#METABOLOMICS WORKBENCH araskind_20170619_113136 DATATRACK_ID:998 STUDY_ID:ST000702 ANALYSIS_ID:AN001091 PROJECT_ID:PR000499 VERSION 1 CREATED_ON July 1, 2017, 5:22 pm #PROJECT PR:PROJECT_TITLE NIH 3T3 fibroblast cells with Ceftriaxone Treatment PR:PROJECT_TYPE MS analysis PR:PROJECT_SUMMARY 3T3 cells treated with 50uM Ceftriaxone for 24hrs, 1hr, or 0 hrs. PR:INSTITUTE Vanderbilt University PR:DEPARTMENT Pharmacology PR:LABORATORY Blakely Lab PR:LAST_NAME Blakely PR:FIRST_NAME Randy PR:ADDRESS Nashville, TN PR:EMAIL randy.blakely@vanderbilt.edu PR:PHONE 615-936-1700 #STUDY ST:STUDY_TITLE NIH 3T3 fibroblast cells with Ceftriaxone Treatment ST:STUDY_TYPE MS analysis ST:STUDY_SUMMARY NIH 3T3 cells were plated at 1.5million cells per 10cM dish into 12 10cm dishes. ST:STUDY_SUMMARY 24hrs post plating media was changed on all plates. At the time of media change, ST:STUDY_SUMMARY 4 dishes were treated with a final concentration of 50uM ceftriaxone (24hrs ST:STUDY_SUMMARY ceftriaxone n=4). 23 hrs post media change, 4 separate dishes were treated with ST:STUDY_SUMMARY 50uM ceftriaxone for 1hr (1hr ceftriaxone n=4). 4 dishes were untreated (0hr, ST:STUDY_SUMMARY basal, n=4). 24 hrs post media change, media was quickly removed from all 12 ST:STUDY_SUMMARY plates, each plate was rapidly washed with MilliQ H2O and rapidly frozen with ST:STUDY_SUMMARY liquid nitrogen poured directly into the dishes. All plates were transferred to ST:STUDY_SUMMARY dry ice and stored at -80 until shipped. ST:INSTITUTE University of Michigan ST:DEPARTMENT Biomedical Research Core Facilities ST:LABORATORY Metabolomics core ST:LAST_NAME Kachman ST:FIRST_NAME Maureen ST:ADDRESS 6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714 ST:EMAIL mkachman@med.umich.edu ST:PHONE (734) 232-8175 ST:NUM_GROUPS 3 ST:TOTAL_SUBJECTS 2 #SUBJECT SU:SUBJECT_TYPE Mouse SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10088 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS SU0012748 S00019331 Timepoint:0hr SUBJECT_SAMPLE_FACTORS SU0012748 S00019332 Timepoint:0hr SUBJECT_SAMPLE_FACTORS SU0012748 S00019333 Timepoint:0hr SUBJECT_SAMPLE_FACTORS SU0012748 S00019334 Timepoint:0hr SUBJECT_SAMPLE_FACTORS SU0012748 S00019335 Timepoint:1hr SUBJECT_SAMPLE_FACTORS SU0012748 S00019336 Timepoint:1hr SUBJECT_SAMPLE_FACTORS SU0012748 S00019337 Timepoint:1hr SUBJECT_SAMPLE_FACTORS SU0012748 S00019338 Timepoint:1hr SUBJECT_SAMPLE_FACTORS SU0012748 S00019339 Timepoint:24hr SUBJECT_SAMPLE_FACTORS SU0012748 S00019340 Timepoint:24hr SUBJECT_SAMPLE_FACTORS SU0012748 S00019341 