#METABOLOMICS WORKBENCH TruxalCarlson_20200324_121313 DATATRACK_ID:1951 STUDY_ID:ST001393 ANALYSIS_ID:AN002323 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 HILIC CH:INSTRUMENT_NAME Waters Acquity I-Class CH:COLUMN_NAME SeQuant ZIC- pHILIC (150 x 2.1mm, 5um) #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 Adenine 656.2235799 842.8193033 895.8754086 1613.077559 1640.419812 1623.832335 517.2994652 969.4688967 715.075525 2745.885184 1878.157277 1981.238671 15337 33193 Adenosyl Homocysteine 19895.22622 20454.78048 21312.7461 31987.28603 28839.88212 36229.37398 18734.95117 25427.83744 23606.85742 57740 42362.36209 36937.48943 143135 100394 Adenosyl Methionine 542049.5112 516209.521 535944.6131 783739.0438 781693.4588 663898.0947 286338.3213 465541.9718 461246.3416 1378471.227 1073598.404 1432811.311 481702 199371 Alanine 58424.19534 57571.9447 66974.78812 33549.26764 32969.15362 36444.21585 46480.95841 61719.8195 49849.58008 46791.93341 36999.62952 33716.90355 58318.66966 212799.0712 Aminobutyric Acid 453.4676354 471.9430334 739.8111151 1057.153819 1161.473477 1213.715841 811.914671 919.0170188 684.0697532 2284.728429 801.1605047 1452.285498 17969 39878 Argininosuccinic Acid 4988.75 5481.108491 5487.081366 32360.15583 31963.64182 36895.45563 5724.975587 7470.977113 8090.690163 85656.16363 57118.00469 58664.96073 77344 26111 Asparagine 14981.16399 18456.84309 20651.56279 30308.64249 29922.95195 25687.25898 17852.19247 20476.93069 15642.16952 70341.5554 57160.538 74424.51349 157382.4016 95519.81423 Aspartic acid 110664.465 120854.3614 127908.5652 39594.89316 40345.08944 43260.1546 68361.33457 103558.3685 79630.61771 52270.17532 42914.49531 43066.51964 195400 317589 Carnitine 223468.8137 229273.8128 282495.5341 351820.3617 316291.98 367786.614 133069.7541 195233.2346 147966.314 428658.2094 337400.6492 399906.0761 1103610.271 869589.612 Choline 4952317.503 6257941.836 5556984.299 2131419.026 1917125.759 2489644.214 4087977.47 5248156.225 3734051.242 3736114.584 2601598.278 2760231.258 1088223.994 1971623.867 Citrulline 58898.2893 45804.63353 70840.28333 116319.9056 108580.2089 75224.76211 37511.67635 47538.69131 50843.78239 184703.6645 125726.0446 121466.0544 1183326 1692075 Cystine 766.9649934 469.04209 524.4969125 311.6467949 586.9785958 230.1230267 286.6275285 497.4105047 597.82881 1357.657271 698.4727113 1166.655589 8267 19747 Dimethyl Glycine 179825.5088 172733.4601 219498.1602 203839.7781 182474.6326 240653.5262 245630.1371 298974.9595 244507.0004 419579.6573 319222.1987 353681.2317 2342121.061 3007074.332 Glucosylglycerol 10929.68835 9829.011205 9393.417312 4746.109956 4445.398196 3237.10838 10698.31583 15173.20186 11372.25469 6210.763974 4914.984181 5689.533565 0 349.3680074 Glutathione 2681.266513 7649.189042 8600.283327 1059.493566 12373.81346 34976.00435 14659.93722 4161.056338 