#METABOLOMICS WORKBENCH TruxalCarlson_20200909_105704 DATATRACK_ID:2158 STUDY_ID:ST001480 ANALYSIS_ID:AN002456 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                      	POSITIVE
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
(3-Carboxypropyl)trimethylammonium (TMAB)	0.248815281	0.171516943	0.21151051	0.248663501	0.012909635	0.016212709	0.006140422	0.005179659	0.006493089	0.005116949	0.004738335	0.004687017	0.006743508	0.008753397	0.007055501	0.011375511	0.007684584	0.008104792	0.002429566	0.002042382	0.002961815
(Iso)leucine	17.99796092	14.37141543								1.748252771		1.930261678					2.440341229	2.519345807	6.040888566		5.862310876
Adenosyl Homocysteine	0.075340969	0.00720887	0.056844644	0.0154331						0.002136796	0.000963847		0.000395059	0.000285267		0.007369635	0.00067298		0.024005766	0.017384601	0.019844159
Adenosyl Methionine	5.447321351	3.341175962	5.197879483	3.024898497	1.07887326	0.700995948	0.251151901	0.243854912	0.259607181	0.62938532	0.530829929	0.658134347	0.281100462	0.284509484	0.280094036	2.121457126	0.87611646	0.62265043	0.318922329	0.080702918	0.080227925
Alanine	115.3977767	94.52566536	58.4859535	77.59800842	6.594668311	6.149927843	4.190378107	3.881396317	4.537531383	2.59635995	2.560149947	3.006205982	3.54806395	4.375822542	3.715192458	3.664944081	2.894606598	2.706637102	13.72656816	12.24768314	14.53306748
Aminobutyric Acid																					
Arginine	115.5972202	96.93536426	87.23228276	77.37448095	26.72676214	21.06124364	28.53166674	26.42229246	20.44844147	25.13535503	17.47647729	18.15793294	23.32315028	26.10985085	19.96571758	25.98232417	17.35235917	11.78669513	51.95047188	59.21365383	52.54628171
Arsenobetaine	0.00919615	0.000220524																			
Asparagine	11.16800048	10.31249064	5.817056239	9.150491156	2.130991731	1.979705175	1.302906804	1.426576686	1.755055384	2.984956686	2.266126136	2.757567234	1.743684191	2.220634722	1.925736717	4.266018339	4.203986679	3.699470755	2.745259114	2.740621854	2.697907101
Aspartic acid	77.97242283	84.13548785	85.21973243	97.60524638	23.96253576	28.50587439	10.87992612	11.36971993	14.60663184	5.918365941	4.953371121	6.972848571	8.147541293	10.16563525	9.602003074	7.384798467	5.907500085	6.158585274	6.104857135	7.268865684	6.034089768
Betaine	42.07370373	31.0325419	23.73411343	27.21888387	19.71719408	19.99111709	10.66444189	9.778588711	10.36964232	11.41644942	10.0903652	9.663529023	11.42484794	13.52280456	10.82624944	22.0826484	15.99057315	15.50689458			
Betonicine					5.902125275	5.124310869	4.165029347	5.173092182	3.979797768	2.457236438	2.093817754	1.924276234	3.798892514	3.834748842	3.26939037	3.057889915	2.644434825	2.177696303			
Carnitine	0.429307503	0.29900502	1.659347514	1.406841238	0.230726556	0.277300605	0.062509721	0.059065354	0.074462164	0.113313519	0.105881708	0.111605886	0.033155947	0.047108942	0.035919702	0.15278835	0.112332332	0.120558016	0.033296552	0.024774949	0.031500704
Choline	4.770674115	3.272052321	45.66424781	51.78792922	3.951568521	4.634924283	1.022806608	1.059079221	1.073246777	0.436390809	0.373038052	0.465698129	0.773900582	0.922208944	0.631337117		0.453163013	0.449072608	0.067072296	0.041684836	0.043831705
Dimethyl Glycine																					
Dimethylsulfonioacetate (DMS-Ac)	0.0069948	0.004065706	0.015342538	0.008233684	0.467304331	0.471909286	0.070665143	0.061187255	0.065924711	0.123512351	0.106021	0.141887905	0.076655597	0.088788568	0.083263328	0.253063481	0.178426824	0.157361697			
