#METABOLOMICS WORKBENCH TruxalCarlson_20200324_121313 DATATRACK_ID:1951 STUDY_ID:ST001393 ANALYSIS_ID:AN002331 PROJECT_ID:PR000956 VERSION 1 CREATED_ON June 4, 2020, 4:06 pm #PROJECT PR:PROJECT_TITLE Sea-ice diatom compatible solute shifts PR:PROJECT_TYPE Marine Metabolomics PR:PROJECT_SUMMARY Sea-ice algae provide an important source of primary production in polar PR:PROJECT_SUMMARY regions, yet we have limited understanding of their responses to the seasonal PR:PROJECT_SUMMARY cycling of temperature and salinity. Using a targeted liquid chromatography-mass PR:PROJECT_SUMMARY spectrometry-based metabolomics approach, we found that axenic cultures of the PR:PROJECT_SUMMARY Antarctic sea-ice diatom, Nitzschia lecointei, displayed large differences in PR:PROJECT_SUMMARY their metabolomes when grown in a matrix of conditions that included PR:PROJECT_SUMMARY temperatures of –1 and 4°C, and salinities of 32 and 41, despite relatively PR:PROJECT_SUMMARY small changes in growth rate. Temperature exerted a greater effect than salinity PR:PROJECT_SUMMARY on cellular metabolite pool sizes, though the N- or S-containing compatible PR:PROJECT_SUMMARY solutes, 2,3-dihydroxypropane-1-sulfonate (DHPS), glycine betaine (GBT), PR:PROJECT_SUMMARY dimethylsulfoniopropionate (DMSP), and proline responded strongly to both PR:PROJECT_SUMMARY temperature and salinity, suggesting complexity in their control. We saw the PR:PROJECT_SUMMARY largest (> 4 fold) response to salinity for proline. DHPS, a rarely studied but PR:PROJECT_SUMMARY potential compatible solute, reached the highest intracellular compatible solute PR:PROJECT_SUMMARY concentrations of ~ 85 mM. When comparing the culture findings to natural Arctic PR:PROJECT_SUMMARY sea-ice diatom communities, we found extensive overlap in metabolite profiles, PR:PROJECT_SUMMARY highlighting the relevance of culture-based studies to probe environmental PR:PROJECT_SUMMARY questions. Large changes in sea-ice diatom metabolomes and compatible solutes PR:PROJECT_SUMMARY over a seasonal cycle could be significant components of biogeochemical cycling PR:PROJECT_SUMMARY within sea ice. PR:INSTITUTE University of Washington PR:DEPARTMENT School of Oceanography PR:LABORATORY Ingalls Lab PR:LAST_NAME Dawson PR:FIRST_NAME Hannah PR:ADDRESS 1501 NE Boat Street, Marine Science Building, Room G, Seattle, WA 98195 PR:EMAIL hmdawson@uw.edu PR:PHONE 2062216750 PR:FUNDING_SOURCE Booth Foundation, NSF, UW Graduate Top Scholar Award, Gordon and Betty Moore PR:FUNDING_SOURCE Foundation PR:PUBLICATIONS Dawson et al., Elementa #STUDY ST:STUDY_TITLE Sea-ice diatom compatible solute shifts ST:STUDY_TYPE Compatible solutes were quantified in sea-ice diatoms ST:STUDY_SUMMARY Sea-ice algae provide an important source of primary production in polar ST:STUDY_SUMMARY regions, yet we have limited understanding of their responses to the seasonal ST:STUDY_SUMMARY cycling of temperature and salinity. Using a targeted liquid chromatography-mass ST:STUDY_SUMMARY spectrometry-based metabolomics approach, we found that axenic cultures of the ST:STUDY_SUMMARY Antarctic sea-ice diatom, Nitzschia