#METABOLOMICS WORKBENCH araskind_20170619_113136 DATATRACK_ID:998 STUDY_ID:ST000702 ANALYSIS_ID:AN001092 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 POSITIVE #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 1-METHYLNICOTINAMIDE M+ 784318 852634 266643 30536 307516 188321 5066712 717188 4323379 1026857 1122849 1-METHYLXANTHINE (M+NA)+ 15430 13761 16547 14514 19273 22003 19393 14688 22328 21082 24564 1-OLEOYL-RAC-GLYCEROL (M+H)+[-H2O] 9021 11408 9035 7921 22435 6844 3139 4636 1832 4765 5223 5830 1-OLEOYL-RAC-GLYCEROL (M+NA)+ 358942 341811 193370 154585 173278 156009 66277 71051 54563 34819 186734 69064 3-HYDROXYBENZALDEHYDE (M+H)+[-H2O] 42731 37828 49508 35173 63961 80078 77205 74002 44392 84439 64404 96981 5'-METHYLTHIOADENOSINE (M+NA)+ 17697 16471 15198 12440 18079 13129 23020 12438 17986 21632 23418 20179 5-AMINOPENTANOATE (M+H)+ 2000475 1997973 1994017 2112606 2034592 2100668 2476046 2102519 2193100 2129668 2183036 5-HYDROXYTRYPTOPHAN (M+H)+ 51378 39154 61197 54482 39073 28335 37457 43479 44821 ADENOSINE (M+H)+ 1190000 408000 75500 70200 630000 148000 104000 147000 28100 970000 459000 ADENOSINE 5'-MONOPHOSPHATE (M+H)+ 596355 416429 31912 343478 487641 632940 435617 344511 244635 37260 529985 ALPHA-TOCOPHEROL (M+H)+ 1843 1550 2085 1363 2188 2163 2149 2055 1602 2252 2230 AZELAIC ACID (M+H)+[-H2O] 6689 2470 3400 3308 6169 4014 2300 2343 3220 1044 3894 2330 BILIVERDIN (M+H)+ 2571 2024 3126 3947 3493 1690 4107 3127 5484 BIS(2-ETHYLHEXYL)PHTHALATE (M+NA)+ 53998 63435 54693 51039 126386 65010 71529 35711 31361 32637 62380 35399 BUTYRYLCARNITINE (*) (M+H)+ 359700 727789 742486 807000 649000 430000 836376 962000 680933 1190000 CARNITINE (M+H)+ 539242 521624 1539690 714446 930494 3690024 724180 4064444 836882 644639 CHOLESTEROL (M+H)+[-H2O] 211256 205669 180000 231000 91800 192000 188000 181000 120000 156000 212000 149000 CHOLINE M+ 2202 2130 1581 1068 1919 1126 10926 1395 2736 1095 8679 CHOLINE PHOSPHATE (M+NA)+ 509649 324824 587648 279270 573919 280267 608043 580495 485297 596934 594198 500862 CITRATE (M+NA)+ 40041 73480 85989 39118 75257 79754 81518 85751 44591 96129 54212 91212 CORTISOL (M+H)+ 3322 2766 3127 3711 3444 6504 5817 4252 3254 4088 4249 7056 CORTISOL (M+NA)+ 1216 1073 1426 1363 2712 2830 2595 2894 1084 1921 2656 3061 CREATINE (M+H)+ 6772129 6300756 7183675 6181665 5827977 6871155 6786794 6916897 7246183 5932289 6561347 5840530 CREATININE (M+H)+ 91909 78929 133283 88045 137225 95964 174398 152309 91118 171640 84850 68148 DECANOATE (2M+NA)+ 8320 6390 4630 3570 3940 2480 DGMP (M+H)+ 78635 61306 41145 44445 32151 52425 83015 27549 13957 54699 54658 ELAIDIC ACID (M+H)+ 8839 6844 6528 6149 1642 6366 3539 3429 2890 3478 7743 5453 ETHANOLAMINE PHOSPHATE (M+NA)+ 7540 3070 8800 1760 19600 23300 5080 11400 2590 29200 6790 GAMMA-L-GLUTAMYL-L-CYSTEINE (M+H)+ 92209 110669 92546 53310 62461 68305 57314 58394 55885 34749 79784 48741 GIBBERELLIC ACID [ISTD] (2M+NA)+ 45680 45870 47555 