#METABOLOMICS WORKBENCH wkumler_20230717_155835 DATATRACK_ID:4170 STUDY_ID:ST002788 ANALYSIS_ID:AN004537 PROJECT_ID:PR001738
VERSION             	1
CREATED_ON             	July 20, 2023, 8:22 am
#PROJECT
PR:PROJECT_TITLE                 	The influence of mesoscale eddy features on marine metabolomic variability in
PR:PROJECT_TITLE                 	the North Pacific Subtropical Gyre
PR:PROJECT_TYPE                  	Marine Metabolomics
PR:PROJECT_SUMMARY               	Mesoscale eddies are a dominant source of variability in the ocean's gyres,
PR:PROJECT_SUMMARY               	often analogized to the "weather" of the sea. They alter the balance between
PR:PROJECT_SUMMARY               	light and nutrients, shifting community composition on both the species and
PR:PROJECT_SUMMARY               	molecular scale. Here, we collected metabolomic samples from across eddy dipoles
PR:PROJECT_SUMMARY               	in the North Pacific Subtropical Gyre to detect and quantify these shifts on a
PR:PROJECT_SUMMARY               	chemical level. The data indicate that metabolites dynamically and robustly
PR:PROJECT_SUMMARY               	track with biological community metrics and result in biochemically distinct
PR:PROJECT_SUMMARY               	particulate matter in cyclonic and anticyclonic eddies.
PR:INSTITUTE                     	University of Washington
PR:DEPARTMENT                    	School of Oceanography
PR:LABORATORY                    	Ingalls Lab
PR:LAST_NAME                     	Kumler
PR:FIRST_NAME                    	William
PR:ADDRESS                       	1501 NE Boat St, Seattle, WA, 98105, USA
PR:EMAIL                         	wkumler@uw.edu
PR:PHONE                         	2062216732
PR:FUNDING_SOURCE                	Simons Collaboration on Ocean Processes and Ecology, NSF
#STUDY
ST:STUDY_TITLE                   	Metabolomic analysis of particulate matter in the NPSG during a 2018 cruise on
ST:STUDY_TITLE                   	the R/V Falkor
ST:STUDY_SUMMARY                 	Targeted and untargeted analysis of metabolomics samples from the North Pacific
ST:STUDY_SUMMARY                 	Subtropical Gyre taken during the 2018 March/April SCOPE cruise on the R/V
ST:STUDY_SUMMARY                 	Falkor (FK180310) across a mesoscale eddy dipole. Particulate matter was
ST:STUDY_SUMMARY                 	collected on 0.2um filters and extracted using a modified Bligh & Dyer before
ST:STUDY_SUMMARY                 	analysis on a QE Orbitrap. Results show significant changes in the absolute
ST:STUDY_SUMMARY                 	quantity and relative composition of particles in the gyre between anticyclonic
ST:STUDY_SUMMARY                 	and cyclonic eddies.