lecointei, displayed large differences in ST:STUDY_SUMMARY their metabolomes when grown in a matrix of conditions that included ST:STUDY_SUMMARY temperatures of –1 and 4°C, and salinities of 32 and 41, despite relatively ST:STUDY_SUMMARY small changes in growth rate. Temperature exerted a greater effect than salinity ST:STUDY_SUMMARY on cellular metabolite pool sizes, though the N- or S-containing compatible ST:STUDY_SUMMARY solutes, 2,3-dihydroxypropane-1-sulfonate (DHPS), glycine betaine (GBT), ST:STUDY_SUMMARY dimethylsulfoniopropionate (DMSP), and proline responded strongly to both ST:STUDY_SUMMARY temperature and salinity, suggesting complexity in their control. We saw the ST:STUDY_SUMMARY largest (> 4 fold) response to salinity for proline. DHPS, a rarely studied but ST:STUDY_SUMMARY potential compatible solute, reached the highest intracellular compatible solute ST:STUDY_SUMMARY concentrations of ~ 85 mM. When comparing the culture findings to natural Arctic ST:STUDY_SUMMARY sea-ice diatom communities, we found extensive overlap in metabolite profiles, ST:STUDY_SUMMARY highlighting the relevance of culture-based studies to probe environmental ST:STUDY_SUMMARY questions. Large changes in sea-ice diatom metabolomes and compatible solutes ST:STUDY_SUMMARY over a seasonal cycle could be significant components of biogeochemical cycling ST:STUDY_SUMMARY within sea ice. ST:INSTITUTE University of Washington ST:DEPARTMENT School of Oceanography ST:LABORATORY Ingalls Lab ST:LAST_NAME Dawson ST:FIRST_NAME Hannah ST:ADDRESS 1501 NE Boat Street, Marine Science Building, Room G, Seattle, WA 98195 ST:EMAIL hmdawson@uw.edu ST:PHONE 2062216750 ST:PUBLICATIONS Dawson et al., Elementa #SUBJECT SU:SUBJECT_TYPE Other SU:SUBJECT_SPECIES Nitzschia lecointei SU:TAXONOMY_ID 186028 SU:GENDER Not applicable #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 - 32ppt-1C_A Type:Smp | Salinity:32 | Temp_degC:-1 Replicate=A; RFU=605.6; Vol_L=0.07; RAW_FILE_NAME=170410_Smp_32ppt-1C_A;170413_Smp_40ppt4C_C;170410_Smp_32ppt-1C_A SUBJECT_SAMPLE_FACTORS - 32ppt-1C_B Type:Smp | Salinity:32 | Temp_degC:-1 Replicate=B; RFU=551.2; Vol_L=0.07; RAW_FILE_NAME=170410_Smp_32ppt-1C_B;170413_Smp_32ppt-1C_B;170410_Smp_32ppt-1C_B SUBJECT_SAMPLE_FACTORS - 32ppt-1C_C Type:Smp | Salinity:32 | Temp_degC:-1 Replicate=C; RFU=550.6; Vol_L=0.07; RAW_FILE_NAME=170410_Smp_32ppt-1C_C;170413_Smp_32ppt-1C_C;170410_Smp_32ppt-1C_C SUBJECT_SAMPLE_FACTORS - 32ppt4C_A Type:Smp | Salinity:32 | Temp_degC:4 Replicate=A; RFU=847.1; Vol_L=0.07; RAW_FILE_NAME=170410_Smp_32ppt4C_A;170413_Smp_32ppt4C_B;170410_Smp_32ppt4C_A SUBJECT_SAMPLE_FACTORS - 32ppt4C_B Type:Smp | Salinity:32 | Temp_degC:4 Replicate=B; RFU=967.1; Vol_L=0.07; RAW_FILE_NAME=170410_Smp_32ppt4C_B;170413_Smp_32ppt4C_A;170410_Smp_32ppt4C_B SUBJECT_SAMPLE_FACTORS - 32ppt4C_C Type:Smp | Salinity:32 | Temp_degC:4 Replicate=C; RFU=918.5; Vol_L=0.07; RAW_FILE_NAME=170410_Smp_32ppt4C_C;170413_Smp_32ppt4C_C;170410_Smp_32ppt4C_C SUBJECT_SAMPLE_FACTORS - 40ppt-1C_A Type:Smp | Salinity:40 | Temp_degC:-1 Replicate=A; RFU=860.2; Vol_L=0.07; RAW_FILE_NAME=170410_Smp_40ppt-1C_A;170413_Smp_40ppt-1C_A;170410_Smp_40ppt-1C_A