44596 43886 47245 43288 48965 42743 36377 48089 44281 GLUTAMATE (M+H)+[-H2O] 217696 1813912 1789552 105672 221729 338009 459758 4882307 1091944 2825787 331390 1203260 GLUTATHIONE OXIDIZED (M+H2)+2 2532188 2648220 2262117 3436059 3862078 3225678 5570980 4485398 3233422 6083665 3115336 3703024 GUANINE (M+H)+ 18685 64830 91229 36478 102336 37603 95312 50255 22561 176364 75029 23660 GUANOSINE (M+H)+ 163060 81898 17514 7169 18468 11716 HIPPURATE (M+H)+ 136790 128238 169816 126526 222652 275605 267778 266225 146285 298629 232932 346767 INOSINE 5'-PHOSPHATE (M+H)+ 225770 181976 168417 125084 112905 196417 INOSINE 5'-PHOSPHATE (M+NA)+ 16863 8686 17594 1089 18477 20907 7043 9120 6075 14477 17342 9859 ISOLEUCINE (M+H)+ 51500000 49400000 55100000 50000000 58800000 62100000 62300000 60800000 52100000 63600000 60700000 65300000 ISOVALERYLCARNITINE (*) (M+H)+ 33056 16683 16775 8765 16966 15686 21765 12601 28403 37885 18411 22477 KYNURENINE (M+H)+ 54254 43751 59590 49316 73935 81793 84165 78605 55780 77924 81886 88345 L-ARGININE (M+H)+ 61239 127092 72216 60750 64794 117825 74941 394286 54782 141323 95961 300802 L-ARGININE (M+NA)+ 17552 17053 18490 12091 18425 23142 24657 24240 15015 25271 18552 28787 L-HISTIDINE (M+H)+ 6070 5773 8447 6487 6673 8826 8001 15925 7516 13661 5510 57326 L-TRYPTOPHAN-15N2 [ISTD] (M+H)+ 2340306 2347104 2366425 2363986 2349309 2312309 2289341 2342039 2222003 2155706 2402362 2221321 LYSINE (M+H)+ 131232 116707 127112 101158 120658 138336 138769 147464 121160 136569 133859 157475 LYSINE (M+NA)+ 20119 20204 17102 15635 20019 24427 19749 24710 17994 23658 19048 32026 MANNOSE-6-PHOSPHATE (M+NA)+ 45952 35117 28375 37189 19614 N-ACETYL-DL-METHIONINE (M+H)+[-H2O] 7026 7035 7939 10001 6683 11776 1734 3740 6504 2666 10807 N-ACETYL-DL-SERINE (M+H)+[-H2O] 5429079 2681050 3194757 2622937 4546409 5014384 4239317 4106095 2713545 4578127 4907054 4695828 N-ACETYL-L-ALANINE (M+NA)+ 3334 319 3209 11303 2939 3244 1917 3859 5852 N-ACETYL-L-LEUCINE (M+NA)+ 31836 21549 33369 28359 27655 26291 27707 28345 36312 27164 N-ACETYLNEURAMINATE (M+H)+ 10678 21155 9787 38666 60574 8190 49696 12396 16898 265220 15071 24285 N-METHYL-D-ASPARTIC ACID (M+NA)+ 49125 47882 40289 49946 45865 49808 43486 61421 36233 66698 42298 72030 NAD (M+2H)+2 181904 220878 169828 239704 219290 178566 226099 183273 222241 212802 214712 175710 NAD (M+H)+ 336077 370729 394254 446067 474610 464135 428621 369766 400829 398062 491484 359961 NAD (M+NA)+ 28049 29221 31318 33559 41966 40208 45009 37761 29917 48153 38511 42558 NORADRENALINE (M+H)+ 370618 397795 395733 436278 444710 472128 523343 414835 520898 377861 574123 O-PHOSPHOSERINE (M+NA)+ 29851 39748 30637 37032 39060 41966 40114 34607 37423 37543 33346 OLEAMIDE (M+H)+ 9913 17302 8283 14276 65021 18575 17637 18989 14352 12617 25874 17177 OLEIC ACID (M+NA)+ 23666 15670 32499 26057 21356 16265 16084 6905 14397 6234 29373 ORNITHINE (M+H)+ 4501 3066 3123 3032 3493 4487 3578 3412 2808 3027 