7000.519465 8341.177381 1903.252641 2812.107251 126167 321023 Glutathione Disulfide 75571.92206 95411.582 116094.3117 40071.68457 40500.79206 34303.68645 71321.62753 113748.3216 81431.67874 42587.39773 43309.90904 30192.24169 163083 46081 Guanine 243.2260885 715.4418053 759.3321611 357.2083891 169.8750042 471.7488856 61.11022876 436.0505081 146.9977131 351.6437762 530.9278977 797.2452699 20150.47347 11536.3715 Guanosine 366.1922061 590.9052975 406.0406829 657.2600637 465.8215283 549.0332063 167.2157638 211.0313967 176.641287 324.7920248 464.352993 1025.116314 47913 373 Histidine 586832.5457 534313.4439 630601.826 584084.2625 564905.9776 763622.0205 526365.0222 714217.4963 620753.3986 1426617.475 1152114.671 1008821.71 12918617.93 36146039.8 Homarine 4005.611858 4692.04402 4154.546851 17269.43636 5702.950803 9290.378636 10445.03902 7123.448685 8754.134508 11086.42803 18168.44134 5603.652119 734711.8312 624063.2739 Homoserine 91732.76727 111363.5389 89676.23224 32021.54524 30409.94751 19399.48162 107479.7685 115521.6662 101972.3905 35581.67891 26944.9439 18631.62721 107292.661 152757.5584 Hypotaurine 122.176354 162.9390421 215.8990193 605.464526 649.0931651 608.624932 99.86166008 251.9468897 182.3197839 819.9071846 576.2309272 623.4259819 14437 0 Isoleucine 44698.3735 49269.84942 52018.08883 60081.73042 51217.5747 64647.82 20726.53382 26120.09755 26583.64227 89462.36198 64855.59586 79872.48897 702164.5551 297847.6765 Leucine 39998.04531 51612.62545 46661.39355 68789.92082 58067.74475 98040.86915 28471.1811 35127.34011 29892.06236 96361.93723 89490.59428 72013.71467 169024.3986 313678.5302 Lysine 387394.3461 344367.2859 410163.1093 334967.1302 330294.8609 373891.2749 293181.6973 449726.0563 343552.0865 361035.5449 315076.0798 290941.2205 452213 469028 Methionine 63966.33321 64375.1312 72097.25291 41037.01922 33660.12752 40327.51506 64437.36505 94383.04274 75587.61573 60899.14593 49644.26739 42700.5502 163424.4988 82157.64554 Methionine Sulfoxide 3955.057021 3463.427993 5018.257196 3989.819502 3486.02387 2516.959445 4586.128801 6477.308994 5472.739231 4471.801366 3357.264068 3151.60136 116987.6059 338881.2474 N-Acetyl-Lysine 27049.39729 29260.11248 26173.13658 59480.84996 55701.8054 78671.92161 15707.103 16912.91373 15027.47759 48256.33551 70308.56221 95574.15106 303191 94683 Ornithine 95365.21797 84860.67489 111877.559 82159.25393 88998.31662 64704.93195 66779.18624 99024.9061 79630.27385 44656.3252 42877.30047 44782.35045 614634 542200 Sarcosine 1513.48178 2049.866197 1447.257572 2554.467683 1497.483487 1762.344197 2004.741218 3260.500425 2945.305669 2944.174868 1931.970313 2373.175659 119598.2443 48619.61487 Serine 18137.8319 17006.47496 17924.71304 11476.94487 11351.39179 10122.89167 13027.20763 17548.39495 12537.80548 15211.75146 11725.08656 11363.66616 246825 258782 Thiamine pyrophosphate 8762.319338 