Timepoint:24hr SUBJECT_SAMPLE_FACTORS SU0012748 S00019342 Timepoint:24hr SUBJECT_SAMPLE_FACTORS SU9999996 S00020154 Timepoint:- #COLLECTION CO:COLLECTION_SUMMARY - #TREATMENT TR:TREATMENT_SUMMARY - #SAMPLEPREP SP:SAMPLEPREP_SUMMARY - SP:SAMPLEPREP_PROTOCOL_FILENAME A003 - Untargeted Metabolomics.pdf #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Agilent CH:COLUMN_NAME Waters Acquity HSS T3 (50 x 2.1mm, 1.8um) #ANALYSIS AN:ANALYSIS_TYPE MS AN:ANALYSIS_PROTOCOL_FILE A003 - Untargeted Metabolomics.pdf #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Agilent 6530 QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area normalized MS_METABOLITE_DATA_START Samples S00019331 S00019332 S00019333 S00019334 S00019335 S00019336 S00019337 S00019338 S00019339 S00019340 S00019341 S00019342 Factors Timepoint:0hr Timepoint:0hr Timepoint:0hr Timepoint:0hr Timepoint:1hr Timepoint:1hr Timepoint:1hr Timepoint:1hr Timepoint:24hr Timepoint:24hr Timepoint:24hr Timepoint:24hr (S)-2-AMINO-3-(3-HYDOXY-4-OXO-4H-PYRIDIN-1-YL)PROPANOATE OR L-MIMOSINE (M-H)- 975 14966 15884 19041 10724 8168 6362 10299 13156 6014 8521 5185 (S)-3-METHYL-2-OXOPENTANOATE (M-H)- 503394 521354 695525 569959 940389 1088098 1000138 917488 624009 1352130 989277 1403809 (S)-DIHYDROOROTATE (M-H)- 1988 1327 2762 5038 1053 6990 1-OLEOYL-RAC-GLYCEROL (M+HCOO)- 10796 2845 8839 8538 11690 6826 6167 4426 3176 2140 3028 2116 18:0 LYSO-PE (M-H)- 601806 74361 168181 349344 48286 380019 231574 204170 146635 145695 286622 143590 2-DEOXY-D-GLUCOSE (M+CL)- 170154 150944 54532 2-HYDROXY-3-METHYLBUTYRIC ACID (M-H)- 23601 20097 17288 20582 23177 27320 24462 24490 12898 31669 26144 30324 3-(4-HYDROXYPHENYL)LACTATE (M-H)- 175545 152000 197364 183735 219000 238707 240582 204896 185357 225484 253385 226167 3-HYDROXYBENZOATE (M-H)- 1776 2103 2695 2762 3773 4945 3850 6957 3774 5571 3397 5327 4-HYDROXYBENZOATE (M-H)- 5229 4553 2321 2784 2966 2085 7097 2276 3561 2181 4-METHYL-2-OXOPENTANOATE (M-H)- 590412 585463 789377 659132 963186 1155630 1004670 1003829 710724 1298657 1100460 1323593 5-AMINOPENTANOATE (M-H)- 27726 31064 12392 32744 15149 35316 18018 27657 33692 19275 31856 41826 ADENOSINE (M+HCOO)- 14257 35558 11111 17364 7868 29896 ADP-RIBOSE (M-H)- 61935 14214 8877 5480 33932 34467 43462 8202 5064 42732 47321 12727 ADPGLUCOSE (M-H)- 92305 90529 86347 108606 65584 64163 66693 64800 101964 62450 73732 55545 ASPARTATE (M-H)- 131645 125514 161515 91361 68184 74428 77853 75654 93936 60964 BEHENIC ACID (M-H)- 6491 3629 6812 4658 3788 3612 5114 6651 5943 6704 8138 CIS-VACCENIC ACID (M-H)- 