#METABOLOMICS WORKBENCH spbaba01_20211215_104225 DATATRACK_ID:2986 STUDY_ID:ST002056 ANALYSIS_ID:AN003348 PROJECT_ID:PR001301 VERSION 1 CREATED_ON January 7, 2022, 8:35 am #PROJECT PR:PROJECT_TITLE Integrated multilayer omics reveals the genomic, proteomic and metabolic PR:PROJECT_TITLE influences of the histidyl dipeptides on heart PR:PROJECT_TYPE Triomics of WT and Carnosin synthase transgenic heart MSTFA dervitization PR:PROJECT_SUMMARY Histidyl dipeptides, such as carnosine, present in a micro-millimolar ranges in PR:PROJECT_SUMMARY the heart, are synthesized via the enzyme carnosine synthase (Carns). These PR:PROJECT_SUMMARY dipeptides facilitate glycolysis by proton buffering, form conjugates with PR:PROJECT_SUMMARY reactive aldehydes, such as acrolein, and attenuate ischemia and reperfusion PR:PROJECT_SUMMARY injury. While these dipeptides exhibit multifunctional properties, a composite PR:PROJECT_SUMMARY understanding of their roles in myocardium is lacking. To identify the landscape PR:PROJECT_SUMMARY of histidyl dipeptide mediated responses in the heart, we used a triomics PR:PROJECT_SUMMARY approach of genome wide RNA sequencing, global proteomics and unbiased PR:PROJECT_SUMMARY metabolomics in the cardio specific Carns transgenic (Tg) mice and integrated PR:PROJECT_SUMMARY the three data sets. Our result show higher myocardial levels of histidyl PR:PROJECT_SUMMARY dipeptides lead to extensive changes in several microRNAs, which could target PR:PROJECT_SUMMARY the expression of contractile proteins, beta-fatty acid oxidation and citric PR:PROJECT_SUMMARY acid cycle (TCA) enzymes. Global proteomics shows, expression of contractile PR:PROJECT_SUMMARY proteins, enzymes of beta-fatty acid oxidation and TCA cycle, were enriched in PR:PROJECT_SUMMARY the CarnsTg heart. Under aerobic conditions, the CarnsTg hearts had lower levels PR:PROJECT_SUMMARY of short and long-chain fatty acids and TCA cycle intermediate-succinic acid, PR:PROJECT_SUMMARY whereas, under ischemic conditions the accumulation of fatty acids and TCA cycle PR:PROJECT_SUMMARY intermediates were significantly attenuated in the CarnsTg heart. Integration of PR:PROJECT_SUMMARY multiple data sets suggests that beta-fatty acid oxidation and TCA cycle PR:PROJECT_SUMMARY pathways exhibited correlative changes in the CarnsTg hearts at all three PR:PROJECT_SUMMARY levels. Our triomics approach shows histidyl dipeptides are critical regulators PR:PROJECT_SUMMARY of myocardial structure, function and energetics. PR:INSTITUTE University of Louisville PR:DEPARTMENT Medicine PR:LABORATORY Diabetes and Obesity Center PR:LAST_NAME Baba PR:FIRST_NAME Shahid PR:ADDRESS 580 S. Preston St PR:EMAIL spbaba01@louisville.edu PR:PHONE 5028524274 PR:FUNDING_SOURCE NIH #STUDY ST:STUDY_TITLE Integrated Multilayer Omics Reveals the Genomic, Proteomic and Metabolic ST:STUDY_TITLE Influences of the Histidyl Dipeptides on Heart ST:STUDY_SUMMARY Histidyl dipeptides, such as carnosine, present in a micro-millimolar ranges in ST:STUDY_SUMMARY the heart, are synthesized via the enzyme carnosine synthase (Carns). These ST:STUDY_SUMMARY dipeptides facilitate glycolysis by proton buffering, form conjugates with ST:STUDY_SUMMARY reactive aldehydes, such as acrolein, and attenuate ischemia and reperfusion ST:STUDY_SUMMARY