#METABOLOMICS WORKBENCH jssacks_20220830_132254 DATATRACK_ID:3439 STUDY_ID:ST002292 ANALYSIS_ID:AN003744 VERSION 1 CREATED_ON 09-28-2022 #PROJECT PR:PROJECT_TITLE Quantification of Dissolved Metabolites in Environmental Samples through PR:PROJECT_TITLE Cation-Exchange Solid Phase Extraction (CX-SPE) paired with Liquid PR:PROJECT_TITLE Chromatography-Mass Spectrometry PR:PROJECT_SUMMARY Small, biologically produced, organic molecules called metabolites play key PR:PROJECT_SUMMARY roles in microbial systems where they directly mediate exchanges of nutrients, PR:PROJECT_SUMMARY energy, and information. However, the study of dissolved polar metabolites in PR:PROJECT_SUMMARY seawater and other environmental matrices has been hampered by analytical PR:PROJECT_SUMMARY challenges including high inorganic ion concentrations, low analyte PR:PROJECT_SUMMARY concentrations, and high chemical diversity. Here we show that a cation-exchange PR:PROJECT_SUMMARY solid phase extraction (CX-SPE) sample preparation approach separates positively PR:PROJECT_SUMMARY charged and zwitterionic metabolites from seawater and freshwater samples, PR:PROJECT_SUMMARY allowing their analysis by liquid chromatography-mass spectrometry (LC-MS). We PR:PROJECT_SUMMARY successfully extracted 69 known compounds from an in-house compound collection PR:PROJECT_SUMMARY and evaluated the performance of the method by establishing extraction PR:PROJECT_SUMMARY efficiencies and limits of detection (pM to low nM range) for these compounds. PR:PROJECT_SUMMARY CX-SPE extracted a range of compounds including amino acids and compatible PR:PROJECT_SUMMARY solutes, resulted in very low matrix effects, and performed robustly across PR:PROJECT_SUMMARY large variations in salinity and dissolved organic matter (DOM) concentration. PR:PROJECT_SUMMARY We compared CX-SPE to an established solid phase extraction procedure (PPL-SPE) PR:PROJECT_SUMMARY and demonstrate that these two methods extract fundamentally different fractions PR:PROJECT_SUMMARY of the dissolved metabolite pool with CX-SPE extracting compounds that are on PR:PROJECT_SUMMARY average smaller and more polar. We use CX-SPE to analyze four environmental PR:PROJECT_SUMMARY samples from distinct aquatic biomes, producing some of the first CX-SPE PR:PROJECT_SUMMARY dissolved metabolomes. Quantified compounds ranged in concentration from 0.0093 PR:PROJECT_SUMMARY nM to 49 nM and were composed primarily of amino acids (0.15 – 16 nM) and PR:PROJECT_SUMMARY compatible solutes such as TMAO (0.89 – 49 nM) and glycine betaine (2.8 – PR:PROJECT_SUMMARY 5.2 nM). PR:INSTITUTE University of Washington PR:DEPARTMENT Oceanography PR:LABORATORY Ingalls Lab PR:LAST_NAME Sacks PR:FIRST_NAME Joshua PR:ADDRESS Ocean Sciences Building, 1492 NE Boat St. Seattle, WA 98105 PR:EMAIL jssacks@uw.edu PR:PHONE 4074090052 PR:FUNDING_SOURCE NSF, Simons Foundation PR:PUBLICATIONS Sacks et al., L&O Methods, accepted PR:DOI http://dx.doi.org/10.21228/M8Q99X #STUDY ST:STUDY_TITLE Quantification of Dissolved Metabolites in Environmental Samples through ST:STUDY_TITLE Cation-Exchange Solid Phase Extraction (CX-SPE) paired with Liquid ST:STUDY_TITLE Chromatography-Mass Spectrometry ST:STUDY_TYPE Method Development for Dissolved Metabolomics in Seawater ST:STUDY_SUMMARY Small, biologically produced, organic molecules called metabolites play key ST:STUDY_SUMMARY roles in microbial systems where they directly mediate exchanges of nutrients, ST:STUDY_SUMMARY energy, and information. However, the study of dissolved polar metabolites in ST:STUDY_SUMMARY seawater and other environmental matrices has been hampered by analytical ST:STUDY_SUMMARY challenges including high inorganic ion concentrations, low analyte ST:STUDY_SUMMARY concentrations, and high chemical diversity. Here we show that a cation-exchange ST:STUDY_SUMMARY solid phase extraction (CX-SPE) sample preparation approach separates positively ST:STUDY_SUMMARY charged and zwitterionic metabolites from seawater and freshwater samples, ST:STUDY_SUMMARY allowing their analysis by liquid chromatography-mass spectrometry (LC-MS). We ST:STUDY_SUMMARY successfully extracted 69 known compounds from an in-house compound collection ST:STUDY_SUMMARY and evaluated the performance of the method by establishing extraction ST:STUDY_SUMMARY efficiencies and limits of detection (pM to low nM range) for these compounds. ST:STUDY_SUMMARY CX-SPE extracted a range of compounds including amino acids and compatible ST:STUDY_SUMMARY solutes, resulted in very low matrix effects, and performed robustly across ST:STUDY_SUMMARY large variations in salinity and dissolved organic matter (DOM) concentration. ST:STUDY_SUMMARY We compared CX-SPE to an established solid phase extraction procedure (PPL-SPE) ST:STUDY_SUMMARY and demonstrate that these two methods extract fundamentally different fractions ST:STUDY_SUMMARY of the dissolved metabolite pool with CX-SPE extracting compounds that are on ST:STUDY_SUMMARY average smaller and more polar. We use CX-SPE to analyze four environmental ST:STUDY_SUMMARY samples from distinct aquatic biomes, producing