766664 421234 647139 213594 53272 193473 344376 113874 1087076 189222 420699 540237 CORTISOL (M+CL)- 5836 4032 6247 4872 7285 9265 8694 8764 5302 8385 7766 8959 D-GLUCOSE 6-PHOSPHATE (M-H)- 1405541 165234 532570 26852 90905 199172 12605 70329 7034789 111678 DEOXYURIDINE (M+HCOO)- 50270 26190 18749 20811 7873 16306 4773 19202 35022 35277 DOCOSAHEXAENOIC ACID (M-H)- 145031 27518 44782 34049 57538 38038 63450 27755 47636 37256 118817 50097 EDTA (M-H)- 6101 1954 5273 7502 6075 8325 19037 9712 5527 22364 8950 25747 ERUCIC ACID (M-H)- 33841 29964 14207 15599 19666 18809 9430 11369 6651 6269 20565 5334 FLAVIN ADENINE DINUCLEOTIDE (M-H)- 132377 132039 93591 90688 68363 96261 69639 83987 127932 71137 124868 101860 FUMARATE (M-H)- 174827 188071 230174 224775 163048 143398 134735 124798 240867 215686 169716 196569 GDPMANNOSE (M-H)- 66496 60096 60120 74190 38652 42010 44706 43564 68069 36662 59443 45110 GIBBERELLIC ACID [ISTD] (M+CL)- 782000 785000 784000 786000 786000 854000 790000 815000 763000 803000 803000 815000 GIBBERELLIC ACID [ISTD] (M-H)- 5105730 5043743 5013039 5043920 4878106 4945200 4930695 5059290 4795212 4943929 5099782 4794984 GLUCOSE (M+CL)- 4487240 4631634 5360046 3849820 6178225 5577937 6894640 7767832 4800274 7933188 5405863 7496904 GLUCURONIC ACID (M-H)- 15371 10846 9643 11914 7379 7142 6636 7546 11576 3961 8250 5587 GLUTAMATE (M-H)- 2450000 2450000 2150000 2650000 1940000 1550000 1610000 1740000 2760000 1420000 1940000 1100000 GLUTARATE (M-H)- 15305 8686 9864 8597 12803 10900 12277 11966 10207 11327 11108 10687 GLUTATHIONE (OXIDIZED) (M-H)- 2680000 2970000 2870000 3520000 6030000 6710000 6340000 6250000 2448654 6020000 6310000 5600000 GLYCEROL 2-PHOSPHATE (M-H)- 38997 19034 38412 37505 8327 4621 GLYCOCHOLATE (M-H)- 1194 1378 2067 1933 2488 2738 2809 4177 2889 GUANOSINE 5'-PHOSPHATE (M-H)- 108252 91810 44202 77030 90629 65434 34482 22361 28479 78454 HEPTADECANOATE (M-H)- 141852 69512 75008 53118 36655 65481 40414 47734 32916 36982 105182 49690 HEXACOSANOIC ACID (M-H)- 11136 4639 13795 8502 7626 9904 10118 12256 8605 10948 12173 8130 HYODEOXYCHOLATE (M-H)- 213512 223303 312248 245496 222069 261050 478415 144321 489108 338077 222957 515208 INOSINE (M-H)- 1334455 251610 127816 49840 110265 24178 77305 216135 323043 INOSINE 5'-PHOSPHATE (M-H)- 327368 257566 407070 134773 441925 513720 165545 298393 182004 228057 448679 301245 ISOLEUCINE (M-H)- 717585 713572 694172 776886 477949 1227637 901997 479105 774675 579177 567006 805943 L-TRYPTOPHAN-15N2 [ISTD] (M-H)- 249271 233422 223094 223959 212699 218872 220690 245036 