#METABOLOMICS WORKBENCH shenshuyan_20250918_181231 DATATRACK_ID:6452 STUDY_ID:ST004294 ANALYSIS_ID:AN007140 PROJECT_ID:PR002711 VERSION 1 CREATED_ON October 15, 2025, 10:37 am #PROJECT PR:PROJECT_TITLE Investigation on the spaceflight-induced phenotypic changes and potential PR:PROJECT_TITLE mechanisms of Bacillus Licheniformis PR:PROJECT_SUMMARY Bacillus licheniformis (BL) is a common probiotic clinically used to treat acute PR:PROJECT_SUMMARY and chronic enteritis and diarrhea. The present study aimed to investigate the PR:PROJECT_SUMMARY phenotypic changes and potential mechanisms of BL after a 6-month spaceflight PR:PROJECT_SUMMARY aboard the China Space Station based on characterization test, morphological PR:PROJECT_SUMMARY observation and multi-omics analysis. 6-month spaceflight delayed BL growth PR:PROJECT_SUMMARY curve, reduced its cell wall thickness, and impaired BL biofilm formation PR:PROJECT_SUMMARY ability and adhesion capacity to Caco-2 cells. BL viability at pH 9.0 was PR:PROJECT_SUMMARY decreased by 9.42%. In the 6-month spaceflight BL, proteomics revealed 164 of PR:PROJECT_SUMMARY differentially expressed proteins mainly involved on DNA damage, ATP binding, PR:PROJECT_SUMMARY helicase activity. Metabolomics identified 55 of significantly altered PR:PROJECT_SUMMARY metabolites, and whole-genome sequencing detected five mutated genes. After PR:PROJECT_SUMMARY 6-month spaceflight, physiological characteristics of BL were significantly PR:PROJECT_SUMMARY changed. Various proteins, metabolites and genes were dramatically altered as PR:PROJECT_SUMMARY well. These novel findings may provide new insights into adaptive evolution of PR:PROJECT_SUMMARY BL under long-term space environment, which is expected to benefit for further PR:PROJECT_SUMMARY clinical application of BL. PR:INSTITUTE Beijing Institute of Technology PR:LAST_NAME shen PR:FIRST_NAME shuyan PR:ADDRESS Beijing Institute of Technology, Zhongguancun Campus, beijing, China, 100081, PR:ADDRESS China PR:EMAIL 1819599261@qq.com PR:PHONE 18526251633 #STUDY ST:STUDY_TITLE Investigation on the spaceflight-induced phenotypic changes and potential ST:STUDY_TITLE mechanisms of Bacillus Licheniformis ST:STUDY_SUMMARY Bacillus licheniformis (BL) is a common probiotic clinically used to treat acute ST:STUDY_SUMMARY and chronic enteritis and diarrhea. The present study aimed to investigate the ST:STUDY_SUMMARY phenotypic changes and potential mechanisms of BL after a 6-month spaceflight ST:STUDY_SUMMARY aboard the China Space Station based on characterization test, morphological ST:STUDY_SUMMARY observation and multi-omics analysis. 6-month spaceflight delayed BL growth ST:STUDY_SUMMARY curve, reduced its cell wall thickness, and impaired BL biofilm formation ST:STUDY_SUMMARY ability and adhesion capacity to Caco-2 cells. BL viability at pH 9.0 was ST:STUDY_SUMMARY decreased by 9.42%. In the 6-month spaceflight BL, proteomics revealed 164 of ST:STUDY_SUMMARY differentially expressed proteins mainly involved on DNA damage, ATP binding, ST:STUDY_SUMMARY helicase activity. Metabolomics identified 55 of significantly altered ST:STUDY_SUMMARY metabolites, and whole-genome sequencing detected five mutated genes. After ST:STUDY_SUMMARY 6-month spaceflight, physiological characteristics of BL were significantly ST:STUDY_SUMMARY changed. Various proteins, metabolites and genes were dramatically altered as ST:STUDY_SUMMARY well. These novel findings may provide new insights into adaptive evolution of ST:STUDY_SUMMARY BL under long-term space environment, which is expected to benefit for further ST:STUDY_SUMMARY clinical application of BL. ST:INSTITUTE Beijing Institute of Technology ST:LAST_NAME shen ST:FIRST_NAME shuyan ST:ADDRESS Beijing Institute of Technology, Zhongguancun Campus, beijing, China, 100081, ST:ADDRESS China ST:EMAIL 1819599261@qq.com ST:PHONE 18526251633 #SUBJECT SU:SUBJECT_TYPE Bacteria SU:SUBJECT_SPECIES Bacillus licheniformis SU:TAXONOMY_ID 1402 #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 NEG-G1 NEG-G1 Sample source:Bacillus licheniformis | factor:ground RAW_FILE_NAME(Raw_FILE_NAME)=NEG-G1.raw SUBJECT_SAMPLE_FACTORS NEG-G2 NEG-G2 