#METABOLOMICS WORKBENCH Lxu_20240904_081105 DATATRACK_ID:5162 STUDY_ID:ST003457 ANALYSIS_ID:AN005680 PROJECT_ID:PR002122
VERSION             	1
CREATED_ON             	September 4, 2024, 7:12 pm
#PROJECT
PR:PROJECT_TITLE                 	Early Metabolic Shifts in the Sepsis: Insights from Presymptomatic Patients and
PR:PROJECT_TITLE                 	In Vivo Models.
PR:PROJECT_SUMMARY               	Metabolomics of human serum to support understanding of early metabolic shift in
PR:PROJECT_SUMMARY               	presymptomatic sepsis patients and further confirmed in in vivo model.
PR:INSTITUTE                     	Leibniz Institute for Natural Product Research and Infection Biology Hans Knöll
PR:INSTITUTE                     	Institute
PR:DEPARTMENT                    	Microbiome Dynamics
PR:LAST_NAME                     	XU
PR:FIRST_NAME                    	LINLIN
PR:ADDRESS                       	Beutenbergstraße 11a, Jena, Thuringia, 07745, Germany
PR:EMAIL                         	Lin-Lin.Xu@hki-jena.de
PR:PHONE                         	+4936415321265
#STUDY
ST:STUDY_TITLE                   	Metabolomics of mice heart to support understanding of early metabolic shift in
ST:STUDY_TITLE                   	presymptomatic sepsis patients. (Part3 mice heart metabolite)
ST:STUDY_SUMMARY                 	Sepsis, a life-threatening organ dysfunction caused by a dysregulated response
ST:STUDY_SUMMARY                 	to infection, is a critical medical condition with significant global impact,
ST:STUDY_SUMMARY                 	responsible for 48.9 million cases and 11 million deaths annually. Metabolic
ST:STUDY_SUMMARY                 	dysregulation in sepsis is a critical determinant of disease outcome, yet most
ST:STUDY_SUMMARY                 	studies focus on patients after severe illness onset, comparing septic versus
ST:STUDY_SUMMARY                 	non-septic groups or survivors versus non-survivors. To truly understand this
ST:STUDY_SUMMARY                 	complex disease, it is essential to investigate early metabolic shifts,
ST:STUDY_SUMMARY                 	capturing the transition from simple infection to sepsis. Our untargeted
ST:STUDY_SUMMARY                 	metabolome analysis of heart tissues in mice model revealed early septic
ST:STUDY_SUMMARY                 	biomarkers in heart.
ST:INSTITUTE                     	Leibniz Institute for Natural Product Research and Infection Biology Hans Knöll
ST:INSTITUTE                     	Institute
ST:DEPARTMENT                    	Microbiome Dynamics
ST:LAST_NAME                     	XU
ST:FIRST_NAME                    	LINLIN
ST:ADDRESS                       	Beutenbergstraße 11a, Jena, Thuringia, 07745, Germany
ST:EMAIL                         	Lin-Lin.Xu@hki-jena.de
ST:PHONE                         	+4936415321265
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
#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           	Bl6J_23_0194	SA020913	Sample source:Heart | Group:Sham / 13C-Serin	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-027-SA020913_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-027-SA020913_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0269	SA020994	Sample source:Heart | Group:Sham / 12C-Serin	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-022-SA020994_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-022-SA020994_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0203	SA020966	Sample source:Heart | Group:Sham / 13C-Serin	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-025-SA020966_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-025-SA020966_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0267	SA020980	Sample source:Heart | Group:CLP (24h) / 12C-Serin	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-029-SA020980_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-029-SA020980_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0201	SA020952	Sample source:Heart | Group:Sham / Vehicle	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-021-SA020952_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-021-SA020952_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0271	SA021008	Sample source:Heart | Group:Sham / 12C-Serin	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-019-SA021008_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-019-SA021008_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0199	SA020938	Sample source:Heart | Group:CLP (24h) / 13C-Serin	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-020-SA020938_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-020-SA020938_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0273	SA021022	Sample source:Heart | Group:CLP (24h) / 12C-Serin	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-017-SA021022_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-017-SA021022_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0274	SA021029	Sample source:Heart | Group:CLP (24h) / 12C-Serin	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-013-SA021029_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-013-SA021029_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0268	SA020987	Sample source:Heart | Group:CLP (24h) / 12C-Serin	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-031-SA020987_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-031-SA020987_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0197	SA020927	Sample source:Heart | Group:CLP (24h) / Vehicle	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-018-SA020927_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-018-SA020927_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0204	SA020973	Sample source:Heart | Group:CLP (24h) / Vehicle	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-024-SA020973_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-024-SA020973_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0202	SA020959	Sample source:Heart | Group:Sham / 13C-Serin	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-026-SA020959_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-026-SA020959_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0198	