#METABOLOMICS WORKBENCH samanthahart_20251027_090722 DATATRACK_ID:6599 STUDY_ID:ST004356 ANALYSIS_ID:AN007273 PROJECT_ID:PR002762 VERSION 1 CREATED_ON November 15, 2025, 2:04 pm #PROJECT PR:PROJECT_TITLE Stable Isotope Tracing of Primary Human Peripheral Blood Mononuclear Cells for PR:PROJECT_TITLE STAR Protocols PR:PROJECT_TYPE Protocol Optimization PR:PROJECT_SUMMARY Stable isotope tracing gives direct insight to the rate of metabolic reactions PR:PROJECT_SUMMARY occurring in cells through analyzing the incorporation of labeled carbons into PR:PROJECT_SUMMARY metabolic pathways. Thus far, there is no standardized protocol using stable PR:PROJECT_SUMMARY isotope tracing and gas chromatography mass spectrometry (GC-MS) in primary PR:PROJECT_SUMMARY human immune cells. Here, we establish and validate a technique for culturing PR:PROJECT_SUMMARY and exposing human peripheral blood mononuclear cells (PBMCs) to U13C Glucose & PR:PROJECT_SUMMARY U13C Glutamine, stable isotopes, and subsequent tracing analysis via single PR:PROJECT_SUMMARY quadrupole GC-MS. PR:INSTITUTE University of Kentucky PR:DEPARTMENT Molecular & Cellular Biochemistry PR:LABORATORY Nikolajczyk PR:LAST_NAME Hart PR:FIRST_NAME Samantha PR:ADDRESS 760 Press Avenue, Room 251 Lexington KY 40536 PR:EMAIL snda236@uky.edu PR:PHONE 5165247315 #STUDY ST:STUDY_TITLE Stable Isotope Tracing of Primary Human Peripheral Blood Mononuclear Cells ST:STUDY_TYPE Protocol Optimization ST:STUDY_SUMMARY Thus far, there is no standardized protocol using stable isotope tracing and gas ST:STUDY_SUMMARY chromatography mass spectrometry (GC-MS) in primary human immune cells. The goal ST:STUDY_SUMMARY of this study was to optimize the stable isotope tracing and subsequent GC-MS ST:STUDY_SUMMARY analysis of primary human peripheral blood mononuclear cells (PBMCs). We exposed ST:STUDY_SUMMARY human PBMCs to uniformly labeled 13C glucose, then extracted metabolites from ST:STUDY_SUMMARY each sample, separating polar metabolites from the protein pellet. Polar samples ST:STUDY_SUMMARY were dried and derivatized then analyzed via GCMS. Through this method, we found ST:STUDY_SUMMARY that incubating 10 million PBMCs per sample with 13C glucose or 13C glutamine ST:STUDY_SUMMARY for 2 hours provided optimal results, as this method allowed us to reliably ST:STUDY_SUMMARY detect the highest quantity of metabolites. We conclude that, when using a ST:STUDY_SUMMARY single-quadrupole GCMS to analyze a stable isotope i.e. 13C glucose's ST:STUDY_SUMMARY incorporation into metabolites in primary human PBMCs, a 2-hour incubation is ST:STUDY_SUMMARY recommended for optimal analysis. ST:INSTITUTE University of Kentucky ST:DEPARTMENT Molecular & Cellular Biochemistry ST:LAST_NAME Hart ST:FIRST_NAME Samantha ST:ADDRESS 760 Press Avenue, Room 251 Lexington KY 40536 ST:EMAIL snda236@uky.edu ST:PHONE 5165247315 ST:NUM_GROUPS 