Sample source:Bacillus licheniformis | factor:ground RAW_FILE_NAME(Raw_FILE_NAME)=NEG-G2.raw SUBJECT_SAMPLE_FACTORS NEG-G3 NEG-G3 Sample source:Bacillus licheniformis | factor:ground RAW_FILE_NAME(Raw_FILE_NAME)=NEG-G3.raw SUBJECT_SAMPLE_FACTORS NEG-G4 NEG-G4 Sample source:Bacillus licheniformis | factor:ground RAW_FILE_NAME(Raw_FILE_NAME)=NEG-G4.raw SUBJECT_SAMPLE_FACTORS NEG-G5 NEG-G5 Sample source:Bacillus licheniformis | factor:ground RAW_FILE_NAME(Raw_FILE_NAME)=NEG-G5.raw SUBJECT_SAMPLE_FACTORS NEG-G6 NEG-G6 Sample source:Bacillus licheniformis | factor:ground RAW_FILE_NAME(Raw_FILE_NAME)=NEG-G6.raw SUBJECT_SAMPLE_FACTORS NEG-F1 NEG-F1 Sample source:Bacillus licheniformis | factor:space RAW_FILE_NAME(Raw_FILE_NAME)=NEG-F1.raw SUBJECT_SAMPLE_FACTORS NEG-F2 NEG-F2 Sample source:Bacillus licheniformis | factor:space RAW_FILE_NAME(Raw_FILE_NAME)=NEG-F2.raw SUBJECT_SAMPLE_FACTORS NEG-F3 NEG-F3 Sample source:Bacillus licheniformis | factor:space RAW_FILE_NAME(Raw_FILE_NAME)=NEG-F3.raw SUBJECT_SAMPLE_FACTORS NEG-F4 NEG-F4 Sample source:Bacillus licheniformis | factor:space RAW_FILE_NAME(Raw_FILE_NAME)=NEG-F4.raw SUBJECT_SAMPLE_FACTORS NEG-F5 NEG-F5 Sample source:Bacillus licheniformis | factor:space RAW_FILE_NAME(Raw_FILE_NAME)=NEG-F5.raw SUBJECT_SAMPLE_FACTORS NEG-F6 NEG-F6 Sample source:Bacillus licheniformis | factor:space RAW_FILE_NAME(Raw_FILE_NAME)=NEG-F6.raw SUBJECT_SAMPLE_FACTORS POS-G1 POS-G1 Sample source:Bacillus licheniformis | factor:ground RAW_FILE_NAME(Raw_FILE_NAME)=POS-G1.raw SUBJECT_SAMPLE_FACTORS POS-G2 POS-G2 Sample source:Bacillus licheniformis | factor:ground RAW_FILE_NAME(Raw_FILE_NAME)=POS-G2.raw SUBJECT_SAMPLE_FACTORS POS-G3 POS-G3 Sample source:Bacillus licheniformis | factor:ground RAW_FILE_NAME(Raw_FILE_NAME)=POS-G3.raw SUBJECT_SAMPLE_FACTORS POS-G4 POS-G4 Sample source:Bacillus licheniformis | factor:ground RAW_FILE_NAME(Raw_FILE_NAME)=POS-G4.raw SUBJECT_SAMPLE_FACTORS POS-G5 POS-G5 Sample source:Bacillus licheniformis | factor:ground RAW_FILE_NAME(Raw_FILE_NAME)=POS-G5.raw SUBJECT_SAMPLE_FACTORS POS-G6 POS-G6 Sample source:Bacillus licheniformis | factor:ground RAW_FILE_NAME(Raw_FILE_NAME)=POS-G6.raw SUBJECT_SAMPLE_FACTORS POS-F1 POS-F1 Sample source:Bacillus licheniformis | factor:space RAW_FILE_NAME(Raw_FILE_NAME)=POS-F1.raw SUBJECT_SAMPLE_FACTORS POS-F2 POS-F2 Sample source:Bacillus licheniformis | factor:space RAW_FILE_NAME(Raw_FILE_NAME)=POS-F2.raw SUBJECT_SAMPLE_FACTORS POS-F3 POS-F3 Sample source:Bacillus licheniformis | factor:space RAW_FILE_NAME(Raw_FILE_NAME)=POS-F3.raw SUBJECT_SAMPLE_FACTORS POS-F4 POS-F4 Sample source:Bacillus licheniformis | factor:space RAW_FILE_NAME(Raw_FILE_NAME)=POS-F4.raw SUBJECT_SAMPLE_FACTORS POS-F5 POS-F5 Sample source:Bacillus licheniformis | factor:space RAW_FILE_NAME(Raw_FILE_NAME)=POS-F5.raw SUBJECT_SAMPLE_FACTORS POS-F6 POS-F6 Sample source:Bacillus licheniformis | factor:space RAW_FILE_NAME(Raw_FILE_NAME)=POS-F6.raw #COLLECTION CO:COLLECTION_SUMMARY The spaceflight strain (SS) and ground strain (GS) were inoculated into 30 mL of CO:COLLECTION_SUMMARY LB liquid medium and cultured until the mid-exponential growth phase CO:COLLECTION_SUMMARY (OD₆₀₀=0.6). 300 µL of methanol was added in SS and GS suspension for CO:COLLECTION_SUMMARY protein precipitation. The supernatant was collected after centrifugation at CO:COLLECTION_SUMMARY 12000 rpm for 10 min at 4 °C. The supernatant of SS and GS was freeze-dried CO:COLLECTION_SUMMARY under vacuum. The powder was redissolved with 200 µL of acetonitrile: water CO:COLLECTION_SUMMARY (1:1, v/v), then centrifuged at 12,000 rpm for 5 min. CO:SAMPLE_TYPE Bacterial cells #TREATMENT TR:TREATMENT_SUMMARY Bacillus licheniformis from the ground group and the flight group flown in space TR:TREATMENT_SUMMARY for 6 months, respectively. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY 300 µL of methanol was added in spaceflight strain (SS) and ground strain (GS) SP:SAMPLEPREP_SUMMARY suspension for protein precipitation. The supernatant was collected after SP:SAMPLEPREP_SUMMARY centrifugation at 12000 rpm for 10 min at 4 °C. The supernatant of SS and GS SP:SAMPLEPREP_SUMMARY was freeze-dried under vacuum. The powder was redissolved with 200 µL of SP:SAMPLEPREP_SUMMARY acetonitrile: water (1:1, v/v), then centrifuged at 12,000 rpm for 5 min. The SP:SAMPLEPREP_SUMMARY ultra-high performance liquid chromatography (UHPLC) analysis (Thermo Fisher SP:SAMPLEPREP_SUMMARY Scientific, Q Exactive HF-X, China) was used for metabolites detection, and the SP:SAMPLEPREP_SUMMARY experimental quality was evaluated by quality control (QC) samples. SP:SAMPLEPREP_SUMMARY Chromatographic separation was carried out on an ACQUITY UPLC BEH C18 column SP:SAMPLEPREP_SUMMARY (2.1×100 mm, 1.7 µm, Thermo Fisher Scientific, America). The mobile phase flow SP:SAMPLEPREP_SUMMARY rate was set at 0.3 mL/min with the injection volume of 2 µL and the column SP:SAMPLEPREP_SUMMARY temperature was maintained at 45 ℃. The mobile phase consisted of 0.1% formic SP:SAMPLEPREP_SUMMARY acid in water (A) and acetonitrile (B). The linear elution program was set at SP:SAMPLEPREP_SUMMARY 0-0.5 min, 1% B; 0.5-2 min, 1%-50% B; 2-9 min, 50%-99% B; 9-10 min, 99% B; SP:SAMPLEPREP_SUMMARY 10-10.5 min, 99%-1% B and 10.5-13 min, 1% B. MS analysis was performed in SP:SAMPLEPREP_SUMMARY positive ion equipped with an electrospray spray ionization (ESI) source. MS SP:SAMPLEPREP_SUMMARY tuning parameters were set as follows: Spray voltage 3.9 kV, the capillary SP:SAMPLEPREP_SUMMARY temperature 320 ℃, the sampling cone 40 V, the source offset 80 V, the SP:SAMPLEPREP_SUMMARY desolvation temperature 450 ℃, the cone gas flow 50 L/h, the desolvation gas SP:SAMPLEPREP_SUMMARY flow 800 L/h. The data were imported into Masslynx V4.1 data analysis software SP:SAMPLEPREP_SUMMARY (Nonlinear Dynamics, Newcastle, UK) for peak alignment, picking, and SP:SAMPLEPREP_SUMMARY normalization, resulting in peak intensities for retention time (TR) and m/z SP:SAMPLEPREP_SUMMARY value. The obtained data were imported into SIMCA-P14.0 statistical software SP:SAMPLEPREP_SUMMARY (UMETRICS, Sweden) for statistical analysis. The criteria of P value was less SP:SAMPLEPREP_SUMMARY than 0.05 in One-way analysis of variance (ANOVA). S plots and VIP values were SP:SAMPLEPREP_SUMMARY used to assess the importance of each variable in class separation. The SP:SAMPLEPREP_SUMMARY unsupervised principal component analysis (PCA) and supervised orthogonal SP:SAMPLEPREP_SUMMARY partial least squares discriminant analysis (OPLS-DA) models were established. SP:SAMPLEPREP_SUMMARY To prevent overfitting, a 200 permutations test was conducted on the OPLS-DA SP:SAMPLEPREP_SUMMARY model. Different metabolites were screened out according to the variable SP:SAMPLEPREP_SUMMARY importance in the projection (VIP) > 1.5 in the OPLS-DA analysis, and the SP:SAMPLEPREP_SUMMARY criteria of P value < 0.05 in One-way ANOVA. Finally, those features were SP:SAMPLEPREP_SUMMARY further annotated by KEGG database to explore the most relevant the metabolic SP:SAMPLEPREP_SUMMARY pathways. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Ion pair CH:INSTRUMENT_NAME Thermo Fisher Scientific, Q Exactive HF-X, CH:COLUMN_NAME Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um) CH:SOLVENT_A 100% water; 0.1% formic acid CH:SOLVENT_B 100% acetonitrile CH:FLOW_GRADIENT The linear elution program was set at 0-0.5 min, 1% B; 0.5-2 min, 1%-50% B; 2-9 CH:FLOW_GRADIENT min, 50%-99% B; 9-10 min, 99% B; 10-10.5 min, 99%-1% B and 10.5-13 min, 1% B. CH:FLOW_RATE 0.3 mL/min CH:COLUMN_TEMPERATURE 60°C #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive HF-X Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS MS tuning parameters were set as follows: Spray voltage 3.9 kV, the capillary MS:MS_COMMENTS temperature 320 °C, the sampling cone 40 V, the source offset 80 V, the MS:MS_COMMENTS desolvation temperature 450 °C, the cone gas flow 50 L/h, the desolvation gas MS:MS_COMMENTS flow 800 L/h. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak area MS_METABOLITE_DATA_START Samples NEG-G1 NEG-G2 NEG-G3 NEG-G4 NEG-G5 NEG-G6 NEG-F1 NEG-F2 NEG-F3 NEG-F4 NEG-F5 NEG-F6 Factors Sample source:Bacillus licheniformis | factor:ground Sample source:Bacillus licheniformis | factor:ground Sample source:Bacillus licheniformis | factor:ground Sample source:Bacillus licheniformis | factor:ground Sample source:Bacillus licheniformis | factor:ground Sample source:Bacillus licheniformis | factor:ground Sample source:Bacillus licheniformis | factor:space Sample source:Bacillus licheniformis | factor:space Sample source:Bacillus licheniformis | factor:space Sample source:Bacillus licheniformis | factor:space Sample source:Bacillus licheniformis | factor:space Sample source:Bacillus licheniformis | factor:space (15Z)-9,12,13-Trihydroxy-15-octadecenoic acid_1 6071114.246 4872705.759 6447262.1 7253829.329 6710730.553 3897200.923 (E)-6-hydroxyoctadec-4-enoic acid 3799392.928 4329185.957 4205190.652 4613666.616 8879087.315 4850223.983 12-Hydroxydodecanoic acid 6534101.512 6984475.045 6646927.05 7186260.116 7960262.545 8279532.669 13S-hydroxy-9Z,11E,15Z-octadecatrienoic acid 3446558.94 3476527.835 3067199.636 2958492.234 3538674.111 3916815.787 16-Hydroxyhexadecanoic acid 3833628.813 3034391.397 3513400.529 2093384.346 4133958.943 4247604.393 2-(acetylamino)-4-(methylthio)butanoic acid_1 54066163.72 264003066.6 244728302.8 335315818.6 287301920.2 2195288.084 2-(acetylamino)-4-(methylthio)butanoic acid_2 295358395.9 203930916.1 26723710.14 29167102.97 34899367.21 158813148.9 2-(acetylamino)-4-(methylthio)butanoic acid_3 58721478.47 36473113.39 1167229.105 1160192.516 4401682.176 189176131.2 2,5-di-tert-Butylhydroquinone 41347228.68 26098919.12 37917464.67 36494941.77 39857203.64 25665124.92 2-Aminooctanedioic acid_1 136156431.6 144501032.5 136172930.8 139471175.7 150143083 163987156.8 2-Aminooctanedioic acid_2 70676341.39 76752033.86 93978259.32 88159436.16 82351339.45 57517969.88 2-Hydroxycaproic acid 27848119.52 66840727.49 155098847.5 168041344.5 166326549.4 21410352.73 2-Hydroxyvaleric acid 395864859 427100316.5 427567566.8 415438421.1 416990787.4 450003111.1 3-(Cyclohexylamino)-2-hydroxy-1-propanesulfonic acid 7225906.778 5992369.751 15092220.72 6100043.208 11318417.06 8491427.109 3,4-Dihydroxybenzenesulfonic acid 873430.2947 29729515.01 43035991.9 41064606.1 69212943.72 907958.7598 3,4-Dihydroxyphenylpropionic acid_1 386559912.1 405568172.4 367801809.4 365914289.2 372992824.2 433936655.3 3,4-Dihydroxyphenylpropionic acid_2 26787615.66 30811695.19 35198443.55 38194478.49 34529185.47 11052203.27 3-Hydroxy-3-methylglutaric acid 122080744.5 217660258 126877159 139158013.7 138858669.5 199313815.9 3-Hydroxydecanoic acid 24595622.58 24333504.94 23395578.39 32769067.27 25053397.22 27451736.93 4-{[3-(Diethylamino)propyl]amino}-4-oxobut-2-enoic acid 86186582.52 96477941.42 97946956.2 115324776.5 94512342.99 100280828.2 4-{[5-(6-Hydroxy-5,5,8a-trimethyl-2-methylenedecahydro-1-naphthalenyl)-3-methylpentyl]oxy}-4-oxobutanoic acid 66356983.52 117160798.6 81984274.33 107250423.9 121576002.6 144759253 4-Cyanoindole 4201465.854 4541220.059 3475606.725 5409679.787 5661188.501 4572684.318 4-Indolecarbaldehyde_1 202312363 197641451.9 88829898.52 176751645.3 198276871.7 156467568 4-Indolecarbaldehyde_2 78727998.08 109383335.2 93341770.07 110511035.9 115270770.9 132956264.9 4-Oxoproline_1 24636519469 24256137642 24948787889 26297832618 25960516551 23941394845 4-Oxoproline_2 1649331697 1639282726 1415097931 1594472325 1636522119 1427222773 5-(2-Methylpropyl)-3,6-dioxo-2-piperazinepropanoic acid_1 8196920737 7149218503 5987559516 6304794336 6669389779 10624165425 5-(2-Methylpropyl)-3,6-dioxo-2-piperazinepropanoic acid_2 1769675386 1826050417 2226201224 2195331026 2069465911 2136552334 5-(2-Methylpropyl)-3,6-dioxo-2-piperazinepropanoic acid_3 1946106955 1841012674 1605390793 1630030054 1854552173 2131457423 5-(2-Methylpropyl)-3,6-dioxo-2-piperazinepropanoic acid_4 69589029.5 1336398785 1272921147 1112095454 1020461656 85989431.79 5-(2-Methylpropyl)-3,6-dioxo-2-piperazinepropanoic acid_5 63322932.86 70824634.68 65943416.22 71359899.25 68638816.04 105169897.8 5-(2-Methylpropyl)-3,6-dioxo-2-piperazinepropanoic acid_6 3848678.535 68282337.48 102457971.4 98314645.26 91339504.83 10393744.84 5-(2-Methylpropyl)-3,6-dioxo-2-piperazinepropanoic acid_7 60363835.27 66396665.31 52372225.64 59705210.4 66998919.25 73114318.87 5-Chloro-4-fluoro-2-nitroaniline 66776156.09 70358411.8 6452834.302 10753028.93 11197088.51 4901797.739 