#METABOLOMICS WORKBENCH Deepshikha_20211013_204226 DATATRACK_ID:2888 STUDY_ID:ST001956 ANALYSIS_ID:AN003188 PROJECT_ID:PR001243 VERSION 1 CREATED_ON October 29, 2021, 11:50 am #PROJECT PR:PROJECT_TITLE Footprint dynamics study PR:PROJECT_TYPE GC-MS quantitative analysis PR:PROJECT_SUMMARY Data analysis at three time points- exponential, early and late stationary phase PR:PROJECT_SUMMARY uncovered dynamic metabolite abundance implying metabolic rewiring of Rubrivivax PR:PROJECT_SUMMARY benzoatilyticus JA2 cells, in response to glucose. To study dynamic changes in PR:PROJECT_SUMMARY the metabolome, footprint analysis (exometabolome extracted from the spent media PR:PROJECT_SUMMARY of glucose grown Rubrivivax benzoatilyticus cells), using GC-MS, was carried out PR:PROJECT_SUMMARY at three time points- exponential phase (G3), early (G9) and late (G18) PR:PROJECT_SUMMARY stationary phase. Metabolites were extracted in methanol, derivatized by adding PR:PROJECT_SUMMARY BSTFA-TCMS to protect the functional groups and analysed by GC-MS. The analysis PR:PROJECT_SUMMARY listed metabolic features at each time point, of which 149 metabolites were PR:PROJECT_SUMMARY identified, based on the mass spectra comparison in the database (NIST PR:PROJECT_SUMMARY similarity >700, Golm database), at one and/or other time point, while other PR:PROJECT_SUMMARY metabolites remained unidentified. Identified metabolic features along with PR:PROJECT_SUMMARY their respective peak area at G3, G9 and G18 were recorded and submitted to PR:PROJECT_SUMMARY MetaboAnalyst 4.0 online software to identify significant metabolic pattern and PR:PROJECT_SUMMARY variation. The result of the Hierarchical Clustering Analysis (HCA) shows that PR:PROJECT_SUMMARY metabolites clustered into five groups based on the response pattern specifying PR:PROJECT_SUMMARY the metabolic dissimilarity between the three samples. Group I, II and V PR:PROJECT_SUMMARY comprises metabolites with high concentration in G18, G9 and G3 samples PR:PROJECT_SUMMARY respectively, group III and IV includes metabolites whose concentration was high PR:PROJECT_SUMMARY in two of the three samples. Pairwise score plot of principal component analysis PR:PROJECT_SUMMARY (PCA) provided an overview of the separation pattern amongst the most PR:PROJECT_SUMMARY significant principal components (PCs). To assess the significance of class PR:PROJECT_SUMMARY discrimination, partial least squares - discriminant analysis (PLS-DA) was PR:PROJECT_SUMMARY performed. The exometabolome samples were seen clearly separated by PLS-DA PR:PROJECT_SUMMARY analysis with the R2 and Q2 value of 0.95 and 0.4 respectively indicating the PR:PROJECT_SUMMARY goodness of fit and predictability, suggesting representative model for the PR:PROJECT_SUMMARY difference in metabolomes. The Variable Importance in Projection (VIP scores) PR:PROJECT_SUMMARY derived from PLS-DA model was used to ascertain key metabolic features PR:PROJECT_SUMMARY significant for group separation. Metabolites with VIP score >1 were considered PR:PROJECT_SUMMARY to have statistically contributed to the model. Forty metabolites were PR:PROJECT_SUMMARY identified as statistically significant contributors to the model and were PR:PROJECT_SUMMARY mainly accountable for group separation in the model. Metabolites were PR:PROJECT_SUMMARY classified based on their chemical structure as alkanes (20%), sugars (28%), PR:PROJECT_SUMMARY organic acid (17%), amino acid (10%), fatty acid (8%), nucleotide (3%) and PR:PROJECT_SUMMARY