219507 238434 239991 221933 L-[15N]ANTHRANILIC ACID [ISTD] (M-H)- 138810 133565 121585 129269 118537 117764 115579 127333 115048 119075 124355 104404 LAURIC ACID (M-H)- 5702 4650 4856 3146 3692 2648 3460 3011 2874 4244 4567 3161 LEUCINE (M-H)- 705752 597757 1398063 665742 1260733 1126587 1308625 686373 671722 1654990 394676 361182 LIGNOCERIC ACID (M-H)- 11514 7210 12075 8457 7425 8264 6484 10691 12475 10845 11067 12656 LINOLEATE (M-H)- 10187601 54204 229872 1570110 41666 5693794 167847 975936 4423403 427397 LUMICHROME (M-H)- 2162 4006 3817 5471 8624 5767 2037 4080 3997 3893 LYSINE (M-H)- 5905 5587 5087 5249 5719 7462 8102 6552 4922 6033 5902 8445 MANNITOL (M-H)- 119809 125019 81186 103115 46628 72361 82620 71539 86053 56715 109168 57932 N-ACETYL-D-GLUCOSAMINE (M+CL)- 15822 11101 13720 9971 10814 17563 9807 9550 13296 8130 13303 13509 N-ACETYL-DL-METHIONINE (M-H)- 79669 64375 77407 55722 70587 78268 70509 65324 56028 66501 88187 64319 N-ACETYL-L-ALANINE (M-H)- 167208 140913 191627 137535 182784 173218 171926 167582 145690 164756 173213 139524 N-ACETYL-L-PHENYLALANINE (M-H)- 11860 12583 6390 15614 9362 20813 15957 13875 12406 17082 15803 17138 N-ACETYLNEURAMINATE (M-H)- 116054 157045 113430 213861 202803 116238 167878 130861 154688 256319 157523 136837 OROTATE (M-H)- 37035 24469 19148 22563 10176 11727 11425 10242 8830 7790 21082 20980 OXALOACETATE (M-H)-[-H2O] 103047 125939 93939 126771 135265 109546 138586 129288 88794 136131 77929 137465 PALMITATE (M-H)- 438059 153360 408194 136143 959502 106839 385333 133588 153395 258566 98824 72084 PALMITOLEIC ACID (M-H)- 140740 318016 431217 173095 234016 336250 271798 236740 267016 206137 364350 PHTHALATE (M-H)- 13842 14009 7205 3724 3388 2584 2316 3466 2601 7676 1786 PIMELIC ACID (M-H)- 14079 4551 6915 10962 5140 6725 4397 8192 4316 6535 4165 PROSTAGLANDIN E2 (M+CL)- 14721 11119 9305 9712 4857 7820 6249 8688 7945 5882 9422 8449 PROSTAGLANDIN E2 (M-H)-[-H2O] 6717 6135 5896 5589 1978 3056 2667 3911 4115 2619 5269 3861 PURINE (M-H)- 22300 18362 21562 40040 49645 21429 5958 18811 13504 PYRUVATE (M-H)- 30236 32538 39495 29427 31136 33106 32162 30928 32485 34394 35554 22734 RETINOATE (M-H)- 19940 17675 16295 13126 21041 16434 14491 12183 9461 10299 20343 13994 SALICYLATE (M-H)- 31845 20512 23075 19879 29400 34400 33000 28700 23120 34500 30300 34000 SEBACIC ACID (M-H)- 18303 13149 13034 14356 18197 15898 13807 12881 13508 13357 15599 13347 SN-GLYCERO-3-PHOSPHOCHOLINE (M+HCOO)- 37065 32454 34525 45171 29144 38090 32287 32411 42180 24844 35434 11675 STEARATE (M-H)- 255371 296375 402530 337301 