SUBJECT_SAMPLE_FACTORS - 40ppt-1C_B Type:Smp | Salinity:40 | Temp_degC:-1 Replicate=B; RFU=681.6; Vol_L=0.07; RAW_FILE_NAME=170410_Smp_40ppt-1C_B;170413_Smp_40ppt4C_B;170410_Smp_40ppt-1C_B SUBJECT_SAMPLE_FACTORS - 40ppt-1C_C Type:Smp | Salinity:40 | Temp_degC:-1 Replicate=C; RFU=814.3; Vol_L=0.07; RAW_FILE_NAME=170410_Smp_40ppt-1C_C;170413_Smp_40ppt-1C_C;170410_Smp_40ppt-1C_C SUBJECT_SAMPLE_FACTORS - 40ppt4C_A Type:Smp | Salinity:40 | Temp_degC:4 Replicate=A; RFU=581.8; Vol_L=0.07; RAW_FILE_NAME=170410_Smp_40ppt4C_A;170413_Smp_40ppt4C_A;170410_Smp_40ppt4C_A SUBJECT_SAMPLE_FACTORS - 40ppt4C_B Type:Smp | Salinity:40 | Temp_degC:4 Replicate=B; RFU=681.6; Vol_L=0.07; RAW_FILE_NAME=170410_Smp_40ppt4C_B;170413_Smp_40ppt-1C_B;170410_Smp_40ppt4C_B SUBJECT_SAMPLE_FACTORS - 40ppt4C_C Type:Smp | Salinity:40 | Temp_degC:4 Replicate=C; RFU=662; Vol_L=0.07; RAW_FILE_NAME=170410_Smp_40ppt4C_C;170413_Smp_32ppt-1C_A;170410_Smp_40ppt4C_C SUBJECT_SAMPLE_FACTORS - ASWFilterBlk_1 Type:Blk | Salinity:NA | Temp_degC:NA Replicate=1; RFU=NA; Vol_L=0.3; RAW_FILE_NAME=170612_Blk_ASWFilterBlk_1;170615_Blk_ASWFilterBlk_1;170612_Blk_ASWFilterBlk_1 SUBJECT_SAMPLE_FACTORS - ASWFilterBlk_2 Type:Blk | Salinity:NA | Temp_degC:NA Replicate=2; RFU=NA; Vol_L=0.3; RAW_FILE_NAME=170612_Blk_ASWFilterBlk_2;170615_Blk_ASWFilterBlk_2;170612_Blk_ASWFilterBlk_2 SUBJECT_SAMPLE_FACTORS - ASWFilterBlk_3 Type:Blk | Salinity:NA | Temp_degC:NA Replicate=3; RFU=NA; Vol_L=0.3; RAW_FILE_NAME=170612_Blk_ASWFilterBlk_3;170615_Blk_ASWFilterBlk_3;170612_Blk_ASWFilterBlk_3 SUBJECT_SAMPLE_FACTORS - MediaBlk_ppt32 Type:Blk | Salinity:32 | Temp_degC:NA Replicate=ppt32; RFU=1; Vol_L=0.07; RAW_FILE_NAME=170410_Blk_MediaBlk_ppt32;170413_Blk_MediaBlk_ppt32;170410_Blk_MediaBlk_ppt32 SUBJECT_SAMPLE_FACTORS - MediaBlk_ppt40 Type:Blk | Salinity:40 | Temp_degC:NA Replicate=ppt40; RFU=1; Vol_L=0.07; RAW_FILE_NAME=170410_Blk_MediaBlk_ppt40;170413_Blk_MediaBlk_ppt40;170410_Blk_MediaBlk_ppt40 SUBJECT_SAMPLE_FACTORS - S2C_4 Type:Smp | Salinity:NA | Temp_degC:NA Replicate=4; RFU=NA; Vol_L=0.1671; RAW_FILE_NAME=170612_Smp_S2C_4;170615_Smp_S2C_4;170612_Smp_S2C_4 SUBJECT_SAMPLE_FACTORS - S2C_5 Type:Smp | Salinity:NA | Temp_degC:NA Replicate=5; RFU=NA; Vol_L=0.2486; RAW_FILE_NAME=170612_Smp_S2C_5;170615_Smp_S2C_5;170612_Smp_S2C_5 SUBJECT_SAMPLE_FACTORS - S2C_6 Type:Smp | Salinity:NA | Temp_degC:NA Replicate=6; RFU=NA; Vol_L=0.2049; RAW_FILE_NAME=170612_Smp_S2C_6;170615_Smp_S2C_6;170612_Smp_S2C_6 #COLLECTION CO:COLLECTION_SUMMARY Cultured diatom cells at different salinities and temperatures grown to CO:COLLECTION_SUMMARY exponential phase were filtered onto 0.2-micron filters and extracted for CO:COLLECTION_SUMMARY metabolites as described in methods. Three dedicated ice cores were sampled from CO:COLLECTION_SUMMARY the Chukchi Sea near Utqiaġvik, AK. The bottom 5-cm sections were placed in CO:COLLECTION_SUMMARY polycarbonate tubs, allowed to melt at 4°C in artificial seawater, and filtered CO:COLLECTION_SUMMARY onto 0.2-micron filters. Filters were extracted for metabolites as described in CO:COLLECTION_SUMMARY methods. All