others (5%). Amongst these forty metabolites, a total of 19, 25 and 33 were PR:PROJECT_SUMMARY detected in G3, G9 and G18 samples respectively. PR:INSTITUTE University of Hyderabad PR:DEPARTMENT Department of Plant sciences PR:LABORATORY Bacterial discovery and metabolomics laboratory PR:LAST_NAME Gupta PR:FIRST_NAME Deepshikha PR:ADDRESS Dept. of Plant Sciences, University of Hyderabad, Hyderabad, India. PR:EMAIL deepshikha@uohyd.ac.in PR:PHONE +918985420802 PR:FUNDING_SOURCE Department of Science and Technology, Government of India #STUDY ST:STUDY_TITLE Timecourse exometabolome analysis of glucose grown Rubrivivax benzoatilyticus ST:STUDY_TITLE cells ST:STUDY_TYPE Timecourse experiment ST:STUDY_SUMMARY Bacterial cells were grown on glucose under photoheterotrophic conditions for 18 ST:STUDY_SUMMARY days. Spent media of cells, harvested at 3rd, 9th and 18th day of growth, was ST:STUDY_SUMMARY vacuum dried and the metabolome was extracted in methanol. The extracted ST:STUDY_SUMMARY metabolites were derivatized and analyzed using GC-MS. ST:INSTITUTE University of Hyderabad ST:LAST_NAME Gupta ST:FIRST_NAME Deepshikha ST:ADDRESS Dept. of Plant Sciences, ST:EMAIL deepshikha@uohyd.ac.in ST:PHONE +918985420802 #SUBJECT SU:SUBJECT_TYPE Bacteria SU:SUBJECT_SPECIES Rubrivivax benzoatilyticus SU:TAXONOMY_ID 987059 SU:GENOTYPE_STRAIN JA2 SU:AGE_OR_AGE_RANGE 3, 9 and 18 days #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - G3-01 Label:1-Exponential phase | Days:3 SUBJECT_SAMPLE_FACTORS - G3-02 Label:1-Exponential phase | Days:3 SUBJECT_SAMPLE_FACTORS - G3-03 Label:1-Exponential phase | Days:3 SUBJECT_SAMPLE_FACTORS - G3-04 Label:1-Exponential phase | Days:3 SUBJECT_SAMPLE_FACTORS - G3-05 Label:1-Exponential phase | Days:3 SUBJECT_SAMPLE_FACTORS - G3-06 Label:1-Exponential phase | Days:3 SUBJECT_SAMPLE_FACTORS - G9-01 Label:2-Early stationary phase | Days:9 SUBJECT_SAMPLE_FACTORS - G9-02 Label:2-Early stationary phase | Days:9 SUBJECT_SAMPLE_FACTORS - G9-03 Label:2-Early stationary phase | Days:9 SUBJECT_SAMPLE_FACTORS - G9-04 Label:2-Early stationary phase | Days:9 SUBJECT_SAMPLE_FACTORS - G9-05 Label:2-Early stationary phase | Days:9 SUBJECT_SAMPLE_FACTORS - G9-06 Label:2-Early stationary phase | Days:9 SUBJECT_SAMPLE_FACTORS - G18-01 Label:3-Late stationary phase | Days:18 SUBJECT_SAMPLE_FACTORS - G18-02 Label:3-Late stationary phase | Days:18 SUBJECT_SAMPLE_FACTORS - G18-03 Label:3-Late stationary phase | Days:18 SUBJECT_SAMPLE_FACTORS - G18-04 Label:3-Late stationary phase | Days:18 SUBJECT_SAMPLE_FACTORS - G18-05 Label:3-Late stationary phase | Days:18 SUBJECT_SAMPLE_FACTORS - G18-06 Label:3-Late stationary phase | Days:18 #COLLECTION CO:COLLECTION_SUMMARY Cells were harvested by centrifugation and the spent media was used for the CO:COLLECTION_SUMMARY metabolome analysis. CO:SAMPLE_TYPE Bacterial cells #TREATMENT TR:TREATMENT_SUMMARY Bacterial cells were grown on D-glucose (22 mM) for 18 days under TR:TREATMENT_SUMMARY photoheterotrophic conditions. Cells were harvested at 3rd, 9th and 18th day of TR:TREATMENT_SUMMARY growth by centrifugation. The spent media of the bacterial cell culture was TR:TREATMENT_SUMMARY evaporated to dryness and used for the footprint analysis using GC-MS. TR:TREATMENT_COMPOUND D-glucose (dextrose) TR:TREATMENT_DOSE 22 mM TR:TREATMENT_DOSEDURATION 18 days #SAMPLEPREP SP:SAMPLEPREP_SUMMARY metabolites were extracted from dried culture supernatant in 1 ml methanol. SP:SAMPLEPREP_SUMMARY Methanol extract (100 µl) was vacuum dried. The dried exometabolome