injury. While these dipeptides exhibit multifunctional properties, a composite ST:STUDY_SUMMARY understanding of their roles in myocardium is lacking. To identify the landscape ST:STUDY_SUMMARY of histidyl dipeptide mediated responses in the heart, we used a triomics ST:STUDY_SUMMARY approach of genome wide RNA sequencing, global proteomics and unbiased ST:STUDY_SUMMARY metabolomics in the cardio specific Carns transgenic (Tg) mice and integrated ST:STUDY_SUMMARY the three data sets. Our result show higher myocardial levels of histidyl ST:STUDY_SUMMARY dipeptides lead to extensive changes in several microRNAs, which could target ST:STUDY_SUMMARY the expression of contractile proteins, beta-fatty acid oxidation and citric ST:STUDY_SUMMARY acid cycle (TCA) enzymes. Global proteomics shows, expression of contractile ST:STUDY_SUMMARY proteins, enzymes of beta-fatty acid oxidation and TCA cycle, were enriched in ST:STUDY_SUMMARY the CarnsTg heart. Under aerobic conditions, the CarnsTg hearts had lower levels ST:STUDY_SUMMARY of short and long-chain fatty acids and TCA cycle intermediate-succinic acid, ST:STUDY_SUMMARY whereas, under ischemic conditions the accumulation of fatty acids and TCA cycle ST:STUDY_SUMMARY intermediates were significantly attenuated in the CarnsTg heart. Integration of ST:STUDY_SUMMARY multiple data sets suggests that beta-fatty acid oxidation and TCA cycle ST:STUDY_SUMMARY pathways exhibited correlative changes in the CarnsTg hearts at all three ST:STUDY_SUMMARY levels. Our triomics approach shows histidyl dipeptides are critical regulators ST:STUDY_SUMMARY of myocardial structure, function and energetics. ST:INSTITUTE University of Louisville ST:LAST_NAME Baba ST:FIRST_NAME Shahid ST:ADDRESS 580 S. Preston St ST:EMAIL spbaba01@louisville.edu ST:NUM_GROUPS 4 ST:STUDY_TYPE Triomics ST:PHONE 5028524274 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:AGE_OR_AGE_RANGE 8-12 weeks SU:GENDER Male #FACTORS #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 - CarnsTG1 Genotype:CarnsTG SUBJECT_SAMPLE_FACTORS - CarnsTG2 Genotype:CarnsTG SUBJECT_SAMPLE_FACTORS - CarnsTG3 Genotype:CarnsTG SUBJECT_SAMPLE_FACTORS - CarnsTG4 Genotype:CarnsTG SUBJECT_SAMPLE_FACTORS - WT1 Genotype:Wild-type SUBJECT_SAMPLE_FACTORS - WT2 Genotype:Wild-type SUBJECT_SAMPLE_FACTORS - WT3 Genotype:Wild-type SUBJECT_SAMPLE_FACTORS - WT4 Genotype:Wild-type #COLLECTION CO:COLLECTION_SUMMARY To determine the effects of the Carns overexpression on the global cardio CO:COLLECTION_SUMMARY metabolomic profile under the basal conditions, hearts were collected from the CO:COLLECTION_SUMMARY WT and CarnsTg mice upon sacrifice and snap-frozen in liquid nitrogen. CO:COLLECTION_PROTOCOL_ID 18404 CO:COLLECTION_PROTOCOL_FILENAME Metabolomics CO:COLLECTION_PROTOCOL_COMMENTS Effect of Carns overexpression CO:SAMPLE_TYPE Heart CO:COLLECTION_LOCATION University of Louisville CO:STORAGE_CONDITIONS -80℃ CO:COLLECTION_TUBE_TEMP -80 #TREATMENT TR:TREATMENT_SUMMARY To determine the changes in the cardio metabolomic profile associated with the TR:TREATMENT_SUMMARY ischemic injury and whether Carns overexpression influences the cardiac TR:TREATMENT_SUMMARY metabolism, isolated hearts from the WT and CarnsTg mice hearts were perfused in TR:TREATMENT_SUMMARY the Langendorff mode for 35 min, perfused