filters were frozen in liquid nitrogen immediately after filtration CO:COLLECTION_SUMMARY and stored in a -80 C freezer until extraction. CO:SAMPLE_TYPE Diatom cells/Particulate matter from sea ice cores CO:STORAGE_CONDITIONS Described in summary #TREATMENT TR:TREATMENT_SUMMARY Diatom cells were cultured in a matrix of two temperatures (–1°C and 4°C) TR:TREATMENT_SUMMARY and two salinities (32 and 40) in triplicate. There was no treatment for the sea TR:TREATMENT_SUMMARY ice cores – this was a study of how the cultured diatoms compare to the TR:TREATMENT_SUMMARY diatom-dominated Arctic sea-ice communities. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Each sample was extracted using a modified Bligh-Dyer extraction. Briefly, SP:SAMPLEPREP_SUMMARY filters were cut up and put into 15 mL teflon centrifuge tubes containing a SP:SAMPLEPREP_SUMMARY mixture of 100 µm and 400 µm silica beads. Heavy isotope-labeled internal SP:SAMPLEPREP_SUMMARY standards were added along with ~2 mL of cold aqueous solvent (50:50 SP:SAMPLEPREP_SUMMARY methanol:water) and ~3 mL of cold organic solvent (dichloromethane). The samples SP:SAMPLEPREP_SUMMARY were shaken on a FastPrep-24 Homogenizer for 30 seconds and chilled in a -20 °C SP:SAMPLEPREP_SUMMARY freezer repeatedly for three cycles of bead-beating and a total of 30 minutes of SP:SAMPLEPREP_SUMMARY chilling. The organic and aqueous layers were separated by spinning samples in a SP:SAMPLEPREP_SUMMARY centrifuge at 4,300 rpm for 2 minutes at 4 °C. The aqueous layer was removed to SP:SAMPLEPREP_SUMMARY a new glass centrifuge tube. The remaining organic fraction was rinsed three SP:SAMPLEPREP_SUMMARY more times with additions of 1 to 2 mL of 50:50 methanol:water. All aqueous SP:SAMPLEPREP_SUMMARY rinses were combined for each sample and dried down under N2 gas. The remaining SP:SAMPLEPREP_SUMMARY organic layer was transferred into a clean glass centrifuge tube and the SP:SAMPLEPREP_SUMMARY remaining bead beating tube was rinsed two more times with cold organic solvent. SP:SAMPLEPREP_SUMMARY The combined organic rinses were centrifuged, transferred to a new tube, and SP:SAMPLEPREP_SUMMARY dried under N2 gas. Dried aqueous fractions were re-dissolved in 380 µL of SP:SAMPLEPREP_SUMMARY water. Dried organic fractions were re-dissolved in 380 µL of 1:1 SP:SAMPLEPREP_SUMMARY water:acetonitrile. 20 µL of isotope-labeled injection standards in water were SP:SAMPLEPREP_SUMMARY added to both fractions. Blank filters were extracted alongside samples as SP:SAMPLEPREP_SUMMARY methodological blanks. SP:PROCESSING_STORAGE_CONDITIONS On ice SP:EXTRACTION_METHOD Bligh-Dyer SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY See attached summary CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity I-Class CH:COLUMN_NAME Waters Acquity UPLC HSS Cyano (100 x 2.1mm, 1.8um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Waters Xevo-TQ-S MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS See protocol, data from culture samples #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Normalized Peak Area Per RFU MS_METABOLITE_DATA_START Samples 