samples SP:SAMPLEPREP_SUMMARY were derivatized, to protect their functional groups, with 40 µl BSTFA SP:SAMPLEPREP_SUMMARY (N,O-bis(trimethylsilyl)trifluoroacetamide) and TMCS (trimethylchlorosilane); SP:SAMPLEPREP_SUMMARY incubated at 70 ○C for 4 h in dry bath and immediately analysed by GC-MS. SP:PROCESSING_STORAGE_CONDITIONS -20℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Derivatized sample (1 µl) was analysed using HP-5 column (30 m, thickness 0.25 CH:CHROMATOGRAPHY_SUMMARY µm) in spitless mode with helium as carrier gas at a constant flow of 1.5 CH:CHROMATOGRAPHY_SUMMARY ml.min-1. Oven temperature was initially held at 70○C for 2 min: ramped to 250 CH:CHROMATOGRAPHY_SUMMARY ○C by 10○C min-1; held for 1 min finally ramped to 280 ○C by 5○C min-1 CH:CHROMATOGRAPHY_SUMMARY and isocratic hold for 15 min (280 ○C). CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent 7890A CH:COLUMN_NAME Agilent HP5-MS (30m x 0.25mm, 0.25 um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 7890A MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE UNSPECIFIED MS:MS_COMMENTS Metabolic features were identified based on mass spectra comparison in the MS:MS_COMMENTS database (NIST similarity >700, Golm database). #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples G3-01 G3-02 G3-03 G3-04 G3-05 G3-06 G9-01 G9-02 G9-03 G9-04 G9-05 G9-06 G18-01 G18-02 G18-03 G18-04 G18-05 G18-06 Factors Label:1-Exponential phase | Days:3 Label:1-Exponential phase | Days:3 Label:1-Exponential phase | Days:3 Label:1-Exponential phase | Days:3 Label:1-Exponential phase | Days:3 Label:1-Exponential phase | Days:3 Label:2-Early stationary phase | Days:9 Label:2-Early stationary phase | Days:9 Label:2-Early stationary phase | Days:9 Label:2-Early stationary phase | Days:9 Label:2-Early stationary phase | Days:9 Label:2-Early stationary phase | Days:9 Label:3-Late stationary phase | Days:18 Label:3-Late stationary phase | Days:18 Label:3-Late stationary phase | Days:18 Label:3-Late stationary phase | Days:18 Label:3-Late stationary phase | Days:18 Label:3-Late stationary phase | Days:18 Label 1-Exponential phase 1-Exponential phase 1-Exponential phase 1-Exponential phase 1-Exponential phase 1-Exponential phase 2-Early stationary phase 2-Early stationary phase 2-Early stationary phase 2-Early stationary phase 2-Early stationary phase 2-Early stationary phase 3-Late stationary phase 3-Late stationary phase 3-Late stationary phase 3-Late stationary phase 3-Late stationary phase 3-Late stationary phase Days 3 3 3 3 3 3 9 9 9 9 9 9 18 18 18 18 18 18 1,2-Bishydroxyethane 23810737 26616571 23229295 21311105 47678484 23893807 36128680 35203886 48417692 24278008 34688673 22235388 36810506 35620346 23131458 31040848 37705504 27188723 11-Methyldodecanol 0 0 0 0 0 0 455572 0 252463 1884654 0 0 764721 0 0 0 0 0 1-Hexene, 3,5,5-trimethyl- 0 0 0 0 0 1149674 0 0 0 0 0 0 0 0 0 0 0 0 1-Hexene, 3,5-dimethyl- 0 0 0 0 0 59441 0 0 0 0 0 0 0 0 0 0 0 0 1-Hexene, 5-methyl- 0 0 0 470593 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1-hydroxy-2-aminoethane 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1140156 2815563 0 1-Iodo-2-methylundecane 3622506 1018634 0 0 0 0 0 0 426599 0 0 281255 631033 0 0 0 0 0 1-Nonene, 4,6,8-trimethyl- 25415197 33517149 11335548 17298670 36264533 7618196 34150455 40922886 10796307 33179119 52717448 30928006 24011623 35979825 6306352 27941085 20884622 46933655 1-Octene, 3,7-dimethyl- 3169562 2220564 4415526 0 5290301 0 4497036 0 2622697 0 8282173 0 10303496 0 2221688 3852048 3222087 0 