ST:INSTITUTE                     	University of Washington
ST:DEPARTMENT                    	School of Oceanography
ST:LABORATORY                    	Ingalls Lab
ST:LAST_NAME                     	Kumler
ST:FIRST_NAME                    	William
ST:ADDRESS                       	1501 NE Boat St, Seattle, WA 98105
ST:EMAIL                         	wkumler@uw.edu
ST:PHONE                         	2062216732
#SUBJECT
SU:SUBJECT_TYPE                  	Other organism
SU:SUBJECT_SPECIES               	Natural mixed marine microbial community
#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           	190715_Smp_FK180310S77C1-25m_A.mzML	190715_Smp_FK180310S77C1-25m_A.mzML	Sample_type:Sample | Station:77 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:20.8044	Time_sampled=2018-04-07 22:57:24; Latitude=24.5306; Longitude=160.787; Temperature=22.8817; Salinity=35.0647; Particulate_carbon=2.1415
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S77C1-25m_B.mzML	190715_Smp_FK180310S77C1-25m_B.mzML	Sample_type:Sample | Station:77 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:20.8044	Time_sampled=2018-04-07 22:57:24; Latitude=24.5306; Longitude=160.787; Temperature=22.8817; Salinity=35.0647; Particulate_carbon=2.1415
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S77C1-25m_C.mzML	190715_Smp_FK180310S77C1-25m_C.mzML	Sample_type:Sample | Station:77 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:20.8044	Time_sampled=2018-04-07 22:57:24; Latitude=24.5306; Longitude=160.787; Temperature=22.8817; Salinity=35.0647; Particulate_carbon=2.1415
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S77C1-DCM_A.mzML	190715_Smp_FK180310S77C1-DCM_A.mzML	Sample_type:Sample | Station:77 | Depth:DCM | Absolute_depth:125 | Sea_level_anomaly:19.87	Time_sampled=2018-04-07 22:57:24; Latitude=24.512; Longitude=160.507; Temperature=22.579; Salinity=35.161; Particulate_carbon=1.48794
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S77C1-DCM_B.mzML	190715_Smp_FK180310S77C1-DCM_B.mzML	Sample_type:Sample | Station:77 | Depth:DCM | Absolute_depth:125 | Sea_level_anomaly:19.87	Time_sampled=2018-04-07 22:57:24; Latitude=24.512; Longitude=160.507; Temperature=22.579; Salinity=35.161; Particulate_carbon=1.48794
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S77C1-DCM_C.mzML	190715_Smp_FK180310S77C1-DCM_C.mzML	Sample_type:Sample | Station:77 | Depth:DCM | Absolute_depth:125 | Sea_level_anomaly:19.87	Time_sampled=2018-04-07 22:57:24; Latitude=24.512; Longitude=160.507; Temperature=22.579; Salinity=35.161; Particulate_carbon=1.48794
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S80C1-25m_A.mzML	190715_Smp_FK180310S80C1-25m_A.mzML	Sample_type:Sample | Station:80 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:20.8044	Time_sampled=2018-04-08 11:14:18; Latitude=24.5306; Longitude=160.787; Temperature=22.8817; Salinity=35.0647; Particulate_carbon=2.1415
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S80C1-25m_B.mzML	190715_Smp_FK180310S80C1-25m_B.mzML	Sample_type:Sample | Station:80 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:20.8044	Time_sampled=2018-04-08 11:14:18; Latitude=24.5306; Longitude=160.787; Temperature=22.8817; Salinity=35.0647; Particulate_carbon=2.1415
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S80C1-25m_C.mzML	190715_Smp_FK180310S80C1-25m_C.mzML	Sample_type:Sample | Station:80 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:20.8044	Time_sampled=2018-04-08 11:14:18; Latitude=24.5306; Longitude=160.787; Temperature=22.8817; Salinity=35.0647; Particulate_carbon=2.1415
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S80C1-DCM_A.mzML	190715_Smp_FK180310S80C1-DCM_A.mzML	Sample_type:Sample | Station:80 | Depth:DCM | Absolute_depth:120 | Sea_level_anomaly:21.03	Time_sampled=2018-04-08 11:14:18; Latitude=24.582; Longitude=160.882; Temperature=22.614; Salinity=35.158; Particulate_carbon=1.59016