3616 3908 PANTOTHENATE (M+H)+ 993000 887000 1160000 1000000 1340000 1560000 1540000 1490000 1010000 1500000 1380000 1630000 PANTOTHENATE (M+NA)+ 1160000 1040000 1310000 1120000 1560000 1750000 1780000 1670000 1130000 1660000 1580000 1870000 PHENYLALANINE (M+H)+ 134000 138000 107000 73700 103000 72786 132755 146000 86900 239000 87100 87700 PROGESTERONE (M+H)+ 8757 8574 8435 8881 9953 8516 7711 8856 7252 8311 7400 9660 PROSTAGLANDIN E2 (M+H)+ 8009 4918 8299 14149 10281 10465 7644 10517 9726 13709 PROSTAGLANDIN E2 (M+H)+[-H2O] 10665 20944 9340 9302 4233 25634 4616 5860 6106 4960 9417 6841 PYRIDOXAMINE (M+H)+ 16694 16679 15506 13647 17560 25646 18098 23052 16886 21769 17294 28925 RAC-GLYCEROL 1-MYRISTATE (M+H)+[-H2O] 14268 16405 10445 12020 10395 14272 3911 1756 2002 3480 4404 6829 RAC-GLYCEROL 1-MYRISTATE (M+NA)+ 177203 197754 176884 140591 171660 158643 89256 90654 77888 55479 130937 96590 S-ADENOSYL-L-HOMOCYSTEINE (M+H)+ 79961 77687 70017 97604 91416 99351 112288 84931 64159 103773 102910 98273 SERINE (M+H)+ 29879 38300 26604 49156 32308 47799 33477 SN-GLYCERO-3-PHOSPHOCHOLINE (M+H)+ 2800932 2704309 2685280 1598378 2260334 2439966 2292599 2408865 3207969 2015767 2552721 1965200 SPERMIDINE (M+H)+ 85024 82240 34978 101883 68602 390634 90292 139310 145076 50281 132235 95300 STEAROYLCARNITINE (*) (M+H)+ 6660 19500 9570 2700 2760 SUBERIC ACID (M+NA)+ 14114 5359 10254 16611 3751 4423 3386 7372 2248 11524 TAURINE (M+NA)+ 28805 19709 28651 11336 23860 15757 28198 71134 26517 21385 25537 43450 TETRADECENOYLCARNITINE (*) M+ 2588 6239 8026 2852 8205 7551 4946 7226 3322 2373 6143 3682 THIAMIN M+[-C6H9NOS] 3107 1974 8748 2011 13418 19423 4612 103504 2337 21011 6831 80252 TRYPTOPHAN (M+H)+ 5327914 4628206 6120538 5068228 7478120 8449014 8513297 8164652 5681219 8428224 8119062 9347235 TYROSINE (M+H)+ 12469652 11820134 14294874 12558345 15775818 17298700 17277514 16675719 13173503 17295272 16541841 18336420 UDP-N-ACETYL-D-GALACTOSAMINE (M+NA)+ 58454 56823 57215 64579 52610 46442 67410 52816 58573 71871 72870 50828 URIDINE-5-MONOPHOSPHATE (M+H)+ 18258 7873 6286 7555 7043 22915 19973 4160 16722 5203 14565 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 1-METHYLNICOTINAMIDE M+ 137.07 0.67 457 1-METHYLXANTHINE (M+NA)+ 166.0478 2.514 80220 1-OLEOYL-RAC-GLYCEROL (M+H)+[-H2O] 356.2918 22.66 5283468 1-OLEOYL-RAC-GLYCEROL (M+NA)+ 356.2929 22.738 5283468 3-HYDROXYBENZALDEHYDE (M+H)+[-H2O] 122.0368 4.795 101 5'-METHYLTHIOADENOSINE (M+NA)+ 297.0902 4.512 439176 5-AMINOPENTANOATE (M+H)+ 117.0783 0.733 138 5-HYDROXYTRYPTOPHAN (M+H)+ 220.0847 1.801 439280 ADENOSINE (M+H)+ 267.0964 1.803 60961 ADENOSINE 5'-MONOPHOSPHATE (M+H)+ 347.0599 1.014 6083 ALPHA-TOCOPHEROL (M+H)+ 430.3775 24.537 14985 AZELAIC ACID (M+H)+[-H2O] 188.1035 10.751 2266 BILIVERDIN (M+H)+ 582.2518 18.657 5280353 BIS(2-ETHYLHEXYL)PHTHALATE (M+NA)+ 390.2776 23.13 8343 BUTYRYLCARNITINE (*) (M+H)+ 231.147 2.962 213144 CARNITINE (M+H)+ 161.1032 0.677 