2,3,4-Trihydroxybutyric acid deriv., 0 0 0 0 0 0 0 0 0 0 0 0 946305 1785126 0 0 0 922528 2,4,6,8-Tetramethyl-1-undecene 0 0 0 0 0 0 889269 0 0 0 0 0 0 0 0 0 0 0 2,4,6-Trimethyl-1-nonene 2617776 4598243 4946910 985515 5029394 0 0 4657552 0 0 380931 2616924 0 0 0 0 0 0 2,4a-Methanonaphthalen-74aH-one, 1,2,3,4,5,6-hexahydro-1,1,5,5-tetramethyl-, 2s-cis- 0 0 0 2498867 0 0 0 0 0 0 4384357 0 0 0 0 0 0 0 2-Butenedioic acid , ester 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 194431 2-Deoxy ribose O,O',O''-- 0 0 0 0 0 0 0 0 30142589 0 0 0 0 0 0 0 0 0 3,6-1,4-cyclohexadiene 0 0 0 0 264579 0 0 0 0 0 0 0 0 0 0 0 0 0 3,6--1,4-cyclohexadiene 3519.8 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3,6-Dioxa-2,7-disilaoctane, 2,2,4,7,7-pentamethyl- 0 1399794 0 2126216 3757987 0 9090055 5423556 3094273 557916 0 562039 4096533 5629001 2723137 5326441 17973450 10004154 3,7-Dioxa-2,8-disilanonan-5-one, 2,2,8,8-tetramethyl- 0 0 0 0 0 0 639278 0 0 0 0 0 0 117178 0 409695 148395 342454 3-Mannobiose, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 12950962 37291170 0 3-Octene, 2,6-dimethyl- 0 0 0 0 0 0 0 0 3699368 0 0 0 0 4476446 0 0 4284423 8231248 3-Pyridinecarboxylic acid, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetic acid, hydroxy-, 0 0 0 263537 0 0 1838176 1653122 0 0 0 818253 7626981 6975614 4843634 4213797 7489641 12272129 Ala-Gly 0 0 0 0 0 0 0 0 0 24278008 0 0 0 0 0 0 0 0 Azulene 65486461 0 0 64997646 68295367 0 0 0 0 0 70196207 0 0 0 0 0 0 77529684 Benzaldehyde, 4-propyl- 15355457 13599533 0 15467364 9633495 0 0 0 0 0 0 0 0 0 0 0 0 685284 Benzene, dichloro- 1112084 2449592 0 1321825 1061446 312293 350353 0 192020 121600 432029 0 503637 0 31677 280357 262296 59581 Benzoic acid 0 543262 18440 1375995 909080 2723.3 4865110 1347301 0 84044 390142 0 2621514 4695581 0 2508543 0 5327818 Benzoic acid, 2-hydroxy-, methyl ester 26352824 29265288 14051369 23536225 30999251 3604417 28873387 31189124 4482776 24714291 33801377 25883254 24606477 32028446 11230329 30736737 34553360 44634474 Benzoic acid, 3,5-bis1,1-dimethylethyl-4-hydroxy-, ethyl ester 0 0 0 10510272 0 0 0 0 0 0 0 0 16478721 0 0 14614961 0 0 Benzoic acid, 4-hydroxy-, 125423 1160550 425658 2116343 3550273 0 6560110 1717298 1069437 0 597327 232339 0 6602587 223463 410186 0 4729752 Bis-1,2-hydroxyethylene 0 0 0 0 0 0 750640 1558173 0 0 0 1543305 0 0 0 0 0 0 borate 954642 727776 0 159273 0 0 0 0 2532494 41334 0 210587 0 0 0 0 0 0 Butanedioic acid, ester 4042458 10104873 1618552 25119634 43713586 5824425 113308696 80302958 24957278 22372037 29143289 27685488 82501008 87527644 47261572 189941730 37201371 0 Butanedioic acid, hydroxy-, ester 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1693361 2355026 749600 Butanoic acid, 2-hydroxy-, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2152273 Butanoic acid, 3-hydroxy-, 0 0 0 0 0 0 226164 87313 0 0 0 0 0 0 0 403413 2115549 2183763 carbamate 0 24560335 27216680 25522209 0 22004479 37317258 0 26125907 33596432 37935685 0 0 0 23694202 32912828 39154790 24634547 D- Erythrofuranose, ether isomer 2 0 0 0 0 0 0 0 345058390 0 0 0 24619133 0 29714990 0 69736127 13026410 22952606 D-Allofuranose, ether 0 0 0 0 0 0 0 0 0 0 0 0 70964832 0 0 0 0 0 D-Altrose, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 17838672 0 D-Cellobiose, 0 0 0 0 0 0 535500 0 0 0 0 0 0 0 0 0 0 0 D-Citramalic acid, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Decane 4495202 0 4237720.6 249166 9245770 1086981 3299109.29 2510081 3076678 3173414 5102935 8367653.8 5791897 3946934 1555652 4536173 1370631 