16 ST:TOTAL_SUBJECTS 16 ST:DISEASE_ASSOCIATION obesity #SUBJECT SU:SUBJECT_TYPE Human SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #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 1 1 Sample source:Blood | Treatment:13C Glc RAW_FILE_NAME(Raw File Name)=1.CDF SUBJECT_SAMPLE_FACTORS 2 2 Sample source:Blood | Treatment:13C Gln RAW_FILE_NAME(Raw File Name)=2.CDF SUBJECT_SAMPLE_FACTORS 3 3 Sample source:Blood | Treatment:13C Glc RAW_FILE_NAME(Raw File Name)=3.CDF SUBJECT_SAMPLE_FACTORS 4 4 Sample source:Blood | Treatment:13C Glc RAW_FILE_NAME(Raw File Name)=4.CDF SUBJECT_SAMPLE_FACTORS 5 5 Sample source:Blood | Treatment:13C Gln RAW_FILE_NAME(Raw File Name)=5.CDF SUBJECT_SAMPLE_FACTORS 6 6 Sample source:Blood | Treatment:13C Gln RAW_FILE_NAME(Raw File Name)=6.CDF SUBJECT_SAMPLE_FACTORS 7 7 Sample source:Blood | Treatment:13C Glc RAW_FILE_NAME(Raw File Name)=7.CDF SUBJECT_SAMPLE_FACTORS 8 8 Sample source:Blood | Treatment:13C Gln RAW_FILE_NAME(Raw File Name)=8.CDF SUBJECT_SAMPLE_FACTORS 9 9 Sample source:Blood | Treatment:13C Glc RAW_FILE_NAME(Raw File Name)=9.CDF SUBJECT_SAMPLE_FACTORS 10 10 Sample source:Blood | Treatment:13C Glc RAW_FILE_NAME(Raw File Name)=10.CDF SUBJECT_SAMPLE_FACTORS 11 11 Sample source:Blood | Treatment:13C Glc RAW_FILE_NAME(Raw File Name)=11.CDF SUBJECT_SAMPLE_FACTORS 12 12 Sample source:Blood | Treatment:13C Glc RAW_FILE_NAME(Raw File Name)=12.CDF SUBJECT_SAMPLE_FACTORS 13 13 Sample source:Blood | Treatment:13C Gln RAW_FILE_NAME(Raw File Name)=13.CDF SUBJECT_SAMPLE_FACTORS 14 14 Sample source:Blood | Treatment:13C Glc RAW_FILE_NAME(Raw File Name)=14.CDF SUBJECT_SAMPLE_FACTORS 15 15 Sample source:Blood | Treatment:13C Gln RAW_FILE_NAME(Raw File Name)=15.CDF SUBJECT_SAMPLE_FACTORS 16 16 Sample source:Blood | Treatment:13C Gln RAW_FILE_NAME(Raw File Name)=16.CDF SUBJECT_SAMPLE_FACTORS 17 17 Sample source:Blood | Treatment:13C Gln RAW_FILE_NAME(Raw File Name)=17.CDF SUBJECT_SAMPLE_FACTORS 18 18 Sample source:Blood | Treatment:13C Gln RAW_FILE_NAME(Raw File Name)=18.CDF #COLLECTION CO:COLLECTION_SUMMARY Primary human peripheral blood mononuclear cells collected from fasted obese CO:COLLECTION_SUMMARY subjects. Cells were isolated from whole blood per the Sigma Aldrich PBMC CO:COLLECTION_SUMMARY isolation method, then stored in liquid nitrogen. The protocol for cell CO:COLLECTION_SUMMARY isolation can be found at the following link: 3. CO:COLLECTION_SUMMARY https://www.sigmaaldrich.com/US/en/technical-documents/protocol/clinical-testing-and-diagnostics-manufacturing/hematology/recommended-standard-method?srsltid=AfmBOorOZE_VCvATbRMpNmG_A1C_bYgCvl_sTC4xgriCvR3yIXAS3_jd CO:SAMPLE_TYPE Mononuclear cells #TREATMENT TR:TREATMENT_SUMMARY Cells were thawed from liquid Nitrogen and cultured at 37 degrees Celsius for 40 TR:TREATMENT_SUMMARY hours in RPMI media, then treated with either 13C glucose or 13C glutamine for TR:TREATMENT_SUMMARY two hours. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY To extract metabolites after stable isotope exposure, cells were pelleted and SP:SAMPLEPREP_SUMMARY washed in NaCl then resuspended in 50% MeOH and frozen at -80 degrees Celsius. SP:SAMPLEPREP_SUMMARY Samples were then thawed, centrifuged, ad dried. Dried pellets were then SP:SAMPLEPREP_SUMMARY resuspended in Methoxyamine HCL + pyridine. MSTFA was subsequently added and SP:SAMPLEPREP_SUMMARY samples were run on GC-MS. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY GC methods used in this study were in accordance with our previous publications, CH:CHROMATOGRAPHY_SUMMARY which can be found at: Williams HC, Piron MA, Nation GK, Walsh AE, Young LEA, CH:CHROMATOGRAPHY_SUMMARY Sun RC, Johnson LA. Oral Gavage Delivery of Stable Isotope Tracer for In Vivo CH:CHROMATOGRAPHY_SUMMARY Metabolomics. Metabolites. 2020 Dec 8;10(12):501. doi: 10.3390/metabo10120501. CH:CHROMATOGRAPHY_SUMMARY PMID: 33302448; PMCID: PMC7764755. CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent 8890C CH:COLUMN_NAME Agilent J&W HP-5MS (30m x 0.25mm, 0.25um) CH:SOLVENT_A none CH:SOLVENT_B none CH:FLOW_GRADIENT none CH:FLOW_RATE 25 mL/min CH:COLUMN_TEMPERATURE Initial temperature 60C, hold for 1 minute. Temperature was then increased by CH:COLUMN_TEMPERATURE 10deg/min to 325C, then held for 10 minutes. #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 5977C MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:ION_MODE POSITIVE MS:MS_COMMENTS DExSI software was used to analyze raw .CDF files #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS area under the curve MS_METABOLITE_DATA_START Samples 1 10 11 12 13 14 15 16 17 18 2 3 4 5 6 7 8 9 Factors Sample source:Blood | Treatment:13C Glc Sample source:Blood | Treatment:13C Glc Sample source:Blood | Treatment:13C Glc Sample source:Blood | Treatment:13C Glc Sample source:Blood | Treatment:13C Gln Sample source:Blood | Treatment:13C Glc Sample source:Blood | Treatment:13C Gln Sample source:Blood | Treatment:13C Gln Sample source:Blood | Treatment:13C Gln Sample source:Blood | Treatment:13C Gln Sample source:Blood | Treatment:13C Gln Sample source:Blood | Treatment:13C Glc Sample source:Blood | Treatment:13C Glc Sample source:Blood | Treatment:13C Gln Sample source:Blood | Treatment:13C Gln Sample source:Blood | Treatment:13C Glc Sample source:Blood | Treatment:13C Gln Sample source:Blood | Treatment:13C Glc pyruvate_0 3392411 2251736 2549080 2453742 2755868 2510765 2562860 2359592 2605575 2590146 3296592 3885310 2760262 1990758 2527416 2148947 2597409 2688266 pyruvate_1 1093750 750184 824736 800996 881896 800923 816804 753571 847593 808924 1067166 1268322 879362 659022 830668 701558 854609 876606 pyruvate_2 91355 63108 68371 74315 80142 68293 71241 68549 65067 70670 39246 105719 65424 58275 71024 59471 70590 