SA020932	Sample source:Heart | Group:Sham / Vehicle	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-033-SA020932_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-033-SA020932_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0200	SA020945	Sample source:Heart | Group:CLP (24h) / 13C-Serin	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-028-SA020945_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-028-SA020945_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0190	SA020892	Sample source:Heart | Group:CLP (24h) / 13C-Serin	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-032-SA020892_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-032-SA020892_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0272	SA021015	Sample source:Heart | Group:Sham / 12C-Serin	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-015-SA021015_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-015-SA021015_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0192	SA020899	Sample source:Heart | Group:CLP (24h) / Vehicle	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-010-SA020899_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-010-SA020899_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0189	SA020885	Sample source:Heart | Group:CLP (24h) / 13C-Serin	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-034-SA020885_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-034-SA020885_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0193	SA020906	Sample source:Heart | Group:Sham / 13C-Serin	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-012-SA020906_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-012-SA020906_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0270	SA021001	Sample source:Heart | Group:Sham / 12C-Serin	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-011-SA021001_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-011-SA021001_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	Bl6J_23_0196	SA020920	Sample source:Heart | Group:Sham / Vehicle	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-014-SA020920_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-014-SA020920_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	NA	Blank_PB	Sample source:blank | Group:Blank	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-045-Blank_PB_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-045-Blank_PB_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	NA	Blank_004	Sample source:blank | Group:Blank	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-044-Blank_004_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-044-Blank_004_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=230704-EXP-PAN-1-044-Blank_004_20230706031901_pos.mzML; RAW_FILE_NAME(Raw file name negative ion repeat1)=230704-EXP-PAN-1-044-Blank_004_20230706031901_neg.mzML
SUBJECT_SAMPLE_FACTORS           	NA	QC_001	Sample source:Heart | Group:QC	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-006-QC_001_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-006-QC_001_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	NA	QC_005	Sample source:Heart | Group:QC	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-016-QC_005_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-016-QC_005_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	NA	Blank_001	Sample source:blank | Group:Blank	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-003-Blank_001_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-003-Blank_001_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	NA	QC_007	Sample source:Heart | Group:QC	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-030-QC_007_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-030-QC_007_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	NA	Blank_002	Sample source:blank | Group:Blank	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-005-Blank_002_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-005-Blank_002_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	NA	Blank_003	Sample source:blank | Group:Blank	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-039-Blank_003_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-039-Blank_003_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	NA	QC_006	Sample source:Heart | Group:QC	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-023-QC_006_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-023-QC_006_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	NA	QC_008	Sample source:Heart | Group:QC	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-035-QC_008_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-035-QC_008_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	NA	QC_004	Sample source:Heart | Group:QC	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-009-QC_004_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-009-QC_004_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
SUBJECT_SAMPLE_FACTORS           	NA	QC_009	Sample source:Heart | Group:QC	RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-041-QC_009_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-041-QC_009_pos.mzML; RAW_FILE_NAME(Raw file name positive ion repeat1)=NA; RAW_FILE_NAME(Raw file name negative ion repeat1)=NA
#COLLECTION
CO:COLLECTION_SUMMARY            	Mice tissue samples were collected from our in vivo mice model and frozen at -80
CO:COLLECTION_SUMMARY            	°C. Samples were shipped to MS-OMICS for metabolomic analysis.
CO:SAMPLE_TYPE                   	Heart
#TREATMENT
TR:TREATMENT_SUMMARY             	Approximately 30% of the cecum was ligated and punctured twice with a 23 Gauge
TR:TREATMENT_SUMMARY             	(G) needle. A small amount of faeces was extruded and the cecum was placed back
TR:TREATMENT_SUMMARY             	into the abdominal cavity. All animals received 40 mL/kg saline s.c. directly
TR:TREATMENT_SUMMARY             	after the procedure and once 25mg/kg s.c. Imipenem/Cilastatin after 6 hours.