some of the first CX-SPE ST:STUDY_SUMMARY dissolved metabolomes. Quantified compounds ranged in concentration from 0.0093 ST:STUDY_SUMMARY nM to 49 nM and were composed primarily of amino acids (0.15 – 16 nM) and ST:STUDY_SUMMARY compatible solutes such as TMAO (0.89 – 49 nM) and glycine betaine (2.8 – ST:STUDY_SUMMARY 5.2 nM). ST:INSTITUTE University of Washington ST:DEPARTMENT Oceanography ST:LABORATORY Ingalls Lab ST:LAST_NAME Sacks ST:FIRST_NAME Joshua ST:ADDRESS Ocean Sciences Building, 1492 NE Boat St. Seattle, WA 98105 ST:EMAIL jssacks@uw.edu ST:PHONE 4074090052 ST:SUBMIT_DATE 2022-08-30 #SUBJECT SU:SUBJECT_TYPE Water sample #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - LW_A Extraction_Approach:CX-SPE | Location:Lake Washington Replicate=A; Type=Smp; Depth_m=1; Vol_mL=40; Latitude=47.633; Longitude=-122.3; Date=2020-06-20; RAW_FILE_NAME=210322_Smp_LW_A; RAW_FILE_NAME=210322_Smp_LW_A; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - LW_B Extraction_Approach:CX-SPE | Location:Lake Washington Replicate=B; Type=Smp; Depth_m=1; Vol_mL=40; Latitude=47.633; Longitude=-122.3; Date=2020-06-20; RAW_FILE_NAME=210322_Smp_LW_B; RAW_FILE_NAME=210322_Smp_LW_B; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - LW_C Extraction_Approach:CX-SPE | Location:Lake Washington Replicate=C; Type=Smp; Depth_m=1; Vol_mL=40; Latitude=47.633; Longitude=-122.3; Date=2020-06-20; RAW_FILE_NAME=210322_Smp_LW_C; RAW_FILE_NAME=210322_Smp_LW_C; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - Blank1_A Extraction_Approach:CX-SPE | Location:NA Replicate=A; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_Blank1_A; RAW_FILE_NAME=210322_Blk_Blank1_A; RAW_FILE_NAME=210329_Blk_Blank1_A SUBJECT_SAMPLE_FACTORS - Blank1_B Extraction_Approach:CX-SPE | Location:NA Replicate=B; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_Blank1_B; RAW_FILE_NAME=210322_Blk_Blank1_B; RAW_FILE_NAME=210329_Blk_Blank1_B SUBJECT_SAMPLE_FACTORS - Blank1_C Extraction_Approach:CX-SPE | Location:NA Replicate=C; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_Blank1_C; RAW_FILE_NAME=210322_Blk_Blank1_C; RAW_FILE_NAME=210329_Blk_Blank1_C SUBJECT_SAMPLE_FACTORS - Blank2_A Extraction_Approach:CX-SPE | Location:NA Replicate=A; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_Blank2_A; RAW_FILE_NAME=210322_Blk_Blank2_A; RAW_FILE_NAME=210329_Blk_Blank2_A SUBJECT_SAMPLE_FACTORS - Blank2_B Extraction_Approach:CX-SPE | Location:NA Replicate=B; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_Blank2_B; RAW_FILE_NAME=210322_Blk_Blank2_B; RAW_FILE_NAME=210329_Blk_Blank2_B SUBJECT_SAMPLE_FACTORS - Blank2_C Extraction_Approach:CX-SPE | Location:NA Replicate=C; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_Blank2_C; RAW_FILE_NAME=210322_Blk_Blank2_C; RAW_FILE_NAME=210329_Blk_Blank2_C SUBJECT_SAMPLE_FACTORS - Blank3_A Extraction_Approach:CX-SPE | Location:NA Replicate=A; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_Blank3_A; RAW_FILE_NAME=210322_Blk_Blank3_A; RAW_FILE_NAME=210329_Blk_Blank3_A SUBJECT_SAMPLE_FACTORS - Blank3_B Extraction_Approach:CX-SPE | Location:NA Replicate=B; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_Blank3_B; RAW_FILE_NAME=210322_Blk_Blank3_B; RAW_FILE_NAME=210329_Blk_Blank3_B SUBJECT_SAMPLE_FACTORS - Blank3_C Extraction_Approach:CX-SPE | Location:NA Replicate=C; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_Blank3_C; RAW_FILE_NAME=210322_Blk_Blank3_C; RAW_FILE_NAME=210329_Blk_Blank3_C SUBJECT_SAMPLE_FACTORS - Blank4_A Extraction_Approach:CX-SPE | Location:NA Replicate=A; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_Blank4_A; RAW_FILE_NAME=210322_Blk_Blank4_A; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - Blank4_B Extraction_Approach:CX-SPE | Location:NA Replicate=B; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_Blank4_B; RAW_FILE_NAME=210322_Blk_Blank4_B; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - Blank4_C Extraction_Approach:CX-SPE | Location:NA Replicate=C; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_Blank4_C; RAW_FILE_NAME=210322_Blk_Blank4_C; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - Blank5_A Extraction_Approach:CX-SPE | Location:NA Replicate=A; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_Blank5_A; RAW_FILE_NAME=210322_Blk_Blank5_A; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - Blank5_B Extraction_Approach:CX-SPE | Location:NA Replicate=B; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_Blank5_B; RAW_FILE_NAME=210322_Blk_Blank5_B; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - Blank5_C Extraction_Approach:CX-SPE | Location:NA Replicate=C; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_Blank5_C; RAW_FILE_NAME=210322_Blk_Blank5_C; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - NP_A Extraction_Approach:CX-SPE | Location:North Pacific Replicate=A; Type=Smp; Depth_m=15; Vol_mL=40; Latitude=41.4; Longitude=-158; Date=2016-04-26; RAW_FILE_NAME=210322_Smp_NP_A; RAW_FILE_NAME=210322_Smp_NP_A; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - NP_B Extraction_Approach:CX-SPE | Location:North Pacific