71146 pyruvate_3 10382 3954 7964 3200 4315 2939 6554 3169 4557 4769 5993 10060 2871 1628 6751 3466 5624 2403 Lactate_0 386418 171015 225240 324090 3812183 201929 332447 218351 5096950 201402 6645527 183548 426793 1997488 2739809 127196 3079897 279364 Lactate_1 165717 76826 103811 137314 731962 90321 114878 76137 986786 73756 1284833 66918 174565 384057 518470 54153 605212 129694 Lactate_2 432836 171940 270543 345275 352260 237693 237601 160010 456270 165170 608817 150729 459700 196943 248626 123085 292518 317468 Lactate_3 5005715 2060559 3154366 4066126 69091 2587149 2697396 1786312 113425 1786028 129246 1633553 5323104 51178 56788 1412934 72492 3758523 alanine_0 331744 143299 200937 256346 608745 163868 208523 132271 813189 134084 1039628 117296 338927 330142 456377 107991 517261 254540 alanine_1 7355977 3428646 4596544 6362093 54565480 4088191 5963361 3947781 71731472 3667964 91087872 3291699 8215759 29457092 39124384 2598868 44751108 5613870 alanine_2 3524272 1551549 2265238 2947380 6059962 1902117 2221939 1493764 8021350 1482298 10170382 1269092 3729007 3224967 4367986 1105358 5006086 2704302 alanine_3 68440416 30409746 44886944 57284036 3015348 37063492 39014028 26950706 4214146 26294508 5044795 22903016 72910056 1747268 2247214 21173218 2584733 53585532 Acetohydroxamic acid_0 1042 773 68462 69578 20265 65821 72281 0 64878 69700 56812 13729 75320 0 54238 0 41299 0 Acetohydroxamic acid_1 0 0 18983 15599 1905 13285 17195 0 19152 18390 15179 1069 15267 0 11784 0 0 6502 Acetohydroxamic acid_2 842 0 7832 6185 2407 7615 5807 0 0 0 0 0 8185 0 5166 0 0 0 Oxalic acid_0 66105 94690 136417 146006 108152 136126 145944 134448 121296 173782 231794 156777 172552 23777 115646 73567 128219 121794 Oxalic acid_1 27141 14953 27017 36447 24878 32641 31578 24729 31190 40909 57334 35237 39540 5872 26302 13199 28674 20921 Oxalic acid_2 7596 8129 11861 16403 9427 13213 11847 9993 10249 13915 19791 14330 16565 0 10386 4937 10247 10592 Oxalic acid_3 0 1607 5370 6314 0 5545 4898 764 1402 2455 2549 7197 11929 0 955 0 0 5844 Oxalic acid_4 0 0 0 0 0 0 0 0 0 0 0 0 687 0 0 0 0 0 leucine_0 205794 310402 473128 550918 359707 458247 516031 459852 414324 666890 766690 573076 587880 66615 396275 242419 417909 404722 leucine_1 37031 47161 78114 85473 60869 76806 69960 70747 60282 92542 115795 84807 84075 13086 57590 34685 57839 69324 leucine_2 72501 90386 152561 168149 126476 155439 151971 144073 128403 209676 234237 177849 180365 21205 117717 79248 126521 122797 leucine_3 297037 403812 573775 656056 453436 576255 626806 587276 547257 776630 880077 663729 715279 103864 471527 292378 514489 512968 leucine_4 30803 36845 57081 62713 44804 54298 63082 53697 54453 70186 87424 58544 68746 10053 46778 27005 46181 43437 leucine_5 0 25697 36155 41047 27290 34816 37865 35009 29898 45855 51751 43781 43231 12089 29160 14516 31038 27138 leucine_6 0 2978 851 1117 992 2238 0 1544 0 