TR:TREATMENT_SUMMARY             	This model is associated with a 14-day mortality of 30%, closely mimicking the
TR:TREATMENT_SUMMARY             	clinical situation. Animals were orally gavaged with 500mg/kg 12C-Serin (Sigma)
TR:TREATMENT_SUMMARY             	or 13C-Serin (CLM-1574-H-0.25 Serine-L (13C3 99%13C), Euroisotope, Germany) in
TR:TREATMENT_SUMMARY             	10mL/kg drinking water or drinking water only (vehicle) 6 and 18 hours after
TR:TREATMENT_SUMMARY             	CLP98,99. After 24 hours, the animals are then perfused with ice cold PBS and
TR:TREATMENT_SUMMARY             	organs were harvested, snap frozen and stored at -80° until further analysis.
TR:TREATMENT_SUMMARY             	Sham animals underwent exactly to the same procedures except for the CLP.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	To prepare samples for semi-polar metabolites extraction, the different tissue
SP:SAMPLEPREP_SUMMARY            	samples were weighted and transferred to Eppendorf tubes. Ceramic beads and
SP:SAMPLEPREP_SUMMARY            	precooled methanol/water (1:2) were added. All tubes were placed in a pre-cooled
SP:SAMPLEPREP_SUMMARY            	(-20°C) bead beater and homogenized (4 x 30 sec., 30 Hz) followed by
SP:SAMPLEPREP_SUMMARY            	ultrasonication (5 min). After centrifugation (18000 RCF, 5 min, 4°C), the
SP:SAMPLEPREP_SUMMARY            	supernatant was collected. The sample pellet was reextracted as described above.
SP:SAMPLEPREP_SUMMARY            	The two extract supernatants were pooled and passed through a phosphor removal
SP:SAMPLEPREP_SUMMARY            	cartridge (Phree filter plates). A precise aliquot of the extract was evaporated
SP:SAMPLEPREP_SUMMARY            	to dryness under a gentle stream of nitrogen, before reconstitution with 10%
SP:SAMPLEPREP_SUMMARY            	Eluent B in Eluent A. All Samples were diluted 5 times in mobile phase eluent A
SP:SAMPLEPREP_SUMMARY            	and stable isotope labelled standards were added before analysis. Mouse serum
SP:SAMPLEPREP_SUMMARY            	(50 μl) was diluted with a mixture of acetonitrile and methanol (200 μl 1:1
SP:SAMPLEPREP_SUMMARY            	v/v). The extract was then passed through a phosphor removal cartridge (Phree,
SP:SAMPLEPREP_SUMMARY            	Phenomenex). An aliquot (100 μl) was transferred into a high recovery HPLC vial
SP:SAMPLEPREP_SUMMARY            	and the solvent was removed under a gentle flow of nitrogen. Extracts were
SP:SAMPLEPREP_SUMMARY            	reconstituted in a mobile phase mixture (100 μl, 10%B in 90%A). To ensure
SP:SAMPLEPREP_SUMMARY            	high-quality sample preparation, a quality control sample (QC sample) was
SP:SAMPLEPREP_SUMMARY            	prepared by pooling small equal aliquots from each sample, to create a
SP:SAMPLEPREP_SUMMARY            	representative average of the entire set.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um)
CH:SOLVENT_A                     	100% water; 10 mM ammonium formate; 0.1% formic acid
CH:SOLVENT_B                     	100% methanol; 10 mM ammonium formate; 0.1% formic acid
CH:FLOW_GRADIENT                 	0-2 min: 0 % B, 4 min 35% B, 6-14 min 90% B, 14.1-15 min 0% B.
CH:FLOW_RATE                     	0.3mL/min
CH:COLUMN_TEMPERATURE            	30
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
AN:LABORATORY_NAME               	MS-Omics, Denmark
AN:ANALYSIS_PROTOCOL_FILE        	Doneanu_etal.pdf
#MS
MS:INSTRUMENT_NAME               	Orbitrap Exploris 240
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	Semi-polar metabolite profiling was performed by MS-Omics (Vedbæk, Denmark).
