#METABOLOMICS WORKBENCH TruxalCarlson_20200909_105704 DATATRACK_ID:2158 STUDY_ID:ST001480 ANALYSIS_ID:AN002457 PROJECT_ID:PR001004 VERSION 1 CREATED_ON September 14, 2020, 4:17 pm #PROJECT PR:PROJECT_TITLE Large diversity in nitrogen- and sulfur-containing compatible solute profiles in PR:PROJECT_TITLE polar and temperate diatoms PR:PROJECT_TYPE Marine Metabolomics PR:PROJECT_SUMMARY Intense bottom-ice algal blooms, often dominated by diatoms, are an important PR:PROJECT_SUMMARY source of food for grazers, organic matter for export during sea ice melt, and PR:PROJECT_SUMMARY dissolved organic carbon. Sea-ice diatoms have a number of adaptations, PR:PROJECT_SUMMARY including accumulation of compatible solutes, that allow them to inhabit this PR:PROJECT_SUMMARY highly variable environment characterized by extremes in temperature, salinity, PR:PROJECT_SUMMARY and light. In addition to protecting them from extreme conditions, these PR:PROJECT_SUMMARY compounds present a labile, nutrient-rich source of organic matter and include PR:PROJECT_SUMMARY precursors to climate active compounds (e.g. DMS), which are likely regulated PR:PROJECT_SUMMARY with environmental change. Here, intracellular concentrations of 45 metabolites PR:PROJECT_SUMMARY were quantified in three sea-ice diatom species and were compared to two PR:PROJECT_SUMMARY temperate diatom species, with a focus on compatible solutes and free amino acid PR:PROJECT_SUMMARY pools. There was a large diversity of metabolite concentrations between diatoms PR:PROJECT_SUMMARY with no clear pattern identifiable for sea-ice species. Concentrations of some PR:PROJECT_SUMMARY compatible solutes (isethionic acid, homarine) approached 1 M in the sea-ice PR:PROJECT_SUMMARY diatoms, Fragilariopsis cylindrus and Navicula cf. perminuta, but not in the PR:PROJECT_SUMMARY larger sea-ice diatom, Nitzschia lecointei or in the temperate diatom species. PR:PROJECT_SUMMARY The differential use of compatible solutes in sea-ice diatoms suggest different PR:PROJECT_SUMMARY adaptive strategies and highlights which small organic compounds may be PR:PROJECT_SUMMARY important in polar biogeochemical cycles. PR:INSTITUTE University of Washington PR:DEPARTMENT 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, USA PR:EMAIL hmdawson@uw.edu PR:PHONE 206-543-0744 PR:PUBLICATIONS Dawson et al, 2020, Integrative and Comparative Biology #STUDY ST:STUDY_TITLE Large diversity in nitrogen- and sulfur-containing compatible solute profiles in ST:STUDY_TITLE polar and temperate diatoms ST:STUDY_TYPE Intracellular metabolites were quantified in diatom species ST:STUDY_SUMMARY Intense bottom-ice algal blooms, often dominated by diatoms, are an important ST:STUDY_SUMMARY source of food for grazers, organic matter for export during sea ice melt, and ST:STUDY_SUMMARY dissolved organic carbon. Sea-ice diatoms have a number of adaptations, ST:STUDY_SUMMARY including accumulation of compatible solutes, that allow them to inhabit this ST:STUDY_SUMMARY highly variable environment characterized by extremes in temperature, salinity, ST:STUDY_SUMMARY and light. In addition to protecting them from extreme conditions, these ST:STUDY_SUMMARY compounds present a labile, nutrient-rich source of organic matter and include ST:STUDY_SUMMARY precursors to climate active compounds (e.g. DMS), which are likely regulated