987 3635 1435 1549 1272 0 0 819 2704 Isoleucine_0 479629 0 722439 1068304 897193 659250 789509 255346 776840 758637 1838016 875210 1458194 0 638110 0 597116 739414 Isoleucine_1 39626 0 46075 82710 67724 49554 60987 20054 53444 57352 120632 67937 114426 0 52503 0 49976 48274 Isoleucine_2 2765 0 682 1087 3115 2252 1892 0 463 955 9219 2558 6172 0 2687 0 714 2344 Isoleucine_3 9666 0 7259 14640 6030 16864 12131 7821 7411 13779 19777 15768 15030 0 10788 0 10785 8387 Isoleucine_4 696 0 0 823 0 0 1720 0 0 0 0 454 746 0 0 0 0 723 Isoleucine_5 20746 0 9534 15069 12217 9064 14619 6297 4693 11607 23930 21926 24063 0 14991 0 8863 6688 Isoleucine_6 3012 0 1498 0 869 0 705 764 0 946 4175 922 2655 0 773 0 0 714 succinic acid_0 25997 0 161741 206142 104591 193138 251445 1699 320241 214770 465171 117273 350820 0 81958 0 222376 232867 succinic acid_1 141474 0 120415 191332 112138 144787 208057 101321 178554 178293 344759 206799 348131 0 78494 0 118837 188445 succinic acid_2 25559 0 32566 49177 30809 37656 57639 20436 58748 43828 120785 44595 87898 0 21264 0 34595 50611 succinic acid_3 15381 0 8851 16825 10621 10662 20236 7744 14899 15021 34568 21482 31342 0 8496 0 15361 16853 succinic acid_4 682 0 0 0 0 0 0 0 778 1092 3426 0 2750 0 0 0 0 0 glycine_0 20959 0 51837 61915 0 35604 48611 0 54631 33643 168811 46764 225681 0 13247 0 4895 20226 glycine_1 3836 0 11612 13496 0 10877 13918 0 13836 7920 47503 10726 58712 0 2344 0 0 3176 glycine_2 769 0 6518 6382 0 2866 5115 0 3703 2606 21946 5874 36767 0 0 0 0 1620 fumarate_0 188144 51209 92847 94668 28977 72363 76472 41225 46035 63239 80783 71102 158214 9280 27227 0 27960 109799 fumarate_1 41868 6570 20406 19823 7926 16486 17753 8955 11502 13647 20970 16202 37681 0 7334 0 4789 24209 fumarate_2 49245 8055 22553 28054 9561 15932 23991 12854 21963 21464 36285 19845 49970 705 14830 0 14238 24529 fumarate_3 14022 3480 7757 16144 13302 4490 11650 4067 32781 4703 41917 2951 26545 3880 16370 0 20491 9192 fumarate_4 9587 0 2235 5613 54702 892 0 755 109139 705 153109 987 12158 21903 65374 0 69201 1494 asparatate_0 0 0 0 0 0 0 923 0 0 0 751 0 1674 0 0 0 0 0 asparatate_1 384909 130271 205318 236033 79591 174638 204968 135557 137429 174792 209580 196394 466508 29714 75942 34747 80043 290716 asparatate_2 113085 32595 58320 79639 32935 48486 80715 48499 60258 59868 99386 62716 158097 12965 31994 10731 37507 85266 asparatate_3 87247 23894 45469 74467 47524 42011 63136 44717 98947 44794 136109 53668 137783 20896 49358 6517 53115 68421 asparatate_4 28066 8776 12728 27965 175831 13797 24120 13403 376572 13057 469213 14526 56735 88315 184279 0 196947 21423 Malate_0 384909 130271 205318 236033 79862 174638 204968 135557 137429 174792 206799 196394 466508 29714 75942 34747 80043 290716 Malate_1 113085 32595 58331 79639 32938 48486 80715 48499 60253 59868 99543 62636 158097 12965 31999 10731 37507 85266 Malate_2 87247 23894 45908 74467 