17626.45811 19024.1069 3219.516641 8410.676343 11166.24389 14443.03326 6878.933132 14800.12265 11304.26269 7035.836124 10361.1312 942238.0242 325148.351 Threonine 41082.29855 36513.16038 45544.2826 36531.79082 32988.12532 39265.27599 38606.38456 44704.777 37200.92595 54665.99862 42407.40317 31995.0423 13647 270448 trans Hydroxyl proline 39210.12219 43500.93614 43591.66364 33158.67548 33297.24124 40140.92107 95227.94699 121825.6808 101066.5455 120245.5002 87991.13263 86452.1571 108161 1083440 Trigonelline 2850.031374 3131.311684 3134.723938 3467.667336 2567.493537 2662.629287 3673.85608 3308.80135 2829.075279 7757.542111 7030.497359 2315.932024 575293 296431 Tyrosine 31360.74837 33970.48015 28967.91053 117312.7419 136734.3416 212595.51 12190.72788 13940.53016 16840.96521 263203.8785 160766.4646 160722.668 86963.58954 111608.3212 Valine 38446.99837 39621.00493 42916.15826 30842.65231 26333.49683 34866.82357 38363.75809 44718.22245 37982.21713 63347.19558 46405.19079 43910.58035 79785.65577 57606.03044 Vitamin B3 106776.9155 113473.9405 99767.22121 116840.0519 99923.25096 151400.0131 62061.08347 106753.9906 76946.05919 126421.6707 138047.5352 135608.2538 2852186 1864412 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name quantitated m/z CHEBI KEGGNAME MS_method KEGG_ID Adenine 136.06232 CHEBI:16708 Adenine; 6-Aminopurine HILIC_TQS_Pos C00147 Adenosyl Homocysteine 385.129416 CHEBI:16680 S-Adenosyl-L-homocysteine; S-Adenosylhomocysteine HILIC_TQS_Pos C00021 Adenosyl Methionine 399.145066 CHEBI:15414 S-Adenosyl-L-methionine; S-Adenosylmethionine; AdoMet; SAM HILIC_TQS_Pos C00019 Alanine 90.055504 CHEBI:16977 L-Alanine; L-2-Aminopropionic acid; L-alpha-Alanine HILIC_TQS_Pos C00041 Aminobutyric Acid 104.071154 CHEBI:16865 4-Aminobutanoate; 4-Aminobutanoic acid; 4-Aminobutyrate; 4-Aminobutyric acid; gamma-Aminobutyric acid; GABA HILIC_TQS_Pos C00334 Argininosuccinic Acid 291.13046 CHEBI:15682 N-(L-Arginino)succinate; 2-(Nomega-L-Arginino)succinate; L-Argininosuccinate; L-Argininosuccinic acid; L-Arginosuccinic acid HILIC_TQS_Pos C03406 Asparagine 133.061318 CHEBI:17196 L-Asparagine; 2-Aminosuccinamic acid HILIC_TQS_Pos C00152 Aspartic acid 134.045334 CHEBI:17053 L-Aspartate; L-Aspartic acid; 2-Aminosuccinic acid; L-Asp HILIC_TQS_Pos C00049 Carnitine 162.113019 CHEBI:17126 Carnitine; gamma-Trimethyl-hydroxybutyrobetaine; 3-Hydroxy-4-trimethylammoniobutanoate HILIC_TQS_Pos C00487 Choline 104.107539 CHEBI:15354 Choline; Bilineurine HILIC_TQS_Pos C00114 Citrulline 176.103517 CHEBI:16349 L-Citrulline; 2-Amino-5-ureidovaleric acid; Citrulline HILIC_TQS_Pos C00327 Cystine 241.031677 CHEBI:16283 L-Cystine; L-Dicysteine; L-alpha-Diamino-beta-dithiolactic acid HILIC_TQS_Pos C00491 Dimethyl Glycine 104.071154 CHEBI:17724 N,N-Dimethylglycine; Dimethylglycine HILIC_TQS_Pos C01026 Glucosylglycerol 255.107995 