#METABOLOMICS WORKBENCH amitch_20160728_135939 DATATRACK_ID:689 STUDY_ID:ST000446 ANALYSIS_ID:AN000697 PROJECT_ID:PR000344 VERSION 1 CREATED_ON August 10, 2016, 11:18 am #PROJECT PR:PROJECT_TITLE Cloning, expression and derivatization of cytotoxic marine natural product PR:PROJECT_TITLE Apratoxin PR:PROJECT_SUMMARY Apratoxin is a hybrid PKS/NRPS secondary metabolite showing potent anticancer PR:PROJECT_SUMMARY activity by inducing G1 phase cell cycle arrest and apoptosis. There have been PR:PROJECT_SUMMARY reported a series of natural apratoxin derivatives with defined chemical PR:PROJECT_SUMMARY modifications in their structures that alter their biological activity. Looking PR:PROJECT_SUMMARY for a better activity, total synthesis of apratoxin has been achieved and a PR:PROJECT_SUMMARY series of synthetic congeners showed increased activity with subnanomolar PR:PROJECT_SUMMARY potency. From the total synthesis point of view, the modification of the PR:PROJECT_SUMMARY polyketide backbone presents a burden for a radical derivatization of PR:PROJECT_SUMMARY apratoxins. We hypothesized that supply the polyketide part naturally by PR:PROJECT_SUMMARY overexpression the corresponding genes it will facilitate the generation of PR:PROJECT_SUMMARY apratoxin congeners by decorating the backbone with a variety of amino acids PR:PROJECT_SUMMARY improving the antitumor activity.Threfore, the apratoxin gene cluster was PR:PROJECT_SUMMARY recovered from a fosmid DNA library. The gene set responsible for the PR:PROJECT_SUMMARY biosynthesis of the polyketide backbone was introduced in E. coli BAP strain and PR:PROJECT_SUMMARY expressed at 30C for 24 hours. Two sets of control and experimental samples with PR:PROJECT_SUMMARY culture broth pH adjusted at 8 and 12 respectively was extracted with ethyl PR:PROJECT_SUMMARY acetate and dried. PR:INSTITUTE University of Florida PR:DEPARTMENT Medicinal Chemistry PR:LAST_NAME Luesch PR:FIRST_NAME Hendrik PR:ADDRESS Medical Sciences Building, Rm P5-26, 1600 SW Archer Rd., FL32610 PR:EMAIL luesch@cop.ufl.edu PR:PHONE none #STUDY ST:STUDY_TITLE Heterologous expression and detection of Apratoxins in E. coli ST:STUDY_TYPE Organic extraction of E coli cultures harboring apratoxin gene cluster ST:STUDY_SUMMARY The apratoxin gene cluster was recovered from fosmid DNA library. The gene set ST:STUDY_SUMMARY responsible for the biosynthesis of the polyketide backbone was introduced in E. ST:STUDY_SUMMARY coli BAP strain and expressed at 30C for 24 hours. Two sets of control and ST:STUDY_SUMMARY experimental samples with culture broth pH adjusted at 8 and 12 respectively was ST:STUDY_SUMMARY extracted with ethyl acetate and dried. ST:INSTITUTE University of Florida ST:DEPARTMENT SECIM ST:LAST_NAME Kallifidas ST:FIRST_NAME Dimitris ST:ADDRESS Medical Sciences Building, Rm P5-26, 1600 SW Archer Rd., FL32610 ST:EMAIL dkallifidas@ufl.edu ST:PHONE None ST:NUM_GROUPS 1 ST:TOTAL_SUBJECTS 12 #SUBJECT SU:SUBJECT_TYPE bacteria cells SU:SUBJECT_SPECIES Escherichia coli SU:TAXONOMY_ID 562 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - E.coli_control_pH8 pH:8 | treatment:control SUBJECT_SAMPLE_FACTORS - E.coli_control_pH12 pH:12 | treatment:control SUBJECT_SAMPLE_FACTORS - E.coli+bAprat14_pH8 pH:8 | treatment:bAprat14 SUBJECT_SAMPLE_FACTORS - E.coli+bAprat14_pH12 pH:12 | treatment:bAprat14 SUBJECT_SAMPLE_FACTORS - E.coli_control_pH8_2 pH:8 | treatment:control SUBJECT_SAMPLE_FACTORS - E.coli_control_pH12_2 pH:12 | treatment:control SUBJECT_SAMPLE_FACTORS - E.coli+bAprat14_pH8_2 pH:8 | treatment:bAprat14 SUBJECT_SAMPLE_FACTORS - E.coli+bAprat14_pH12_2 pH:12 | treatment:bAprat14 