for 20 min followed by 5 min of TR:TREATMENT_SUMMARY ischemia and perfused for 20 min followed by 15 min of ischemia as described TR:TREATMENT_SUMMARY previousl #SAMPLEPREP SP:SAMPLEPREP_SUMMARY To extract polar metabolites, 800 µL of methanol was added to 200 µL of SP:SAMPLEPREP_SUMMARY homogenized heart sample. Mixture was vortexed for 2 min and then placed on ice SP:SAMPLEPREP_SUMMARY for 10 min, followed by another 2 min of vortex mixing, and centrifuged at SP:SAMPLEPREP_SUMMARY 15,000 rpm for 20 min at 4 °C. Supernatant was transferred into a glass vial SP:SAMPLEPREP_SUMMARY and dried in a SpeedVac evaporator to remove methanol, followed by SP:SAMPLEPREP_SUMMARY lyophilization to remove water. The dried metabolite extract was dissolved with SP:SAMPLEPREP_SUMMARY 30 µL of 20 mg/mL methoxyamine hydrochloride pyridine solution followed by SP:SAMPLEPREP_SUMMARY vigorous vortex mixing for 1 min. Methoxylation was performed by sonicating the SP:SAMPLEPREP_SUMMARY sample for 20 min and incubation at 60 °C for 1 h. Derivatization was performed SP:SAMPLEPREP_SUMMARY by adding 20 µL of N-trimethylsilyl-N-methyl trifluoroacetamide (MSTFA) or SP:SAMPLEPREP_SUMMARY N-Methyl-N-tert-butyldimethysilyltrifuoroacetamide (MTBSTFA) with 1% SP:SAMPLEPREP_SUMMARY trimethylchlorosilane to the glass vial. Samples were incubated for 1 h at 60 SP:SAMPLEPREP_SUMMARY °C and the mixture was transferred to a GC vial for analysis. Pooled samples SP:SAMPLEPREP_SUMMARY were prepared by mixing 30 µL of derivatized metabolite extract from each SP:SAMPLEPREP_SUMMARY sample to monitor the instrumental variations during the course of GC×GC-MS SP:SAMPLEPREP_SUMMARY analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY The extracted and derivatized samples were analyzed on a LECO Pegasus GC×GC-TOF CH:CHROMATOGRAPHY_SUMMARY MS instrument (LECO Corp., St. Joseph, MI, USA) coupled to an Agilent 6890 gas CH:CHROMATOGRAPHY_SUMMARY chromatography and a Gerstel MPS2 autosampler (GERSTEL Inc., Linthicum, MD, CH:CHROMATOGRAPHY_SUMMARY USA), featuring a LECO two-stage cryogenic modulator and secondary oven. The CH:CHROMATOGRAPHY_SUMMARY primary column was a 60 m × 0.25 mm 1dc × 0.25 µm 1df DB-5 ms GC capillary CH:CHROMATOGRAPHY_SUMMARY column (phenyl arylene polymer virtually equivalent to CH:CHROMATOGRAPHY_SUMMARY (5%-phenyl)-methylpolysiloxane). The secondary GC column 1 m × 0.25 mm 2dc × CH:CHROMATOGRAPHY_SUMMARY 0.25 µm 2df, DB-17 ms ((50% phenyl)-methylpolysiloxane) was placed inside the CH:CHROMATOGRAPHY_SUMMARY secondary GC oven following the thermal modulator. Both columns were obtained CH:CHROMATOGRAPHY_SUMMARY from Agilent Technologies (Agilent Technologies J&W, Santa Clara, CA, USA) and CH:CHROMATOGRAPHY_SUMMARY were connected through a press fit connector. The helium carrier gas (99.999% CH:CHROMATOGRAPHY_SUMMARY purity) flow rate was set to 2.0 mL/min at a corrected constant flow via CH:CHROMATOGRAPHY_SUMMARY pressure ramps. The inlet temperature was set at 280 °C. The primary column CH:CHROMATOGRAPHY_SUMMARY temperature was programmed with an initial temperature of 60 °C for 0.5 min, CH:CHROMATOGRAPHY_SUMMARY then ramped at 5°C/min to 270 °C, and maintained for 15 min. The secondary CH:CHROMATOGRAPHY_SUMMARY column temperature program was set to an initial temperature of 70 °C for 0.5 