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S80C1-DCM_B.mzML	190715_Smp_FK180310S80C1-DCM_B.mzML	Sample_type:Sample | Station:80 | Depth:DCM | Absolute_depth:120 | Sea_level_anomaly:21.03	Time_sampled=2018-04-08 11:14:18; Latitude=24.582; Longitude=160.882; Temperature=22.614; Salinity=35.158; Particulate_carbon=1.59016
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S80C1-DCM_C.mzML	190715_Smp_FK180310S80C1-DCM_C.mzML	Sample_type:Sample | Station:80 | Depth:DCM | Absolute_depth:120 | Sea_level_anomaly:21.03	Time_sampled=2018-04-08 11:14:18; Latitude=24.582; Longitude=160.882; Temperature=22.614; Salinity=35.158; Particulate_carbon=1.59016
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S62C1-25m_A.mzML	190715_Smp_FK180310S62C1-25m_A.mzML	Sample_type:Sample | Station:62 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:-12.48	Time_sampled=2018-04-03 22:32:50; Latitude=22.2318; Longitude=156.855; Temperature=23.0028; Salinity=35.1175; Particulate_carbon=2.35172
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S62C1-25m_B.mzML	190715_Smp_FK180310S62C1-25m_B.mzML	Sample_type:Sample | Station:62 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:-12.48	Time_sampled=2018-04-03 22:32:50; Latitude=22.2318; Longitude=156.855; Temperature=23.0028; Salinity=35.1175; Particulate_carbon=2.35172
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S62C1-25m_C.mzML	190715_Smp_FK180310S62C1-25m_C.mzML	Sample_type:Sample | Station:62 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:-12.48	Time_sampled=2018-04-03 22:32:50; Latitude=22.2318; Longitude=156.855; Temperature=23.0028; Salinity=35.1175; Particulate_carbon=2.35172
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S62C1-DCM_A.mzML	190715_Smp_FK180310S62C1-DCM_A.mzML	Sample_type:Sample | Station:62 | Depth:DCM | Absolute_depth:98 | Sea_level_anomaly:-13.6	Time_sampled=2018-04-03 22:32:50; Latitude=22.367; Longitude=156.57; Temperature=21.264; Salinity=35.176; Particulate_carbon=2.31849
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S62C1-DCM_B.mzML	190715_Smp_FK180310S62C1-DCM_B.mzML	Sample_type:Sample | Station:62 | Depth:DCM | Absolute_depth:98 | Sea_level_anomaly:-13.6	Time_sampled=2018-04-03 22:32:50; Latitude=22.367; Longitude=156.57; Temperature=21.264; Salinity=35.176; Particulate_carbon=2.31849
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S62C1-DCM_C.mzML	190715_Smp_FK180310S62C1-DCM_C.mzML	Sample_type:Sample | Station:62 | Depth:DCM | Absolute_depth:98 | Sea_level_anomaly:-13.6	Time_sampled=2018-04-03 22:32:50; Latitude=22.367; Longitude=156.57; Temperature=21.264; Salinity=35.176; Particulate_carbon=2.31849
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S64C1-25m_A.mzML	190715_Smp_FK180310S64C1-25m_A.mzML	Sample_type:Sample | Station:64 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:-12.48	Time_sampled=2018-04-04 10:50:59; Latitude=22.2318; Longitude=156.855; Temperature=23.0028; Salinity=35.1175; Particulate_carbon=2.35172
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S64C1-25m_B.mzML	190715_Smp_FK180310S64C1-25m_B.mzML	Sample_type:Sample | Station:64 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:-12.48	Time_sampled=2018-04-04 10:50:59; Latitude=22.2318; Longitude=156.855; Temperature=23.0028; Salinity=35.1175; Particulate_carbon=2.35172
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S64C1-25m_C.mzML	190715_Smp_FK180310S64C1-25m_C.mzML	Sample_type:Sample | Station:64 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:-12.48	Time_sampled=2018-04-04 10:50:59; Latitude=22.2318; Longitude=156.855; Temperature=23.0028; Salinity=35.1175; Particulate_carbon=2.35172
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S64C1-DCM_A.mzML	190715_Smp_FK180310S64C1-DCM_A.mzML	Sample_type:Sample | Station:64 | Depth:DCM | Absolute_depth:115 | Sea_level_anomaly:-12.1225	Time_sampled=2018-04-04 10:50:59; Latitude=22.1359; Longitude=156.984; Temperature=21.1728; Salinity=35.1679; Particulate_carbon=1.7286
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S64C1-DCM_B.mzML	190715_Smp_FK180310S64C1-DCM_B.mzML	Sample_type:Sample | Station:64 | Depth:DCM | Absolute_depth:115 | Sea_level_anomaly:-12.1225	Time_sampled=2018-04-04 10:50:59; Latitude=22.1359; Longitude=156.984; Temperature=21.1728; Salinity=35.1679; Particulate_carbon=1.7286