SUBJECT_SAMPLE_FACTORS - E.coli_control_pH8_3 pH:8 | treatment:control SUBJECT_SAMPLE_FACTORS - E.coli_control_pH12_3 pH:12 | treatment:control SUBJECT_SAMPLE_FACTORS - E.coli+bAprat14_pH8_3 pH:8 | treatment:bAprat14 SUBJECT_SAMPLE_FACTORS - E.coli+bAprat14_pH12_3 pH:12 | treatment:bAprat14 #COLLECTION CO:COLLECTION_SUMMARY 50 ml of LB broth inoculated with the appropriate E. coli strains were extracted CO:COLLECTION_SUMMARY twice with equal volume of etheyl acetate and dried. CO:SAMPLE_TYPE Dried ethyl acetate cell extracts #TREATMENT TR:TREATMENT_SUMMARY The set of genes responsible for the biosynthesis of the Apratoxin polyketide TR:TREATMENT_SUMMARY backbone was introduced in E. coli BAP strain and expressed at 30C for 24 hours. TR:TREATMENT_SUMMARY Two sets of control and experimental samples with culture broth pH adjusted at 8 TR:TREATMENT_SUMMARY and 12 respectively was extracted with ethyl acetate and dried. TR:CELL_MEDIA Luria Bertani (LB) broth #SAMPLEPREP SP:SAMPLEPREP_SUMMARY none SP:SAMPLEPREP_PROTOCOL_FILENAME Consolidated_Cellular_Metabolomics_SOP.pdf SP:SAMPLE_SPIKING Appendix A - Internal Standard Prep GLCMS.pdf #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Scientific-Dionex Ultimate 3000 CH:COLUMN_NAME ACE Excel 2 C18-PFP (100 x 2.1mm, 2um) CH:METHODS_FILENAME Metabolomics_LCMSProtocol.pdf #ANALYSIS AN:ANALYSIS_TYPE MS AN:ANALYSIS_PROTOCOL_FILE Metabolomics_LCMSProtocol.pdf #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS HESI #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples E.coli+bAprat14_pH12 E.coli_control_pH12 E.coli_control_pH12_2 E.coli_control_pH12_3 E.coli+bAprat14_pH12_2 E.coli+bAprat14_pH12_3 E.coli+bAprat14_pH8 E.coli+bAprat14_pH8_2 E.coli_control_pH8 E.coli_control_pH8_2 E.coli_control_pH8_3 E.coli+bAprat14_pH8_3 Factors pH:12 | treatment:bAprat14 pH:12 | treatment:control pH:12 | treatment:control pH:12 | treatment:control pH:12 | treatment:bAprat14 pH:12 | treatment:bAprat14 pH:8 | treatment:bAprat14 pH:8 | treatment:bAprat14 pH:8 | treatment:control pH:8 | treatment:control pH:8 | treatment:control pH:8 | treatment:bAprat14 1,2-Benzisothiazolin-3-one 334000.0000 400000.0000 356000.0000 379000.0000 333000.0000 360000.0000 940000.0000 901000.0000 940000.0000 1070000.0000 1090000.0000 974000.0000 2,4-DIHYDROXYPTERIDINE 16400.0000 3410.0000 3670.0000 0.0000 13300.0000 13400.0000 432000.0000 439000.0000 1320000.0000 1300000.0000 1340000.0000 374000.0000 2-AMINOPHENOL 97100000.0000 130000000.0000 131000000.0000 146000000.0000 93300000.0000 96800000.0000 565000000.0000 547000000.0000 836000000.0000 856000000.0000 884000000.0000 556000000.0000 3,5-Di-tert-butylbenzaldehyde 2130000.0000 2150000.0000 2020000.0000 2170000.0000 2170000.0000 2240000.0000 2180000.0000 2370000.0000 2260000.0000 2190000.0000 2150000.0000 2260000.0000 3'-CMP 11700000.0000 6570000.0000 6560000.0000 6980000.0000 11800000.0000 11900000.0000 2800000.0000 2780000.0000 759000.0000 757000.0000 709000.0000 2710000.0000 3-HYDROXY-3-METHYLGLUTARATE 3590000.0000 3120000.0000 3200000.0000 2900000.0000 3770000.0000 3690000.0000 8330000.0000 9600000.0000 6760000.0000 7290000.0000 6850000.0000 8440000.0000 3-METHOXYTYRAMINE 934000.0000 1180000.0000 1240000.0000 1380000.0000 927000.0000 841000.0000 15900000.0000 3020000.0000 6730000.0000 6720000.0000 6880000.0000 15300000.0000 3-METHYL-2-OXINDOLE 1740000.0000 2260000.0000 2190000.0000 2290000.0000 1790000.0000 1850000.0000 2850000.0000 2870000.0000 3840000.0000 3440000.0000 3810000.0000 2760000.0000 3-UREIDOPROPIONATE 816000.0000 554000.0000 735000.0000 653000.0000 824000.0000 756000.0000 591000.0000 874000.0000 400000.0000 542000.0000 