772685 Decane, 2,3,5,8-tetramethyl- 0 0 0 0 0 0 763663 0 0 996664 2995116 0 0 1273130 0 0 1285877 0 Decane, 6-ethyl-2-methyl- 0 0 116964 0 123328 22624 0 0 0 0 801950 0 924689 0 130934 0 0 0 Deoxyribopyranose, ether 0 0 0 0 0 0 0 67324512 0 0 0 23453591 0 0 0 0 0 0 D-Erythronic acid delta-lactone, ether 0 0 0 0 134458 0 292392 0 0 0 0 0 0 135277 0 1175994 0 3666196 D-Erythro-Pentofuranose, 2-deoxy- 0 458956 0 0 0 0 26978737 0 0 0 0 0 0 0 0 0 0 0 D-Erythro-Pentopyranose, 2-deoxy 0 0 0 0 0 0 32075092 0 0 0 0 0 0 0 0 163851722 0 0 d-Erythrotetrofuranose, 87832996 201341493 0 199018123 151961384 0 0 0 0 157239941 123508139 0 0 0 37382698 0 0 0 D-Fructofuranose, ether isomer 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 12461483 5376010 D-Glucose, 0 0 0 0 0 0 1055758 0 0 0 0 0 0 0 0 0 0 0 Dibutyl phthalate 66606679 274344 29456 0 39486774 0 0 0 0 0 888808 0 0 0 0 0 0 0 Diethyl Phthalate 97908219 87001775 49022452 110989869 47391451 0 135275281 269964249 32466037 68044918 0 110473153 153885856 37583658 16639576 112781530 0 95775062 DL-Phenylalanine 23810737 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 D-Lyxofuranose, 0 1370668 28845138 940761 0 0 142827 0 0 0 4080691 0 2437065 0 0 0 0 0 D-Mannopyranose, 0 0 0 0 0 27706486 0 0 0 60842949 0 0 0 0 0 0 0 0 Dodecane, 4,6-dimethyl- 5292986 1186598 0 315239 1012432 0 3083341 1044554 0 4622064 7685554 0 3422371 3416934 244989 2285873 3686413 395228 Dodecanoic acid, 0 0 0 0 4555.6 0 0 92646 0 0 0 0 0 0 0 0 0 0 Dodecanoic acid, 1-methylethyl ester 1907477 0 238659 0 278628 0 0 0 0 394100 0 0 0 1002161 0 0 0 0 Dodecanoic acid, propyl ester 0 0 0 1242104 0 0 0 0 0 0 0 0 0 0 0 76606 0 0 D-Ornithine 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 802087 d-Proline, N-methoxycarbonyl-, heptyl ester 0 0 0 15234443 0 0 0 0 0 0 0 0 0 21065481 0 0 0 0 d-Proline, N-methoxycarbonyl-, nonyl ester 0 0 0 0 0 0 0 0 17404930 0 0 0 0 0 0 0 0 0 d-Proline, N-methoxycarbonyl-, undecyl ester 0 0 0 0 18871454 0 0 0 0 0 21218461 0 0 0 0 0 0 0 D-Talofuranose, 35232392 67190183 16122826 93260473 191397991 13154894 0 0 125150080 22649803 26495533 143593033 1276188 181443907 18492658 0 0 0 D-Talopyranose, 0 191271391 0 104193224 0 57422792 38643230 12749500 0 0 0 16314544 0 0 23218657 45852166 595920 0 D-Turanose, 0 0 0 0 1210664 0 15896907 869070 0 0 0 0 0 0 1499148 0 0 0 D-Xylopyranose, 188058946 0 4268802 116263684 0 0 6228477 0 0 31509054 0 0 0 0 0 4211525 0 0 D-Xylopyranose, ether 0 0 0 0 0 0 12119719 0 0 0 19896087 0 0 174059737 0 0 0 0 D-Xylose, 0 0 59677464 0 0 0 0 0 0 0 0 0 0 0 0 0 1522653 0 D-Xylulose 0 0 0 0 539399 0 0 0 0 0 0 0 0 0 0 0 0 0 Eicosanoic acid, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 858341 Ethanedioic acid, ester 4083606 5277976 3775380 0 5500735 0 28465175 0 0 0 0 0 0 0 0 0 0 0 Ethanol, 0 0 0 15234443 0 0 0 0 17404930 0 21218461 0 0 0 94.281 0 0 0 Ethylamine 0 0 0 0 0 0 2275786 0 0 0 2332288 0 0 0 0 0 0 0 D-Galactopyranose, 0 50178267 0 0 0 0 0 0 0 0 0 0 4041721 0 0 0 0 2630070 Glucofuranoside, methyl -, D- 70557376 0 5009382 0 0 40707083 0 0 16616197 0 0 0 0 0 3115546 0 0 0 Glucopyranose, 0 0 50424909 0 0 13605863 783166 0 20358051 142595033 0 0 0 0 0 163851722 0 0 Glucopyranose, phosphate 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 884011 Glycerol, ether 3202042 363670 331704 299269 4926768 0 4975482 4186741 531846 597466 0 1914698 36474993 6142454 2814934 11554757 9526669 26030342 Heptacosane 236142 0 0 0 2274111 749307 0 0 0 0 6891250 0 284802 0 0 4920545 8582716 14722876 Hexadecane 12102865 