SUBJECT_SAMPLE_FACTORS           	190715_Smp_FK180310S64C1-DCM_C.mzML	190715_Smp_FK180310S64C1-DCM_C.mzML	Sample_type:Sample | Station:64 | Depth:DCM | Absolute_depth:115 | Sea_level_anomaly:-12.1225	Time_sampled=2018-04-04 10:50:59; Latitude=22.1359; Longitude=156.984; Temperature=21.1728; Salinity=35.1679; Particulate_carbon=1.7286
SUBJECT_SAMPLE_FACTORS           	190715_Blk_KM1906U14-Blk_C.mzML	190715_Blk_KM1906U14-Blk_C.mzML	Sample_type:Blank | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
SUBJECT_SAMPLE_FACTORS           	190715_Poo_TruePooFK180310_Full1.mzML	190715_Poo_TruePooFK180310_Full1.mzML	Sample_type:QC Pooled | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
SUBJECT_SAMPLE_FACTORS           	190715_Poo_TruePooFK180310_Full2.mzML	190715_Poo_TruePooFK180310_Full2.mzML	Sample_type:QC Pooled | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
SUBJECT_SAMPLE_FACTORS           	190715_Poo_TruePooFK180310_Full3.mzML	190715_Poo_TruePooFK180310_Full3.mzML	Sample_type:QC Pooled | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
SUBJECT_SAMPLE_FACTORS           	190715_Poo_TruePooFK180310_Half1.mzML	190715_Poo_TruePooFK180310_Half1.mzML	Sample_type:QC Pooled | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
SUBJECT_SAMPLE_FACTORS           	190715_Poo_TruePooFK180310_Half2.mzML	190715_Poo_TruePooFK180310_Half2.mzML	Sample_type:QC Pooled | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
SUBJECT_SAMPLE_FACTORS           	190715_Poo_TruePooFK180310_Half3.mzML	190715_Poo_TruePooFK180310_Half3.mzML	Sample_type:QC Pooled | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
SUBJECT_SAMPLE_FACTORS           	190715_Std_4uMStdsMix1InH2O_1.mzML	190715_Std_4uMStdsMix1InH2O_1.mzML	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
SUBJECT_SAMPLE_FACTORS           	190715_Std_4uMStdsMix1InH2O_2.mzML	190715_Std_4uMStdsMix1InH2O_2.mzML	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
SUBJECT_SAMPLE_FACTORS           	190715_Std_4uMStdsMix1InMatrix_1.mzML	190715_Std_4uMStdsMix1InMatrix_1.mzML	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
SUBJECT_SAMPLE_FACTORS           	190715_Std_4uMStdsMix1InMatrix_2.mzML	190715_Std_4uMStdsMix1InMatrix_2.mzML	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
SUBJECT_SAMPLE_FACTORS           	190715_Std_4uMStdsMix2InH2O_1.mzML	190715_Std_4uMStdsMix2InH2O_1.mzML	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
SUBJECT_SAMPLE_FACTORS           	190715_Std_4uMStdsMix2InH2O_2.mzML	190715_Std_4uMStdsMix2InH2O_2.mzML	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
SUBJECT_SAMPLE_FACTORS           	190715_Std_4uMStdsMix2InMatrix_1.mzML	190715_Std_4uMStdsMix2InMatrix_1.mzML	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
SUBJECT_SAMPLE_FACTORS           	190715_Std_4uMStdsMix2InMatrix_2.mzML	190715_Std_4uMStdsMix2InMatrix_2.mzML	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
SUBJECT_SAMPLE_FACTORS           	190715_Std_H2OinMatrix_1.mzML	190715_Std_H2OinMatrix_1.mzML	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
SUBJECT_SAMPLE_FACTORS           	190715_Std_H2OinMatrix_2.mzML	190715_Std_H2OinMatrix_2.mzML	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Time_sampled=NA; Latitude=NA; Longitude=NA; Temperature=NA; Salinity=NA; Particulate_carbon=NA
#COLLECTION
CO:COLLECTION_SUMMARY            	Samples were collected aboard the R/V Falkor in March and April 2018. Water was
CO:COLLECTION_SUMMARY            	collected with Niskin bottles attached to the CTD from the surface and the deep
CO:COLLECTION_SUMMARY            	chlorophyll maximum (DCM) in both the morning (~6AM) and evening (~6PM). Water
CO:COLLECTION_SUMMARY            	was filtered onto a 0.2um pore size filter under gentle vacuum pressure prior to
CO:COLLECTION_SUMMARY            	flash-freezing in LN2 before storage at -80 in precombusted aluminum foil.