MS:MS_COMMENTS                   	The analysis was carried out using a Vanquish LC (Thermo Scientific) coupled to
MS:MS_COMMENTS                   	a Orbitrap Exploris 240 MS (Thermo Scientific). The UHPLC was performed using an
MS:MS_COMMENTS                   	adapted version of the protocol described by Doneanu et al . An electrospray
MS:MS_COMMENTS                   	ionization interface was used as ionization source. Analysis was performed in
MS:MS_COMMENTS                   	positive and negative ionization mode under polarity switching. Metabolomics
MS:MS_COMMENTS                   	processing was performed untargeted using Compound Discoverer 3.3 (Thermo
MS:MS_COMMENTS                   	Scientific) and Skyline 22.2 (MacCoss Lab Software) for peak picking and feature
MS:MS_COMMENTS                   	grouping, followed by an in-house annotation and curation pipeline written in
MS:MS_COMMENTS                   	MatLab (2022b, MathWorks). Identification of compounds were performed at four
MS:MS_COMMENTS                   	levels; Level 1: identification by retention times (compared against in-house
MS:MS_COMMENTS                   	authentic standards), accurate mass (with an accepted deviation of 3ppm), and
MS:MS_COMMENTS                   	MS/MS spectra, Level 2a: identification by retention times (compared against
MS:MS_COMMENTS                   	in-house authentic standards), accurate mass (with an accepted deviation of
MS:MS_COMMENTS                   	3ppm). Level 2b: identification by accurate mass (with an accepted deviation of
MS:MS_COMMENTS                   	3ppm), and MS/MS spectra, Level 3: identification by accurate mass alone (with
MS:MS_COMMENTS                   	an accepted deviation of 3ppm). The data have been normalized to the QC samples
MS:MS_COMMENTS                   	to remove systematic drift over the analysis sequence. The normalization is
MS:MS_COMMENTS                   	performed using linear regression for the QC’s within each analytical batch
MS:MS_COMMENTS                   	with signal = a*injection number – b. Each value is then normalized with:
MS:MS_COMMENTS                   	normSignal = average(all QC samples)*(signal – injection number*a)/b. This is
MS:MS_COMMENTS                   	done independently for each compound.
MS:MS_RESULTS_FILE               	ST003457_AN005680_Results.txt	UNITS:Normalized peak area	Has m/z:Neutral masses	Has RT:Yes	RT units:Minutes
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Normalized peak area
MS_METABOLITE_DATA_START
Samples	SA020885	SA020892	SA020899	SA020906	SA020913	SA020920	SA020927	SA020932	SA020938	SA020945	SA020952	SA020959	SA020966	SA020973	SA020980	SA020987	SA020994	SA021001	SA021008	SA021015	SA021022	SA021029
Factors	Sample source:Heart | Group:CLP (24h) / 13C-Serin	Sample source:Heart | Group:CLP (24h) / 13C-Serin	Sample source:Heart | Group:CLP (24h) / Vehicle	Sample source:Heart | Group:Sham / 13C-Serin	Sample source:Heart | Group:Sham / 13C-Serin	Sample source:Heart | Group:Sham / Vehicle	Sample source:Heart | Group:CLP (24h) / Vehicle	Sample source:Heart | Group:Sham / Vehicle	Sample source:Heart | Group:CLP (24h) / 13C-Serin	Sample source:Heart | Group:CLP (24h) / 13C-Serin	Sample source:Heart | Group:Sham / Vehicle	Sample source:Heart | Group:Sham / 13C-Serin	Sample source:Heart | Group:Sham / 13C-Serin	Sample source:Heart | Group:CLP (24h) / Vehicle	Sample source:Heart | Group:CLP (24h) / 12C-Serin	Sample source:Heart | Group:CLP (24h) / 12C-Serin	Sample source:Heart | Group:Sham / 12C-Serin	Sample source:Heart | Group:Sham / 12C-Serin	Sample source:Heart | Group:Sham / 12C-Serin	Sample source:Heart | Group:Sham / 12C-Serin	Sample source:Heart | Group:CLP (24h) / 12C-Serin	Sample source:Heart | Group:CLP (24h) / 12C-Serin