47516 42011 62663 44717 98947 44794 134107 53668 137783 20896 49358 6517 53115 68421 Malate_3 28066 8776 12730 27965 175831 13797 24120 13403 376572 13057 469213 14526 56735 88315 184279 0 196947 21423 Malate_4 5079 0 2903 5788 28051 1373 3588 2841 62794 3981 83764 3835 15881 10758 36965 0 34533 4495 pyroglutamic acid_0 1907985 153699 1330951 1217294 290659 1337407 1314534 579062 327025 1240248 690630 1451158 2140800 13504 213767 92527 232536 1562101 pyroglutamic acid_1 425212 31813 305585 278488 76754 292671 297217 135134 83626 291389 182568 334655 485456 2442 67370 18033 63607 353623 pyroglutamic acid_2 322321 22002 241880 270806 52445 234545 315889 132330 73021 307071 122642 352353 459355 778 41087 14035 49138 248040 pyroglutamic acid_3 89365 3948 55646 77301 125006 56777 83702 34276 186418 87286 223803 99906 134118 7424 122479 783 125037 49669 pyroglutamic acid_4 43355 0 22964 49987 87888 29095 54112 19260 127599 47955 204219 58088 88667 2088 72907 0 82242 24427 pyroglutamic acid_5 13705 0 9308 16133 1363318 9111 17883 5356 1973938 17013 3231008 19027 31324 85578 1063986 0 1208116 5022 Cysteine_0 309977 29691 190582 198005 133278 222433 182010 117592 169793 206805 301999 231942 308414 10321 94797 17592 108026 236967 Cysteine_1 128491 12690 72836 75866 273055 91067 74791 49050 361324 91581 559506 99514 125928 21869 195985 6386 209291 72307 Cysteine_2 84303 7891 50058 52582 107005 61742 51088 29562 141634 58656 242078 70304 88974 8260 75387 2056 90797 56558 Cysteine_3 149574 21277 85872 93605 109782 106505 80868 50517 120744 106321 226600 128182 146869 11766 74522 10121 85509 82435 Creatinine_0 141253 15724 118181 122972 225617 112544 196510 54433 344259 194741 588896 147864 216432 21969 216357 6543 234164 84840 Creatinine_1 115042 12327 307156 329128 235828 317800 539796 25231 364738 535610 700592 404099 601134 8587 256074 5418 286874 298271 Creatinine_2 245336 21390 314791 333293 310024 317288 563567 88424 528772 530619 890371 401385 599074 22799 337294 18969 339839 300259 Creatinine_3 53765 2258 123591 166431 485787 131522 261796 14813 864466 242065 1488108 192320 295361 5812 503024 969 587791 108335 Creatinine_4 76149 5745 82395 108920 259227 90293 162116 20924 472674 152650 790602 118439 185426 3414 273410 1043 314134 73266 Threonic acid_0 80791 24560 36247 26756 49608 32545 23497 14483 55125 18794 132907 22127 47652 14248 44336 12272 53829 45603 Threonic acid_1 22416 6117 9777 9193 13621 9240 6262 6651 14881 5312 37577 6981 14462 2260 11268 2578 13898 13012 Threonic acid_2 73592 19776 27439 26783 5958 29599 23569 20198 3209 19156 13624 25982 42885 0 5105 10050 4375 35756 Threonic acid_3 20038 3013 8689 4218 0 7313 3765 2775 0 4710 1733 4818 13285 0 0 705 0 7344 Threonic acid_4 6626 2340 1737 2118 0 0 2530 851 0 0 0 1553 3526 0 0 0 0 3436 2IPM 448028 117558 253182 250093 265149 351597 455023 281461 264931 442976 687603 502596 