12473154 1696376 6732692 26573903 6696377 1700622 12665319 3417415 14527868 23695308 6618754 6487061 17678081 2139609 19364178.4 26377486 27094143 Hexadecanoic acid, 0 0 43109447 71622344 0 0 0 0 0 50960980 0 0 0 0 0 0 0 0 Hexane, 3,3-dimethyl- 0 78247 0 0 30928301 11885 0 500249 215843 219820 0 0 0 0 0 0 0 0 Hydroxyisobutyric acid 0 0 0 0 0 0 0 0 134784 0 0 0 0 0 0 0 0 0 Hydroxypyruvic acid, , 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 5871463 l-2-Aminobutyric acid, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 754792 L-2-Aminobutyric acid, N-dimethylaminomethylene-, butyl ester 0 0 0 0 0 0 0 0 0 0 0 0 0 0 116810 0 0 0 l-Alanine, 0 0 0 0 193419 0 35034637 22098425 785011 0 10775586 1176673 11452106 12446345 4592830 38073642 62900427 28795200 l-Aspartic acid, ester 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Levoglucosan, 0 15188913 0 28896371 13344079 14764901 0 0 0 45298913 24469675 0 73226937 0 0 0 93896358 0 l-Galactopyranoside, methyl 6-deoxy-2,3,4- 0 0 0 21042153 0 0 0 0 0 0 0 0 0 0 0 0 0 0 L-Isoleucine, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 23312492 L-Lactic acid, 0 0 0 0 0 0 0 0 0 0 0 66743184 0 0 0 0 0 0 l-Leucine, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1610410 0 3511001 L-Norleucine, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 L-Norvaline, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 476471 2993860 0 0 L-Valine, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3383677 5122738 Lys-Met-Arg 0 0 0 0 0 0 1679422 0 0 0 0 0 0 0 0 0 0 0 Lyxose, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 77798 0 584140 Malonic acid, bis2-ethyl ester 0 0 0 0 0 0 1354980 0 0 0 0 0 0 0 0 691044 0 0 Maltose, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3846369 Mannose, 6-deoxy,L- 0 0 0 0 0 0 0 1653743 0 0 0 0 0 0 0 10133610 20998765 0 Melibiose, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 829034 0 meso-Erythritol, ether 0 0 0 0 0 0 0 280253 0 0 0 0 0 0 0 0 16380651 52054047 N,O-valine 0 0 0 0 0 0 0 0 109.94 0 0 0 0 0 0 0 0 0 N-Acetyl-D-glucosamine, ether 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1013158 620058 Naphthalene 0 73584796 45672447 0 0 38168316 34740044 43931806 0 48748943 0 39638687 50307084 54915281 27856467 52633466 63308833 0 Nonadecane 8397951 9179007 539913 2903963 10605716 751094 0 3035580 122084 909861 9894997 554399 6406357 1863375 0 5804974 6907544 0 Nonane, 2,6-dimethyl- 0 0 2034696 0 0 3071738 2491190 3536824 2896512 0 0 0 2807544 4550048 1881479 4525203 6655142 4885289 Octadecanoic acid, 71566892 0 0 103483485 141070288 15882824 15331022 122655785 42985549 16472491 0 47853409 103142214 0 25665771 0 0 0 Octane, 2,3,6,7-tetramethyl- 0 0 0 59868 1188184 0 0 0 0 0 26436 0 0 0 1412197 0 1506657 0 Octane, 3,3-dimethyl- 1983274 1925530 2150024 0 3660527 0 2801935 3327060 0 1740629 2500073 0 2316184 372219 2546619 1288048 0 2652851 Octane, 4-ethyl- 289185 0 433660 994021 0 726130 1272118 661972 0 1298905 1139187 943220 911738 1047473 428073 818592 929053 0 Oxalic acid, allyl dodecyl ester 0 0 0 0 0 0 0 0 0 0 0 972766 0 0 0 0 0 0 Oxalic acid, allyl hexadecyl ester 0 0 0 0 0 811487 0 0 0 20583 0 0 198275 0 0 17205946 0 0 Oxalic acid, allyl octadecyl ester 0 0 0 0 0 0 0 0 0 0 0 0 0 222820 0 1170566 0 0 Oxalic acid, allyl pentadecyl ester 0 0 0 52019 0 0 0 0 0 0 0 0 398825 0 0 0 0 0 Oxalic acid, allyl tetradecyl ester 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 40430 0 Oxalic acid, allyl tridecyl ester 0 710122 0 0 0 184350 0 0 0 364174 0 1234842 0 0 1158523 0 0 221648 