5-Hydroxyindole-3-acetic acid 63423948.51 57319727.66 7435442.339 65978612.59 73150932.33 58777185.03 5-Hydroxytryptophan_1 160863371 154828957.9 138550515 134094806.4 143755124.5 189097690.7 5-Hydroxytryptophan_2 102834600.3 97261182.88 110038403.6 110086814.8 114410959.4 109955271 5-Hydroxytryptophan_3 91390429.03 103013882.4 104022909 105499496.8 92353235.97 88388702.2 5-Hydroxytryptophan_4 48129032.4 57155188.51 52242226.78 75767595.88 86315509.21 97617102.29 6-Hydroxycaproic acid_1 356934657.9 563000856.3 152926588.4 157988792.6 270559577.9 617521183.2 6-Hydroxycaproic acid_2 177923368 170221352.6 444763255 587034585.3 419299206.7 209704112.1 Adenine_1 294308237.7 295266656.6 272203431.4 281727772.9 252018890.8 334608339.7 Adenine_2 47376283.04 43128385.59 41141529.95 36979759.28 35325342.12 89578101.56 Adenosine 3'5'-cyclic monophosphate 1216065616 1185719527 1126330052 1221203288 1129579189 1345582660 Adenosine 5'-monophosphate_1 813360209.4 854616090.3 799096878.4 835095214 806743008.3 883467622.6 Adenosine 5'-monophosphate_2 313210616.4 309340164.6 345652795.8 317844374.5 301450648.3 364441468.6 Adenosine 5'-monophosphate_3 315129958.6 313601308.4 264257130 283728850.9 301450648.3 362015815.5 Adenosine 5'-monophosphate_4 287405176.7 280813417 251371151.2 245460720 265554999.4 333688609.1 Biotin 55579438.22 60061101.2 66009080.1 63015514.79 61692745.87 61631860.03 Capryloylglycine 244078668.8 250897298.2 243360654.9 253160333.1 260058816.4 244531039.7 Cholic acid 59525748.58 54091536.15 54876945.4 65114652.28 59428977.11 66499161.35 Citric acid_1 3697430041 3972734494 5793098094 5587067717 5130373902 1397345282 Citric acid_2 2126415947 2194097958 2265892723 2213570160 2264760562 2419695775 D-(+)-Mannose 159395016.3 165142735 165956973 153520044.4 179661993 69269001.37 D-(+)-Tryptophan_1 8363843465 8574080416 7399937635 6285392589 5115190469 9659995191 D-(+)-Tryptophan_2 6270201012 8574080416 1113111860 1330253378 2806202592 9657952556 Deoxycholic acid 3012695.992 3050438.385 6731196.128 6373389.211 3924143.052 4338624.108 Dibutyl phthalate 15105139.81 15776630.86 13197091.13 17474406.59 15982403.58 13494490.96 DL-β-Leucine 6568627428 6141131215 5481726138 5501591239 4199251414 7315259307 Dodecyl sulfate_1 1840449.744 516502700.8 367687303.8 1647659250 900116409.2 637728287 Dodecyl sulfate_2 406785915.1 504871142.9 356923287.2 1632163475 893401680.8 634945580.7 D-α-Hydroxyglutaric acid 221418185 234622911 229422238.3 247567179 246439052.6 187641400.2 Glutaric acid 42540747.52 45241641.96 44690659.32 44314661.91 51742055.21 36381674.85 Glycerol 3-phosphate 179934768.8 162827750.9 235583723.2 210351186.7 177749940.7 181947907.9 Glycocholic acid 14194813.44 13194650.24 12798647.98 13301389.37 12809594.6 10190070.98 Glycodeoxycholic acid 3794169.993 3243914.882 3553675.381 3715225.017 4232937.923 4831987.468 Guanine 503128167.7 508438126.2 507490770.4 490228584.6 494639936.3 547730792 Guanosine monophosphate (GMP)_1 740739937.2 797522216.3 811875687.4 903215901.7 743503418.9 849931291.6 Guanosine monophosphate (GMP)_2 510450464.9 546875623.9 522962970.8 565524976 517393841.4 509734955.5 Guanosine monophosphate (GMP)_3 154473561.5 128823051 132809409.5 135369423 138659175.7 129699051.3 Guanosine monophosphate (GMP)_4 114921886.4 81943451.36 46796933.98 34491316.61 125063737.9 112574601.8 Guanosine monophosphate (GMP)_5 108863127.8 101264352.5 108409840 108015697.8 119566355.2 116067054.7 Hexadecanedioic acid 4973825.391 5302241.249 6177702.215 6206496.356 5781015.979 5427446.659 Hypoxanthine 506199877.6 523454516.6 458719798.1 496658473.6 510024755.7 540541007.7 Indole-3-acetic acid_1 56012574.99 55755544.71 8948087.842 64875009.69 59360314.65 64396067.78 Indole-3-acetic acid_2 34984658.52 45229098.23 38244979.87 37724610.88 39774972.93 41319099.86 L-(-)-3-Phenyllactic acid 1719107401 1788929514 139650091.9 664356009.4 1869769129 1834147649 L-Aspartic acid 31341796.15 23024552.45 41488538.2 30261356.16 23876827.72 30895949.28 L-Dopa 64137384.16 55187499.92 75663762.89 50983118.7 74966644.73 82404749.31 L-Histidine_1 130801714 119394592.1 156646935.4 134542516.3 152339653.5 194943230.5 L-Histidine_2 84282709.09 