402427 32223 227702 63448 250992 324111 Hypotaurine_0 31026 9444 18093 19081 17002 23487 31583 21422 18328 26740 43616 29530 27810 0 19852 3238 21474 23945 Hypotaurine_1 71725 25255 49205 48304 58216 67187 92417 55742 65513 91020 113505 85201 74380 5936 50614 11182 52403 63276 Hypotaurine_2 26969 9847 16397 17436 23252 25638 30455 19493 19195 27719 47756 29862 25225 1137 16961 4389 16789 21593 Glycerol-1-phosphate_0 31984 9535 12608 17058 11213 14758 13835 5294 12928 12062 29943 18620 35583 0 10326 0 11672 21714 Glycerol-1-phosphate_1 9903 1215 3073 5625 1570 2120 3293 1959 1732 2582 9231 3114 10845 0 1507 0 2528 6202 Glycerol-1-phosphate_2 11066 1942 0 4660 2728 4210 3244 0 4404 2059 8775 1457 9596 0 1557 0 1151 6495 Glycerol-1-phosphate_3 719 0 0 0 0 855 0 0 0 0 1327 0 1574 0 0 0 0 0 o-phosphocolamine_0 31984 9535 12607 17058 11213 14640 13835 5294 12928 12062 29943 18620 35583 0 10326 0 11672 21714 o-phosphocolamine_1 9903 1215 3073 5625 1570 2120 3293 1959 1732 2582 9231 3114 10909 0 1507 0 2528 6202 o-phosphocolamine_2 11066 1942 0 4660 2728 4108 3244 0 4404 2059 8775 1457 9596 0 1557 0 1151 6495 citrate_0 58853 0 98404 104868 31501 88879 75723 37600 39434 64872 121952 84838 232546 0 35591 0 35981 131170 citrate_1 16769 0 29168 40166 14706 26834 22154 7777 16931 19194 54229 23455 82083 0 16249 0 18160 41100 citrate_2 72995 0 90263 196614 13974 93778 116397 57760 17709 94158 48390 121690 408676 0 16586 0 15792 129402 citrate_3 24299 0 33438 76254 37143 32727 40071 22588 66031 32367 150659 46152 160004 0 53035 0 54022 43276 citrate_4 19672 0 27112 61563 9748 25219 33622 16373 17094 27053 31046 34223 131412 0 13239 0 11768 29025 citrate_5 5424 0 8336 26338 20694 6802 11320 5254 49347 6942 80043 11247 55149 0 26168 0 34099 6494 citrate_6 723 0 0 5274 1702 0 1979 0 6141 0 13751 1224 14086 0 5189 0 2951 755 Tyrosine_0 542482 262732 220640 172519 53303 406559 164958 106866 52330 385345 89685 446724 249882 8275 32658 88664 45265 260772 Tyrosine_1 231839 108808 82682 73318 19671 155967 66280 47060 17090 145987 30033 181964 99395 3642 14918 43330 16432 99447 Tyrosine_2 54449 23451 22715 17246 4634 42451 11726 8082 2068 33302 7211 46037 22546 0 842 7877 2712 23655 Tyrosine_3 15327 5352 4696 2195 882 7547 1982 0 733 6926 0 10537 4103 0 0 0 0 5849 Tyrosine_4 0 728 0 0 14563 0 887 0 18788 0 33126 2397 0 1001 10502 0 16663 0 Tyrosine_5 3779 0 0 0 2772 0 0 0 6124 0 11732 0 0 1548 1065 0 2450 0 Tyrosine_6 3133 833 0 0 46436 737 1010 0 49037 2062 87842 2544 732 7348 25062 0 37836 892 Tyrosine_7 0 0 0 0 11018 787 0 0 14496 0 23693 0 0 0 7340 0 8717 0 Tyrosine_8 3238 687 0 0 35395 0 0 1165 46278 0 65369 1892 0 5768 20803 0 33432 0 Tyrosine_9 3584 1083 0 0 30118 969 0 0 28233 2736 52909 869 751 3726 20729 0 23258 768 Palmitate 59558 17941 49190 67556 51806 52290 51181 25153 67729 117831 295535 70706 71257 0 43386 5116 51086 54039 Myo-inositol_0 11182469 