CO:COLLECTION_SUMMARY            	Samples were collected from two locations, one in a cyclonic eddy (-13cm,
CO:COLLECTION_SUMMARY            	stations 62 and 64) and one in an anticyclonic eddy (+20cm, stations 77 and 80)
CO:COLLECTION_SUMMARY            	three days apart.
CO:SAMPLE_TYPE                   	Suspended Marine Particulate Matter
CO:COLLECTION_METHOD             	CTD Niskin Bottle
CO:COLLECTION_LOCATION           	North Pacific
CO:VOLUMEORAMOUNT_COLLECTED      	10L
CO:STORAGE_CONDITIONS            	-80℃
#TREATMENT
TR:TREATMENT_SUMMARY             	No experimental manipulations were performed on the samples.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Polar metabolites were extracted using a modified Bligh−Dyer extraction using
SP:SAMPLEPREP_SUMMARY            	1:1 methanol/water and dichloromethane. A methodological blank was extracted and
SP:SAMPLEPREP_SUMMARY            	analyzed alongside. Isotope-labeled internal standards were added both before
SP:SAMPLEPREP_SUMMARY            	and after the extraction for all samples, blanks, and pooled samples. To
SP:SAMPLEPREP_SUMMARY            	evaluate the effect of obscuring variation due to different matrix strengths and
SP:SAMPLEPREP_SUMMARY            	analytical drift, pooled samples were run at both full and half concentration
SP:SAMPLEPREP_SUMMARY            	(diluted with water) three times during the run.
SP:PROCESSING_STORAGE_CONDITIONS 	On ice
SP:EXTRACTION_METHOD             	Bligh-Dyer
SP:EXTRACT_STORAGE               	-80℃
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	A SeQuant ZIC-pHILIC column (5 um particle size, 2.1 mm x 150 mm, from
CH:CHROMATOGRAPHY_SUMMARY        	Millipore) was used with 10 mM ammonium carbonate in 85:15 acetonitrile to water
CH:CHROMATOGRAPHY_SUMMARY        	(Solvent A) and 10 mM ammonium carbonate in 85:15 water to acetonitrile (Solvent
CH:CHROMATOGRAPHY_SUMMARY        	B) at a flow rate of 0.15 mL/min. This column was compared with a Waters UPLC
CH:CHROMATOGRAPHY_SUMMARY        	BEH amide and a Millipore cHILIC column; the pHILIC showed superior
CH:CHROMATOGRAPHY_SUMMARY        	reproducibility and peak shapes. The column was held at 100% A for 2 minutes,
CH:CHROMATOGRAPHY_SUMMARY        	ramped to 64% B over 18 minutes, ramped to 100% B over 1 minute, held at 100% B
CH:CHROMATOGRAPHY_SUMMARY        	for 5 minutes, and equilibrated at 100% A for 25 minutes (50 minutes total). The
CH:CHROMATOGRAPHY_SUMMARY        	column was maintained at 30 C. The injection volume was 2 µL for samples and
CH:CHROMATOGRAPHY_SUMMARY        	standard mixes. When starting a batch, the column was equilibrated at the
CH:CHROMATOGRAPHY_SUMMARY        	starting conditions for at least 30 minutes. To improve the performance of the
CH:CHROMATOGRAPHY_SUMMARY        	HILIC column, we maintained the same injection volume, kept the instrument
CH:CHROMATOGRAPHY_SUMMARY        	running water blanks between samples as necessary, and injected standards in a
CH:CHROMATOGRAPHY_SUMMARY        	representative matrix in addition to standards in water. After each batch, the
CH:CHROMATOGRAPHY_SUMMARY        	column was flushed with 10 mM ammonium carbonate in 85:15 water to acetonitrile
CH:CHROMATOGRAPHY_SUMMARY        	for 20 to 30 minutes.