106397987.6 96485709.84 99757929.92 104189490.5 102765575.9 L-Iditol 68509511.14 77103321.73 74978400.62 71706709.15 84692996.14 113702679.1 L-Phenylalanine_1 8607831776 7855893400 6095757732 6349785316 7512651950 9945508786 L-Phenylalanine_2 1341803261 1289774928 1023417621 1186417382 1834456125 1432468833 L-Threonic acid 128089337.2 130231626.7 124863708 116291906.4 108469888.2 173062771.6 L-Tyrosine_1 918225572.6 827820086.9 1097861159 823731941.8 1125256062 1426155644 L-Tyrosine_2 709956898.5 663876523.6 561283697.5 562086670.6 634577452.8 864421600.5 L-Tyrosine_3 99444827.25 95232399.86 107284216.2 108040705.4 94324234.05 104774831.9 Mevalonic acid 164133175.3 139691325.1 140021808.3 174476328.3 152258962.8 187209765.4 Myriocin 8153866.927 9184335.331 8417919.626 12240714.03 9002066.469 14226366.2 Myristyl sulfate 33278310.06 50572106.61 14784734.34 6230021.099 16972395.1 4109645.212 N1-tetrahydrofuran-2-ylmethyl-2-cyanoacetamide 57835378.94 47579971.48 49986442.16 51993411.49 50850363.07 64084340.01 N-Acetylalanine 44033664.66 48851726.39 104333032.5 59563090.07 62440998.87 84909277.01 N-Acetyl-D-alloisoleucine_1 677336864 486755298.9 306428638 462882560.1 488104977.9 699713711.1 N-Acetyl-D-alloisoleucine_2 651351460.8 484610156.5 512277107.5 509937783.7 486693358.8 667934118.2 N-Acetyl-D-alloisoleucine_3 242818527.9 237627771.1 127810829.2 208370326.2 212688546 226509102.3 N-Acetyl-D-alloisoleucine_4 233951072.9 223156572.4 217519949.8 239325645.6 219354077.4 54948969.18 N-Acetyl-DL-glutamic acid 482420650.3 488177910.9 489742834.8 507135405.6 471557139.5 474679378.6 N-Acetyl-L-phenylalanine 120088747.6 163183153.5 195449441.5 172414061.9 148693256.7 123190912.3 N-Acetyl-L-tyrosine_1 52714720.62 109756500.1 75203638 99758475.6 101218981.5 2224969.495 N-Acetyl-L-tyrosine_2 16054577.09 16530181.34 3231845.757 16516909.53 14148083.76 60034452.92 N-Acetylornithine 295387410.7 284311456 302699578.4 332471003.8 302957483.7 277989858.6 Nateglinide 7097817.942 6650744.323 11225124.07 6955803.078 73013.18266 8701883.542 N-Formylmethionine 59857014.32 58894438.83 57332359.44 65971384.27 61427596.66 47294169.18 Nicotinic acid 171852719.2 169615877.9 153871100.1 183090825.7 173661793.7 193323300 N-Isovalerylglycine 608364511.7 459831424.6 506493623.7 479394583.7 446567317.4 448952443.4 N-Isovalerylglycine_1 348340531.3 295916937.3 300017264.7 286610479.4 383231349 305396368.9 N-Isovalerylglycine_2 268787644.3 268750296.9 206626352.5 287487714.8 194995539.9 286550570.2 N-Tigloylglycine 387195207.4 418950115.6 358601774.8 345334869.6 363108765.1 348668601.5 O-Acetylserine 231358231.5 377968191.2 374358432.5 265779098.7 450501726.3 342746300.3 Ornithine_1 38898637.14 37307017.85 45026816.82 39819079.8 45697343.13 55721073.85 Ornithine_2 24537137.58 23639231.91 22950513.46 27134748.36 21547531.64 27123764.76 Pantothenic acid_1 956543402.2 1089455863 1033144815 1029733482 1044353232 2250865818 Pantothenic acid_2 111339510.6 110615669 32154882.6 26355296.68 103412162.8 140814944.6 Pantothenic acid_3 11724764.1 6672542.962 6638707.263 5555524.682 6154897.464 121289846.7 Phenylacetaldehyde_1 1708191830 1788929514 2210236843 1988062606 1845300296 1809048191 Phenylacetaldehyde_2 124745028.5 122106329.3 7124610.503 2411040.789 131993554.4 132714353.5 Porphobilinogen_1 7032408270 6480805355 6338162122 6438464765 6692628727 7915077753 Porphobilinogen_2 794810938.5 1022709180 724561118.5 718367191.5 867565704.6 2180620293 Porphobilinogen_3 875554220 929665718.8 759004717.4 788683601.2 935652794.9 804143885.5 Riboflavin 19265194.23 22387132.4 22989918.91 34208726.99 29474487.59 28233487.37 Salicylic acid 62615830.58 46318361.69 64420548.98 54810997.65 72053329.73 15840633.67 Succinic acid_1 7722441220 7423115767 7015994609 6957504083 6987890906 12974083496 Succinic acid_2 1284537767 1382113038 1596289995 1556152118 1479077064 1248451870 Succinic acid_3 1012396493 950169076.3 932926912.4 964619042.3 926904476.9 1061810450 Threonine 129940936.1 154077959.1 144581254.9 140680555.2 147195352.4 169176982.1 Thymidine 5'-monophosphate 44843508.65 47601423.2 45172707.73 41114040.06 41205796.38 50160950.27 trans-Cinnamic acid_1 339809064.8 