Replicate=B; Type=Smp; Depth_m=15; Vol_mL=40; Latitude=41.4; Longitude=-158; Date=2016-04-26; RAW_FILE_NAME=210322_Smp_NP_B; RAW_FILE_NAME=210322_Smp_NP_B; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - NP_C Extraction_Approach:CX-SPE | Location:North Pacific Replicate=C; Type=Smp; Depth_m=15; Vol_mL=40; Latitude=41.4; Longitude=-158; Date=2016-04-26; RAW_FILE_NAME=210322_Smp_NP_C; RAW_FILE_NAME=210322_Smp_NP_C; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - PS_A Extraction_Approach:CX-SPE | Location:Puget Sound Replicate=A; Type=Smp; Depth_m=8; Vol_mL=40; Latitude=47.683; Longitude=-122.416; Date=2017-05-07; RAW_FILE_NAME=210322_Smp_PS_A; RAW_FILE_NAME=210322_Smp_PS_A; RAW_FILE_NAME=210329_Smp_PS_A SUBJECT_SAMPLE_FACTORS - PS_B Extraction_Approach:CX-SPE | Location:Puget Sound Replicate=B; Type=Smp; Depth_m=8; Vol_mL=40; Latitude=47.683; Longitude=-122.416; Date=2017-05-07; RAW_FILE_NAME=210322_Smp_PS_B; RAW_FILE_NAME=210322_Smp_PS_B; RAW_FILE_NAME=210329_Smp_PS_B SUBJECT_SAMPLE_FACTORS - PS_C Extraction_Approach:CX-SPE | Location:Puget Sound Replicate=C; Type=Smp; Depth_m=8; Vol_mL=40; Latitude=47.683; Longitude=-122.416; Date=2017-05-07; RAW_FILE_NAME=210322_Smp_PS_C; RAW_FILE_NAME=210322_Smp_PS_C; RAW_FILE_NAME=210329_Smp_PS_C SUBJECT_SAMPLE_FACTORS - Aloha_A Extraction_Approach:CX-SPE | Location:Station ALOHA Replicate=A; Type=Smp; Depth_m=15; Vol_mL=40; Latitude=22.75; Longitude=-158; Date=2016-04-20; RAW_FILE_NAME=210322_Smp_Aloha_A; RAW_FILE_NAME=210322_Smp_Aloha_A; RAW_FILE_NAME=210329_Smp_Aloha_A SUBJECT_SAMPLE_FACTORS - Aloha_B Extraction_Approach:CX-SPE | Location:Station ALOHA Replicate=B; Type=Smp; Depth_m=15; Vol_mL=40; Latitude=22.75; Longitude=-158; Date=2016-04-20; RAW_FILE_NAME=210322_Smp_Aloha_B; RAW_FILE_NAME=210322_Smp_Aloha_B; RAW_FILE_NAME=210329_Smp_Aloha_B SUBJECT_SAMPLE_FACTORS - Aloha_C Extraction_Approach:CX-SPE | Location:Station ALOHA Replicate=C; Type=Smp; Depth_m=15; Vol_mL=40; Latitude=22.75; Longitude=-158; Date=2016-04-20; RAW_FILE_NAME=210322_Smp_Aloha_C; RAW_FILE_NAME=210322_Smp_Aloha_C; RAW_FILE_NAME=210329_Smp_Aloha_C SUBJECT_SAMPLE_FACTORS - TruePoo_Full1 Extraction_Approach:NA | Location:NA Replicate=1; Type=Poo; Depth_m=NA; Vol_mL=NA; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Poo_TruePoo_Full1; RAW_FILE_NAME=210322_Poo_TruePoo_Full1; RAW_FILE_NAME=210329_Poo_TruePoo_Full1 SUBJECT_SAMPLE_FACTORS - TruePoo_Full2 Extraction_Approach:NA | Location:NA Replicate=2; Type=Poo; Depth_m=NA; Vol_mL=NA; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Poo_TruePoo_Full2; RAW_FILE_NAME=210322_Poo_TruePoo_Full2; RAW_FILE_NAME=210329_Poo_TruePoo_Full2 SUBJECT_SAMPLE_FACTORS - TruePoo_Full3 Extraction_Approach:NA | Location:NA Replicate=3; Type=Poo; Depth_m=NA; Vol_mL=NA; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Poo_TruePoo_Full3; RAW_FILE_NAME=210322_Poo_TruePoo_Full3; RAW_FILE_NAME=210329_Poo_TruePoo_Full3 SUBJECT_SAMPLE_FACTORS - TruePoo_Half1 Extraction_Approach:NA | Location:NA Replicate=1; Type=Poo; Depth_m=NA; Vol_mL=NA; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Poo_TruePoo_Half1; RAW_FILE_NAME=210322_Poo_TruePoo_Half1; RAW_FILE_NAME=210329_Poo_TruePoo_Half1 SUBJECT_SAMPLE_FACTORS - TruePoo_Half2 Extraction_Approach:NA | Location:NA Replicate=2; Type=Poo; Depth_m=NA; Vol_mL=NA; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Poo_TruePoo_Half2; RAW_FILE_NAME=210322_Poo_TruePoo_Half2; RAW_FILE_NAME=210329_Poo_TruePoo_Half2 SUBJECT_SAMPLE_FACTORS - TruePoo_Half3 Extraction_Approach:NA | Location:NA Replicate=3; Type=Poo; Depth_m=NA; Vol_mL=NA; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Poo_TruePoo_Half3; RAW_FILE_NAME=210322_Poo_TruePoo_Half3; RAW_FILE_NAME=210329_Poo_TruePoo_Half3 SUBJECT_SAMPLE_FACTORS - PPL_Blank_A Extraction_Approach:PPL-SPE | Location:NA Replicate=A; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_PPL_Blank_A; RAW_FILE_NAME=210322_Blk_PPL_Blank_A; RAW_FILE_NAME=210329_Blk_PPL_Blank_A SUBJECT_SAMPLE_FACTORS - PPL_Blank_B Extraction_Approach:PPL-SPE | Location:NA Replicate=B; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_PPL_Blank_B; RAW_FILE_NAME=210322_Blk_PPL_Blank_B; RAW_FILE_NAME=210329_Blk_PPL_Blank_B SUBJECT_SAMPLE_FACTORS - PPL_Blank_C Extraction_Approach:PPL-SPE | Location:NA Replicate=C; Type=Blk; Depth_m=NA; Vol_mL=40; Latitude=NA; Longitude=NA; Date=NA; RAW_FILE_NAME=210322_Blk_PPL_Blank_C; RAW_FILE_NAME=210322_Blk_PPL_Blank_C; RAW_FILE_NAME=210329_Blk_PPL_Blank_C SUBJECT_SAMPLE_FACTORS - PPL_PS_A Extraction_Approach:PPL-SPE | Location:Puget Sound Replicate=A; Type=Smp; Depth_m=8; Vol_mL=40; Latitude=47.683; Longitude=-122.416; Date=2017-05-07; RAW_FILE_NAME=210322_Smp_PPL_PS_A; RAW_FILE_NAME=210322_Smp_PPL_PS_A; RAW_FILE_NAME=210329_Smp_PPL_PS_A SUBJECT_SAMPLE_FACTORS - PPL_PS_B Extraction_Approach:PPL-SPE | Location:Puget Sound Replicate=B; Type=Smp; Depth_m=8; Vol_mL=40; Latitude=47.683; Longitude=-122.416; Date=2017-05-07; RAW_FILE_NAME=210322_Smp_PPL_PS_B; RAW_FILE_NAME=210322_Smp_PPL_PS_B; RAW_FILE_NAME=210329_Smp_PPL_PS_B SUBJECT_SAMPLE_FACTORS - PPL_PS_C Extraction_Approach:PPL-SPE | Location:Puget Sound