417000.0000 798000.0000 4-AMINOBUTANOATE (GABA) 4480000.0000 2220000.0000 2570000.0000 2780000.0000 3210000.0000 1730000.0000 3570000.0000 4190000.0000 6020000.0000 5800000.0000 5760000.0000 3680000.0000 4-GUANIDINOBUTANOATE 48300000.0000 4800000.0000 4790000.0000 5300000.0000 46500000.0000 47700000.0000 138000000.0000 134000000.0000 13900000.0000 14300000.0000 14300000.0000 139000000.0000 4-HYDROXYPROLINE 1000000.0000 710000.0000 1700000.0000 849000.0000 2440000.0000 919000.0000 2500000.0000 1730000.0000 1700000.0000 1710000.0000 2570000.0000 1770000.0000 4-IMIDAZOLEACETIC ACID 5130000.0000 3450000.0000 3480000.0000 4360000.0000 4150000.0000 4560000.0000 11200000.0000 12200000.0000 10800000.0000 13300000.0000 10200000.0000 12100000.0000 5,6-DIHYDROURACIL 3830000.0000 3120000.0000 3100000.0000 7700000.0000 3610000.0000 6790000.0000 23700000.0000 25500000.0000 46600000.0000 48000000.0000 47600000.0000 25300000.0000 5'-DEOXYADENOSINE 105000.0000 4830000.0000 4700000.0000 4720000.0000 86000.0000 90400.0000 8480000.0000 7920000.0000 4940000.0000 4640000.0000 4960000.0000 8320000.0000 5-METHYLCYTOSINE HYDROCLORIDE 1200000.0000 1750000.0000 1650000.0000 1770000.0000 1320000.0000 1240000.0000 905000.0000 4510000.0000 4390000.0000 4510000.0000 4610000.0000 4600000.0000 6-DEOXY-HEXOSE 640000.0000 982000.0000 808000.0000 828000.0000 613000.0000 670000.0000 741000.0000 822000.0000 959000.0000 1200000.0000 1170000.0000 2100000.0000 Acetaminophen/Paracetamol 1060000.0000 1420000.0000 2070000.0000 1480000.0000 1010000.0000 1060000.0000 7980000.0000 7880000.0000 13900000.0000 14800000.0000 14300000.0000 8180000.0000 Acyl-Carnitine(4-OH) 516000.0000 294000.0000 210000.0000 207000.0000 556000.0000 574000.0000 1670000.0000 1840000.0000 4920000.0000 5020000.0000 5000000.0000 1740000.0000 ADENINE 176000000.0000 257000000.0000 254000000.0000 273000000.0000 170000000.0000 176000000.0000 851000000.0000 825000000.0000 574000000.0000 579000000.0000 594000000.0000 839000000.0000 ADENOSINE 5'-MONOPHOSPHATE 4560000.0000 4240000.0000 4510000.0000 4330000.0000 4540000.0000 4590000.0000 1140000.0000 1160000.0000 519000.0000 496000.0000 515000.0000 1180000.0000 ALANINE/SARCOSINE 960000.0000 4340000.0000 4680000.0000 1070000.0000 3060000.0000 1010000.0000 18200000.0000 17900000.0000 22300000.0000 23000000.0000 23500000.0000 18700000.0000 ALDO/KETO-HEXOSE 22100000.0000 18600000.0000 19200000.0000 20000000.0000 19400000.0000 22100000.0000 8880000.0000 9650000.0000 6010000.0000 5590000.0000 5510000.0000 9780000.0000 ALDOPENTOSE 1720000.0000 1530000.0000 1650000.0000 1600000.0000 1830000.0000 2000000.0000 2030000.0000 2960000.0000 3270000.0000 3190000.0000 3230000.0000 1970000.0000 Allopurinol 47800.0000 365000.0000 378000.0000 344000.0000 39400.0000 28600.0000 4880000.0000 4600000.0000 6030000.0000 6250000.0000 6640000.0000 4940000.0000 ALPHA-AMINOADIPATE/N-METHYL-L-GLUTAMATE 96800000.0000 71900000.0000 72000000.0000 72200000.0000 96500000.0000 101000000.0000 279000000.0000 272000000.0000 164000000.0000 167000000.0000 163000000.0000 264000000.0000 Anthranilate 8330000.0000 9650000.0000 9910000.0000 10000000.0000 8260000.0000 8920000.0000 12300000.0000 11300000.0000 24800000.0000 26000000.0000 25500000.0000 12000000.0000 ANTHRANILATE/4-AMINOBENZOATE 761000.0000 1560000.0000 1450000.0000 1520000.0000 1010000.0000 843000.0000 10800000.0000 11100000.0000 18500000.0000 19000000.0000 18100000.0000 10900000.0000 ASPARTATE 960000.0000 1640000.0000 1650000.0000 1660000.0000 1010000.0000 2000000.0000 3400000.0000 3350000.0000 2290000.0000 2470000.0000 2450000.0000 5120000.0000 BETAINE 1120000000.0000 920000000.0000 895000000.0000 1010000000.0000 