311040655.6 235649676.9 243986379.8 303392275.8 393428000.5 trans-Cinnamic acid_2 167632052 161659368.3 205122372.3 196886588.9 178620562.1 172934056.7 Uracil_1 134561142 143197184.4 129413908.8 135387644.6 87584707.12 144825073.1 Uracil_2 95417022.99 98900814.63 93065905.07 108022178.3 120258909.1 110643416.6 Uridine monophosphate (UMP)_1 741061349.9 781538643.3 769325347.5 895913205.3 829297615.2 711721670.3 Uridine monophosphate (UMP)_2 737766660.5 839713901.8 655581780.8 635810153.2 747289375.3 842316879.7 Uridine monophosphate (UMP)_3 337942981.2 296469955.1 258863109.3 275738733.4 295488781.3 240239259.1 Uridine monophosphate (UMP)_4 216494970.7 222807265.4 294924401.8 190662263.5 317666364.9 418649875.9 Xanthine 887835901.7 916101933 863394823.8 980273620.1 971415554.1 991825150.5 Xanthosine 43886002.34 42668627.25 44155366.04 44873279.6 55007432.03 46946778.03 α-Aspartylphenylalanine 43718328.87 27300209.97 116444790.9 71564683.44 49507123.45 2907727.673 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name m/z RT [min] Formula (15Z)-9,12,13-Trihydroxy-15-octadecenoic acid_1 329.23333 17.599 C18 H34 O5 (E)-6-hydroxyoctadec-4-enoic acid 297.2434 18.046 C18 H34 O3 12-Hydroxydodecanoic acid 215.16525 15.731 C12 H24 O3 13S-hydroxy-9Z,11E,15Z-octadecatrienoic acid 293.21225 15.682 C18 H30 O3 16-Hydroxyhexadecanoic acid 271.22776 17.803 C16 H32 O3 2-(acetylamino)-4-(methylthio)butanoic acid_1 190.05439 6.359 C7 H13 N O3 S 2-(acetylamino)-4-(methylthio)butanoic acid_2 190.05435 6.537 C7 H13 N O3 S 2-(acetylamino)-4-(methylthio)butanoic acid_3 190.05435 6.687 C7 H13 N O3 S 2,5-di-tert-Butylhydroquinone 221.15466 16.561 C14 H22 O2 2-Aminooctanedioic acid_1 188.09277 2.302 C8 H15 N O4 2-Aminooctanedioic acid_2 188.09292 8.772 C8 H15 N O4 2-Hydroxycaproic acid 131.0714 9.391 C6 H12 O3 2-Hydroxyvaleric acid 117.05573 4.35 C5 H10 O3 3-(Cyclohexylamino)-2-hydroxy-1-propanesulfonic acid 236.09632 14.672 C9 H19 N O4 S 3,4-Dihydroxybenzenesulfonic acid 188.98637 5.616 C6 H6 O5 S 3,4-Dihydroxyphenylpropionic acid_1 181.05063 5.461 C9 H10 O4 3,4-Dihydroxyphenylpropionic acid_2 181.05075 5.983 C9 H10 O4 3-Hydroxy-3-methylglutaric acid 161.04559 1.406 C6 H10 O5 3-Hydroxydecanoic acid 187.13403 16.009 C10 H20 O3 4-{[3-(Diethylamino)propyl]amino}-4-oxobut-2-enoic acid 227.14016 5.03 C11 H20 N2 O3 4-{[5-(6-Hydroxy-5,5,8a-trimethyl-2-methylenedecahydro-1-naphthalenyl)-3-methylpentyl]oxy}-4-oxobutanoic acid 407.28051 16.68 C24 H40 O5 4-Cyanoindole 141.04599 14.899 C9 H6 N2 4-Indolecarbaldehyde_1 144.04551 6.44 C9 H7 N O 4-Indolecarbaldehyde_2 144.04552 7.709 C9 H7 N O 4-Oxoproline_1 128.03528 1.257 C5 H7 N O3 4-Oxoproline_2 128.03533 1.697 C5 H7 N O3 5-(2-Methylpropyl)-3,6-dioxo-2-piperazinepropanoic acid_1 241.11938 10.783 C11 H18 N2 O4 5-(2-Methylpropyl)-3,6-dioxo-2-piperazinepropanoic acid_2 241.11941 9.939 C11 H18 N2 O4 5-(2-Methylpropyl)-3,6-dioxo-2-piperazinepropanoic acid_3 241.11939 9.377 C11 H18 N2 O4 5-(2-Methylpropyl)-3,6-dioxo-2-piperazinepropanoic acid_4 241.11943 9.057 C11 H18 N2 O4 5-(2-Methylpropyl)-3,6-dioxo-2-piperazinepropanoic acid_5 241.11945 8.14 C11 H18 N2 O4 5-(2-Methylpropyl)-3,6-dioxo-2-piperazinepropanoic acid_6 241.11937 8.871 C11 H18 N2 O4 5-(2-Methylpropyl)-3,6-dioxo-2-piperazinepropanoic acid_7 241.11938 5.752 C11 H18 N2 O4 5-Chloro-4-fluoro-2-nitroaniline 188.98633 5.796 C6 H4 Cl F N2 O2 5-Hydroxyindole-3-acetic acid 190.05099 11.82 C10 H9 N O3 5-Hydroxytryptophan_1 219.07751 5.424 C11 H12 N2 O3 5-Hydroxytryptophan_2 219.07753 4.221 C11 H12 N2 O3 5-Hydroxytryptophan_3 219.07746 3.579 C11 H12 N2 O3 5-Hydroxytryptophan_4 219.07745 3.893 C11 H12 N2 O3 6-Hydroxycaproic acid_1 131.07137 9.998 C6 H12 O3 6-Hydroxycaproic acid_2 131.07142 9.715 C6 H12 O3 Adenine_1 134.04722 1.683 C5 H5 N5 Adenine_2 134.04723 1.399 C5 H5 N5 Adenosine 3'5'-cyclic monophosphate 328.04507 1.682 C10 H12 N5 O6 P Adenosine 5'-monophosphate_1 346.0556 1.533 C10 H14 N5 O7 P Adenosine 5'-monophosphate_2 346.05564 1.433 C10 H14 N5 O7 P Adenosine 5'-monophosphate_3 346.05557 1.383 C10 H14 N5 O7 P Adenosine 5'-monophosphate_4 346.05555 1.75 C10 H14 N5 O7 P Biotin 243.0808 10.466 C10 H16 N2 O3 S Capryloylglycine 200.12932 15.066 C10 