Aspartic acid	3194127	8257600	3565857	5783815	2598755	5474726	4869402	8781291	6940603	4257762	6925213	9253670	6046142	4791860	2779856	4064410	9565317	7601916	9403899	3558387	1537074	1569599
Citrulline	2954355.59217538	750981.944720902	920179.765354499	1187708.18336284	669823.33368318	666785.03298736	795772.13931291	1143745.15020581	765462.292892528	736217.15222007	826430.258934598	1088117.68036852	1161884.08545802	1436283.85079404	1582227.99197746	2082980.9186339	1222953.62944925	869301.588949547	1098340.14795001	484172.314218772	1114944.27127952	727889.291034475
Glutamine	60553164	27453650	36910568	52152132	40563760	33839416	38759180	41579056	34516756	31850668	29664204	48806276	42418808	71784872	53341880	67181992	42810696	32981246	39104904	17817124	25272096	26933498
Glutathione	1569938.36970927	1645613.18258347	4220116.14586208	1472044.52937048	856784.344644862	1905092.53078924	2590835.87893214	2394464.51950749	1980923.03661457	3754168.16901896	3065548.22313292	1021803.81202275	127025.113694033	98295.2772859119	106822.861978574	959637.821954571	760329.165408606	451398.738057934	754676.669141905	381134.365789493	178592.6364604	426078.441848416
Hexoses	4933142.08273854	645429.784363229	3658978.98716716	4402484.70365246	469311.039169041	1194722.34659164	13225964.9250885	2261179.09212655	1261649.93745468	6874290.48759732	2949841.12329405	8722454.95957981	6496960.75249785	2608733.18253199	12301897.909226	15546396.2146444	5203191.24486247	1547520.98836379	3522513.69005361	1462170.92251567	1140734.32586681	1216351.3211848
Lactic acid	381181715.600674	161362613.051459	375883376.466261	355003880.387721	122011730.93742	422526861.646716	240153538.011469	428798694.877233	208004549.622317	365943455.369135	288509692.760601	314189202.034327	351993832.715191	265844305.596396	267282938.661409	338411766.193246	450226054.729898	407728881.760418	401342455.443276	151381569.032258	206944911.215926	231285035.354985
Malic acid	54998340	48143736	40136628	74732888	30030604	52179132	31413714	97300784	44669036	42925416	60059928	81693528	62314796	64789548	68771496	76802864	94166296	79473536	75883872	23143242	33667284	42065556
N-Acetylneuraminic acid	508404.63593237	580628.27568965	652898.952418725	472667.403192035	259589.598556719	443217.581604198	557050.381461112	485559.832216101	654384.82702227	457824.363626759	482073.215160534	527891.231335198	589446.48700663	407470.987457496	608407.978648592	739718.228354441	522254.588460066	429860.128645748	575659.091620135	153470.132830151	213529.577632535	203138.806104379
Ornithine	2790735.38993302	752693.734454179	856304.935407388	1020077.53814778	656856.722289784	654667.606178282	780267.289702341	965313.426335271	761668.255257901	754581.745184214	858247.081585002	1029958.06965852	1100253.76134868	1351312.709475	1404151.6777914	1718353.88952626	1252648.06867246	862019.366131415	1009863.0528193	394655.162390401	1036199.60859686	787110.468141323
Ribulose 5-phosphate	17649896.775164202	13676781.01531	16089408.7140854	16705682.352025	8005178.68811737	16054983.0090313	14366312.9310987	17290123.4059972	13662434.5205348	14479976.0119557	12964046.4186109	22263875.5795054	30377761.4354316	25839441.1851351	30133624.2113714	28099659.7308005	22382302.7617953	18969671.7508123	17226880.5427338	7488202.30968239	12908474.2072758	13617366.5176434
Succinic acid	259843.960479367	885167.628551808	243000.322831148	294965.369909343	368955.680843839	475674.469257543	239124.860400902	410754.722837804	704195.060226855	241354.250353213	501942.834448341	604029.978524178	244786.536940913	286756.45138567	356329.456258109	301891.422463851	232831.344744383	162627.671838552	187790.342573688	142909.263196445	129433.157475458	115772.818603137