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Waters Acquity I-Class
CH:COLUMN_NAME                   	SeQuant ZIC-HILIC (150 x 2.1mm, 5um)
CH:SOLVENT_A                     	85% acetonitrile/15% water
CH:SOLVENT_B                     	15% acetonitrile/85% water
CH:FLOW_GRADIENT                 	100% A for 2 minutes, ramped to 64% B over 18 minutes, ramped to 100% B over 1
CH:FLOW_GRADIENT                 	minute, held at 100% B for 5 minutes, and equilibrated at 100% A for 25 minutes
CH:FLOW_GRADIENT                 	(50 minutes total)
CH:FLOW_RATE                     	0.15 mL/min
CH:COLUMN_TEMPERATURE            	30
#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                   	Metabolomic data was collected on a Thermo Q Exactive HF hybrid Orbitrap (QE)
MS:MS_COMMENTS                   	mass spectrometer. The capillary and auxiliary gas heater temperatures were
MS:MS_COMMENTS                   	maintained at 320°C and 100°C, respectively. The S-lens RF level was kept at
MS:MS_COMMENTS                   	65, the H-ESI voltage was set to 3.3 kV and sheath gas, auxiliary gas, and sweep
MS:MS_COMMENTS                   	gas flow rates were set at 16, 3, and 1, respectively. Polarity switching was
MS:MS_COMMENTS                   	used with a scan range of 60 to 900 m/z and a resolution of 60,000. Calibration
MS:MS_COMMENTS                   	was performed every 3-4 days at a target mass of 200 m/z. All files were then
MS:MS_COMMENTS                   	converted to an open-source mzML format and centroided via Proteowizard’s
MS:MS_COMMENTS                   	msConvert tool. Skyline was used to for manual integration and QC of quantified
MS:MS_COMMENTS                   	data, while XCMS was used with a custom QC method for peak area data with the
MS:MS_COMMENTS                   	model trained on positive mode data applied to these negative mode results.
MS:MS_RESULTS_FILE               	ST002788_AN004537_Results.txt	UNITS:normalized peak area per liter of seawater filtered	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	nM
MS_METABOLITE_DATA_START
Samples	190715_Std_4uMStdsMix1InH2O_1.mzML	190715_Std_4uMStdsMix1InH2O_2.mzML	190715_Std_4uMStdsMix1InMatrix_1.mzML	190715_Std_4uMStdsMix1InMatrix_2.mzML	190715_Std_4uMStdsMix2InH2O_1.mzML	190715_Std_4uMStdsMix2InH2O_2.mzML	190715_Std_4uMStdsMix2InMatrix_1.mzML	190715_Std_4uMStdsMix2InMatrix_2.mzML	190715_Std_H2OinMatrix_2.mzML	190715_Std_H2OinMatrix_1.mzML	190715_Blk_KM1906U14-Blk_C.mzML	190715_Poo_TruePooFK180310_Full1.mzML	190715_Poo_TruePooFK180310_Full2.mzML	190715_Poo_TruePooFK180310_Full3.mzML	190715_Poo_TruePooFK180310_Half1.mzML	190715_Poo_TruePooFK180310_Half2.mzML	190715_Poo_TruePooFK180310_Half3.mzML	190715_Smp_FK180310S62C1-25m_A.mzML	190715_Smp_FK180310S62C1-25m_B.mzML	190715_Smp_FK180310S62C1-25m_C.mzML	190715_Smp_FK180310S62C1-DCM_A.mzML	190715_Smp_FK180310S62C1-DCM_B.mzML	190715_Smp_FK180310S62C1-DCM_C.mzML	190715_Smp_FK180310S64C1-25m_A.mzML	190715_Smp_FK180310S64C1-25m_B.mzML	190715_Smp_FK180310S64C1-25m_C.mzML	190715_Smp_FK180310S64C1-DCM_A.mzML	190715_Smp_FK180310S64C1-DCM_B.mzML	190715_Smp_FK180310S64C1-DCM_C.mzML	190715_Smp_FK180310S77C1-25m_A.mzML	190715_Smp_FK180310S77C1-25m_B.mzML	190715_Smp_FK180310S77C1-25m_C.mzML	190715_Smp_FK180310S77C1-DCM_A.mzML	190715_Smp_FK180310S77C1-DCM_B.mzML	190715_Smp_FK180310S77C1-DCM_C.mzML	190715_Smp_FK180310S80C1-25m_A.mzML	190715_Smp_FK180310S80C1-25m_B.mzML	190715_Smp_FK180310S80C1-25m_C.mzML	190715_Smp_FK180310S80C1-DCM_A.mzML	190715_Smp_FK180310S80C1-DCM_B.mzML	190715_Smp_FK180310S80C1-DCM_C.mzML