CH:CHROMATOGRAPHY_SUMMARY min and then ramped at the same temperature gradient employed in the first CH:CHROMATOGRAPHY_SUMMARY column to 280 °C, accordingly. The thermal modulator was set to 15 °C relative CH:CHROMATOGRAPHY_SUMMARY to the primary oven and a modulation time was 2s. The mass range was set as CH:CHROMATOGRAPHY_SUMMARY 29-800 m/z with an acquisition rate of 200 mass spectra per second. The ion CH:CHROMATOGRAPHY_SUMMARY source chamber was 230°C with the transfer line temperature of 280°C, and the CH:CHROMATOGRAPHY_SUMMARY detector voltage was 1440 V with electron energy of 70 eV. The acceleration CH:CHROMATOGRAPHY_SUMMARY voltage was turned on after a solvent delay of 544 s, the split ratio was set at CH:CHROMATOGRAPHY_SUMMARY 10:1. CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent 6890N CH:COLUMN_NAME Agilent DB5-MS (60m × 0.25mm, 0.25um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Leco Pegasus III GC TOF MS:INSTRUMENT_TYPE GC x GC-TOF MS:MS_TYPE EI MS:ION_MODE UNSPECIFIED MS:MS_COMMENTS None #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS intensity MS_METABOLITE_DATA_START Samples CarnsTG1 CarnsTG2 CarnsTG3 CarnsTG4 WT1 WT2 WT3 WT4 Factors Genotype:CarnsTG Genotype:CarnsTG Genotype:CarnsTG Genotype:CarnsTG Genotype:Wild-type Genotype:Wild-type Genotype:Wild-type Genotype:Wild-type (Aminooxy)acetic acid, 2TBDMS derivative 1015646 1040729 910757 653449 1178562 1285733 1214331 1017974 (2R)-Pyrrolidine-1,2-dicarboxylic acid, bis(tert-butyldimethylsilyl) ester 39593 37984 92149 53183 48812 35911 42588 63643 1,3-Dihydro-1,3-dioxo-5-isobenzofurancarboxylic acid 45412 53629 34607 21753 61366 57328 61507 89971 1-Butanol, TBDMS derivative 2040435 2195037 2076864 2236466 2086849 2033440 1807911 1679078 1-Octadecanoyl-2-(5Z,8Z,11Z,14Z-eicosatetraenoyl)-sn-glycero-3-phosphoethanolamine 11074 11547 12381 14925 12369 12314 13288 11491 2-Bromotetradecane 0 11930 0 10169 7083 0 0 6502 2'-Deoxyguanosine 5'-monophosphate 53295 63793 39198 38833 40958 59342 55739 54224 2-Hydroxy-3-methylbutyric acid, 2TBDMS derivative 4801 6711 11994 5199 3672 5386 0 0 2-Hydroxybutanoic acid, 2TBDMS derivative 6537 0 7122 5329 5702 0 0 6458 Pentanedioic acid, 2-[(tert-butyldimethylsilyl)oxy]-, bis(tert-butyldimethylsilyl) ester 21772 15017 15482 18448 20452 0 30672 17674 2'-Methoxy-à-naphthoflavone 19490 17110 13216 0 12319 21976 0 4833 2-Methoxyethanol, TBDMS derivative 43330 47257 41619 44238 41263 41003 40520 43502 2-Pentanol, TBDMS derivative 34197 43125 43835 38990 36771 37166 33609 35402 2-Tridecen-1-ol, (E)- 4120 0 4267 6169 5421 5772 5172 0 3,3-Dimethyl-2-butanol, TBDMS derivative 17541 20745 20748 18633 16795 18332 11720 18254 3,4-hydroxyphenylpyruvic acid MTBSTFA Deri 2845443 2685942 3020573 3082536 2995567 2872074 2415158 3040883 3-Aminobutyric acid, 2TBDMS derivative 49362 54970 58499 54560 65058 67360 56314 73327 3-Hydroxy-3-phenylpropionic acid, 2TBDMS derivative 37644 0 40264 38051 41115 43237 37187 41878 3-Hydroxybutyric acid, (R)-, 2TBDMS derivative 34828 15981 51836 55852 38900 19310 59513 42465 Propanoic acid, 3-[(tert-butyldimethylsilyl)oxy]-2-methyl-, tert-butyldimethylsilyl ester 17484 8666 33971 26335 21449 10361 21910 21106 3-Pyridinol, TBDMS derivative 44712 46240 26968 25135 30711 44469 37591 29765 5-Aminolevulinic acid 22517 21868 