Oxalic acid, isobutyl nonyl ester 0 0 0 0 0 0 0 0 0 0 273180 0 0 0 0 0 0 0 Oxalic acid, isobutyl octyl ester 0 0 0 0 350612 0 0 0 0 0 0 0 0 0 0 0 0 0 Pentanoic acid, 3-methylbutyl ester 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1854703 0 0 Pentanoic acid, 4-methyl-2-hydroxy-, 0 0 0 0 0 0 0 0 0 0 0 0 0 62820 0 0 0 176350 Phytol 328704 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Propanetriol, 2-methyl-, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 460157 0 Propanoic acid, 2,3-bishydroxy-, 0 0 0 0 0 0 0 0 0 0 0 0 0 1001864 0 3015703 4009519 10528002 Propanoic acid, 2-hydroxy-, 62618197 72155888 39900657 114299212 160878916 49824101 131931010 155400195 102021813 63007693 75426435 66743184 130001048 133264244 74193024 119778354 42450316 40930269 Propanoic acid, 2-methyl-2,3-bishydroxy-, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Propanoic acid, 2-oxo-, 77481 0 0 0 0 0 0 0 0 0 0 0 653126 0 0 0 0 0 Pyrimidine, 5-methyl-2,4-bishydroxy- 0 0 0 0 0 0 1893944 2381398 0 0 0 0 486185 0 0 0 2554357 2820286 Ribitol, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 347065 7094327 0 S-4-Chlorophenyl 3-4-ethoxyphenyl-2-propenethioate 0 0 131376 0 0 19180 0 0 0 0 0 0 0 0 0 6994.3 0 0 Ser-Pro 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1644304 0 Silane, 1-cyclohexen-1-yloxytrimethyl- 4744482 5071821 13799186 5432268 4828088 12692719 22845151 8355515 5282907 22970846 0 16248092 22059982 23190040 12735313 14713460 18255280 5988621 Silane, 2-cyclohexen-1-yloxytrimethyl- 0 0 0 0 0 0 0 0 0 0 21035295 0 0 0 0 0 0 0 Silane, 2-furanylmethoxytrimethyl- 0 0 762428 410122 2584470 1386092 1921823 1225110 186388 1211633 1957686 0 1764649 1374603 0 1107650 1161099 0 Silane, cyclohexyloxytrimethyl- 3571272 5975517 5310551 4074084 3641084 5057880 8021817 6928786 5550095 5064146 6426377 5030769 6058554 6524869 4529887 5148606 6601126 3642490 Silanol, trimethyl-, carbonate 21 16703766 13394107 0 15234443 18871454 0 0 0 4885510 0 5539412 0 1964184 7300815 0 0 0 0 Silanol, trimethyl-, phosphate 31 152226993 26346399 17718201 0 17772631 0 3028356 0 0 0 0 0 0 0 3522723 22290528 0 0 Tetradecane, 1-iodo- 114731 0 0 0 0 0 0 0 0 0 796216 72108 274876 44823 0 0 0 277038 Tetradecanoic acid, 492268 5463094 405220 1714118 1306980 152183 35520908 0 0 163295 0 0 0 0 0 0 0 0 Tetrasiloxane, decamethyl- 0 0 201632 0 0 0 2769178 1469314 0 0 0 0 0 1971.2 0 65786 0 0 Tetratetracontane 0 0 0 0 0 0 1415901 0 0 1195981 1017880 250642 0 268339 0 787524 1993408 0 trehalose 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 26681284 Tridecane 0 1036333 602552 2503271 0 528250 0 0 696397 0 0 0 0 911343 5149402 0 1136706 0 Tridecane, 3-methyl- 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Trifluoromethyl-methyl ketone 88828963 0 0 0 107332851 0 70885821 83049454 0 17679071 65646527 50371695 0 0 0 74389241 1645352 0 Trishydroxymethylaminomethane, O,O',O''- ether 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 149819 Trisiloxane, octamethyl- 0 0 0 0 0 15461079 17143071 15466047 0 0 17954343 0 36575024 39852472 1468131 0 0 0 Undecane 0 0 0 0 0 0 0 112290 0 1519499 0 1003920 0 0 0 1854703 0 0 Undecane, 4,7-dimethyl- 136224 4429968 0 0 0 14891 0 0 54151 1780255 0 0 21440103 574675 0 0 0 0 Urea 0 0 2034696 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Label Days 1,2-Bishydroxyethane 11-Methyldodecanol 1-Hexene, 3,5,5-trimethyl- 1-Hexene, 3,5-dimethyl- 1-Hexene, 5-methyl- 1-hydroxy-2-aminoethane 1-Iodo-2-methylundecane 1-Nonene, 4,6,8-trimethyl- 1-Octene, 3,7-dimethyl- 2,3,4-Trihydroxybutyric