ST:STUDY_SUMMARY with environmental change. Here, intracellular concentrations of 45 metabolites ST:STUDY_SUMMARY were quantified in three sea-ice diatom species and were compared to two ST:STUDY_SUMMARY temperate diatom species, with a focus on compatible solutes and free amino acid ST:STUDY_SUMMARY pools. There was a large diversity of metabolite concentrations between diatoms ST:STUDY_SUMMARY with no clear pattern identifiable for sea-ice species. Concentrations of some ST:STUDY_SUMMARY compatible solutes (isethionic acid, homarine) approached 1 M in the sea-ice ST:STUDY_SUMMARY diatoms, Fragilariopsis cylindrus and Navicula cf. perminuta, but not in the ST:STUDY_SUMMARY larger sea-ice diatom, Nitzschia lecointei or in the temperate diatom species. ST:STUDY_SUMMARY The differential use of compatible solutes in sea-ice diatoms suggest different ST:STUDY_SUMMARY adaptive strategies and highlights which small organic compounds may be ST:STUDY_SUMMARY important in polar biogeochemical cycles. ST:INSTITUTE University of Washington ST:DEPARTMENT 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, USA ST:EMAIL hmdawson@uw.edu ST:PHONE 206-543-0744 ST:PUBLICATIONS Dawson et al, 2020, Integrative and Comparative Biology #SUBJECT SU:SUBJECT_TYPE Other SU:SUBJECT_SPECIES Nitzschia lecointei;Fragilariopsis cylindrus;Navicula cf. perminuta;Navicula pelliculosa #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 - Nl_32ppt-1C_1 Species:Nitzschia lecointei | Salinity:32 | Temp_degC:-1 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=1.2756163; Vol_filtered_mL=70; Replicate=1; Type=Smp; Strain=NA; RAW_FILE_NAME=200309_Smp_32ppt-1C_A 200309_Smp_32ppt-1C_A SUBJECT_SAMPLE_FACTORS - Nl_32ppt-1C_2 Species:Nitzschia lecointei | Salinity:32 | Temp_degC:-1 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=1.174789; Vol_filtered_mL=70; Replicate=2; Type=Smp; Strain=NA; RAW_FILE_NAME=200309_Smp_32ppt-1C_B 200309_Smp_32ppt-1C_ SUBJECT_SAMPLE_FACTORS - Nl_32ppt-1C_3 Species:Nitzschia lecointei | Salinity:32 | Temp_degC:-1 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=1.1610501; Vol_filtered_mL=70; Replicate=3; Type=Smp; Strain=NA; RAW_FILE_NAME=200309_Smp_32ppt-1C_C 200309_Smp_32ppt-1C_C SUBJECT_SAMPLE_FACTORS - Nl_32ppt4C_1 Species:Nitzschia lecointei | Salinity:32 | Temp_degC:4 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=1.532752987; Vol_filtered_mL=70; Replicate=1; Type=Smp; Strain=NA; RAW_FILE_NAME=200309_Smp_32ppt4C_A 200309_Smp_32ppt4C_A SUBJECT_SAMPLE_FACTORS - Nl_32ppt4C_2 Species:Nitzschia lecointei | Salinity:32 | Temp_degC:4 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=1.75542558; Vol_filtered_mL=70; Replicate=2; Type=Smp; Strain=NA; RAW_FILE_NAME=200309_Smp_32ppt4C_B 200309_Smp_32ppt4C_B SUBJECT_SAMPLE_FACTORS - Nl_32ppt4C_3 Species:Nitzschia lecointei | Salinity:32 | Temp_degC:4 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=1.66524318; Vol_filtered_mL=70; Replicate=3; Type=Smp; Strain=NA; RAW_FILE_NAME=200309_Smp_32ppt4C_C 200309_Smp_32ppt4C_C SUBJECT_SAMPLE_FACTORS - Nl_41ppt-1C_1 Species:Nitzschia lecointei | Salinity:41 | Temp_degC:-1 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=1.746388756; Vol_filtered_mL=70; Replicate=1; Type=Smp; Strain=NA; RAW_FILE_NAME=200309_Smp_41ppt-1C_A 200309_Smp_41ppt-1C_A SUBJECT_SAMPLE_FACTORS - Nl_41ppt-1C_2 Species:Nitzschia lecointei | Salinity:41 | Temp_degC:-1 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=1.399316304; Vol_filtered_mL=70; Replicate=2; Type=Smp; Strain=NA; RAW_FILE_NAME=200309_Smp_41ppt-1C_B 200309_Smp_41ppt-1C_B SUBJECT_SAMPLE_FACTORS - Nl_41ppt-1C_3 Species:Nitzschia lecointei | Salinity:41 | Temp_degC:-1 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=1.657191524; Vol_filtered_mL=70; Replicate=3; Type=Smp; Strain=NA; RAW_FILE_NAME=200309_Smp_41ppt-1C_C 200309_Smp_41ppt-1C_C SUBJECT_SAMPLE_FACTORS - Nl_41ppt4C_1 Species:Nitzschia lecointei | Salinity:41 | Temp_degC:4 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=1.11758985; Vol_filtered_mL=70; Replicate=1; Type=Smp; Strain=NA; RAW_FILE_NAME=200309_Smp_41ppt4C_A 200309_Smp_41ppt4C_A SUBJECT_SAMPLE_FACTORS - Nl_41ppt4C_2 Species:Nitzschia lecointei | Salinity:41 | Temp_degC:4 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=1.310557636; Vol_filtered_mL=70; Replicate=2; Type=Smp; Strain=NA; RAW_FILE_NAME=200309_Smp_41ppt4C_B 200309_Smp_41ppt4C_B SUBJECT_SAMPLE_FACTORS - Nl_41ppt4C_3 Species:Nitzschia lecointei | Salinity:41 | Temp_degC:4 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=1.272660153; Vol_filtered_mL=70; Replicate=3; Type=Smp; Strain=NA; RAW_FILE_NAME=200309_Smp_41ppt4C_C 200309_Smp_41ppt4C_C SUBJECT_SAMPLE_FACTORS - Fc_1 Species:Fragilariopsis cylindrus | Salinity:31 | Temp_degC:-1 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=0.232254; Vol_filtered_mL=69; Replicate=1; Type=Smp; Strain=CCMP1102; RAW_FILE_NAME=200309_Smp_Fc_1 200309_Smp_Fc_1 SUBJECT_SAMPLE_FACTORS - Fc_2 Species:Fragilariopsis cylindrus | Salinity:31 | Temp_degC:-1 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=0.23529; Vol_filtered_mL=69; Replicate=2; Type=Smp; Strain=CCMP1102; RAW_FILE_NAME=200309_Smp_Fc_2 200309_Smp_Fc_2 SUBJECT_SAMPLE_FACTORS - Nl_1 Species:Nitzschia lecointei | Salinity:31 | Temp_degC:-1 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=0.51129; Vol_filtered_mL=69; Replicate=1; Type=Smp; Strain=NA; RAW_FILE_NAME=200309_Smp_Nl_1 200309_Smp_Nl_1 SUBJECT_SAMPLE_FACTORS - Nl_2 Species:Nitzschia lecointei | Salinity:31 | Temp_degC:-1 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=0.490314; Vol_filtered_mL=69; Replicate=2; Type=Smp; Strain=NA; RAW_FILE_NAME=200309_Smp_Nl_2 200309_Smp_Nl_2 SUBJECT_SAMPLE_FACTORS - Nperm_1 Species:Navicula cf. perminuta | Salinity:31 | Temp_degC:-1 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=0.203895; Vol_filtered_mL=69; Replicate=1; Type=Smp; Strain=NA; RAW_FILE_NAME=200309_Smp_Np_1 200309_Smp_Np_1 SUBJECT_SAMPLE_FACTORS - Nperm_2 Species:Navicula cf. perminuta | Salinity:31 | Temp_degC:-1 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=0.193545; Vol_filtered_mL=69; Replicate=2; Type=Smp; Strain=NA; RAW_FILE_NAME=200309_Smp_Np_2 200309_Smp_Np_2 SUBJECT_SAMPLE_FACTORS - Npell_1 Species:Navicula pelliculosa | Salinity:35 | Temp_degC:13 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=0.6318675; Vol_filtered_mL=69; Replicate=1; Type=Smp; Strain=CCMP543; RAW_FILE_NAME=200309_Smp_NpB12SL_AB 200309_Smp_NpB12SL_AB SUBJECT_SAMPLE_FACTORS - Npell_2 Species:Navicula pelliculosa | Salinity:35 | Temp_degC:13 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=0.6090975; Vol_filtered_mL=69; Replicate=2; Type=Smp; Strain=CCMP543; RAW_FILE_NAME=200309_Smp_NpB12SL_.D 200309_Smp_NpB12SL_CD