. Data were acquired in positive electrospray ionization (ESI) mode with a full scan from m/z 50 to 1000 at a rate of 1 spectra·s-1. The resulting CE-TOF/MS data files were cleaned of background noise and unrelated ions by the Batch Recursive Feature Extraction tool with Agilent MassHunter Profinder version B.06.00 software. Data were extracted using a data mining algorithm based on the software.","CAPILLARY_VOLTAGE":"3.5 kV","DRY_GAS_FLOW":"10 L·min-1","OCTPOLE_VOLTAGE":"750 V","MS_RESULTS_FILE":"ST001328_AN002214_Results.txt UNITS:Abundance Has m/z:Neutral masses Has RT:Yes RT units:Minutes"}

}