18136 18488 23946 0 0 17471 6-Hydroxy-7-methoxycoumarin 34016 31819 34257 0 37025 16911 0 26187 9-Hexadecenoic acid, (Z)-, TBDMS derivative 24319 33653 99881 119347 105500 37494 36260 48496 á-Alanine, 2TBDMS derivative 3867 4679 7798 11574 8643 5372 11001 5475 N,N'-Bis(tert-butyldimethylsilyl)acetamidine 2838142 1601202 2529695 2588729 1626151 2744498 1552623 2921222 Adenine, 2TBDMS derivative 14668 7833 5955 4930 8016 8029 8077 7075 Alanylglycine, 2TBDMS derivative 18210 21068 13062 15361 19394 22399 17726 19289 Arachidonic Acid, TBDMS derivative 120568 158304 114621 144805 140952 134065 139887 105728 Tris(tert-butyldimethylsilyloxy)arsane 21518 21354 0 0 20296 22645 24991 21378 Benzoic Acid, TBDMS derivative 5525 6118 0 6276 6012 0 0 6202 Carbonic acid, 2-dimethylaminoethyl 2-methoxyethyl ester 57524 70897 35803 16074 79428 86890 77881 111559 Cholesta-4,6-dien-3-one 0 939 0 848 771 793 0 0 Cholesterol, TBDMS derivative 115874 85090 0 110014 90435 110794 0 126274 Citric acid, 4TBDMS derivative 1295932 671558 609507 549969 698407 948368 627269 1010913 Danthron 23205 20590 0 24139 21831 24052 26916 27349 Di-n-octyl phthalate 0 27231 28606 0 25104 26197 24674 25423 DL-Glyceraldehyde, 2TBDMS derivative 872897 825853 1253102 944032 952974 765261 793545 988399 Dodecanal, O-methyloxime 0 2076 2420 1811 3276 3818 3178 2771 Dodecanoic acid, TBDMS derivative 21683 16525 11126 16088 16188 15805 17968 19385 D-Pipecolic acid, TBDMS derivative 13372 0 6776 0 0 19616 0 15097 D-Pyroglutamic acid, 2TBDMS derivative 720176 621109 341084 311425 549439 650093 788416 552703 Erythrono-1,4-lactone, 2TBDMS derivative 663857 625590 684930 728802 801161 674108 724082 1065082 Ethanolamine, 2TBDMS derivative 35831 38318 42646 51302 43322 31008 42231 32789 Ethanone, 1-(2-pyridinyl)- 0 46648 32433 58292 38646 0 33448 24142 Ethyl (tert-butyldimethylsilyl)oxycarbamate 49900 43211 81309 28211 0 63651 31104 0 2-(Ethylamino)ethanol, tert-butyldimethylsilyl) ether 735245 0 817704 1103542 828401 719056 0 603716 Ethylene glycol, 2TBDMS derivative 1314244 2010005 1483131 1511534 1919050 1869206 1864590 1416521 2-Butenedioic acid, 2TBDMS derivative 38692 32151 33049 28257 35776 25378 52592 45000 Glycerol, 3TBDMS derivative 252102 166485 409163 339841 390665 203877 285211 222842 Glycine, 2TBDMS derivative 130618 132131 48270 35444 82570 128937 142702 97979 Glycolic acid, 2TBDMS derivative 29390 30613 33692 26497 27254 33135 26511 33360 Heptadecanoic acid, TBDMS derivative 20818 23419 16664 16893 18355 21566 20396 18072 Hydroquinone, TBDMS derivative 19961 18615 25949 0 21794 25395 22623 29027 Hypoxanthine, 2TBDMS derivative 163796 158747 180256 155381 191765 126169 187168 148404 Ibutilide 0 0 25401 35273 29324 21001 23474 23761 Isocitric acid lactone, 2TBDMS derivative 51957 62891 79575 45721 72354 74163 32443 56970 Isoleucine, 2TBDMS derivative 77825 49374 60868 60347 55081 44143 62753 56544 Isopropylamine, N-tert-butyldimethylsilyl- 91434 114428 100315 119323 94812 80542 82778 87309 Lactic acid, 2TBDMS derivative 0 3430 2264 3165 2659 2947 5685 0 L-Alanine, 2TBDMS derivative 700737 577106 660937 618116 615899 515352 728468 648104 L-Asparagine, 3TBDMS derivative 11286 10460 8337 7182 9208 10682 12058 11173 L-Aspartic acid, 3TBDMS derivative 861412 967279 417902 604015 636342 798224 781937 725064 L-Cysteine, 3TBDMS derivative 0 11107 18408 29040 30469 0 24168 0 L-Glutamic acid, 3TBDMS derivative 1704092 1553984 1194134 1212009 1463575 1470795 1567068 1366577 L-Glutamine, 3TBDMS derivative 425494 310051 114857 138211 255508 326584 381571 285847 L-Histidine, N,1-bis(tert-butyldimethylsilyl)-, tert-butyldimethylsilyl ester 47993 23155 0 0 15737 0 23005 0 Linoelaidic acid, tert.