1100000000.0000 1110000000.0000 2900000000.0000 2900000000.0000 2040000000.0000 2030000000.0000 2100000000.0000 2940000000.0000 Bis(2-ethylhexyl) phthalate 11700000.0000 22600000.0000 12200000.0000 9780000.0000 23300000.0000 15400000.0000 16600000.0000 28400000.0000 31000000.0000 24700000.0000 10600000.0000 6890000.0000 C5-SUGAR ALCOHOL 3430000.0000 3000000.0000 3060000.0000 3240000.0000 3030000.0000 3400000.0000 1260000.0000 1210000.0000 1360000.0000 1140000.0000 1420000.0000 1170000.0000 Choline 434000000.0000 370000000.0000 382000000.0000 387000000.0000 413000000.0000 440000000.0000 623000000.0000 644000000.0000 462000000.0000 488000000.0000 483000000.0000 651000000.0000 CREATINE 339000.0000 264000.0000 239000.0000 258000.0000 352000.0000 315000.0000 2230000.0000 2060000.0000 993000.0000 1410000.0000 1340000.0000 2250000.0000 CREATININE 1110000.0000 1170000.0000 895000.0000 1130000.0000 1020000.0000 1040000.0000 2860000.0000 2600000.0000 3450000.0000 3500000.0000 3730000.0000 2770000.0000 CYTIDINE 2730000.0000 2150000.0000 2100000.0000 2140000.0000 2790000.0000 2760000.0000 2190000.0000 1740000.0000 9400000.0000 9510000.0000 1050000.0000 1960000.0000 CYTOSINE 1260000.0000 1470000.0000 1420000.0000 1360000.0000 1190000.0000 1300000.0000 3150000.0000 3190000.0000 3280000.0000 3300000.0000 3380000.0000 3110000.0000 Dibutyl phthalate 2630000.0000 2950000.0000 2960000.0000 3120000.0000 2630000.0000 2810000.0000 3170000.0000 3110000.0000 2650000.0000 2520000.0000 2700000.0000 3240000.0000 Dihydroxyquinoine 335000.0000 482000.0000 410000.0000 460000.0000 287000.0000 333000.0000 1470000.0000 1350000.0000 2590000.0000 2270000.0000 2560000.0000 1350000.0000 DIMETHYLBENZIMIDAZOLE 3550000.0000 642000.0000 6030000.0000 709000.0000 3480000.0000 3560000.0000 1730000.0000 4390000.0000 2600000.0000 2550000.0000 2920000.0000 1680000.0000 Dipropylene glycol 1260000.0000 639000.0000 1850000.0000 1710000.0000 1230000.0000 1400000.0000 1050000.0000 1980000.0000 3650000.0000 4480000.0000 3600000.0000 1720000.0000 Erucamide 1150000.0000 2840000.0000 1510000.0000 1390000.0000 1470000.0000 1540000.0000 1590000.0000 3660000.0000 3230000.0000 1270000.0000 1350000.0000 1400000.0000 Garcinia lactone dibutyl ester 11100000.0000 19200000.0000 19500000.0000 21200000.0000 9870000.0000 10600000.0000 6060000.0000 5730000.0000 9150000.0000 9490000.0000 9840000.0000 6540000.0000 GLUTARATE 4970000.0000 5610000.0000 6090000.0000 5470000.0000 5120000.0000 5030000.0000 9280000.0000 10500000.0000 8020000.0000 7580000.0000 7340000.0000 10200000.0000 Glutathione Disulfide 1560000.0000 1340000.0000 1340000.0000 1330000.0000 1490000.0000 1610000.0000 15600000.0000 16000000.0000 8740000.0000 9120000.0000 8670000.0000 15400000.0000 GLYCINE 315000.0000 504000.0000 606000.0000 341000.0000 760000.0000 640000.0000 624000.0000 1350000.0000 1350000.0000 1440000.0000 1490000.0000 1340000.0000 Glycyl-L-leucine 7730000.0000 6520000.0000 6400000.0000 6370000.0000 7310000.0000 7640000.0000 2300000.0000 2320000.0000 4250000.0000 4540000.0000 4460000.0000 2160000.0000 GUANINE 3630000.0000 6830000.0000 6810000.0000 7300000.0000 3820000.0000 4020000.0000 21000000.0000 19800000.0000 16300000.0000 15700000.0000 17100000.0000 20100000.0000 HEXOSE-DISACCHARIDE 1610000.0000 1260000.0000 1420000.0000 1410000.0000 1340000.0000 1570000.0000 7560000.0000 7020000.0000 741000.0000 1040000.0000 1040000.0000 7190000.0000 Hippuric Acid 946000.0000 689000.0000 798000.0000 712000.0000 902000.0000 878000.0000 1930000.0000 2170000.0000 1100000.0000 1240000.0000 1360000.0000 1970000.0000 HOMOCYSTEINE 2170000.0000 1660000.0000 1600000.0000 834000.0000 2330000.0000 