32ppt-1C_A 32ppt-1C_B 32ppt-1C_C 32ppt4C_A 32ppt4C_B 32ppt4C_C 40ppt-1C_A 40ppt-1C_B 40ppt-1C_C 40ppt4C_A 40ppt4C_B 40ppt4C_C MediaBlk_ppt32 MediaBlk_ppt40 Factors Type:Smp | Salinity:32 | Temp_degC:-1 Type:Smp | Salinity:32 | Temp_degC:-1 Type:Smp | Salinity:32 | Temp_degC:-1 Type:Smp | Salinity:32 | Temp_degC:4 Type:Smp | Salinity:32 | Temp_degC:4 Type:Smp | Salinity:32 | Temp_degC:4 Type:Smp | Salinity:40 | Temp_degC:-1 Type:Smp | Salinity:40 | Temp_degC:-1 Type:Smp | Salinity:40 | Temp_degC:-1 Type:Smp | Salinity:40 | Temp_degC:4 Type:Smp | Salinity:40 | Temp_degC:4 Type:Smp | Salinity:40 | Temp_degC:4 Type:Blk | Salinity:32 | Temp_degC:NA Type:Blk | Salinity:40 | Temp_degC:NA Aminobenzoic Acid 222.1994716 195.2086357 245.0472212 166.8504309 175.5464792 139.7735438 87.5029063 204.4175469 154.0881739 311.5383293 943.4624413 300.2205438 13684 14116 Carotene 733.6459709 67.85195936 116.1823465 89.02845001 158.0074449 1237.301034 1185.173216 1696.026995 430.0994719 217.0660021 215.5780516 157.9697885 1164 114 Coenzyme B12 187.9219898 193.8499784 266.5913551 34.32993541 70.03574606 70.65753103 125.7051633 249.8146046 122.2676461 158.1988687 86.66613549 141.9749015 2526.080228 989.2758583 Cyano B12 4458.010627 5560.687567 6769.70982 2621.998238 2176.576899 1614.125794 2973.909964 3947.959083 3194.75807 3425.820143 3661.970899 3365.663566 147843.7859 151212.7562 Hydroxo B12 21514.91778 18998.48752 31154.68068 7507.03584 6863.050979 8644.76825 17297.7342 13747.66191 14954.36746 12437.05931 9801.4477 10141.50298 38511.2911 41602.56383 Kynurenine 267.3778071 217.8592163 303.5851798 1065.606186 762.3999586 1054.648884 252.5029063 322.1038732 253.4987105 3062.454452 1865.233275 2093.291541 34030 18016 Methyl B12 149.0108983 293.684688 191.8852161 47.73344351 41.81056768 44.02286336 114.2664497 280.1672535 241.4282206 182.9030595 101.9322183 128.9199396 5558 3961 Methylthioadenosine 362136.0263 381847.4049 477074.549 731308.5991 686672.1102 870397.9544 301695.4125 490383.6677 333768.551 1242380.882 1011336.815 972165.7548 85903.42157 75850.0716 Phenylacetic Acid 54.26519155 59.62082729 59.68579731 42.39759178 35.16699411 49.15187806 38.20390607 48.21449531 40.35736215 90.07047095 62.90492958 61.79154079 6076 10921 Phenylalanine 754523.0949 792938.5864 768163.913 1012231.482 896853.1129 1132130.267 532303.9944 707996.3195 590574.6367 1563871.077 1269217.067 1240483.018 2615390.709 4890494.387 Pyridoxal 1803.502325 1761.249789 1982.724604 1129.708948 1358.391012 1908.285821 1256.666702 2672.731313 1698.319075 2682.222317 1723.398046 1507.376037 9529.022866 11526.18837 Pyridoxal Phosphate 64916.216 52246.10896 52261.66306 41894.74709 34785.96038 49084.12181 34784.69112 53669.00458 40703.60713 55473.17688 56796.07567 43039.79018 173786.5798 152996.8212 Thiamine monophosphate 343.4008022 245.1907192 193.6580859 541.7024336 250.4245734 425.2635488 110.9457601 190.8977053 138.0602021 363.7904491 353.963326 307.3150769 2614.526152 3196.701168 Tocopherol (Vit E) 399.4633421 225.8164006 133.0148928 164.0455672 224.6313721 355.9129015 402.9190886 302.0319836 345.5618322 512.2189756 161.907277 166.7009063 