H19 N O3 Cholic acid 407.28037 16.043 C24 H40 O5 Citric acid_1 191.01969 1.215 C6 H8 O7 Citric acid_2 191.01976 1.655 C6 H8 O7 D-(+)-Mannose 179.05624 1.176 C6 H12 O6 D-(+)-Tryptophan_1 203.08264 5.416 C11 H12 N2 O2 D-(+)-Tryptophan_2 203.08262 5.643 C11 H12 N2 O2 Deoxycholic acid 391.28543 17.385 C24 H40 O4 Dibutyl phthalate 277.1445 18.107 C16 H22 O4 DL-β-Leucine 130.08733 1.796 C6 H13 N O2 Dodecyl sulfate_1 265.14791 16.255 C12 H26 O4 S Dodecyl sulfate_2 265.14792 16.027 C12 H26 O4 S D-α-Hydroxyglutaric acid 147.0299 1.239 C5 H8 O5 Glutaric acid 131.03501 3.172 C5 H8 O4 Glycerol 3-phosphate 171.00642 1.326 C3 H9 O6 P Glycocholic acid 464.30199 15.261 C26 H43 N O6 Glycodeoxycholic acid 448.30704 16.358 C26 H43 N O5 Guanine 150.04212 1.788 C5 H5 N5 O Guanosine monophosphate (GMP)_1 362.0509 1.551 C10 H14 N5 O8 P Guanosine monophosphate (GMP)_2 362.05096 1.879 C10 H14 N5 O8 P Guanosine monophosphate (GMP)_3 362.05053 1.731 C10 H14 N5 O8 P Guanosine monophosphate (GMP)_4 362.05059 1.442 C10 H14 N5 O8 P Guanosine monophosphate (GMP)_5 362.05064 2.093 C10 H14 N5 O8 P Hexadecanedioic acid 285.20728 15.855 C16 H30 O4 Hypoxanthine 135.03126 1.531 C5 H4 N4 O Indole-3-acetic acid_1 174.05614 12.302 C10 H9 N O2 Indole-3-acetic acid_2 174.05605 3.42 C10 H9 N O2 L-(-)-3-Phenyllactic acid 165.05575 11.851 C9 H10 O3 L-Aspartic acid 132.03027 1.049 C4 H7 N O4 L-Dopa 196.06155 1.608 C9 H11 N O4 L-Histidine_1 154.06237 1.167 C6 H9 N3 O2 L-Histidine_2 154.06228 1.031 C6 H9 N3 O2 L-Iditol 181.07197 1.184 C6 H14 O6 L-Phenylalanine_1 164.07177 2.922 C9 H11 N O2 L-Phenylalanine_2 164.07185 10.897 C9 H11 N O2 L-Threonic acid 135.03 1.2 C4 H8 O5 L-Tyrosine_1 180.06667 1.606 C9 H11 N O3 L-Tyrosine_2 180.06666 2.902 C9 H11 N O3 L-Tyrosine_3 180.06668 2.546 C9 H11 N O3 Mevalonic acid 147.06634 2.214 C6 H12 O4 Myriocin 400.2708 15.321 C21 H39 N O6 Myristyl sulfate 293.17929 19.616 C14 H30 O4 S N1-tetrahydrofuran-2-ylmethyl-2-cyanoacetamide 167.08264 2.17 C8 H12 N2 O2 N-Acetylalanine 130.05102 1.172 C5 H9 N O3 N-Acetyl-D-alloisoleucine_1 172.09802 11.336 C8 H15 N O3 N-Acetyl-D-alloisoleucine_2 172.09808 11.148 C8 H15 N O3 N-Acetyl-D-alloisoleucine_3 172.09795 10.616 C8 H15 N O3 N-Acetyl-D-alloisoleucine_4 172.09801 10.476 C8 H15 N O3 N-Acetyl-DL-glutamic acid 188.05645 1.282 C7 H11 N O5 N-Acetyl-L-phenylalanine 206.08234 12.507 C11 H13 N O3 N-Acetyl-L-tyrosine_1 222.07719 6.432 C11 H13 N O4 N-Acetyl-L-tyrosine_2 222.07712 6.712 C11 H13 N O4 N-Acetylornithine 173.09318 1.203 C7 H14 N2 O3 Nateglinide 316.19172 17.422 C19 H27 N O3 N-Formylmethionine 176.03877 5.053 C6 H11 N O3 S Nicotinic acid 122.02476 1.527 C6 H5 N O2 N-Isovalerylglycine 158.0823 6.225 C7 H13 N O3 N-Isovalerylglycine_1 158.08232 10.324 C7 H13 N O3 N-Isovalerylglycine_2 158.08233 9.576 C7 H13 N O3 N-Tigloylglycine 156.06669 3.177 C7 H11 N O3 O-Acetylserine 146.04592 1.196 C5 H9 N O4 Ornithine_1 131.08268 1.201 C5 H12 N2 O2 Ornithine_2 131.08262 0.988 C5 H12 N2 O2 Pantothenic acid_1 218.10334 4.041 C9 H17 N O5 Pantothenic acid_2 218.10335 4.367 C9 H17 N O5 Pantothenic acid_3 218.1033 4.525 C9 H17 N O5 Phenylacetaldehyde_1 165.05577 11.578 C8 H8 O Phenylacetaldehyde_2 119.05025 11.778 C8 H8 O Porphobilinogen_1 225.08805 3.702 C10 H14 N2 O4 Porphobilinogen_2 225.08802 3.958 C10 H14 N2 O4 Porphobilinogen_3 225.08805 3.104 C10 H14 N2 O4 Riboflavin 375.13092 10.914 C17 H20 N4 O6 Salicylic acid 137.0245 5.456 C7 H6 O3 Succinic acid_1 117.01929 1.36 C4 H6 O4 Succinic acid_2 117.01933 1.574 C4 H6 O4 Succinic acid_3 117.01933 1.865 C4 H6 O4 Threonine 118.05107 1.191 C4 H9 N O3 Thymidine 5'-monophosphate 321.04926 1.893 C10 H15 N2 O8 P trans-Cinnamic acid_1 147.04523 2.903 C9 H8 O2 trans-Cinnamic acid_2 147.04526 11.58 C9 H8 O2 Uracil_1 111.01999 1.409 C4 H4 N2 O2 Uracil_2 111.01996 1.539 C4 H4 N2 O2 Uridine monophosphate (UMP)_1 323.02838 1.54 C9 H13 N2 O9 P Uridine monophosphate (UMP)_2 323.02843 1.414 C9 H13 N2 O9 P Uridine monophosphate (UMP)_3 323.02842 1.746 C9 H13 N2 O9 P Uridine monophosphate (UMP)_4 323.02842 1.613 C9 H13 N2 O9 P Xanthine 151.02615 1.55 C5 H4 N4 O2 Xanthosine 283.06833 2.059 C10 H12 N4 O6 α-Aspartylphenylalanine 279.09865 6.487 C13 H16 N2 O5 METABOLITES_END #END