705635 331218 740523 285772 348512 557732 168831 320916 SUCCINATE (M-H)- 425387 377960 400110 228453 357708 348042 331692 259033 206398 360374 288656 409405 THYMIDINE (M+CL)- 23681 12385 8451 7461 11594 8346 6549 11045 17863 THYMIDINE 5'-MONOPHOSPHATE (M-H)- 10540 2503 8275 7495 11769 13910 7412 2377 6839 TRANS-ACONITATE (M-H)- 265182 427496 151059 287290 110154 420177 178077 200910 112795 200531 147930 322526 TRICARBALLYLIC ACID (M-H)-[-H2O] 50329 51314 58369 52508 31411 35272 37065 61807 54708 21101 24472 86940 TYROSINE (M-H)- 538012 475307 609154 547608 610685 621731 654201 683164 575926 738381 635802 638079 UDP-N-ACETYL-D-GALACTOSAMINE (M-H)+[-H2O] 83286 90813 82285 109571 61353 71838 62679 61589 89919 59928 67557 58867 URIDINE (M-H)- 170383 64382 46794 53166 13510 32891 23545 37808 27520 11500 58238 86207 URIDINE-5-MONOPHOSPHATE (M-H)- 298196 252653 212914 95371 238360 343795 261177 137402 159216 115355 241800 VALINE (M-H)- 27587 31137 12091 34083 14682 29261 17694 25413 27952 27497 31500 42042 XANTHINE (M-H)- 135634 23970 47849 34379 2930 3645 2092 4281 1687 35842 49415 XANTHOSINE 5'-PHOSPHATE (M-H)- 3292 2690 11635 2501 9150 9629 3463 10540 5342 4260 7848 6730 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name moverz_quant ri ri_type pubchem_id inchi_key kegg_id other_id other_id_type (S)-2-AMINO-3-(3-HYDOXY-4-OXO-4H-PYRIDIN-1-YL)PROPANOATE OR L-MIMOSINE (M-H)- 198.0628 0.679 440473 (S)-3-METHYL-2-OXOPENTANOATE (M-H)- 130.0632 4.253 47 (S)-DIHYDROOROTATE (M-H)- 158.0344 0.712 1149667 1-OLEOYL-RAC-GLYCEROL (M+HCOO)- 356.2902 22.701 5283468 18:0 LYSO-PE (M-H)- 481.3164 22.679 9547068 2-DEOXY-D-GLUCOSE (M+CL)- 164.068 0.882 439268 2-HYDROXY-3-METHYLBUTYRIC ACID (M-H)- 118.0626 3.456 853180 3-(4-HYDROXYPHENYL)LACTATE (M-H)- 182.0577 3.802 9378 3-HYDROXYBENZOATE (M-H)- 138.0316 5.05 7420 4-HYDROXYBENZOATE (M-H)- 138.0315 4.099 135 4-METHYL-2-OXOPENTANOATE (M-H)- 130.063 4.658 70 5-AMINOPENTANOATE (M-H)- 117.0782 0.74 138 ADENOSINE (M+HCOO)- 267.0962 1.794 60961 ADP-RIBOSE (M-H)- 559.0684 0.734 33576 ADPGLUCOSE (M-H)- 589.0807 1.025 16500 ASPARTATE (M-H)- 133.0378 0.661 5960 BEHENIC ACID (M-H)- 340.3335 24.09 8215 CIS-VACCENIC ACID (M-H)- 282.2562 22.966 5282761 CORTISOL (M+CL)- 362.2094 13.789 5754 D-GLUCOSE 6-PHOSPHATE (M-H)- 260.0302 0.712 5958 DEOXYURIDINE (M+HCOO)- 228.0733 1.648 13712 DOCOSAHEXAENOIC ACID (M-H)- 328.2408 22.706 445580 EDTA (M-H)- 292.0908 1.205 6049 ERUCIC ACID (M-H)- 338.3181 23.668 5281116 FLAVIN ADENINE DINUCLEOTIDE (M-H)- 785.1568 6.348 643975 FUMARATE (M-H)- 116.0111 1.265 444972 