Taurine	197240000	192885648	193446960	203220624	111787528	215644208	181211712	203823088	194687568	182904928	190671648	225006416	225226160	194583008	201732400	241020672	247222592	217342368	243678992	126445992	152586384	162467808
Threonine	4280083	2460555	2650532	3498190	1381323	2456652	3026617	2193932	2366448	2537221	2307336	2856556	2661759	3555067	2322729	3326048	2370343	1891134	1933867	805454	1254048	1356435
Uracil	1380651.08338571	1870255.37786015	1182345.50327611	1607634.53717363	660281.26226443	1307840.50942172	974807.36224128	1121749.26287642	1601455.53167978	1052581.35489861	1245659.75198914	2288293.75096234	1499711.75774418	1304839.609206	1628200.92681748	1625275.8683348	1614248.12984614	1099719.93055325	1861399.29442407	797266.957721783	991375.205798688	1301423.36078831
Uric acid	1741812.41403756	2149347.70189727	2227873.55654318	1674135.54676395	589206.553220056	1458039.83802131	1517344.70141222	1299109.34598892	2516791.89543615	1862148.29906499	1396211.05854766	1353925.97425435	331008.148737938	829691.164407029	380536.146205267	2014704.63590333	2276558.74681377	1432491.57163113	698411.602994132	997448.279307231	795576.156436958	833323.718429698
Uridine	43039969.2818705	13670676.0884966	24179807.9173788	26932205.9260228	8707576.06757622	17976853.4926035	28761128.0470555	23323772.4892813	14733003.2723101	25491014.1701082	15464030.7653074	36633948.0709802	27436581.623975102	23612118.7985697	37257114.3217715	50603709.5601989	31531710.5903483	19216449.0014867	24191734.1060993	8479241.67070693	13560399.918777	10975948.2839945
Xanthine	8622600	13572537	7904971	9031720	5884739	9314543	6842603	9045490	13768719	7626777	11702587	13800909	10082927	10389795	9074956	10861711	12374976	9531107	13353958	5399935	5304910	5561692
2-C-Methyl-erythritol-4-phosphoric acid	27038375.4130528	3691766.81941367	15116793.0248615	18455001.450381	2068114.19422893	6774765.49006656	42081029.9265904	11626281.7100894	6297660.37788743	26739962.7549561	15377089.0092405	32040663.6570658	25907316.2582093	14513111.4861039	48329169.6370701	56141021.3589994	20933745.992252	7643223.055542	14721616.5050186	7490512.18621817	4923061.89742213	5941201.68647402
2-Hydroxybutyric acid/ 3-Hydroxybutyric acid	4655873	798536	1031875	6038795	1383010	1641968	290528	6628573	720417	916258	1031506	3880773	1041934	14305071	5552848	3209271	1316928	1397553	1345237	682825	1673028	2724037
Allantoin	2069159	736271	654724	934612	327077	403328	459941	978422	732634	540464	597371	905911	791750	1185805	2781750	3672546	672649	503808	827214	438190	846241	513787
Argininosuccinic acid	231260	304026	314254	393167	300023	220844	271596	450618	404937	317053	413986	444904	428268	563280	442110	445757	514591	347716	323869	121694	167120	223492
Fumaric acid_1	2051294	413632	757960	732965	221076	660418	861821	861982	646009	690891	394600	663782	842329	1466683	2083683	2492686	872594	776880	579742	219818	750957	870227
cis-Aconitic acid	2729624	1418012	2314774	1780514	642070	1468947	1216795	1445313	1506244	2417285	1603229	3114287	2840470	2841034	2564558	3023632	2645835	1728503	2343108	840796	1081444	1614631
Fumaric acid_2	587543	565596	455718	868403	399730	606741	406402	1060492	563756	470765	697374	719234	631276	743717	692396	810309	816906	791608	774105	352489	403854	512692
Galactonic acid/ Gluconic acid	7089466	2838364	3042359	3176814	1336263	2968913	2747800	4051463	2329601	2611317	4383852	5549138	3545926	4632362	7017946	11963117	5296717	3068660	4873524	1187851	1438516	1638289
Glutamic acid/ N-Methylaspartic acid	13006336	19691620	16431368	15238150	7264837	20832264	21002682	28860044	22850516	17573362	27223738	31019514	26521144	14460569	14384101	19430566	36377288	29125262	41771632	13221808	5650201	8382089
Glyceraldehyde 3-phosphate	465604	262287	643417	474100	113798	258789	1049074	339183	377272	875934	391302	739840	868014	426524	1138886	1912731	510109	259752	663603	53736	207653	228525