3193927 4307608 4643063 5078200 4649359 5296248 3635530 5320193 4409060 11748214 5103413 7653240 1376695 4041728 2004760 4424122 4778158 Myo-inositol_1 3297451 933300 1269151 1342967 1460925 1369233 1548450 1044661 1556364 1315141 3455982 1497758 2194103 403654 1147572 583982 1293712 1363619 Myo-inositol_2 1653875 463133 623228 664975 709021 669618 771275 520869 747329 652165 1765590 757385 1130299 205891 566513 292924 644810 659540 Myo-inositol_3 346707 98325 127643 142221 146245 140099 170140 119146 164350 134319 351810 163370 239984 42578 123503 66735 130394 145470 Myo-inositol_4 89505 26250 37371 35305 37363 43585 44948 26893 44028 38808 91318 38831 56221 9396 29595 12413 39454 38957 Myo-inositol_5 9304 3759 3078 4604 4965 3858 4187 2734 4453 3874 12897 4756 9274 0 2905 0 4744 2381 Myo-inositol_6 955 0 0 0 0 0 0 0 0 0 946 0 0 0 728 0 0 0 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name product m/z pyruvate_0 174 pyruvate_1 175 pyruvate_2 176 pyruvate_3 177 Lactate_0 219 Lactate_1 220 Lactate_2 221 Lactate_3 222 alanine_0 116 alanine_1 117 alanine_2 118 alanine_3 119 Acetohydroxamic acid_0 249 Acetohydroxamic acid_1 250 Acetohydroxamic acid_2 251 Oxalic acid_0 190 Oxalic acid_1 191 Oxalic acid_2 192 Oxalic acid_3 193 Oxalic acid_4 194 leucine_0 86 leucine_1 87 leucine_2 88 leucine_3 89 leucine_4 90 leucine_5 91 leucine_6 92 Isoleucine_0 86 Isoleucine_1 87 Isoleucine_2 88 Isoleucine_3 89 Isoleucine_4 90 Isoleucine_5 91 Isoleucine_6 92 succinic acid_0 218 succinic acid_1 219 succinic acid_2 220 succinic acid_3 221 succinic acid_4 222 glycine_0 276 glycine_1 277 glycine_2 278 fumarate_0 245 fumarate_1 246 fumarate_2 247 fumarate_3 248 fumarate_4 249 asparatate_0 232 asparatate_1 233 asparatate_2 234 asparatate_3 235 asparatate_4 236 Malate_0 233 Malate_1 234 Malate_2 235 Malate_3 236 Malate_4 237 pyroglutamic acid_0 258 pyroglutamic acid_1 259 pyroglutamic acid_2 260 pyroglutamic acid_3 261 pyroglutamic acid_4 262 pyroglutamic acid_5 263 Cysteine_0 100 Cysteine_1 101 Cysteine_2 102 Cysteine_3 103 Creatinine_0 115 Creatinine_1 116 Creatinine_2 117 Creatinine_3 118 Creatinine_4 119 Threonic acid_0 292 Threonic acid_1 293 Threonic acid_2 294 Threonic acid_3 295 Threonic acid_4 296 2IPM 377 Hypotaurine_0 100 Hypotaurine_1 101 Hypotaurine_2 102 Glycerol-1-phosphate_0 387 Glycerol-1-phosphate_1 388 Glycerol-1-phosphate_2 389 Glycerol-1-phosphate_3 390 o-phosphocolamine_0 387 o-phosphocolamine_1 388 o-phosphocolamine_2 389 citrate_0 273 citrate_1 274 citrate_2 275 citrate_3 276 citrate_4 277 citrate_5 278 citrate_6 279 Tyrosine_0 280 Tyrosine_1 281 Tyrosine_2 282 Tyrosine_3 283 Tyrosine_4 284 Tyrosine_5 285 Tyrosine_6 286 Tyrosine_7 287 Tyrosine_8 288 Tyrosine_9 289 Palmitate 313 Myo-inositol_0 305 Myo-inositol_1 306 Myo-inositol_2 307 Myo-inositol_3 308 Myo-inositol_4 309 Myo-inositol_5 310 Myo-inositol_6 311 METABOLITES_END #END