-butyldimerthylsilyl ester 374049 401817 425588 382325 392786 408445 386616 364943 L-Leucine, 2TBDMS derivative 108959 80135 105166 99638 77641 66645 89503 93501 L-Lysine, 3TBDMS derivative 141863 111442 130156 88240 142235 115227 129422 139252 L-Methionine, 2TBDMS derivative 60249 58057 69849 43866 66608 60493 62902 91561 L-Ornithine, 3TBDMS derivative 29003 24455 27790 21240 26397 23051 25237 32991 L-Phenylalanine, 2TBDMS derivative 65213 48951 71205 56112 48442 52171 56864 65434 L-Proline, 2TBDMS derivative 15421 15128 18869 14479 18931 16281 16637 23296 L-Pyroglutamic acid, 2TBDMS derivative 63550 46136 56500 35401 17522 18678 19457 32064 L-Serine, 3TBDMS derivative 364352 340200 383404 352428 369597 408589 397365 586398 L-Threonine, 3TBDMS derivative 71094 123397 137209 175353 179271 135777 182707 165316 L-Tyrosine, 3TBDMS derivative 102841 78357 112185 90602 110716 61024 106066 134004 L-Valine, 2TBDMS derivative 57276 44520 56915 53120 50355 46837 52814 54413 Malic acid, 3TBDMS derivative 608911 1006062 730804 642589 943665 1256162 1045251 1459135 Metharbital 9085 0 13319 26723 6075 0 6899 0 Methoxyacetic acid, TBDMS derivative 8454 0 8113 9390 9145 9167 9101 10566 Methyl lactate, TBDMS derivative 22644 15375 37560 28498 30088 15398 19153 15660 Methylmalonic acid, 2TBDMS derivative 73721 43263 105560 89473 0 51037 70684 0 Myristic acid, TBDMS derivative 12776 12274 26530 29888 24867 13989 14393 17207 Naphtho[2,3-b]furan-4,9-dione 15055 24490 0 30501 0 22717 37219 0 Neopentyl alcohol, TBDMS derivative 0 0 8124 7389 7902 7442 0 7005 Niacinamide, TBDMS derivative 317721 313037 224293 205082 278074 329190 368321 277898 Nizatidine 29338 40612 117273 101417 86357 57068 42659 59799 N-Methyldodecanamide 0 8996 0 7042 8725 7180 0 6888 Nonanoic acid, TBDMS derivative 6748 10185 7642 7002 6852 5825 8032 0 Nonanoyl chloride 11832 13539 8254 7267 10183 12434 12502 9690 Oleamide, TBDMS derivative 28381 27225 23135 39436 0 0 22284 19802 Oleic Acid, (E)-, TBDMS derivative 269133 321352 426556 391122 396500 344515 322088 307342 Palmitic Acid, TBDMS derivative 886509 1002090 933112 1096470 942557 962561 1002395 859595 Pentadecanoic acid, TBDMS derivative 5685 6019 5393 6966 6754 5564 4743 5187 Phosphoric acid, tris(tert-butyldimethylsilyl) ester 1266385 645129 1239495 1420191 1481795 1437524 1203259 1819480 Phthalic acid, hept-3-yl isohexyl ester 0 44671 356964 0 357088 0 0 371904 Pipecolic acid, 2TBDMS derivative 37336 39002 22301 44618 31610 36609 28584 23536 Propanoic acid, TBDMS derivative 118147 117749 98937 127904 127129 136475 146791 158029 Propargyl alcohol, TBDMS derivative 73261 80361 53985 0 62057 74731 74454 60423 Stearic acid, TBDMS derivative 880103 997638 830587 821693 849467 959665 915407 856349 Butanedioic acid, 2TBDMS derivative 4529 5353 5276 0 6090 6025 0 7178 Sulfurous acid, 2-ethylhexyl hexyl ester 0 4533 0 3754 3997 4037 3307 4369 Taurine, 2TBDMS derivative 3647183 3437993 