2110000.0000 6860000.0000 7220000.0000 6050000.0000 6110000.0000 5810000.0000 6750000.0000 Homocystine 4370000.0000 3100000.0000 2850000.0000 3250000.0000 4320000.0000 4210000.0000 13900000.0000 14600000.0000 11300000.0000 11100000.0000 11000000.0000 13900000.0000 HYPOXANTHINE 70500000.0000 67600000.0000 68400000.0000 71600000.0000 67600000.0000 69100000.0000 691000000.0000 654000000.0000 729000000.0000 753000000.0000 742000000.0000 674000000.0000 INDOLE-3-ACETATE 45800000.0000 82600000.0000 88900000.0000 90900000.0000 41000000.0000 44900000.0000 276000000.0000 272000000.0000 411000000.0000 443000000.0000 449000000.0000 276000000.0000 Isocytosine 4040000.0000 3140000.0000 3210000.0000 3160000.0000 4180000.0000 4650000.0000 2490000.0000 2450000.0000 5190000.0000 5230000.0000 1420000.0000 2240000.0000 Kynurenic Acid 1270000.0000 978000.0000 1020000.0000 969000.0000 1260000.0000 1270000.0000 4290000.0000 4440000.0000 1350000.0000 1390000.0000 1400000.0000 4830000.0000 KYNURENINE 966000.0000 443000.0000 492000.0000 438000.0000 880000.0000 887000.0000 2890000.0000 2780000.0000 4440000.0000 4150000.0000 4690000.0000 3010000.0000 L-2-PHOSPHOGLYCERIC ACID 954000.0000 553000.0000 693000.0000 423000.0000 1000000.0000 732000.0000 5020000.0000 4550000.0000 2270000.0000 2640000.0000 2190000.0000 4590000.0000 L-Acetylcarnitine 3890000.0000 5420000.0000 5550000.0000 5690000.0000 3600000.0000 3840000.0000 5280000.0000 5330000.0000 5840000.0000 5820000.0000 5980000.0000 5440000.0000 L-ARGININE 1500000.0000 1330000.0000 1460000.0000 1570000.0000 1620000.0000 1690000.0000 3370000.0000 3210000.0000 2780000.0000 3070000.0000 2640000.0000 3390000.0000 L-ASPARAGINE 1010000.0000 851000.0000 868000.0000 862000.0000 1020000.0000 1110000.0000 2020000.0000 1840000.0000 1490000.0000 1720000.0000 1730000.0000 2070000.0000 L-CARNITINE 3280000.0000 2390000.0000 2270000.0000 2580000.0000 3080000.0000 3200000.0000 19400000.0000 18500000.0000 7260000.0000 7160000.0000 7310000.0000 17400000.0000 Leu Pro 7940000.0000 3710000.0000 4730000.0000 8810000.0000 2740000.0000 10200000.0000 5890000.0000 4870000.0000 3590000.0000 3610000.0000 5200000.0000 4730000.0000 L-GLUTAMIC ACID 14300000.0000 11200000.0000 11200000.0000 11700000.0000 13600000.0000 14600000.0000 11900000.0000 11900000.0000 4660000.0000 4810000.0000 4870000.0000 12400000.0000 L-GLUTAMINE 6870000.0000 6230000.0000 8590000.0000 6380000.0000 6780000.0000 7100000.0000 2280000.0000 2400000.0000 4510000.0000 4560000.0000 4740000.0000 2350000.0000 L-HISTIDINE 25500000.0000 20700000.0000 22800000.0000 21700000.0000 23000000.0000 31800000.0000 188000000.0000 186000000.0000 136000000.0000 155000000.0000 140000000.0000 187000000.0000 L-HOMOCYSTEINE THIOLACTONE 94300.0000 114000.0000 27700.0000 109000.0000 44500.0000 72200.0000 3920000.0000 4140000.0000 8460000.0000 8630000.0000 8960000.0000 4010000.0000 L-Isoleucine 2920000000.0000 2390000000.0000 2420000000.0000 2480000000.0000 2900000000.0000 2940000000.0000 7040000000.0000 7000000000.0000 4890000000.0000 4990000000.0000 5040000000.0000 7140000000.0000 L-METHIONINE 614000000.0000 395000000.0000 403000000.0000 393000000.0000 613000000.0000 624000000.0000 1900000000.0000 1890000000.0000 1210000000.0000 1210000000.0000 1230000000.0000 1940000000.0000 L-Tryptophan 13C11 15000.0000 369000.0000 446000.0000 445000.0000 28800.0000 32300.0000 557000.0000 221000.0000 952000.0000 9520000.0000 8930000.0000 0.0000 L-TYROSINE 239000000.0000 193000000.0000 193000000.0000 199000000.0000 237000000.0000 242000000.0000 749000000.0000 752000000.0000 522000000.0000 535000000.0000 534000000.0000 749000000.0000 L-VALINE/5-AMINOPENTANOATE/L-NORVALINE 