Factors	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:Standard mix | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:Blank | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:QC Pooled | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:QC Pooled | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:QC Pooled | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:QC Pooled | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:QC Pooled | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:QC Pooled | Station:NA | Depth:NA | Absolute_depth:NA | Sea_level_anomaly:NA	Sample_type:Sample | Station:62 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:-12.48	Sample_type:Sample | Station:62 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:-12.48	Sample_type:Sample | Station:62 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:-12.48	Sample_type:Sample | Station:62 | Depth:DCM | Absolute_depth:98 | Sea_level_anomaly:-13.6	Sample_type:Sample | Station:62 | Depth:DCM | Absolute_depth:98 | Sea_level_anomaly:-13.6	Sample_type:Sample | Station:62 | Depth:DCM | Absolute_depth:98 | Sea_level_anomaly:-13.6	Sample_type:Sample | Station:64 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:-12.48	Sample_type:Sample | Station:64 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:-12.48	Sample_type:Sample | Station:64 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:-12.48	Sample_type:Sample | Station:64 | Depth:DCM | Absolute_depth:115 | Sea_level_anomaly:-12.1225	Sample_type:Sample | Station:64 | Depth:DCM | Absolute_depth:115 | Sea_level_anomaly:-12.1225	Sample_type:Sample | Station:64 | Depth:DCM | Absolute_depth:115 | Sea_level_anomaly:-12.1225	Sample_type:Sample | Station:77 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:20.8044	Sample_type:Sample | Station:77 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:20.8044	Sample_type:Sample | Station:77 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:20.8044	Sample_type:Sample | Station:77 | Depth:DCM | Absolute_depth:125 | Sea_level_anomaly:19.87	Sample_type:Sample | Station:77 | Depth:DCM | Absolute_depth:125 | Sea_level_anomaly:19.87	Sample_type:Sample | Station:77 | Depth:DCM | Absolute_depth:125 | Sea_level_anomaly:19.87	Sample_type:Sample | Station:80 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:20.8044	Sample_type:Sample | Station:80 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:20.8044	Sample_type:Sample | Station:80 | Depth:Surface | Absolute_depth:25 | Sea_level_anomaly:20.8044	Sample_type:Sample | Station:80 | Depth:DCM | Absolute_depth:120 | Sea_level_anomaly:21.03	Sample_type:Sample | Station:80 | Depth:DCM | Absolute_depth:120 | Sea_level_anomaly:21.03	Sample_type:Sample | Station:80 | Depth:DCM | Absolute_depth:120 | Sea_level_anomaly:21.03