acid deriv., 2,4,6,8-Tetramethyl-1-undecene 2,4,6-Trimethyl-1-nonene 2,4a-Methanonaphthalen-74aH-one, 1,2,3,4,5,6-hexahydro-1,1,5,5-tetramethyl-, 2s-cis- 2-Butenedioic acid , ester 2-Deoxy ribose O,O',O''-- 3,6-1,4-cyclohexadiene 3,6--1,4-cyclohexadiene 3,6-Dioxa-2,7-disilaoctane, 2,2,4,7,7-pentamethyl- 3,7-Dioxa-2,8-disilanonan-5-one, 2,2,8,8-tetramethyl- 3-Mannobiose, 3-Octene, 2,6-dimethyl- 3-Pyridinecarboxylic acid, Acetic acid, hydroxy-, Ala-Gly Azulene Benzaldehyde, 4-propyl- Benzene, dichloro- Benzoic acid Benzoic acid, 2-hydroxy-, methyl ester Benzoic acid, 3,5-bis1,1-dimethylethyl-4-hydroxy-, ethyl ester Benzoic acid, 4-hydroxy-, Bis-1,2-hydroxyethylene borate Butanedioic acid, ester Butanedioic acid, hydroxy-, ester Butanoic acid, 2-hydroxy-, Butanoic acid, 3-hydroxy-, carbamate D- Erythrofuranose, ether isomer 2 D-Allofuranose, ether D-Altrose, D-Cellobiose, D-Citramalic acid, Decane Decane, 2,3,5,8-tetramethyl- Decane, 6-ethyl-2-methyl- Deoxyribopyranose, ether D-Erythronic acid delta-lactone, ether D-Erythro-Pentofuranose, 2-deoxy- D-Erythro-Pentopyranose, 2-deoxy d-Erythrotetrofuranose, D-Fructofuranose, ether isomer 1 D-Glucose, Dibutyl phthalate Diethyl Phthalate DL-Phenylalanine D-Lyxofuranose, D-Mannopyranose, Dodecane, 4,6-dimethyl- Dodecanoic acid, Dodecanoic acid, 1-methylethyl ester Dodecanoic acid, propyl ester D-Ornithine d-Proline, N-methoxycarbonyl-, heptyl ester d-Proline, N-methoxycarbonyl-, nonyl ester d-Proline, N-methoxycarbonyl-, undecyl ester D-Talofuranose, D-Talopyranose, D-Turanose, D-Xylopyranose, D-Xylopyranose, ether D-Xylose, D-Xylulose Eicosanoic acid, Ethanedioic acid, ester Ethanol, Ethylamine D-Galactopyranose, Glucofuranoside, methyl -, D- Glucopyranose, Glucopyranose, phosphate Glycerol, ether Heptacosane Hexadecane Hexadecanoic acid, Hexane, 3,3-dimethyl- Hydroxyisobutyric acid Hydroxypyruvic acid, , l-2-Aminobutyric acid, L-2-Aminobutyric acid, N-dimethylaminomethylene-, butyl ester l-Alanine, l-Aspartic acid, ester Levoglucosan, l-Galactopyranoside, methyl 6-deoxy-2,3,4- L-Isoleucine, L-Lactic acid, l-Leucine, L-Norleucine, L-Norvaline, L-Valine, Lys-Met-Arg Lyxose, Malonic acid, bis2-ethyl ester Maltose, Mannose, 6-deoxy,L- Melibiose, meso-Erythritol, ether N,O-valine N-Acetyl-D-glucosamine, ether Naphthalene Nonadecane Nonane, 2,6-dimethyl- Octadecanoic acid, Octane, 2,3,6,7-tetramethyl- Octane, 3,3-dimethyl- Octane, 4-ethyl- Oxalic acid, allyl dodecyl ester Oxalic acid, allyl hexadecyl ester Oxalic acid, allyl octadecyl ester Oxalic acid, allyl pentadecyl ester Oxalic acid, allyl tetradecyl ester Oxalic acid, allyl tridecyl ester Oxalic acid, isobutyl nonyl ester Oxalic acid, isobutyl octyl ester Pentanoic acid, 3-methylbutyl ester Pentanoic acid, 4-methyl-2-hydroxy-, Phytol Propanetriol, 2-methyl-, Propanoic acid, 2,3-bishydroxy-, Propanoic acid, 2-hydroxy-, Propanoic acid, 2-methyl-2,3-bishydroxy-, Propanoic acid, 2-oxo-, Pyrimidine, 5-methyl-2,4-bishydroxy- Ribitol, S-4-Chlorophenyl 3-4-ethoxyphenyl-2-propenethioate Ser-Pro Silane, 1-cyclohexen-1-yloxytrimethyl- Silane, 2-cyclohexen-1-yloxytrimethyl- Silane, 2-furanylmethoxytrimethyl- Silane, cyclohexyloxytrimethyl- Silanol, trimethyl-, carbonate 21 Silanol, trimethyl-, phosphate 31 Tetradecane, 1-iodo- Tetradecanoic acid, Tetrasiloxane, decamethyl- Tetratetracontane trehalose Tridecane Tridecane, 3-methyl- Trifluoromethyl-methyl ketone Trishydroxymethylaminomethane, O,O',O''- ether Trisiloxane, octamethyl- Undecane Undecane, 4,7-dimethyl- Urea METABOLITES_END #END