2150000000.0000 1760000000.0000 1750000000.0000 1810000000.0000 2090000000.0000 2130000000.0000 4700000000.0000 4680000000.0000 3190000000.0000 3310000000.0000 3310000000.0000 4670000000.0000 LysoPE (18:1) 2240000.0000 31700000.0000 29600000.0000 30700000.0000 2760000.0000 1960000.0000 23200.0000 27500.0000 39800.0000 11500.0000 9990.0000 14700.0000 Methionine Sulfoxide 74000000.0000 96800000.0000 99200000.0000 101000000.0000 73200000.0000 78400000.0000 55000000.0000 54600000.0000 77500000.0000 80300000.0000 81100000.0000 56700000.0000 METHYL INDOLE-3-ACETATE 78400000.0000 101000000.0000 100000000.0000 99600000.0000 75700000.0000 81100000.0000 392000000.0000 418000000.0000 421000000.0000 447000000.0000 453000000.0000 390000000.0000 Monobutyl phthalate 418000.0000 560000.0000 606000.0000 630000.0000 377000.0000 384000.0000 798000.0000 751000.0000 576000.0000 632000.0000 642000.0000 828000.0000 N-ACETYL-DL-GLUTAMIC ACID 1980000.0000 1590000.0000 1610000.0000 1750000.0000 1860000.0000 1990000.0000 3950000.0000 3860000.0000 2370000.0000 2430000.0000 2480000.0000 4050000.0000 N-ACETYL-DL-SERINE 3570000.0000 3190000.0000 3390000.0000 3480000.0000 3630000.0000 3860000.0000 6320000.0000 6330000.0000 4890000.0000 5130000.0000 5170000.0000 5980000.0000 N-ACETYL-L-ASPARTIC ACID 461000.0000 559000.0000 416000.0000 604000.0000 497000.0000 517000.0000 4810000.0000 4930000.0000 3890000.0000 4030000.0000 4070000.0000 5130000.0000 N-ACETYL-L-LEUCINE 15500000.0000 35300000.0000 35800000.0000 39200000.0000 14800000.0000 16100000.0000 2410000.0000 2680000.0000 3120000.0000 3180000.0000 3080000.0000 2540000.0000 N-ACETYLNEURAMINATE 8480000.0000 5630000.0000 5650000.0000 5890000.0000 7810000.0000 8570000.0000 31800000.0000 32000000.0000 15500000.0000 17200000.0000 16000000.0000 32000000.0000 N-ACETYLPUTRESCINE 20500000.0000 12100000.0000 11900000.0000 13800000.0000 18900000.0000 19900000.0000 44000000.0000 41800000.0000 12600000.0000 12100000.0000 13200000.0000 42500000.0000 N-ALPHA-ACETYL-L-LYSINE 11900000.0000 10900000.0000 10900000.0000 10800000.0000 12200000.0000 12300000.0000 26300000.0000 26300000.0000 14300000.0000 13400000.0000 13700000.0000 26400000.0000 N-Butylbenzenesulfonamide 314000.0000 304000.0000 303000.0000 333000.0000 252000.0000 338000.0000 1110000.0000 1100000.0000 365000.0000 448000.0000 356000.0000 1170000.0000 Orthophosphate 5230000.0000 5200000.0000 5180000.0000 5170000.0000 5670000.0000 5170000.0000 27300000.0000 27600000.0000 26000000.0000 25800000.0000 25500000.0000 27300000.0000 OxPC (20:3(KeKe)) 1330000.0000 1130000.0000 1360000.0000 1350000.0000 1290000.0000 1280000.0000 12600000.0000 12800000.0000 8420000.0000 8860000.0000 8320000.0000 12200000.0000 Pantothenic Acid 13800000.0000 14400000.0000 14600000.0000 14500000.0000 13800000.0000 14400000.0000 78400000.0000 76500000.0000 49000000.0000 49400000.0000 52000000.0000 77500000.0000 PEG n5 628000.0000 2540000.0000 2710000.0000 1950000.0000 563000.0000 979000.0000 15200000.0000 16100000.0000 31700000.0000 32900000.0000 30100000.0000 15700000.0000 PHENETHYLAMINE 2450000.0000 2990000.0000 3140000.0000 3430000.0000 2350000.0000 2100000.0000 620000000.0000 4730000.0000 15000000.0000 15900000.0000 15800000.0000 615000000.0000 Phenylalanine 3330000000.0000 2860000000.0000 2870000000.0000 2940000000.0000 3240000000.0000 3280000000.0000 9460000000.0000 9340000000.0000 7320000000.0000 7500000000.0000 7560000000.0000 9290000000.0000 PHOSPHOCHOLINE 2130000.0000 1400000.0000 1420000.0000 1530000.0000 2280000.0000 2440000.0000 10500000.0000 9040000.0000 3510000.0000 3610000.0000 3910000.0000 10300000.0000 PICOLINIC ACID 49500000.0000 39700000.0000 