Replicate=C; Type=Smp; Depth_m=8; Vol_mL=40; Latitude=47.683; Longitude=-122.416; Date=2017-05-07; RAW_FILE_NAME=210322_Smp_PPL_PS_C; RAW_FILE_NAME=210322_Smp_PPL_PS_C; RAW_FILE_NAME=210329_Smp_PPL_PS_C SUBJECT_SAMPLE_FACTORS - PPL_Aloha_A Extraction_Approach:PPL-SPE | Location:Station ALOHA Replicate=A; Type=Smp; Depth_m=15; Vol_mL=40; Latitude=22.75; Longitude=-158; Date=2016-04-20; RAW_FILE_NAME=210322_Smp_PPL_Aloha_A; RAW_FILE_NAME=210322_Smp_PPL_Aloha_A; RAW_FILE_NAME=210329_Smp_PPL_Aloha_A SUBJECT_SAMPLE_FACTORS - PPL_Aloha_B Extraction_Approach:PPL-SPE | Location:Station ALOHA Replicate=B; Type=Smp; Depth_m=15; Vol_mL=40; Latitude=22.75; Longitude=-158; Date=2016-04-20; RAW_FILE_NAME=210322_Smp_PPL_Aloha_B; RAW_FILE_NAME=210322_Smp_PPL_Aloha_B; RAW_FILE_NAME=210329_Smp_PPL_Aloha_B SUBJECT_SAMPLE_FACTORS - PPL_Aloha_C Extraction_Approach:PPL-SPE | Location:Station ALOHA Replicate=C; Type=Smp; Depth_m=15; Vol_mL=40; Latitude=22.75; Longitude=-158; Date=2016-04-20; RAW_FILE_NAME=210322_Smp_PPL_Aloha_C; RAW_FILE_NAME=210322_Smp_PPL_Aloha_C; RAW_FILE_NAME=210329_Smp_PPL_Aloha_C #COLLECTION CO:COLLECTION_SUMMARY Samples for dissolved metabolites from four unique environmental locations (Lake CO:COLLECTION_SUMMARY Washington (LW), Puget Sound (PS), Station ALOHA in the North Pacific CO:COLLECTION_SUMMARY Subtropical Gyre (ALOHA), and the North Pacific (NP)) from depths of 15 m for CO:COLLECTION_SUMMARY ALOHA and NP samples, 8 m for the PS sample, and 1 m for the LW sample. Samples CO:COLLECTION_SUMMARY were processed by filtering the water through 147 mm 0.2 μm PTFE filters using CO:COLLECTION_SUMMARY peristaltic pumps (ALOHA, NP, PS samples) and or glass vacuum filtration onto a CO:COLLECTION_SUMMARY 47 mm 0.2 μm PTFE filter. The filtrated was collected and stored in 2 L CO:COLLECTION_SUMMARY acid-washed polycarbonate bottles (ALOHA, NP, PS samples) or 45 mL acid-washed CO:COLLECTION_SUMMARY polypropylene falcon tubes at -20 C for between 1 month and 4 years until CO:COLLECTION_SUMMARY analysis. CO:SAMPLE_TYPE seawater CO:STORAGE_CONDITIONS -20℃ #TREATMENT TR:TREATMENT_SUMMARY Not Applicable #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Cation-Exchange Solid Phase Extraction (CX-SPE) Sample Volume: 40 mLs Solid SP:SAMPLEPREP_SUMMARY Phase Resin/Column: Strong cation-exchange resin (Dowex 50WX8; H+ form, 100-200 SP:SAMPLEPREP_SUMMARY mesh, Sigma-Aldrich, Vienna, Austria) Solvents/Reagents: 1 M NH3, 3 M HNO3 Brief SP:SAMPLEPREP_SUMMARY Procedure: 35 g of resin was added to a glass chromatography column with a SP:SAMPLEPREP_SUMMARY fritted disk and a PTFE stopcock. The resin was equilibrated with 50 mL water, SP:SAMPLEPREP_SUMMARY 100 mL of 1M NH3, 50 mL water, 100 mL 3M HNO3, and 50 mL H2O. The samples were SP:SAMPLEPREP_SUMMARY acidified with HNO3 to pH 2 and heavy isotope-labeled internal standards added. SP:SAMPLEPREP_SUMMARY The sample was loaded onto column, allowed to stand for 5 minutes, and then SP:SAMPLEPREP_SUMMARY drained from the column. The column was then rinsed with 50 mL water. SP:SAMPLEPREP_SUMMARY Approximately 200 mL of 1M NH3 was added to column. Ammonia eluted from column SP:SAMPLEPREP_SUMMARY in 10 mL fractions. The pH of each fraction checked by dabbing a small drop of SP:SAMPLEPREP_SUMMARY sample onto a pH strip with a combusted glass Pasteur pipette. The alkaline SP:SAMPLEPREP_SUMMARY front (the 10 mL fraction where the pH increases from approximately 2-4 to SP:SAMPLEPREP_SUMMARY 9-11), the fraction before, and the two fractions after were collected, SP:SAMPLEPREP_SUMMARY combined, and dried down under N2 gas in a water bath of approximately 32 SP:SAMPLEPREP_SUMMARY degrees C. Dried fractions were redissolved in 380 uL of water. 20 uL of SP:SAMPLEPREP_SUMMARY isotope-labeled injection standards in water were added to the fractions. SP:SAMPLEPREP_SUMMARY Columns were regenerated for reuse through the addition of 50 mL of water, 100 SP:SAMPLEPREP_SUMMARY mL 3M HNO3, and 50 mL of water. When not in use, columns were stored completely SP:SAMPLEPREP_SUMMARY filled with 0.01 M HNO3. Water was extracted alongside samples as methodological SP:SAMPLEPREP_SUMMARY blanks. PPL-Solid Phase Extraction (PPL-SPE) Sample Volume: 40 mLs Solid Phase SP:SAMPLEPREP_SUMMARY Resin/Column: Agilent Bond Elut PPL cartridge, 1 g bed mass, 6 mL volume SP:SAMPLEPREP_SUMMARY Solvents/Reagents: methanol, 0.01 M HCl, Brief Procedure: Sample acidified with SP:SAMPLEPREP_SUMMARY HCl to pH 2 and heavy isotope-labeled internal standards added. Column prepped SP:SAMPLEPREP_SUMMARY by adding 1 cartridge volumes of each of methanol followed by 0.01 M HCl. Sample SP:SAMPLEPREP_SUMMARY loaded onto columns using a peristaltic pump at a flow rate of 10 mL/min Column SP:SAMPLEPREP_SUMMARY rinsed with 2 cartridge volumes 0.01 M HCl. Sample eluted with 1 cartridge SP:SAMPLEPREP_SUMMARY volume methanol and dried down under N2 gas. Dried fractions were redissolved in SP:SAMPLEPREP_SUMMARY 380 uL of water. 20 uL of isotope-labeled injection standards in water were SP:SAMPLEPREP_SUMMARY added to the fractions. Water was extracted alongside samples as methodological SP:SAMPLEPREP_SUMMARY blanks. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY See attached summary CH:METHODS_FILENAME Ingalls_Lab_LC_Methods_2.txt CH:INSTRUMENT_NAME Waters Acquity I-Class CH:COLUMN_NAME SeQuant ZIC- pHILIC (150 x 2.1mm, 5um) CH:CHROMATOGRAPHY_TYPE HILIC #ANALYSIS AN:ANALYSIS_TYPE MS AN:ANALYSIS_PROTOCOL_FILE CXSPE_Method_Paper_MS_Methods.txt #MS MS:INSTRUMENT_NAME Thermo Q Exactive HF hybrid Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:MS_COMMENTS See Protocol MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak Area MS_METABOLITE_DATA_START Samples LW_A LW_B LW_C NP_A NP_B NP_C PS_A PS_B PS_C Aloha_A Aloha_B Aloha_C TruePoo_Full1 TruePoo_Full2 TruePoo_Full3 TruePoo_Half1 TruePoo_Half2 TruePoo_Half3 PPL_PS_A PPL_PS_B PPL_PS_C PPL_Aloha_A PPL_Aloha_B PPL_Aloha_C Factors Extraction_Approach:CX-SPE | Location:Lake Washington Extraction_Approach:CX-SPE | Location:Lake Washington Extraction_Approach:CX-SPE | Location:Lake Washington Extraction_Approach:CX-SPE | Location:North Pacific Extraction_Approach:CX-SPE | Location:North Pacific Extraction_Approach:CX-SPE | Location:North Pacific Extraction_Approach:CX-SPE | Location:Puget Sound Extraction_Approach:CX-SPE | Location:Puget Sound Extraction_Approach:CX-SPE | Location:Puget Sound Extraction_Approach:CX-SPE | Location:Station ALOHA Extraction_Approach:CX-SPE | Location:Station ALOHA Extraction_Approach:CX-SPE | Location:Station ALOHA Extraction_Approach:NA | Location:NA Extraction_Approach:NA | Location:NA Extraction_Approach:NA | Location:NA Extraction_Approach:NA | Location:NA Extraction_Approach:NA | Location:NA Extraction_Approach:NA | Location:NA Extraction_Approach:PPL-SPE | Location:Puget Sound Extraction_Approach:PPL-SPE | Location:Puget Sound Extraction_Approach:PPL-SPE | Location:Puget Sound Extraction_Approach:PPL-SPE | Location:Station ALOHA Extraction_Approach:PPL-SPE | Location:Station ALOHA Extraction_Approach:PPL-SPE | Location:Station ALOHA (3-Carboxypropyl)trimethylammonium (TMAB) 563234.0000 534058.0000 719264.0000 1802639.0000 2163733.0000 1867806.0000 923077.0000 786690.0000 857221.0000 92533184.0000 100536680.0000 98479808.0000 85869456.0000 101739176.0000 93118728.0000 4-Hydroxyisoleucine 2620344.7438 1555153.3511 1662899.4967 716529.9228 249826.1547 474788.2371 817494.6024 934565.2842 1209032.7031 5833898.6512 5437889.5654 5909376.9252 4427130.8659 5537350.8457 6269549.2020 5-Methylcytosine 389502.0000 503177.0000 534559.0000 58489864.0000 62346292.0000 63613780.0000 57097900.0000 63228220.0000 65095056.0000 6-Methyladenine 239109.0000 311823.0000 668560.0000 199084.0000 147242.0000 283401.0000 84493592.0000 90392080.0000 95696648.0000 82102632.0000 91874560.0000 93012952.0000 417820.0000 567269.0000 838311.0000 247266.0000 282884.0000 283536.0000 Acetylcholine 188816.0000 76709.0000 62081.0000 25161706.0000 26444346.0000 23080344.0000 22823404.0000 27965752.0000 23167868.0000 Acetylglutamic acid 848939.0000 966831.0000 900651.0000 789382.0000 949428.0000 359200.0000 Acetyl-L-carnitine 147809.0000 73048.0000 77010.0000 1846661.0000 1421521.0000 1535400.0000 108401456.0000 109626384.0000 107246568.0000 101561776.0000 113863816.0000 106604608.0000 Adenine 4991580.7860 7103718.5430 10722982.3234 168892722.0555 175441612.3702 177605734.4340 174133354.0459 179688893.4665 177228542.7727 4902608.4224 2345691.6878 5477442.7801 2335590.0754 1941088.0389 1687647.1289 Adenosine 3145363.8469 4666103.3849 5633273.2025 53440145.1228 53795500.4534 59817455.4138 55679160.1783 57853724.3532 57468265.4966 1429975.9381 1371219.0747 1605774.9863 271498.5760 204476.0160 223984.9900 Adenosyl Homocysteine 8345872.8404 7956935.4234 7942396.2705 8633666.3139 7656910.7599 8813643.8508 Adenosyl Methionine 731725.0000 560443.0000 616097.0000 458548.0000 201196.0000 341704.0000 Alanine 2907819.3786 3237557.0719 2877771.4444 2380347.8686 1989414.7528 2658838.6296 4847921.0124 6104435.8025 5082855.6257 6978174.1416 7056826.7116 6713119.1825 7043190.5951 7280956.2872 7464729.4184 Aminobutyric Acid 11262943.6426 10100266.0052 9607525.0755 10978604.1459 13271247.0301 11542315.4729 Anserine 7411294.5428 6682006.5757 7637062.3491 7621788.9646 7251995.6394 7430497.1467 Arginine 7501959.0000 7561426.0000 5905970.0000 7945394.0000 8456192.0000 6465646.0000 Argininosuccinic Acid 980169.0000 963085.0000 775673.0000 830058.0000 832316.0000 716288.0000 Arsenobetaine 1955464.0000 1612009.0000 1804554.0000 114620304.0000 115199280.0000 110786840.0000 103271680.0000 121784136.0000 111424848.0000 Asparagine 695353.4546 916202.7915 787232.5608 1352770.8244 1264799.9838 1469009.4710 2072190.0344 2196889.8139 2455015.7021 1997358.0468 2145936.6859 2220370.5202 Aspartic acid 757223.0000 1532888.0000 1236328.0000 869742.0000 918007.0000 2293642.0000 1220802.0000 1575528.0000 1105729.0000 1685533.0000 1862695.0000 1858950.0000 1963934.0000 2052020.0000 2025096.0000 beta-Alanine 1439365.9214 1176089.7756 1292349.6994 1882400.0968 1304473.3267 1346902.1028 2722287.6879 1632443.8712 2167542.6931 2071832.4991 1993813.6581 1459619.1474 5746420.4860 5950150.1667 5306391.2791 