CHEBI:82766 2-O-(alpha-D-Glucopyranosyl)glycerol; 2-O-alpha-D-Glucosylglycerol HILIC_TQS_Pos C11546 Glutathione 308.091634 CHEBI:16856 Glutathione; 5-L-Glutamyl-L-cysteinylglycine; N-(N-gamma-L-Glutamyl-L-cysteinyl)glycine; gamma-L-Glutamyl-L-cysteinyl-glycine; GSH; Reduced glutathione HILIC_TQS_Pos C00051 Glutathione Disulfide 613.159793 CHEBI:17858 Glutathione disulfide; GSSG; Oxiglutatione; Oxidized glutathione HILIC_TQS_Pos C00127 Guanine 152.057235 CHEBI:16235 Guanine; 2-Amino-6-hydroxypurine HILIC_TQS_Pos C00242 Guanosine 284.099495 CHEBI:16750 Guanosine HILIC_TQS_Pos C00387 Histidine 156.077302 CHEBI:15971 L-Histidine; (S)-alpha-Amino-1H-imidazole-4-propionic acid HILIC_TQS_Pos C00135 Homarine 138.055503 CHEBI:69061 HILIC_TQS_Pos Homoserine 120.066069 CHEBI:15699 L-Homoserine; 2-Amino-4-hydroxybutyric acid HILIC_TQS_Pos C00263 Hypotaurine 110.027576 CHEBI:16668 Hypotaurine; 2-Aminoethanesulfinic acid HILIC_TQS_Pos C00519 Isoleucine 132.102454 CHEBI:17191 L-Isoleucine; 2-Amino-3-methylvaleric acid HILIC_TQS_Pos C00407 Leucine 132.102454 CHEBI:15603 L-Leucine; 2-Amino-4-methylvaleric acid; (2S)-alpha-2-Amino-4-methylvaleric acid; (2S)-alpha-Leucine HILIC_TQS_Pos C00123 Lysine 147.113353 CHEBI:18019 L-Lysine; Lysine acid; 2,6-Diaminohexanoic acid HILIC_TQS_Pos C00047 Methionine 150.058876 CHEBI:16811 L-Methionine; Methionine; L-2-Amino-4methylthiobutyric acid HILIC_TQS_Pos C00073 Methionine Sulfoxide 166.053791 CHEBI:17016 L-Methionine S-oxide HILIC_TQS_Pos C02989 N-Acetyl-Lysine 189.123918 CHEBI:17752 N6-Acetyl-L-lysine HILIC_TQS_Pos C02727 Ornithine 133.097703 CHEBI:15729 L-Ornithine; (S)-2,5-Diaminovaleric acid; (S)-2,5-Diaminopentanoic acid; (S)-2,5-Diaminopentanoate HILIC_TQS_Pos C00077 Sarcosine 90.055504 CHEBI:15611 Sarcosine; N-Methylglycine HILIC_TQS_Pos C00213 Serine 106.050419 CHEBI:17115 L-Serine; L-2-Amino-3-hydroxypropionic acid; L-3-Hydroxy-alanine; Serine HILIC_TQS_Pos C00065 Thiamine pyrophosphate 425.044974 CHEBI:9532 Thiamin diphosphate; Thiamine diphosphate; Thiamin pyrophosphate; TPP; ThPP HILIC_TQS_Pos C00068 Threonine 120.066069 CHEBI:16857 L-Threonine; 2-Amino-3-hydroxybutyric acid HILIC_TQS_Pos C00188 trans Hydroxyl proline 132.066069 CHEBI:18095 Hydroxyproline; L-Hydroxyproline; trans-4-Hydroxy-L-proline HILIC_TQS_Pos C01157 Trigonelline 138.055503 CHEBI:18123 N-Methylnicotinate; Trigonelline; Trigenelline; 1-Methylpyridinio-3-carboxylate; Betaine nicotinate; Caffearin; Gynesine HILIC_TQS_Pos C01004 Tyrosine 182.081719 CHEBI:17895 L-Tyrosine; (S)-3-(p-Hydroxyphenyl)alanine; (S)-2-Amino-3-(p-hydroxyphenyl)propionic acid; Tyrosine HILIC_TQS_Pos C00082 Valine 118.086804 CHEBI:16414 L-Valine; 2-Amino-3-methylbutyric acid HILIC_TQS_Pos C00183 Vitamin B3 124.039854 CHEBI:15940 Nicotinate; Nicotinic acid; Niacin; 3-Pyridinecarboxylic acid HILIC_TQS_Pos C00253 METABOLITES_END #END