41400000.0000 40600000.0000 51100000.0000 51600000.0000 137000000.0000 136000000.0000 93400000.0000 94500000.0000 94600000.0000 137000000.0000 PIPECOLATE/L-PIPECOLIC ACID 45400000.0000 36900000.0000 39000000.0000 37700000.0000 47400000.0000 44600000.0000 4780000.0000 118000000.0000 9010000.0000 10000000.0000 8930000.0000 4900000.0000 Proprionylcarnitine 3850000.0000 5120000.0000 5160000.0000 5460000.0000 3820000.0000 4070000.0000 3510000.0000 3630000.0000 4900000.0000 4910000.0000 5040000.0000 1430000.0000 PYRIDOXAL 666000.0000 1280000.0000 1010000.0000 945000.0000 688000.0000 737000.0000 1620000.0000 1710000.0000 1220000.0000 2360000.0000 1330000.0000 1840000.0000 Pyroglutamic Acid 1190000000.0000 960000000.0000 966000000.0000 1020000000.0000 1150000000.0000 1180000000.0000 2590000000.0000 2570000000.0000 1770000000.0000 1830000000.0000 1840000000.0000 2590000000.0000 Resorcinol monoacetate 337000.0000 688000.0000 767000.0000 707000.0000 306000.0000 369000.0000 127000.0000 143000.0000 4600000.0000 4280000.0000 4590000.0000 113000.0000 SEROTONIN 2070000.0000 2660000.0000 2170000.0000 2770000.0000 1890000.0000 1850000.0000 2080000.0000 2080000.0000 1590000.0000 2310000.0000 2040000.0000 2140000.0000 Stachydrine 920000.0000 698000.0000 441000.0000 830000.0000 937000.0000 831000.0000 3500000.0000 29100000.0000 3650000.0000 5650000.0000 3880000.0000 30300000.0000 Sulcatol 977000.0000 1030000.0000 1020000.0000 918000.0000 940000.0000 852000.0000 1060000.0000 988000.0000 1500000.0000 1310000.0000 1460000.0000 909000.0000 THREONINE/HOMOSERINE 3830000.0000 3240000.0000 3330000.0000 3430000.0000 3650000.0000 4020000.0000 4210000.0000 4330000.0000 4980000.0000 4880000.0000 5030000.0000 4570000.0000 THYMINE 138000000.0000 157000000.0000 160000000.0000 162000000.0000 144000000.0000 144000000.0000 1480000000.0000 1400000000.0000 2090000000.0000 2150000000.0000 2150000000.0000 1370000000.0000 TRANS-CINNAMATE 752000.0000 943000.0000 945000.0000 966000.0000 690000.0000 734000.0000 2240000.0000 2560000.0000 4190000.0000 4270000.0000 4590000.0000 2750000.0000 Triethyl phosphate 4560000.0000 5080000.0000 4930000.0000 5310000.0000 4430000.0000 4570000.0000 4770000.0000 4330000.0000 4300000.0000 4530000.0000 5180000.0000 4700000.0000 Trigonelline 8040000.0000 933000.0000 4000000.0000 4880000.0000 6750000.0000 7110000.0000 35500000.0000 32900000.0000 29200000.0000 30400000.0000 30600000.0000 34700000.0000 Tryptophan 5160000.0000 14600000.0000 15500000.0000 14500000.0000 5260000.0000 5450000.0000 40100000.0000 39400000.0000 54700000.0000 59200000.0000 58100000.0000 38000000.0000 TYRAMINE 2360000.0000 3810000.0000 3880000.0000 3970000.0000 1950000.0000 2780000.0000 3410000.0000 7500000.0000 19600000.0000 20100000.0000 20900000.0000 7980000.0000 URACIL 122000000.0000 117000000.0000 121000000.0000 116000000.0000 99800000.0000 95200000.0000 1010000000.0000 1290000000.0000 1620000000.0000 1400000000.0000 1420000000.0000 1080000000.0000 URIDINE 502000.0000 382000.0000 411000.0000 424000.0000 509000.0000 491000.0000 1660000.0000 1690000.0000 1910000.0000 1870000.0000 1760000.0000 1610000.0000 URIDINE-5-MONOPHOSPHATE 573000.0000 852000.0000 1230000.0000 1310000.0000 876000.0000 1010000.0000 76300.0000 54000.0000 146000.0000 247000.0000 255000.0000 106000.0000 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z PubChem ID KEGG ID 1,2-Benzisothiazolin-3-one 9.96 152.0165 17520 2,4-DIHYDROXYPTERIDINE 3.50 165.0410 10250 2-AMINOPHENOL 1.93 110.0602 5801 3,5-Di-tert-butylbenzaldehyde 14.31 219.1740 6440942 3'-CMP 1.69 324.0590 6131 3-HYDROXY-3-METHYLGLUTARATE 3.04 185.0425 1662 3-METHOXYTYRAMINE 6.35 168.1013 1669 3-METHYL-2-OXINDOLE 