DMSP	89.8777507	68.97266464	140.9467369	153.3157991	88.37325556	88.67025049	37.95663725	37.58829396	38.30212743	23.96755717	21.0584327	22.52772464	30.92430756	37.99172308	30.0755893	43.26257981	30.8138578	31.29516834			
Ectoine	0.01700636	0.016427704	0.007241716	0.008813689	0.001471897	0.001509537	0.000633046	0.00063142	0.000719141	0.000498341	0.000443044	0.000531398	0.000581219	0.000707807	0.000630729	0.000602925	0.000570238	0.000672207	0.001983513	0.002212526	0.002111085
Glucosylglycerol	2.50511671	1.615387733	5.388097342	7.018704633	34.32912157	34.01752345	6.259840228	5.967978802	5.833165142	4.15798683	4.499687999	4.072855777	6.608330458	8.206972403	7.102185871	6.189157499	4.59088179	5.548703417	2.218960031	1.700842918	2.004095066
Glutamic acid	302.0915854	283.7047477	334.0308693	458.8433828	54.57660205	39.64272683	22.90730634	22.46969517	21.72099184	18.72658199	12.88193096	16.19762727	15.61010467	17.57289255	13.72664966	19.13220893	15.08138564	11.82588916	23.6734599	19.80395469	22.58672285
Glutamine	37.66792161	24.32588963	63.28293945	80.81126053	13.35272272	14.45864878	6.126812368	5.695553759	7.844001597	7.768276843	6.789113502	6.713672788	6.018329509	7.202774702	7.220147778	8.210710111	7.068063323	7.127979069	7.92899374	6.22474725	7.233120994
Glycine	2.312670949	2.84999908	2.115711918	3.898350633	0.622083013	0.561763864	0.304464724	0.324036964	0.327732138	0.331086644	0.265585847	0.304253743	0.408906704	0.39128665	0.293033762	0.472216934	0.44709393	0.447376412	1.048009077	1.344349548	1.18601216
Gonyol							0.025207463	0.028342237	0.025346081	0.148702315	0.139213007	0.145548754	0.018689939	0.023795484	0.021106855	0.089444585	0.078051846	0.080833388			
Histidine	2.409738267	2.032875845	1.684686552	2.24359818	0.473453798	0.418023972	0.25147613	0.23692709	0.265473763	0.320857088	0.323532851	0.360210055	0.339005533	0.325008923	0.270828863	0.717360092	0.552913649	0.530830155	0.700732122		0.642637717
Homarine	862.4923308	607.932279	769.6164364	891.0053175	1.887391004	0.650735595				0.033501585	0.012899952	0.041835906	0.016509337	0.015295869	0.029108126	0.021179169	0.022500066	0.024914314			
Homoserine	20.84894987	14.03313276	13.98934095	17.59618644	0.950979511	1.068829361	0.477683081	0.455407817	0.503873199	0.192017224	0.164866499	0.162013411	0.612250067	0.746016956	0.613334974	0.197183856	0.173228022	0.151110414			
Hydroxyectoine					0.00031965	0.000304142	9.69E-05	0.00011249	8.13E-05	8.82E-05	8.04E-05	9.61E-05							8.95E-05	8.04E-05	6.67E-05
Hypotaurine							0.035485301	0.042888439	0.040738839	0.282728821	0.221173046	0.2959834	0.029079225	0.032511409	0.03879985	0.293127136	0.320667948	0.225599825			
Lysine	165.0799719	150.0720932	12.02951646	10.78352973	2.839971135	2.436932997	4.005434477	3.739616272	3.245282109	4.072976772	3.378098117	3.600703221	4.261423804	4.601022557	3.161398756	5.886349176	3.772776569	2.503819484	1.839436502	1.95621987	1.57585151
Methionine	23.75027903	19.22067639	5.296842663	6.991325682	0.359729913	0.31624283	0.149983521	0.150381896	0.157834478	0.111676006	0.091893264	0.108254152	0.162060413	0.218354868	0.201307871	0.135628181	0.108211568	0.104946958	0.290334469	0.280916427	0.234085448
N(e)-Acetyl-Lysine	0.074005409	0.047423564	0.037795946	0.042315962	0.006143427	0.006047415	0.008451573	0.007560621	0.01106451	0.025666574	0.018678883	0.037288553	0.008731437	0.011815273	0.013596832	0.074543768	0.041408519	0.039930865	0.011883518	0.005309388	0.003776951