2225259 711325 2990833 3515210 2519055 3488839 Thymol, TBDMS derivative 64031 82769 31385 17446 44219 74180 88722 58316 Tianeptine 53221 0 60796 54874 60317 52186 66502 48022 trans-1-Chloro-2-trimethylsilyloxycyclohexane 0 39511 37388 41627 40090 42499 0 46932 Undecanamide, 11-bromo- 66094 0 0 82385 0 60665 55696 0 Uracil, 2TBDMS derivative 9302 8527 15519 18429 21782 13222 17271 13921 Urea, 2TBDMS derivative 1235549 1342446 1507491 1398797 1317407 1187888 1115703 1277275 1-(2-Methoxyethoxy)-2-methyl-2-propanol, TMS derivative 0 0 41552 19271 10106 10757 12467 0 6-Methoxy-2-hexanol, TMS derivative 126548 103694 100805 123035 91214 93684 70512 52180 Propanedioic acid, 2TMS derivative 453890 511668 402998 449065 366899 436046 0 342640 Methanesulfonic acid, TMS derivative 58763 73790 61089 39078 78925 70739 70636 110390 1-Methyl-1H-tetrazol-5-amine, TMS derivative 3151841 3169463 2491074 1846127 2317258 3063767 3138475 363728 Methylmalonic acid, 2TMS derivative 11417 0 0 13665 0 10480 11772 12922 2,4,6(1H,3H,5H)-Pyrimidinetrione, 5-ethyl-1,3-dimethyl-5-(1-methylpropyl)- 63289 56027 76531 208330 63776 53240 86068 31340 1-Hexanol, TMS derivative 0 12781 14607 12146 12424 13547 12134 12907 Itaconic acid, 2TMS derivative 16093 21548 24530 39706 27884 21992 21760 15303 2-(Methylamino)ethanol, tert-butyldimethylsilyl ether 14654 16453 0 0 0 16317 15608 11993 5-Nonanol, TMS derivative 0 10085 8445 8799 0 7995 9834 11353 Methyl aminoacetate, 2TMS derivative 77070 118496 41428 28246 42663 110834 56929 67008 Propanoic acid, 3-[methyl[2-oxo-2-[(trimethylsilyl)oxy]ethyl]amino]-3-oxo-, trimethylsilyl ester 30774 0 41686 22646 24290 18989 28056 49233 2-Ethoxyethanol, TMS derivative 18908 20425 19117 18639 18401 19261 19222 20920 2-Butenoic acid, 2-[(trimethylsilyl)oxy]-, trimethylsilyl ester 2325 0 0 1335 1670 1828 1606 1730 3,4-Dihydroxyhydrocinnamic acid, 3TMS derivative 54056 0 85077 0 72112 59468 0 0 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name (Aminooxy)acetic acid, 2TBDMS derivative (2R)-Pyrrolidine-1,2-dicarboxylic acid, bis(tert-butyldimethylsilyl) ester 1,3-Dihydro-1,3-dioxo-5-isobenzofurancarboxylic acid 1-Butanol, TBDMS derivative 1-Octadecanoyl-2-(5Z,8Z,11Z,14Z-eicosatetraenoyl)-sn-glycero-3-phosphoethanolamine 2-Bromotetradecane 2'-Deoxyguanosine 5'-monophosphate 2-Hydroxy-3-methylbutyric acid, 2TBDMS derivative 2-Hydroxybutanoic acid, 2TBDMS derivative Pentanedioic acid, 2-[(tert-butyldimethylsilyl)oxy]-, bis(tert-butyldimethylsilyl) ester 2'-Methoxy-à-naphthoflavone 2-Methoxyethanol, TBDMS derivative 2-Pentanol, TBDMS derivative 2-Tridecen-1-ol, (E)- 3,3-Dimethyl-2-butanol, TBDMS derivative 3,4-hydroxyphenylpyruvic acid MTBSTFA Deri 3-Aminobutyric acid, 2TBDMS derivative 3-Hydroxy-3-phenylpropionic acid, 2TBDMS derivative 3-Hydroxybutyric acid, (R)-, 2TBDMS derivative Propanoic acid, 3-[(tert-butyldimethylsilyl)oxy]-2-methyl-, tert-butyldimethylsilyl ester 3-Pyridinol, TBDMS derivative 5-Aminolevulinic acid 6-Hydroxy-7-methoxycoumarin 9-Hexadecenoic acid, (Z)-, TBDMS derivative á-Alanine, 2TBDMS derivative N,N'-Bis(tert-butyldimethylsilyl)acetamidine Adenine, 2TBDMS derivative Alanylglycine, 2TBDMS derivative Arachidonic Acid, TBDMS derivative Tris(tert-butyldimethylsilyloxy)arsane Benzoic Acid, TBDMS derivative Carbonic acid, 2-dimethylaminoethyl 