30373 33101 Tryptophan 450541.3535 433085.8266 463629.798 1167782.657 1004404.723 1224763.974 311926.4936 397753.3006 400168.5869 2033722.511 1554590.616 1569276.562 916151.8627 1779650.446 Vitamin B2 1035.992362 1018.556749 1199.671822 2405.964454 1301.730488 3478.30864 1076.829302 1027.680963 1079.29212 3809.456946 3076.337645 1879.668219 33242.1693 27384.67206 Xanthine 22840.39346 19561.71665 14437.72612 22620.34747 17909.1042 31033.52285 13780.49208 16385.8382 19801.63679 42899.09936 35594.81178 32598.93189 133250.1629 124755.75 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name quantitated m/z KEGGNAME CHEBI KEGG ID MS_method Aminobenzoic Acid 138.055503 4-Aminobenzoate; ABEE; 4-Aminobenzoic acid; p-Aminobenzoate CHEBI:30753 C00568 RP_TQS_Pos Carotene 536.4382 beta-Carotene; all-trans-beta-Carotene CHEBI:17579 C02094 RP_TQS_Pos Coenzyme B12 790.3367 Cobamide coenzyme; Adenosylcobalamin; Adenosylcob(III)alamin; Deoxyadenosylcobalamin; Cobamamide; Vitamin B12 coenzyme; 5,6-Dimethylbenzimidazolyl-5-deoxyadenosyl-cobamide; (5'-Deoxy-5'-adenosyl)cobamide coenzyme; (5,6-Dimethylbenzimidazolyl)cobamide coenzyme; alpha-(5,6-Dimethylbenzimidazolyl)cobamide coenzyme; 5'-Deoxy-5'-adenosylcobalamin; 5'-Deoxy-5'-adenosyl vitamin B12; 5'-Deoxy-5'-adenosyl-5,6-dimethylbenzimidazolylcobamide; 5,6-Dimethylbenzimidazolyl-Co-5'-deoxy-5'-adenosylcobamide; Calomide; Cobalamin coenzyme; Coenzyme B12; DMBC coenzyme; Dibencozide; Funacomide CHEBI:18408 C00194 RP_TQS_Pos Cyano B12 678.2894 Cyanocobalamin; Cyanocob(III)alamin; Dicopac; Vitamin B12 complex CHEBI:17439 C02823 RP_TQS_Pos Hydroxo B12 664.7861 Hydroxocobalamin; Vitamin B12a; Hydroxycobalamin; Hydroxycob(lll)alamin CHEBI:27786 C08230 RP_TQS_Pos Kynurenine 209.092618 L-Kynurenine; 3-Anthraniloyl-L-alanine CHEBI:16946 C00328 RP_TQS_Pos Methyl B12 672.8011 Methylcobalamin; Methylcob(III)alamin CHEBI:28115 C06453 RP_TQS_Pos Methylthioadenosine 298.097387 5'-Methylthioadenosine; Methylthioadenosine; S-Methyl-5'-thioadenosine; 5-Methylthioadenosine; 5'-Deoxy-5'-(methylthio)adenosine; Thiomethyladenosine; MTA CHEBI:17509 C00170 RP_TQS_Pos Phenylacetic Acid 137.060254 Phenylacetic acid; Benzylformic acid; Phenylacetate; Benzeneacetic acid CHEBI:30745 C07086 RP_TQS_Pos Phenylalanine 166.086804 L-Phenylalanine; (S)-alpha-Amino-beta-phenylpropionic acid CHEBI:17295 C00079 RP_TQS_Pos Pyridoxal 168.066069 Pyridoxal CHEBI:17310 C00250 RP_TQS_Pos Pyridoxal Phosphate 248.032402 Pyridoxal phosphate; Pyridoxal 5-phosphate; Pyridoxal 5'-phosphate; PLP CHEBI:18405 C00018 RP_TQS_Pos Thiamine monophosphate 345.078641 Thiamin monophosphate; Thiamine monophosphate; Thiamin phosphate; Thiamine phosphate; TMP CHEBI:9533 C01081 RP_TQS_Pos Tocopherol (Vit E) 431.388905 alpha-Tocopherol; Vitamin E CHEBI:22470 C02477 RP_TQS_Pos Tryptophan 205.097703 L-Tryptophan; Tryptophan; (S)-alpha-Amino-beta-(3-indolyl)-propionic acid CHEBI:16828 C00078 RP_TQS_Pos Vitamin B2 377.146111 Riboflavin; Lactoflavin; 7,8-Dimethyl-10-ribitylisoalloxazine; Vitamin B2 CHEBI:17015 C00255 RP_TQS_Pos Xanthine 153.041251 Xanthine CHEBI:15318 C00385 RP_TQS_Pos METABOLITES_END #END