(2R)-3-sulfopropanediol	NA	NA	0.07623	0.06059	1.021	1.044	0.4037	0.3864	0.06674	0.08343	NA	0.0575	0.05806	0.05844	0.05493	0.03943	0.04726	0.1094	0.04871	0.06538	0.1132	0.5176	0.1212	0.0424	0.0759	0.05156	0.09176	0.1126	0.1167	0.02705	0.02764	0.01491	0.1526	0.08737	0.05025	0.02736	0.07617	0.05254	0.03782	0.08405	0.1122
Pyroglutamic acid	0.3573	0.3326	0.5999	0.5241	2.127e-05	NA	0.2372	0.2125	0.2072	0.2768	0.05881	0.1624	0.1249	0.1372	0.1536	0.1379	0.1125	0.1474	0.09714	0.308	0.08918	0.1992	0.1919	0.1227	0.1258	0.1714	0.06559	0.1266	0.1122	0.1345	0.1097	0.259	0.1294	0.04499	0.153	0.1677	0.1293	0.1528	0.05499	0.1052	0.1719
AMP	NA	NA	0.001566	0.00237	NA	NA	0.5098	0.999	0.002022	NA	NA	0.03154	0.02532	0.0348	0.02138	0.02489	0.03594	0.06451	0.02268	0.178	0	0.04288	0.01329	0.01969	0.03075	0.2031	0.003622	0.01836	0.01406	0.006448	0	0.006666	0.0005242	0.003922	0.007322	0.002006	0.02362	0.0479	0.001828	0.009006	0.01527
2-Hydroxyethanesulfonate	NA	NA	0.1262	0.1397	NA	NA	0.4871	0.5517	0.1471	0.1399	0.005158	0.05967	0.0517	0.05536	0.05402	0.04879	0.05438	0.0742	0.04614	0.2424	0.03543	0.09519	0.05668	0.07249	0.05386	0.1173	0.0298	0.04616	0.05112	0.04049	0.03997	0.02171	0.02854	0.02014	0.02829	0.03334	0.05085	0.06381	0.01676	0.03756	0.04465
Malic acid	0.3973	0.2848	0.467	0.3248	0.02386	0.006001	0.09729	0.05982	0.06019	0.09163	0.01967	0.05342	0.05203	0.06395	0.06062	0.08843	0.06077	0.07537	0.07326	0.08236	0.05073	0.1045	0.07023	0.06924	0.07189	0.06446	0.05177	0.05414	0.07008	0.0567	0.04606	0.05148	0.05732	0.05353	0.06813	0.06576	0.08195	0.07215	0.03888	0.07578	0.06178
Sucrose	NA	NA	0.5824	0.725	NA	NA	0.1501	0.1387	0.1374	0.1466	0.03394	0.1789	0.1628	0.1738	0.1671	0.1718	0.166	0.1839	0.1357	0.1661	0.02714	0.03843	0.04884	0.318	0.287	0.321	0.1355	0.1584	0.1314	0.1177	0.1174	0.1909	0.0509	0.02666	0.0319	0.211	0.2494	0.2547	0.1457	0.3612	NA
Taurine	NA	NA	0.09289	0.09332	NA	NA	0.3923	0.4295	0.08632	0.08816	0.006899	0.102	0.09377	0.1005	0.1167	0.1107	0.1041	0.1241	0.06072	0.4825	0.05064	0.2302	0.08265	0.09987	0.1325	0.2123	0.0443	0.09777	0.08364	0.05702	0.07094	0.0406	0.05355	0.06292	0.05192	0.04944	0.07572	0.2227	0.03269	0.06618	0.07566
Trehalose	0.0006444	0	0.004686	0.002619	0.3411	0.2368	0.3692	0.2819	0.002212	0.008819	0.001892	0.004014	0.006906	0.006039	0.001572	0.001065	0	0.001233	0	0.007317	0	0.00778	0.0009122	0.01587	0.01598	0.01997	0.003898	0.006521	0.005651	0.006817	0.001312	0.0005974	0.0007464	0	0.0006634	0.00594	0.01515	0.009641	0.001671	0.007191	0.01282
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	formula	stan_mz	stan_rt_expected	which_standard_mix	concentration_added	kegg_id
(2R)-3-sulfopropanediol	C3H8O5S	155.001422	10.67	Mix2	4	cpd:C19675
Pyroglutamic acid	C5H7NO3	128.034768	7.97	Mix1	4	cpd:C01879
AMP	C10H14N5O7P	346.055262	11.585	Mix2	4	cpd:C00020
2-Hydroxyethanesulfonate	C2H6O4S	124.990857	7.2	Mix2	4	cpd:C05123
Malic acid	C4H6O5	133.0137	11.97	Mix1	4	cpd:C00149
Sucrose	C12H22O11	341.10839	11.93	Mix1	5	cpd:C00089
Taurine	C2H7NO3S	124.006841	11.13	Mix2	4	cpd:C00245
Trehalose	C12H22O11	341.10839	13	Mix2	4	cpd:C01083
METABOLITES_END
#END