2-methoxyethyl ester Cholesta-4,6-dien-3-one Cholesterol, TBDMS derivative Citric acid, 4TBDMS derivative Danthron Di-n-octyl phthalate DL-Glyceraldehyde, 2TBDMS derivative Dodecanal, O-methyloxime Dodecanoic acid, TBDMS derivative D-Pipecolic acid, TBDMS derivative D-Pyroglutamic acid, 2TBDMS derivative Erythrono-1,4-lactone, 2TBDMS derivative Ethanolamine, 2TBDMS derivative Ethanone, 1-(2-pyridinyl)- Ethyl (tert-butyldimethylsilyl)oxycarbamate 2-(Ethylamino)ethanol, tert-butyldimethylsilyl) ether Ethylene glycol, 2TBDMS derivative 2-Butenedioic acid, 2TBDMS derivative Glycerol, 3TBDMS derivative Glycine, 2TBDMS derivative Glycolic acid, 2TBDMS derivative Heptadecanoic acid, TBDMS derivative Hydroquinone, TBDMS derivative Hypoxanthine, 2TBDMS derivative Ibutilide Isocitric acid lactone, 2TBDMS derivative Isoleucine, 2TBDMS derivative Isopropylamine, N-tert-butyldimethylsilyl- Lactic acid, 2TBDMS derivative L-Alanine, 2TBDMS derivative L-Asparagine, 3TBDMS derivative L-Aspartic acid, 3TBDMS derivative L-Cysteine, 3TBDMS derivative L-Glutamic acid, 3TBDMS derivative L-Glutamine, 3TBDMS derivative L-Histidine, N,1-bis(tert-butyldimethylsilyl)-, tert-butyldimethylsilyl ester Linoelaidic acid, tert.-butyldimerthylsilyl ester L-Leucine, 2TBDMS derivative L-Lysine, 3TBDMS derivative L-Methionine, 2TBDMS derivative L-Ornithine, 3TBDMS derivative L-Phenylalanine, 2TBDMS derivative L-Proline, 2TBDMS derivative L-Pyroglutamic acid, 2TBDMS derivative L-Serine, 3TBDMS derivative L-Threonine, 3TBDMS derivative L-Tyrosine, 3TBDMS derivative L-Valine, 2TBDMS derivative Malic acid, 3TBDMS derivative Metharbital Methoxyacetic acid, TBDMS derivative Methyl lactate, TBDMS derivative Methylmalonic acid, 2TBDMS derivative Myristic acid, TBDMS derivative Naphtho[2,3-b]furan-4,9-dione Neopentyl alcohol, TBDMS derivative Niacinamide, TBDMS derivative Nizatidine N-Methyldodecanamide Nonanoic acid, TBDMS derivative Nonanoyl chloride Oleamide, TBDMS derivative Oleic Acid, (E)-, TBDMS derivative Palmitic Acid, TBDMS derivative Pentadecanoic acid, TBDMS derivative Phosphoric acid, tris(tert-butyldimethylsilyl) ester Phthalic acid, hept-3-yl isohexyl ester Pipecolic acid, 2TBDMS derivative Propanoic acid, TBDMS derivative Propargyl alcohol, TBDMS derivative Stearic acid, TBDMS derivative Butanedioic acid, 2TBDMS derivative Sulfurous acid, 2-ethylhexyl hexyl ester Taurine, 2TBDMS derivative Thymol, TBDMS derivative Tianeptine trans-1-Chloro-2-trimethylsilyloxycyclohexane Undecanamide, 11-bromo- Uracil, 2TBDMS derivative Urea, 2TBDMS derivative 1-(2-Methoxyethoxy)-2-methyl-2-propanol, TMS derivative 6-Methoxy-2-hexanol, TMS derivative Propanedioic acid, 2TMS derivative Methanesulfonic acid, TMS derivative 1-Methyl-1H-tetrazol-5-amine, TMS derivative Methylmalonic acid, 2TMS derivative 2,4,6(1H,3H,5H)-Pyrimidinetrione, 5-ethyl-1,3-dimethyl-5-(1-methylpropyl)- 1-Hexanol, TMS derivative Itaconic acid, 2TMS derivative 2-(Methylamino)ethanol, tert-butyldimethylsilyl ether 5-Nonanol, TMS derivative Methyl aminoacetate, 2TMS derivative Propanoic acid, 3-[methyl[2-oxo-2-[(trimethylsilyl)oxy]ethyl]amino]-3-oxo-, trimethylsilyl ester 2-Ethoxyethanol, TMS derivative 2-Butenoic acid, 2-[(trimethylsilyl)oxy]-, trimethylsilyl ester 3,4-Dihydroxyhydrocinnamic acid, 3TMS derivative METABOLITES_END #END