SUBJECT_SAMPLE_FACTORS - Npell_3 Species:Navicula pelliculosa | Salinity:35 | Temp_degC:13 Light=Saturating; Cobalamin=Replete; Vol_intracellular_µL=0.59409; Vol_filtered_mL=69; Replicate=3; Type=Smp; Strain=CCMP543; RAW_FILE_NAME=200309_Smp_NpB12SL_EF 200309_Smp_NpB12SL_EF #COLLECTION CO:COLLECTION_SUMMARY Axenic cultures of three Antarctic sea-ice diatoms (N. lecointei, N. cf. CO:COLLECTION_SUMMARY perminuta, and F. cylindrus) and two temperate diatoms (T. pseudonana and N. CO:COLLECTION_SUMMARY pelliculosa) were chosen for study. Cells were harvested during exponential CO:COLLECTION_SUMMARY growth onto 47 mm 0.2 µm PTFE filters (Omnipore) using combusted glassware and CO:COLLECTION_SUMMARY gentle filtration and stored at –80 °C until extraction. For each biological CO:COLLECTION_SUMMARY replicate (n = 2 for Antarctic species, n = 3 for temperate species), two 35 mL CO:COLLECTION_SUMMARY cultures were harvested onto each filter . An un-inoculated media blank was CO:COLLECTION_SUMMARY prepared and treated in the same manner as samples. CO:SAMPLE_TYPE Cultured diatom cells CO:STORAGE_CONDITIONS Described in summary #TREATMENT TR:TREATMENT_SUMMARY Antarctic species were grown at −1°C and a PAR irradiance of 45 𝜇mol TR:TREATMENT_SUMMARY photons m−2 s−1 (16:8 light:dark cycle) using cool white lights. Temperate TR:TREATMENT_SUMMARY species were grown at 13°C and a PAR irradiance of 120 𝜇mol photons m−-2 TR:TREATMENT_SUMMARY s−-1(12:12 light:dark cycle). In both cases, light was saturating. Cultures TR:TREATMENT_SUMMARY were grown in artificial seawater (ESAW, salinity 31, for Antarctic species and TR:TREATMENT_SUMMARY Instant Ocean, salinity ~35 for temperate species). Cobalamin (vitamin B12) was TR:TREATMENT_SUMMARY replete in all cultures. To explore the effect of growth conditions on metabolic TR:TREATMENT_SUMMARY profiles using non-metric dimensional scaling analysis, samples were included of TR:TREATMENT_SUMMARY N. lecointei grown at temperatures of −1 and 4˚C and salinities of 32 and 41. #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. An un-inoculated media blank was prepared and treated SP:SAMPLEPREP_SUMMARY in the same manner as the samples. 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 Thermo Q Exactive HF hybrid Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS See attached protocol. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS mM (intracellular concentration) MS_METABOLITE_DATA_START Samples Fc_1 Fc_2 Nperm_1 Nperm_2 Nl_1 Nl_2 Nl_32ppt-1C_1 Nl_32ppt-1C_2 Nl_32ppt-1C_3 Nl_32ppt4C_1 Nl_32ppt4C_2 Nl_32ppt4C_3 Nl_41ppt-1C_1 Nl_41ppt-1C_2 Nl_41ppt-1C_3 Nl_41ppt4C_1 Nl_41ppt4C_2 Nl_41ppt4C_3 Npell_1 Npell_2 Npell_3 Factors Species:Fragilariopsis cylindrus | Salinity:31 | Temp_degC:-1 Species:Fragilariopsis cylindrus | Salinity:31 | Temp_degC:-1 Species:Navicula cf. perminuta | Salinity:31 | Temp_degC:-1 Species:Navicula cf. perminuta | Salinity:31 | Temp_degC:-1 Species:Nitzschia lecointei | Salinity:31 | Temp_degC:-1 Species:Nitzschia lecointei | Salinity:31 | Temp_degC:-1 Species:Nitzschia lecointei | Salinity:32 | Temp_degC:-1 Species:Nitzschia lecointei | Salinity:32 | Temp_degC:-1 Species:Nitzschia lecointei | Salinity:32 | Temp_degC:-1 Species:Nitzschia lecointei | Salinity:32 | Temp_degC:4 Species:Nitzschia lecointei | Salinity:32 | Temp_degC:4 Species:Nitzschia lecointei | Salinity:32 | Temp_degC:4 Species:Nitzschia lecointei | Salinity:41 | Temp_degC:-1 Species:Nitzschia lecointei | Salinity:41 | Temp_degC:-1 Species:Nitzschia lecointei | Salinity:41 | Temp_degC:-1 Species:Nitzschia lecointei | Salinity:41 | Temp_degC:4 Species:Nitzschia lecointei | Salinity:41 | Temp_degC:4 Species:Nitzschia lecointei | Salinity:41 | Temp_degC:4 Species:Navicula pelliculosa | Salinity:35 | Temp_degC:13 Species:Navicula pelliculosa | Salinity:35 | Temp_degC:13 Species:Navicula pelliculosa | Salinity:35 | Temp_degC:13 Cysteic Acid 0.220308128 0.180471397 38.93058894 47.73955314 1.79630068 2.573834052 0.451618071 0.49157852 0.483654448 0.322380339 0.257077757 0.432169789 0.213342879 0.280651409 0.236813466 0.370011503 0.276918049 0.274553617 DHPS 205.1945008 217.5637618 71.32511869 80.4518594 34.66824326 33.18666562 41.31862866 29.22191195 23.47146157 32.21356354 32.75438582 40.3458025 35.1679633 63.83991191 33.39737247 34.59442417 7.191210725 3.069111334 2.810251636 Isethionic Acid 1036.940614 909.4273807 0.333396755 0.188736694 1.559017281 0.313406235 0.02400944 0.022469527 0.023298774 0.050380358 0.044745878 0.048401826 0.042865604 0.035874663 0.027861386 L-Pyroglutamic acid 2.568606578 2.212950335 10.49756325 13.55793016 1.871343159 2.60337302 0.908882395 0.95367034 1.186045876 0.377215007 0.386287727 0.59671078 0.749597316 0.829637875 0.759777659 0.643583351 0.418551353 0.596552437 21.11499564 13.52200313 12.79691343 Sucrose 0.036684942 0.043287353 0.028184314 0.104058566 0.040666763 0.135520841 0.041580783 0.052382281 0.178537884 0.061821958 0.28590915 Sulfolactic Acid 0.003837326 0.007451488 0.004441515 0.006770601 0.036844077 0.042405272 0.003984052 0.005242126 0.01558195 0.030448103 0.027774478 0.053616041 0.008838588 0.007004662 0.009488644 0.061659192 0.046209417 0.039208448 Taurine 342.9803485 341.5099969 0.53981803 0.122821752 0.088859694 0.104881747 0.105480207 0.378777795 0.360269576 0.347204642 0.092183839 0.119254789 0.104301346 0.43884965 0.351090828 0.278562739 Trehalose 0.0505798 0.064189205 0.00312502 0.023300023 0.007525223 0.005173465 0.012639175 0.010232693 0.0156806 0.013538649 0.015893105 0.012209089 0.01632141 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name quantitated m/z KEGG_ID CHEBI KEGGNAME MS_method Cysteic Acid 167.996671 C00506 CHEBI:17285 L-Cysteate; L-Cysteic acid; 3-Sulfoalanine; 2-Amino-3-sulfopropionic acid HILIC_QE_Neg DHPS 155.001422 C19675 CHEBI:60997 (R)-2,3-Dihydroxypropane-1-sulfonate HILIC_QE_Neg Isethionic Acid 124.990857 C05123 CHEBI:1157 2-Hydroxyethanesulfonate; 2-Hydroxyethanesulfonic acid; 2-Hydroxyethane-1-sulfonic acid; Isethionic acid; Isethionate HILIC_QE_Neg L-Pyroglutamic acid 128.034768 C01879 CHEBI:18183 5-Oxoproline; Pidolic acid; Pyroglutamic acid; 5-Pyrrolidone-2-carboxylic acid; Pyroglutamate; 5-Oxo-L-proline; L-Pyroglutamic acid; L-5-Pyrrolidone-2-carboxylic acid HILIC_QE_Neg Sucrose 341.10839 C00089 CHEBI:17992 Sucrose; Cane sugar; Saccharose; 1-alpha-D-Glucopyranosyl-2-beta-D-fructofuranoside HILIC_QE_Neg Sulfolactic Acid 168.980687 C16069 CHEBI:50519 3-Sulfolactate HILIC_QE_Neg Taurine 124.006841 C00245 CHEBI:15891 Taurine; 2-Aminoethanesulfonic acid; Aminoethylsulfonic acid HILIC_QE_Neg Trehalose 341.10839 C01083 CHEBI:16551 alpha,alpha-Trehalose; alpha,alpha'-Trehalose; Trehalose HILIC_QE_Neg METABOLITES_END #END