8.66 148.0761 150923 3-UREIDOPROPIONATE 1.28 133.0610 111 4-AMINOBUTANOATE (GABA) 0.77 104.0709 119 4-GUANIDINOBUTANOATE 1.51 146.0924 500 4-HYDROXYPROLINE 0.75 132.0657 5810 4-IMIDAZOLEACETIC ACID 0.93 127.0503 96215 5,6-DIHYDROURACIL 1.26 115.0503 649 5'-DEOXYADENOSINE 6.01 252.1094 439182 5-METHYLCYTOSINE HYDROCLORIDE 1.13 126.0664 65040 6-DEOXY-HEXOSE 0.93 187.0577 840 Acetaminophen/Paracetamol 7.01 152.0708 1983 Acyl-Carnitine(4-OH) 5.48 248.1494 ADENINE 1.41 136.0619 190 ADENOSINE 5'-MONOPHOSPHATE 1.59 348.0713 6083 ALANINE/SARCOSINE 0.94 90.0551 ALDO/KETO-HEXOSE 0.75 203.0527 ALDOPENTOSE 0.85 173.0425 Allopurinol 6.16 137.0458 2094 ALPHA-AMINOADIPATE/N-METHYL-L-GLUTAMATE 0.89 162.0764 Anthranilate 8.62 120.0448 227 ANTHRANILATE/4-AMINOBENZOATE 7.99 138.0553 ASPARTATE 0.82 134.0449 5960 BETAINE 0.81 118.0863 247 Bis(2-ethylhexyl) phthalate 16.63 391.2843 8343 C5-SUGAR ALCOHOL 0.74 175.0583 Choline 0.80 104.1071 305 CREATINE 0.96 132.0772 586 CREATININE 0.92 114.0663 588 CYTIDINE 1.50 244.0934 6175 CYTOSINE 0.86 112.0509 597 Dibutyl phthalate 14.71 279.1589 3026 Dihydroxyquinoine 8.10 162.0550 54680871 DIMETHYLBENZIMIDAZOLE 7.34 147.0917 675 Dipropylene glycol 6.46 157.0837 6510 Erucamide 16.88 338.3417 5365371 Garcinia lactone dibutyl ester 8.74 325.1240 GLUTARATE 4.61 155.0314 743 Glutathione Disulfide 3.82 307.0835 65359 GLYCINE 0.80 76.0394 750 Glycyl-L-leucine 6.80 189.1233 92843 GUANINE 1.46 152.0566 764 HEXOSE-DISACCHARIDE 0.80 365.1055 Hippuric Acid 7.94 180.0660 464 HOMOCYSTEINE 1.02 136.0430 778 Homocystine 1.01 269.0619 10100 HYPOXANTHINE 2.01 137.0457 790 INDOLE-3-ACETATE 9.16 176.0711 802 Isocytosine 1.49 112.0505 66950 Kynurenic Acid 7.67 190.0498 3845 KYNURENINE 6.33 209.0917 846 L-2-PHOSPHOGLYCERIC ACID 0.76 187.0002 439278 L-Acetylcarnitine 3.81 204.1232 7045767 L-ARGININE 0.71 175.1189 6322 L-ASPARAGINE 0.71 133.0610 6267 L-CARNITINE 0.90 162.1126 10917 Leu Pro 7.27 229.1552 L-GLUTAMIC ACID 0.76 148.0607 33032 L-GLUTAMINE 0.85 147.0763 5961 L-HISTIDINE 0.71 156.0768 6274 L-HOMOCYSTEINE THIOLACTONE 0.94 118.0325 134505 L-Isoleucine 2.03 132.1020 6306 L-METHIONINE 1.34 150.0588 6137 L-Tryptophan 13C11 7.55 216.1337 L-TYROSINE 3.25 182.0811 6057 L-VALINE/5-AMINOPENTANOATE/L-NORVALINE 1.07 118.0865 LysoPE (18:1) 13.64 480.3075 9547071 Methionine Sulfoxide 0.78 166.0537 847 METHYL INDOLE-3-ACETATE 10.71 190.0866 74706 Monobutyl phthalate 12.41 223.0962 8575 N-ACETYL-DL-GLUTAMIC ACID 2.16 190.0710 185 N-ACETYL-DL-SERINE 1.06 148.0606 352294 N-ACETYL-L-ASPARTIC ACID 1.62 176.0557 65065 N-ACETYL-L-LEUCINE 7.77 174.1122 70912 N-ACETYLNEURAMINATE 0.80 310.1121 439197 N-ACETYLPUTRESCINE 1.07 131.1179 122356 N-ALPHA-ACETYL-L-LYSINE 1.24 189.1234 92907 N-Butylbenzenesulfonamide 11.96 214.0883 19241 Orthophosphate 0.74 98.9844 1061 OxPC (20:3(KeKe)) 7.02 588.2988 Pantothenic Acid 6.62 220.1187 6613 PEG n5 7.07 261.1315 62551 PHENETHYLAMINE 6.83 122.0967 1001 Phenylalanine 6.05 166.0863 6140 PHOSPHOCHOLINE 0.73 184.0734 1014 PICOLINIC ACID 1.31 124.0397 1018 PIPECOLATE/L-PIPECOLIC ACID 1.29 130.0866 439227 Proprionylcarnitine 6.69 218.1389 107738 PYRIDOXAL 1.78 168.0650 1050 Pyroglutamic Acid 1.79 130.0500 7405 Resorcinol monoacetate 8.29 153.0545 5055 SEROTONIN 6.34 177.1025 5202 Stachydrine 1.03 144.1023 115244 Sulcatol 8.97 129.1278 20745 THREONINE/HOMOSERINE 0.73 120.0656 THYMINE 3.24 127.0503 1135 TRANS-CINNAMATE 9.78 149.0595 444539 Triethyl phosphate 9.33 183.0779 6535 Trigonelline 0.90 138.0549 5570 Tryptophan 7.65 205.0976 6305 TYRAMINE 4.47 138.0917 5610 URACIL 1.37 113.0348 1174 URIDINE 2.79 267.0593 6029 URIDINE-5-MONOPHOSPHATE 1.26 325.0438 6030 METABOLITES_END #END