5145087.8415 5626749.6460 5339687.5184 beta-Glutamic acid 6353386.8365 6158070.1428 6342075.2308 6728936.4629 6340589.0190 5540100.5646 Betaine 93646776.0000 153009424.0000 134932880.0000 240724512.0000 298542240.0000 260596160.0000 168642864.0000 159114960.0000 158657904.0000 312345440.0000 326642240.0000 323218016.0000 294913984.0000 334581760.0000 317677760.0000 Betonicine 1084536.0000 1009753.0000 566826.0000 554013.0000 1134534.0000 1166528.0000 956701.0000 802908.0000 605163.0000 3630631.0000 2842177.0000 2705921.0000 134587792.0000 143010416.0000 125540760.0000 127595808.0000 147082464.0000 125770232.0000 Butyryl-L-carnitine 280837952.0000 290113152.0000 275269056.0000 263791280.0000 292328928.0000 275898208.0000 Carnitine 1856920.0000 4744607.0000 4616431.0000 1318565.0000 2456527.0000 1882880.0000 5410541.0000 7324034.0000 5824863.0000 8280793.0000 7836088.0000 7469360.0000 165516176.0000 184250448.0000 178450160.0000 157871712.0000 182322240.0000 173707152.0000 Carnosine 7357503.6906 6862359.0019 7371499.9896 6824220.8292 6596921.2398 6849337.0934 Chitobiose 325450.0000 183484.0000 307977.0000 2212287.0000 1908900.0000 1588248.0000 1327914.0000 1117206.0000 1237002.0000 Choline 288831.0000 176212.0000 168877.0000 52414976.0000 53916608.0000 44653104.0000 47482288.0000 54869196.0000 46171200.0000 Citrulline 1304842.6686 1489513.2051 1029609.2969 1784609.7098 2052176.3965 1024654.0623 6762524.0074 7069767.6655 6766856.3960 6345777.0788 6817765.1329 6920454.4126 Creatine 2324571.0000 4868025.0000 4807911.0000 2294376.0000 3724844.0000 3511658.0000 1340584.0000 1582383.0000 1616444.0000 7159522.0000 7173212.0000 6542656.0000 34698584.0000 38948372.0000 38879852.0000 34528960.0000 38561460.0000 38983408.0000 Cys-Gly oxidized 1936641.9455 1751817.1923 1978223.7736 1889924.1931 1778177.6363 1826937.5855 Cystathionine 751831.0000 779089.0000 755205.0000 572550.0000 636150.0000 625398.0000 Cysteinesulfinic acid 284945.0000 291659.0000 302400.0000 265384.0000 275958.0000 269504.0000 Cystine 611485.0000 578078.0000 640631.0000 940650.0000 645374.0000 603950.0000 Cytidine 152651.7448 287480.8494 420657.7862 68485.6233 86418.8859 77928.9522 8509147.7700 8139515.9433 8670881.1337 8859009.5146 8585680.1332 7757193.3438 123998.8522 88933.5499 79743.9106 45301.5167 45352.6377 59191.1443 Cytosine 1380874.0000 2467779.0000 2726913.0000 636450.0000 1597657.0000 2199661.0000 2010231.0000 3032781.0000 2685437.0000 611542.0000 450496.0000 767444.0000 34161904.0000 37098428.0000 40495920.0000 32446370.0000 38280980.0000 42117540.0000 Deoxyadenosine 35934999.6281 34316386.6400 34732200.3960 33372102.1227 35141092.4682 33707901.2212 Dimethyl Glycine 32792070.5065 28335526.2147 50939602.3207 39235140.8853 38072465.5806 62108033.8589 Dimethylsulfonioacetate (DMS-Ac) 13965831.0000 22220112.0000 19532664.0000 8667720.0000 10308397.0000 9687338.0000 9388103.0000 8628455.0000 8777906.0000 55505144.0000 60379876.0000 52725344.0000 54073532.0000 61383224.0000 53094628.0000 DMSP 136515.0000 201091.0000 260472.0000 77732000.0000 121033104.0000 106582616.0000 31982692.0000 38867632.0000 32170416.0000 54319452.0000 48742296.0000 53409188.0000 87004648.0000 93645096.0000 92354936.0000 82892352.0000 93201328.0000 87540840.0000 57552.0000 75956.0000 77941.0000 218217.0000 86132.0000 235076.0000 Ectoine 1350933.0000 1552985.0000 1255619.0000 1726388.0000 1703042.0000 1499734.0000 361907392.0000 395718880.0000 372017376.0000 361277856.0000 381687680.0000 393069824.0000 Ergothioneine 1373899.0000 1323340.0000 1239868.0000 1449640.0000 1521416.0000 1421910.0000 Glucosamine 122588.7561 178078.1597 169259.5300 6441537.6434 6333782.8458 6523368.9064 6255175.0232 6393608.3217 7042041.2997 Glucosylglycerol 261013.0210 307355.8868 295633.8352 210864.4715 220162.7432 Glutamic acid 2587287.0407 2304087.5764 2254565.3271 1673070.8709 964520.8945 1383463.9288 2001759.9038 1795619.7148 2203549.1820 4379833.6147 4351046.0450 5627666.5673 7810032.2664 7036226.4319 8633044.0473 7924185.8207 8086701.0058 8332393.7973 Glutamine 957213.0000 1740269.0000 1586444.0000 396339.0000 929214.0000 549558.0000 2465370.0000 2654058.0000 2657606.0000 4664820.0000 3957024.0000 3984704.0000 8359966.0000 9490607.0000 9930846.0000 8601728.0000 9200310.0000 10426206.0000 Glutamylphenylalanine 69913.7588 45568.9885 51943.4730 51661.3247 41514.9649 74392.5135 12815259.8275 11403323.6778 10903201.8217 13016237.8543 12134820.7662 12435904.9726 223132.0300 99371.0185 395915.6087 242606.1122 191479.2573 80088.2550 Glutathione Disulfide 349223.0000 324308.0000 320075.0000 299122.0000 354172.0000 243038.0000 Glycerophosphocholine 130633.0000 146326.0000 145614.0000 3839667.0000 4148025.0000 4309581.0000 3737498.0000 4017860.0000 4156930.0000 Glycine 2172765.8495 1944058.1284 1454042.9944 1307659.6295 1229442.2779 2230908.2622 1518849.6255 1943742.2580 2329098.4327 1978702.9121 1905923.7138 2192153.0355 1975879.8477 2277679.9373 2330641.7636 Gonyol 921127.0000 2078233.0000 2046525.0000 440468.0000 857922.0000 840113.0000 2530225.0000 3324228.0000 2794214.0000 1260172.0000 1063635.0000 1247954.0000 5822613.0000 