Ornithine	608.5175125	761.8505989	45.42360635	40.60684423	5.076173009	3.55842725	3.065552103	2.98860457	2.651858049	3.15816805	2.90760239	2.132451257	2.909561933	2.957590214	2.033273698	2.318729867	1.603262813	1.377854968	9.711387222	10.01795102	8.782030411
Proline	142.1000007	109.3283207	507.0386759	746.2533583	49.98175468	48.73741274	15.97365495	18.09035288	18.09414198	10.18484278	9.18333706	12.47604754	36.74672403	47.20485391	37.64533372	34.8523381	25.44475295	24.63783559	96.31382043	75.32686382	85.55505973
Sarcosine	1.99053822	1.371358684	0.734677767	1.141674546	0.093405965	0.119922698	0.030388539	0.037313644	0.040109894	0.066077861	0.061943533	0.063684924	0.05033598	0.067030882	0.050905221	0.06888162	0.084967836	0.068747847			
Serine	15.82638061	15.21094623	7.989346215	12.40826212	2.185259313	2.044081131	0.979578519	0.937879934	1.058747599	0.906645323	0.806381107	0.838532335	1.087841344	0.972108704	0.788941017	1.048539513	0.961472333	0.932628764	1.262154275	1.190246846	1.180419837
Stachydrine hydrochloride	0.007058172	0.005377246	0.007190518	0.007845373	0.001739727	0.002018675				0.000865559	0.000776554		0.001043972		0.000952532	0.001483818	0.001594551				
Threonine	24.02459536	11.72196338	18.03336544	12.32421931	1.511483459	0.795405283	0.485983405	0.41712759	0.320408678	0.441345053	0.395347699	0.284526693	0.474624292	0.443974189	0.260200106	0.893273767	0.458171573	0.338956682	0.644885831	0.485735728	0.575301256
trans Hydroxyl proline	0.323084956	0.218096316	0.617123473	0.655679767	0.061375604	0.072788566	0.027429782	0.028414903	0.029797436	0.026860999	0.025547819	0.028767643	0.066840091	0.081426528	0.065185607	0.092149503	0.067301717	0.069403075	0.241290814	0.163394615	0.212167865
Trigonelline					0.00573808	0.005801546							0.002169511	0.001654133	0.001794472	0.004108829	0.003206721	0.002107947			
Trimethylamine N-oxide	0.042354075	0.034632872																			
Trimethylammonium Propionate (TMAP)					0.363460627	0.377091736	0.045608564	0.049451994	0.048597982	0.047535738	0.041348011	0.046095041	0.027409411	0.032709761	0.031731604	0.042605032	0.04893376	0.042479758			
Tyrosine	35.01637628	30.52153879	1.014429595	1.175633707	0.589623025	0.423731929	0.12870241	0.129554987	0.146958989	0.963184436	0.764916229	1.030313422	0.113933296	0.107692157	0.094317329	1.515959192	1.579190821	1.511412055	0.143140853	0.100319436	0.099308939
Valine																					
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	quantitated m/z	KEGG_ID	CHEBI	KEGGNAME	MS_method
(3-Carboxypropyl)trimethylammonium (TMAB)	146.118104		CHEBI:1941		HILIC_QE_Pos
(Iso)leucine	132.102454	C00407	CHEBI:17191	L-Isoleucine; 2-Amino-3-methylvaleric acid	HILIC_QE_Pos
Adenosyl Homocysteine	385.129416	C00021	CHEBI:16680	S-Adenosyl-L-homocysteine; S-Adenosylhomocysteine	HILIC_QE_Pos
Adenosyl Methionine	399.145066	C00019	CHEBI:15414	S-Adenosyl-L-methionine; S-Adenosylmethionine; AdoMet; SAM	HILIC_QE_Pos
Alanine	90.055504	C00041	CHEBI:16977	L-Alanine; L-2-Aminopropionic acid; L-alpha-Alanine	HILIC_QE_Pos
Aminobutyric Acid	104.071154	C00334	CHEBI:16865	4-Aminobutanoate; 4-Aminobutanoic acid; 4-Aminobutyrate; 4-Aminobutyric acid; gamma-Aminobutyric acid; GABA	HILIC_QE_Pos
Arginine	175.119501	C00062	CHEBI:16467	L-Arginine; (S)-2-Amino-5-guanidinovaleric acid; L-Arg	HILIC_QE_Pos
Arsenobetaine	179.005326	C19331	CHEBI:82392	Arsenobetaine	HILIC_QE_Pos
Asparagine	133.061318	C00152	CHEBI:17196	L-Asparagine; 2-Aminosuccinamic acid	HILIC_QE_Pos
Aspartic acid	134.045334	C00049	CHEBI:17053	L-Aspartate; L-Aspartic acid; 2-Aminosuccinic acid; L-Asp	HILIC_QE_Pos