GDPMANNOSE (M-H)- 605.0748 0.863 18396 GIBBERELLIC ACID [ISTD] (M+CL)- 346.1414 8.49 6466 GIBBERELLIC ACID [ISTD] (M-H)- 346.1421 8.493 6466 GLUCOSE (M+CL)- 180.0651 0.668 5793 GLUCURONIC ACID (M-H)- 194.0406 0.679 94715 GLUTAMATE (M-H)- 147.0525 0.663 33032 GLUTARATE (M-H)- 132.0422 2.036 743 GLUTATHIONE (OXIDIZED) (M-H)- 612.1521 1.36 65359 GLYCEROL 2-PHOSPHATE (M-H)- 172.0146 0.676 2526 GLYCOCHOLATE (M-H)- 465.3081 18.843 10140 GUANOSINE 5'-PHOSPHATE (M-H)- 363.0519 0.996 6804 HEPTADECANOATE (M-H)- 270.2555 23.025 10465 HEXACOSANOIC ACID (M-H)- 396.396 25.642 10469 HYODEOXYCHOLATE (M-H)- 392.295 22.909 5283820 INOSINE (M-H)- 268.0814 1.835 6021 INOSINE 5'-PHOSPHATE (M-H)- 348.0498 0.991 8582 ISOLEUCINE (M-H)- 131.0945 1.514 6306 L-TRYPTOPHAN-15N2 [ISTD] (M-H)- 206.0843 3.633 16217285 L-[15N]ANTHRANILIC ACID [ISTD] (M-H)- 138.0447 5.335 227 LAURIC ACID (M-H)- 200.1779 21.772 3893 LEUCINE (M-H)- 131.0945 1.542 6106 LIGNOCERIC ACID (M-H)- 368.3651 24.748 11197 LINOLEATE (M-H)- 280.2409 22.732 5280450 LUMICHROME (M-H)- 242.0802 10.934 5326566 LYSINE (M-H)- 146.1052 0.585 5962 MANNITOL (M-H)- 182.079 0.674 6251 N-ACETYL-D-GLUCOSAMINE (M+CL)- 221.0916 0.684 446101 N-ACETYL-DL-METHIONINE (M-H)- 191.0613 4.038 448580 N-ACETYL-L-ALANINE (M-H)- 131.0581 1.503 88064 N-ACETYL-L-PHENYLALANINE (M-H)- 207.0889 7.683 74839 N-ACETYLNEURAMINATE (M-H)- 309.1066 0.688 439197 OROTATE (M-H)- 156.0161 0.814 967 OXALOACETATE (M-H)-[-H2O] 132.0038 0.589 970 PALMITATE (M-H)- 256.2405 22.853 985 PALMITOLEIC ACID (M-H)- 254.2251 22.6 445638 PHTHALATE (M-H)- 166.0249 4.792 1017 PIMELIC ACID (M-H)- 160.0733 5.666 385 PROSTAGLANDIN E2 (M+CL)- 352.2251 16.766 5280360 PROSTAGLANDIN E2 (M-H)-[-H2O] 352.2243 16.768 5280360 PURINE (M-H)- 120.0435 1.505 1044 PYRUVATE (M-H)- 88.017 0.741 1060 RETINOATE (M-H)- 300.2098 22.26 444795 SALICYLATE (M-H)- 138.0316 8.214 338 SEBACIC ACID (M-H)- 202.1205 12.94 5192 SN-GLYCERO-3-PHOSPHOCHOLINE (M+HCOO)- 257.1005 0.715 439285 STEARATE (M-H)- 284.2722 23.193 5281 SUCCINATE (M-H)- 118.0269 1.304 1110 THYMIDINE (M+CL)- 242.0898 2.735 5789 THYMIDINE 5'-MONOPHOSPHATE (M-H)- 322.0583 1.529 12803287 TRANS-ACONITATE (M-H)- 174.0165 1.131 444212 TRICARBALLYLIC ACID (M-H)-[-H2O] 176.0302 1.327 14925 TYROSINE (M-H)- 181.0736 1.355 6057 UDP-N-ACETYL-D-GALACTOSAMINE (M-H)+[-H2O] 607.0926 1.025 23724461 URIDINE (M-H)- 244.0716 1.366 6029 URIDINE-5-MONOPHOSPHATE (M-H)- 324.0364 0.857 6030 VALINE (M-H)- 117.0782 0.757 6287 XANTHINE (M-H)- 152.0336 1.264 1188 XANTHOSINE 5'-PHOSPHATE (M-H)- 364.043 1.028 73323 METABOLITES_END #END