6325302.0000 5920218.0000 5404894.0000 6094168.0000 5769370.0000 53701.0000 98529.0000 94296.0000 114437.0000 66874.0000 114248.0000 Guanine 1598479.1129 2906215.1701 3315361.3563 1839326.6232 1685691.5242 1668502.4154 1454857.8869 1259915.5748 1259498.0696 12370343.8477 11841545.4238 11608647.8739 11581717.7826 11293645.8000 12247945.9980 1404096.8794 1068784.3793 1615633.8294 612507.7033 476849.0393 575190.9306 Guanosine 3961846.0000 4962825.0000 3668973.0000 4217896.0000 4207366.0000 3881532.0000 Histidine 6507065.0000 6724905.0000 6024395.0000 4423476.0000 2685565.0000 3977024.0000 18764884.0000 19081874.0000 19381272.0000 11010130.0000 20376684.0000 19450660.0000 Homarine 3846864.0000 6430063.0000 6903652.0000 23262468.0000 36700512.0000 30110544.0000 52072052.0000 67072392.0000 55494532.0000 103556496.0000 93229776.0000 95537488.0000 19075900.0000 20541524.0000 19895636.0000 18313076.0000 21079434.0000 19929922.0000 3203487.0000 1646866.0000 2422138.0000 6062874.0000 5169631.0000 6306368.0000 Homocysteine Thiolactone 1613141.1086 1595590.7617 1667385.2798 1513489.1055 1674486.8475 1572354.6104 Homocystine 424337.0000 481487.0000 483326.0000 281000.0000 367160.0000 349672.0000 Homoserine 871840.0000 555944.0000 442957.0000 3876794.0000 4305045.0000 4771043.0000 3467296.0000 3965018.0000 3440938.0000 Hordenine 57376576.0000 58807860.0000 58760504.0000 53262456.0000 60513460.0000 56111908.0000 Hydroxyectoine 2352423.0000 2525079.0000 1729163.0000 5353278.0000 5621252.0000 6046006.0000 4893386.0000 6073814.0000 6246056.0000 Hydroxylysine 501027.0000 587671.0000 496588.0000 289292.0000 369540.0000 317110.0000 Hypoxanthine 6405165.0000 12675104.0000 15533020.0000 7126579.0000 5416052.0000 5336370.0000 48112224.0000 52522124.0000 50162868.0000 45988248.0000 51481648.0000 52256268.0000 2477790.0000 1652557.0000 2209318.0000 2501752.0000 2521880.0000 2637861.0000 Isoleucine 3756613.1020 6679136.4795 5152703.8568 1945985.4523 2253462.5343 1765551.9664 5263879.2669 5197962.0356 5402683.0880 16297932.3094 17376801.7786 16014829.1853 32474615.9592 35172835.8966 31871699.0936 30996455.1019 37295621.1118 36941203.2205 6814951.2610 6376091.9536 5645609.9045 15296650.4081 14244140.5292 14159783.4389 Leucine 2193478.5259 3944148.5677 3017037.1501 3510962.0022 3705786.9782 4138102.3320 9621975.8390 9839793.2656 9818817.4468 19274499.0933 18307347.9992 18797120.4431 18263449.7545 16174695.8656 18022378.9716 5257776.2241 5358632.8311 5346784.5436 10510593.7426 9938241.2204 8596793.6428 Lysine 507218.0000 506597.0000 614309.0000 1495057.0000 1369941.0000 1079855.0000 1700212.0000 1656498.0000 1147902.0000 Melamine 6425009.5966 5877641.5970 7156961.4648 7095719.6470 4800227.7072 9623859.6018 10487020.2018 10534059.2035 14043667.9198 69218914.8988 71885592.8528 69823829.7631 69316000.5029 76906307.7541 70831367.4500 Methionine 5346421.8309 5173110.3390 5943521.4428 5543558.1090 5281593.3822 5798055.8719 Methionine Sulfoxide 560580.0821 623815.6065 532750.7463 703424.2969 1136917.7472 1336458.4410 2472019.9932 2283287.8166 3058183.9188 20395519.7818 19453807.7683 20481578.2014 19690000.9028 20550235.7166 20400046.3368 Methylmalonyl carnitine 40417444.0000 43026364.0000 43262164.0000 39041640.0000 42775656.0000 43780096.0000 Muramic Acid 5291641.1016 4908350.8697 5158469.2056 5524881.9791 5349753.1965 4882540.7168 N-Acetylglucosamine 495613.0000 456179.0000 478187.0000 334234.0000 491248.0000 465878.0000 N-Acetylmuramic acid 2877188.8976 3182733.6628 2971506.0276 2797509.5986 2684863.8081 3039087.8936 181870.5026 214649.9118 207722.9586 212438.8628 217871.9096 308640.3647 N-Acetyl-Serine 337603.2252 505051.8040 621414.1613 310308.9151 469042.8422 N(e)-Acetyl-Lysine 100016.1627 93133.3742 200608.4902 102485.9012 59440.2962 58619.3156 275908.9545 177609.9193 229487.7428 32085571.7180 34592700.6112 34768968.2242 34773929.4722 34985216.1184 33655837.9755 N-methyltaurine 6302324.4736 5748265.5169 6628493.8194 7416207.3183 6269799.5499 6193947.4103 Ophthalmic Acid 6857834.2362 6573985.6757 6986971.0276 7190599.5523 6702029.3895 7110502.9163 Ornithine 3188180.0000 3586167.0000 2869676.0000 2977601.0000 3279157.0000 3354703.0000 5947816.0000 5720041.0000 5039561.0000 7988970.0000 7027948.0000 6495590.0000 PQS 213471.0000 47007.0000 94830.0000 113904.0000 Proline 7800069.3059 9940884.1239 8570549.4300 3611953.4737 3540396.7423 2922470.5882 3354236.1709 3285262.6696 4468508.5971 17398016.6604 22411771.4870 19988913.4013 45975689.6803 47311382.5478 50397031.9352 47046919.5809 47746943.4671 47535831.2047 Propionyl-L-carnitine 84540.0000 89312.0000 113165.0000 62935.0000 50474.0000 63252.0000 73690360.0000 77331368.0000 73257128.0000 71582320.0000 77938760.0000 75045296.0000 Pyridinedicarboxylic acid 5878847.4072 5414815.6004 5383441.0468 5742919.8090 5631595.1927 5453304.7778 Riboflavin Monophosphate 1517419.0000 1093485.0000 1067933.0000 1323416.0000 1131218.0000 970116.0000 Sarcosine 909724.3613 490957.1230 608395.1409 405015.3204 187317.1051 345895.7771 525240.5726 701314.0172 357451.2084 5918628.3668 6244361.6694 6505756.4343 6266865.8726 5979550.1997 6186648.4903 