Betaine	118.086804	C00719	CHEBI:17750	Betaine; Trimethylaminoacetate; Glycine betaine; N,N,N-Trimethylglycine; Trimethylammonioacetate	HILIC_QE_Pos
Betonicine	160.097369	C08269	CHEBI:85533	(-)-Betonicine; Betonicine; trans-4-Hydroxy-L-proline betaine	HILIC_QE_Pos
Carnitine	162.113019	C00487	CHEBI:17126	Carnitine; gamma-Trimethyl-hydroxybutyrobetaine; 3-Hydroxy-4-trimethylammoniobutanoate	HILIC_QE_Pos
Choline	104.107539	C00114	CHEBI:15354	Choline; Bilineurine	HILIC_QE_Pos
Dimethyl Glycine	104.071154	C01026	CHEBI:17724	N,N-Dimethylglycine; Dimethylglycine	HILIC_QE_Pos
Dimethylsulfonioacetate (DMS-Ac)	121.032327	C03392	CHEBI:4623	Dimethylsulfonioacetate	HILIC_QE_Pos
DMSP	135.047977	C04022	CHEBI:16457	S,S-Dimethyl-beta-propiothetin; S-Dimethylsulfonium propionic acid; Dimethylpropiothetin; DMPT; DMSP	HILIC_QE_Pos
Ectoine	143.082053	C06231	CHEBI:27592	Ectoine; L-Ectoine	HILIC_QE_Pos
Glucosylglycerol	255.107995	C11546	CHEBI:82766	2-O-(alpha-D-Glucopyranosyl)glycerol; 2-O-alpha-D-Glucosylglycerol	HILIC_QE_Pos
Glutamic acid	148.060984	C00025	CHEBI:16015	L-Glutamate; L-Glutamic acid; L-Glutaminic acid; Glutamate	HILIC_QE_Pos
Glutamine	147.076968	C00064	CHEBI:18050	L-Glutamine; L-2-Aminoglutaramic acid	HILIC_QE_Pos
Glycine	76.039854	C00037	CHEBI:15428	Glycine; Aminoacetic acid; Gly	HILIC_QE_Pos
Gonyol	179.074191				HILIC_QE_Pos
Histidine	156.077302	C00135	CHEBI:15971	L-Histidine; (S)-alpha-Amino-1H-imidazole-4-propionic acid	HILIC_QE_Pos
Homarine	138.055503		CHEBI:69061		HILIC_QE_Pos
Homoserine	120.066069	C00263	CHEBI:15699	L-Homoserine; 2-Amino-4-hydroxybutyric acid	HILIC_QE_Pos
Hydroxyectoine	159.076968	C16432	CHEBI:49432	5-Hydroxyectoine	HILIC_QE_Pos
Hypotaurine	110.027576	C00519	CHEBI:16668	Hypotaurine; 2-Aminoethanesulfinic acid	HILIC_QE_Pos
Lysine	147.113353	C00047	CHEBI:18019	L-Lysine; Lysine acid; 2,6-Diaminohexanoic acid	HILIC_QE_Pos
Methionine	150.058876	C00073	CHEBI:16811	L-Methionine; Methionine; L-2-Amino-4methylthiobutyric acid	HILIC_QE_Pos
N(e)-Acetyl-Lysine	189.123918	C02727	CHEBI:17752	N6-Acetyl-L-lysine	HILIC_QE_Pos
Ornithine	133.097703	C00077	CHEBI:15729	L-Ornithine; (S)-2,5-Diaminovaleric acid; (S)-2,5-Diaminopentanoic acid; (S)-2,5-Diaminopentanoate	HILIC_QE_Pos
Proline	116.071154	C00148	CHEBI:17203	L-Proline; 2-Pyrrolidinecarboxylic acid	HILIC_QE_Pos
Sarcosine	90.055504	C00213	CHEBI:15611	Sarcosine; N-Methylglycine	HILIC_QE_Pos
Serine	106.050419	C00065	CHEBI:17115	L-Serine; L-2-Amino-3-hydroxypropionic acid; L-3-Hydroxy-alanine; Serine	HILIC_QE_Pos
Stachydrine hydrochloride	144.102454	C10172	CHEBI:35280	Stachydrine; Proline betaine; L-Proline betaine	HILIC_QE_Pos
Threonine	120.066069	C00188	CHEBI:16857	L-Threonine; 2-Amino-3-hydroxybutyric acid	HILIC_QE_Pos
trans Hydroxyl proline	132.066069	C01157	CHEBI:18095	Hydroxyproline; L-Hydroxyproline; trans-4-Hydroxy-L-proline	HILIC_QE_Pos
Trigonelline	138.055503	C01004	CHEBI:18123	N-Methylnicotinate; Trigonelline; Trigenelline; 1-Methylpyridinio-3-carboxylate; Betaine nicotinate; Caffearin; Gynesine	HILIC_QE_Pos
Trimethylamine N-oxide	76.076239	C01104	CHEBI:15724	Trimethylamine N-oxide; (CH3)3NO	HILIC_QE_Pos
Trimethylammonium Propionate (TMAP)	132.1018	C08263	CHEBI:28825	beta-Alaninebetaine; beta-Alanine betaine; Propiobetaine; Trimethylalanine	HILIC_QE_Pos
Tyrosine	182.081719	C00082	CHEBI:17895	L-Tyrosine; (S)-3-(p-Hydroxyphenyl)alanine; (S)-2-Amino-3-(p-hydroxyphenyl)propionic acid; Tyrosine	HILIC_QE_Pos
Valine	118.086804	C00183	CHEBI:16414	L-Valine; 2-Amino-3-methylbutyric acid	HILIC_QE_Pos
METABOLITES_END
#END