10917 CHOLESTEROL (M+H)+[-H2O] 386.3551 24.632 5997 CHOLINE M+ 104.1065 0.657 305 CHOLINE PHOSPHATE (M+NA)+ 183.0651 0.644 1014 CITRATE (M+NA)+ 192.027 1.053 311 CORTISOL (M+H)+ 362.2067 13.827 5754 CORTISOL (M+NA)+ 362.2067 13.824 5754 CREATINE (M+H)+ 131.0687 0.693 586 CREATININE (M+H)+ 113.0583 0.688 588 DECANOATE (2M+NA)+ 172.1459 22.761 2969 DGMP (M+H)+ 347.0597 1.319 6994968 ELAIDIC ACID (M+H)+ 282.2553 22.95 637517 ETHANOLAMINE PHOSPHATE (M+NA)+ 141.0193 0.759 1015 GAMMA-L-GLUTAMYL-L-CYSTEINE (M+H)+ 250.0601 1.006 123938 GIBBERELLIC ACID [ISTD] (2M+NA)+ 346.1397 8.518 6466 GLUTAMATE (M+H)+[-H2O] 147.0527 0.665 33032 GLUTATHIONE OXIDIZED (M+H2)+2 612.1531 1.364 65359 GUANINE (M+H)+ 151.0507 0.928 764 GUANOSINE (M+H)+ 283.0921 1.86 6802 HIPPURATE (M+H)+ 179.0585 4.796 464 INOSINE 5'-PHOSPHATE (M+H)+ 348.0504 0.991 8582 INOSINE 5'-PHOSPHATE (M+NA)+ 348.0464 0.989 8582 ISOLEUCINE (M+H)+ 131.0942 1.523 6306 ISOVALERYLCARNITINE (*) (M+H)+ 245.1616 4.592 169235 KYNURENINE (M+H)+ 208.083 2.229 161166 L-ARGININE (M+H)+ 174.1103 0.643 6322 L-ARGININE (M+NA)+ 174.111 0.629 6322 L-HISTIDINE (M+H)+ 155.0682 0.647 6274 L-TRYPTOPHAN-15N2 [ISTD] (M+H)+ 206.0849 3.656 16217285 LYSINE (M+H)+ 146.1055 0.563 5962 LYSINE (M+NA)+ 146.1046 0.582 5962 MANNOSE-6-PHOSPHATE (M+NA)+ 260.0279 0.72 439198 N-ACETYL-DL-METHIONINE (M+H)+[-H2O] 191.0599 4.049 448580 N-ACETYL-DL-SERINE (M+H)+[-H2O] 147.0532 0.926 65249 N-ACETYL-L-ALANINE (M+NA)+ 131.0573 1.392 88064 N-ACETYL-L-LEUCINE (M+NA)+ 173.1071 7.15 75619 N-ACETYLNEURAMINATE (M+H)+ 309.1022 0.689 439197 N-METHYL-D-ASPARTIC ACID (M+NA)+ 147.0525 0.655 4376 NAD (M+2H)+2 663.1097 1.057 5893 NAD (M+H)+ 663.1084 1.047 5893 NAD (M+NA)+ 663.1057 1.054 5893 NORADRENALINE (M+H)+ 169.0724 0.721 951 O-PHOSPHOSERINE (M+NA)+ 185.007 0.648 68841 OLEAMIDE (M+H)+ 281.2699 22.643 5283387 OLEIC ACID (M+NA)+ 282.2561 22.528 445639 ORNITHINE (M+H)+ 132.0898 0.562 6262 PANTOTHENATE (M+H)+ 219.1108 3.255 6613 PANTOTHENATE (M+NA)+ 219.1112 3.254 6613 PHENYLALANINE (M+H)+ 165.0787 2.389 6140 PROGESTERONE (M+H)+ 314.2231 19.497 5994 PROSTAGLANDIN E2 (M+H)+ 352.2203 16.772 5280360 PROSTAGLANDIN E2 (M+H)+[-H2O] 352.2238 16.8 5280360 PYRIDOXAMINE (M+H)+ 168.0869 0.582 1052 RAC-GLYCEROL 1-MYRISTATE (M+H)+[-H2O] 302.245 22.147 79050 RAC-GLYCEROL 1-MYRISTATE (M+NA)+ 302.2461 22.171 79050 S-ADENOSYL-L-HOMOCYSTEINE (M+H)+ 384.1191 1.554 439155 SERINE (M+H)+ 105.0415 1.017 617 SN-GLYCERO-3-PHOSPHOCHOLINE (M+H)+ 257.1017 0.671 439285 SPERMIDINE (M+H)+ 145.1577 0.584 1102 STEAROYLCARNITINE (*) (M+H)+ 427.3652 21.855 52922056 SUBERIC ACID (M+NA)+ 174.0917 8.256 10457 TAURINE (M+NA)+ 125.0133 0.654 1123 TETRADECENOYLCARNITINE (*) M+ 370.2918 18.823 22833575 THIAMIN M+[-C6H9NOS] 265.1104 0.659 1130 TRYPTOPHAN (M+H)+ 204.0878 3.655 6305 TYROSINE (M+H)+ 181.0721 1.351 6057 UDP-N-ACETYL-D-GALACTOSAMINE (M+NA)+ 607.0797 0.841 23724461 URIDINE-5-MONOPHOSPHATE (M+H)+ 324.0342 0.871 6030 METABOLITES_END #END