Serine 5144370.0000 8101166.0000 6474962.0000 4515635.0000 7649999.0000 3936649.0000 2902958.0000 3203254.0000 3188674.0000 1829402.0000 2357684.0000 2500650.0000 Stachydrine hydrochloride 1126703.0000 2020271.0000 1678307.0000 2955887.0000 2315316.0000 2460891.0000 7749240.0000 7416006.0000 6918055.0000 31579482.0000 32688604.0000 32756046.0000 30374466.0000 34770116.0000 34014592.0000 Succinylglycine 788377.3139 811108.7677 858929.1966 666388.1115 753885.8144 802912.6451 Thiamine monophosphate 1215332.0000 1305779.0000 1216665.0000 861452.0000 1033246.0000 903594.0000 Threonine 641860.7187 259292.2893 515623.6892 6793765.5685 7276351.7433 8145778.4915 6305828.8730 8385059.8179 6850246.1032 trans Hydroxyl proline 165954.8636 345237.2284 262438.8137 13600170.0328 13157087.6885 12964776.8162 13749291.6810 12941512.2190 13427277.0821 Trigonelline 2447768.0000 5763791.0000 5934156.0000 5277081.0000 8932684.0000 7848948.0000 6289983.0000 7596008.0000 6158276.0000 38983824.0000 31289628.0000 33339078.0000 27044546.0000 30203242.0000 30111786.0000 26383324.0000 30161362.0000 29604300.0000 Trimethylamine 698642.0000 614348.0000 499240.0000 872910.0000 573768.0000 494542.0000 Trimethylamine N-oxide 7272081.0000 9124623.0000 9161857.0000 25966166.0000 43247148.0000 38066496.0000 21369098.0000 23221758.0000 20177242.0000 434063008.0000 403748544.0000 562661120.0000 82024480.0000 85544984.0000 85803032.0000 80862160.0000 90120176.0000 89947456.0000 649619.0000 649069.0000 778138.0000 4703859.0000 2933888.0000 4664000.0000 Trimethylammonium Propionate (TMAP) 699806.0000 1238433.0000 1309503.0000 2226187.0000 3597547.0000 3201003.0000 54574752.0000 68711032.0000 58179572.0000 4679681.0000 4255983.0000 4956806.0000 95561944.0000 104532856.0000 100817384.0000 93838976.0000 100136576.0000 103885128.0000 Trimethyl-L-lysine 8973351.0000 9467279.0000 9114069.0000 8333980.0000 9496176.0000 9207198.0000 Turicine 503080.0000 490834.0000 490280.0000 681990.0000 1276176.0000 768011.0000 835310.0000 958952.0000 627597.0000 3630631.0000 2544792.0000 3090857.0000 134587792.0000 143010416.0000 125540760.0000 127595808.0000 147272592.0000 125770232.0000 Tyrosine 778204.7138 1266627.7083 634455.3353 1696608.1921 1578258.8566 909903.8064 7980953.6049 7990150.6141 8599843.1708 8142728.4332 8393025.7558 8789393.6292 952634.8338 925732.0257 923576.6401 2060784.0112 2361528.5192 1905579.8375 Valine 2152458.9899 5005370.0255 2830312.6077 3708767.9752 30111048.9920 5626180.7278 12927547.1224 12586640.1099 14704670.7538 9485779.4293 14864946.4042 12547689.4732 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name pubchem_id inchi_key kegg_id other_id other_id_type ri ri_type moverz_quant (3-Carboxypropyl)trimethylammonium (TMAB) NA 4-Hydroxyisoleucine cpd:C20802 5-Methylcytosine cpd:C02376 6-Methyladenine cpd:C08434 Acetylcholine cpd:C01996 Acetylglutamic acid cpd:C00624 Acetyl-L-carnitine cpd:C02571 Adenine cpd:C00147 Adenosine cpd:C00212 Adenosyl Homocysteine cpd:C00021 Adenosyl Methionine cpd:C00019 Alanine cpd:C00041 Aminobutyric Acid cpd:C00334 Anserine cpd:C01262 Arginine cpd:C00062 Argininosuccinic Acid cpd:C03406 Arsenobetaine cpd:C19331 Asparagine cpd:C00152 Aspartic acid cpd:C00049 beta-Alanine cpd:C00099 beta-Glutamic acid NA Betaine cpd:C00719 Betonicine cpd:C08269 Butyryl-L-carnitine NA Carnitine cpd:C00487 Carnosine cpd:C00386 Chitobiose cpd:C01674 Choline cpd:C00114 Citrulline cpd:C00327 Creatine cpd:C00300 Cys-Gly oxidized NA Cystathionine cpd:C02291 Cysteinesulfinic acid cpd:C00606 Cystine cpd:C00491 Cytidine cpd:C00475 Cytosine cpd:C00380 Deoxyadenosine cpd:C00559 Dimethyl Glycine cpd:C01026 Dimethylsulfonioacetate (DMS-Ac) cpd:C03392 DMSP cpd:C04022 Ectoine cpd:C06231 Ergothioneine cpd:C05570 Glucosamine NA Glucosylglycerol cpd:C11546 Glutamic acid cpd:C00025 Glutamine cpd:C00064 Glutamylphenylalanine NA Glutathione Disulfide cpd:C00127 Glycerophosphocholine cpd:C00670 Glycine cpd:C00037 Gonyol NA Guanine cpd:C00242 Guanosine cpd:C00387 Histidine cpd:C00135 Homarine NA Homocysteine Thiolactone NA Homocystine cpd:C01817 Homoserine cpd:C00263 Hordenine cpd:C06199 Hydroxyectoine cpd:C16432 Hydroxylysine cpd:C16741 Hypoxanthine cpd:C00262 Isoleucine cpd:C00407 Leucine cpd:C00123 Lysine cpd:C00047 Melamine cpd:C08737 Methionine cpd:C00073 Methionine Sulfoxide cpd:C02989 Methylmalonyl carnitine NA Muramic Acid cpd:C06470 N-Acetylglucosamine cpd:C00140 N-Acetylmuramic acid cpd:C02713 N-Acetyl-Serine cpd:C00979 N(e)-Acetyl-Lysine cpd:C02727 N-methyltaurine NA Ophthalmic Acid NA Ornithine cpd:C00077 PQS cpd:C11848 Proline cpd:C00148 Propionyl-L-carnitine cpd:C03017 Pyridinedicarboxylic acid cpd:C03722 Riboflavin Monophosphate cpd:C00061 Sarcosine cpd:C00213 Serine cpd:C00065 Stachydrine hydrochloride cpd:C10172 Succinylglycine NA Thiamine monophosphate cpd:C01081 Threonine cpd:C00188 trans Hydroxyl proline cpd:C01157 Trigonelline cpd:C01004 Trimethylamine cpd:C00565 Trimethylamine N-oxide cpd:C01104 Trimethylammonium Propionate (TMAP) cpd:C08263 Trimethyl-L-lysine cpd:C03793 Turicine cpd:C21513 Tyrosine cpd:C00082 Valine cpd:C00183 METABOLITES_END #END