Glycylleucine	2486323	5900280	5351050	3372066	2754675	3315906	3393675	2155089	3531449	3883593	4499589	3208983	559439	578001	703725	1083355	948291	425126	557869	145272	233905	212299
Glyoxylic acid	221657	233761	179021	312700	117374	233650	136554	376643	195901	174465	249744	355997	250512	265509	275420	354264	377531	338487	319721	101328	155078	202667
Hexose dimers	514857	331470	652844	653823	213014	276323	5883491	361598	504634	1441814	353167	369238	911183	467811	2102802	4090140	429495	369331	503889	216127	277892	327078
Isocitric acid	1078405	480869	942401	891412	352159	503049	406015	673322	424681	907554	480032	704440	566371	610449	603764	649165	680964	471073	561670	269808	441517	434233
N-Acetylaspartic acid	848831	410335	567221	572017	262145	504214	346598	346949	382639	588157	696507	760014	529355	769914	596864	663209	869271	231702	998843	220220	272324	458116
O-Phosphorylethanolamine	479928	582778	425443	294954	349794	297601	361363	319725	596286	353580	574690	581910	335990	697298	646864	776513	709350	499400	539231	232936	475660	571955
Pentoses	1733027	2378110	1487686	2119496	1023372	2498687	1557535	2222744	2131420	1526632	2224829	3147281	2317330	2081556	2261633	3012175	3305300	2657813	3178444	1097241	986821	956433
Phenylalanine	842314	1373794	1316640	1039448	579519	918466	1039156	896820	948658	994656	1123316	1015000	956872	871900	763127	1015571	945787	668828	681432	276158	424428	417321
Serine	1551891	1342321	1224337	1759982	921409	2602924	1352498	1009488	1135823	1093351	1638813	2264075	1289001	1105811	1222458	1796385	1876821	1892621	1520793	845395	699408	597461
Threonic acid	863746	293644	212874	291341	77763	367518	276923	344472	318670	179281	383945	407115	411988	361015	1029426	1889933	555232	420476	866470	126858	185034	220202
Thymidine	1594704	1850088	1348946	2317710	865295	1098402	1200780	2100186	1737245	1745371	1202469	1605521	1959229	2018885	1540406	1799598	1522167	1026041	1955965	710922	1040101	1047214
UMP	372554	126347	431477	251493	42912	276316	255483	195365	222184	282184	95102	172091	376799	482076	495324	351370	161380	144420	96402	27776	158507	180599
α-aminobutyric acid/ N,N-Dimethylglycine	527737	802452	628758	567481	257046	815065	832680	1161034	887769	681243	1097766	1254921	1061596	567385	463587	732094	1445510	1187403	1676702	509540	225604	302013
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Neutral_masses	Retention_time
Aspartic acid	133.037508	1.14
Citrulline	175.09564	1.3
Glutamine	146.069142	1.2
Glutathione	307.08375	2.49
Hexoses	180.06339	1.233
Lactic acid	90.03169	2.102
Malic acid	134.021523	1.61
N-Acetylneuraminic acid	309.10596	1.278
Ornithine	132.08991	1.299
Ribulose 5-phosphate	230.01913	1.123
Succinic acid	118.02657	4.301
Taurine	125.014664	1.15
Threonine	119.058243	1.21
Uracil	112.02728	2.512
Uric acid	168.02828	3.792
Uridine	244.0695	4.603
Xanthine	152.033425	4.57
2-C-Methyl-erythritol-4-phosphoric acid	216.04005	1.232
2-Hydroxybutyric acid/ 3-Hydroxybutyric acid	104.047344	4.54
Allantoin	158.04399	1.38
Argininosuccinic acid	290.122634	1.39
Fumaric acid_1	446.060382	1.54
cis-Aconitic acid	174.016438	1.56
Fumaric acid_2	116.010959	2.4
Galactonic acid/ Gluconic acid	196.058303	1.19
Glutamic acid/ N-Methylaspartic acid	147.053158	1.23
Glyceraldehyde 3-phosphate	169.998025	1.12
Glycylleucine	188.116092	5.85
Glyoxylic acid	74.000394	1.61
Hexose dimers	342.116212	1.47
Isocitric acid	192.027003	1.57
N-Acetylaspartic acid	175.048072	2.08
O-Phosphorylethanolamine	141.019095	1.13
Pentoses	150.052824	1.34
Phenylalanine	165.078979	5.58
Serine	105.042593	1.14
Threonic acid	136.037173	1.25
Thymidine	242.090272	5.53
UMP	324.035867	1.89
α-aminobutyric acid/ N,N-Dimethylglycine	103.063329	1.24
METABOLITES_END
#END