#METABOLOMICS WORKBENCH dhananjay_shinde5_20240914_131224 DATATRACK_ID:5191 STUDY_ID:ST003482 ANALYSIS_ID:AN005717 PROJECT_ID:PR002139
VERSION             	1
CREATED_ON             	September 14, 2024, 3:09 pm
#PROJECT
PR:PROJECT_TITLE                 	Tissue niche influences immune and metabolic profiles to Staphylococcus aureus
PR:PROJECT_TITLE                 	biofilm infection
PR:PROJECT_TYPE                  	Intracellular data
PR:PROJECT_SUMMARY               	Infection is a devastating post-surgical complication, often requiring
PR:PROJECT_SUMMARY               	additional procedures and prolonged antibiotic therapy. This is especially
PR:PROJECT_SUMMARY               	relevant for craniotomy and prosthetic joint infections (PJI), both of which are
PR:PROJECT_SUMMARY               	characterized by biofilm formation on the bone or implant surface, respectively,
PR:PROJECT_SUMMARY               	with S. aureus representing a primary cause. The local tissue microenvironment
PR:PROJECT_SUMMARY               	likely has profound effects on immune attributes that can influence treatment
PR:PROJECT_SUMMARY               	efficacy, which becomes critical to consider when developing therapeutics for
PR:PROJECT_SUMMARY               	biofilm infections. However, the extent to which distinct tissue niches
PR:PROJECT_SUMMARY               	influence immune function during biofilm development remains relatively unknown.
PR:PROJECT_SUMMARY               	To address this, we compare the metabolomic, transcriptomic, and functional
PR:PROJECT_SUMMARY               	attributes of leukocytes in mouse models of S. aureus craniotomy and PJI
PR:PROJECT_SUMMARY               	complemented with patient samples from both infection modalities, which reveals
PR:PROJECT_SUMMARY               	profound tissue niche-dependent differences in nucleic acid, amino acid, and
PR:PROJECT_SUMMARY               	lipid metabolism with links to immune modulation. These signatures are both
PR:PROJECT_SUMMARY               	spatially and temporally distinct, differing not only between infection sites
PR:PROJECT_SUMMARY               	but evolving over time within a single model. Collectively, this demonstrates
PR:PROJECT_SUMMARY               	that biofilms elicit unique immune and metabolic responses that are heavily
PR:PROJECT_SUMMARY               	influenced by the local tissue microenvironment, which will likely have
PR:PROJECT_SUMMARY               	important implications when designing therapeutic approaches targeting these
PR:PROJECT_SUMMARY               	infections.
PR:INSTITUTE                     	UNMC
PR:DEPARTMENT                    	Department of pathology, microbiology and immunology
PR:LABORATORY                    	Pathology, Microbiology and Immunology
PR:LAST_NAME                     	Dhananjay
PR:FIRST_NAME                    	Shinde
PR:ADDRESS                       	DRC 2, 7066, UNMC, Emily st
PR:EMAIL                         	dhananjay.shinde@unmc.edu
PR:PHONE                         	4025597623
#STUDY
ST:STUDY_TITLE                   	Tissue niche influences immune and metabolic profiles to Staphylococcus aureus
ST:STUDY_TITLE                   	biofilm infection
ST:STUDY_SUMMARY                 	Infection is a devastating post-surgical complication, often requiring
ST:STUDY_SUMMARY                 	additional procedures and prolonged antibiotic therapy. This is especially
ST:STUDY_SUMMARY                 	relevant for craniotomy and prosthetic joint infections (PJI), both of which are
ST:STUDY_SUMMARY                 	characterized by biofilm formation on the bone or implant surface, respectively,
ST:STUDY_SUMMARY                 	with S. aureus representing a primary cause. The local tissue microenvironment
ST:STUDY_SUMMARY                 	likely has profound effects on immune attributes that can influence treatment
ST:STUDY_SUMMARY                 	efficacy, which becomes critical to consider when developing therapeutics for
ST:STUDY_SUMMARY                 	biofilm infections. However, the extent to which distinct tissue niches
ST:STUDY_SUMMARY                 	influence immune function during biofilm development remains relatively unknown.
ST:STUDY_SUMMARY                 	To address this, we compare the metabolomic, transcriptomic, and functional
ST:STUDY_SUMMARY                 	attributes of leukocytes in mouse models of S. aureus craniotomy and PJI
ST:STUDY_SUMMARY                 	complemented with patient samples from both infection modalities, which reveals
ST:STUDY_SUMMARY                 	profound tissue niche-dependent differences in nucleic acid, amino acid, and
ST:STUDY_SUMMARY                 	lipid metabolism with links to immune modulation. These signatures are both
ST:STUDY_SUMMARY                 	spatially and temporally distinct, differing not only between infection sites
ST:STUDY_SUMMARY                 	but evolving over time within a single model. Collectively, this demonstrates
ST:STUDY_SUMMARY                 	that biofilms elicit unique immune and metabolic responses that are heavily
ST:STUDY_SUMMARY                 	influenced by the local tissue microenvironment, which will likely have
ST:STUDY_SUMMARY                 	important implications when designing therapeutic approaches targeting these
ST:STUDY_SUMMARY                 	infections.
ST:INSTITUTE                     	UNMC
ST:DEPARTMENT                    	Department of pathology, microbiology and immunology
ST:LAST_NAME                     	Shinde
ST:FIRST_NAME                    	Dhananjay
ST:ADDRESS                       	DRC 2, 7066, UNMC, Emily st
ST:EMAIL                         	dhananjay.shinde@unmc.edu
ST:PHONE                         	4025597623
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:GENDER                        	Male and female
#FACTORS
#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           	-	d3-Crain_1	Sample source:Galea | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_1.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-Crain_2	Sample source:Galea | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_2.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-Crain_3	Sample source:Galea | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_3.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-Crain_4	Sample source:Galea | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_4.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-Crain_5	Sample source:Galea | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_5.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-PJI_1	Sample source:Tissue (PJI) | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_1.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-PJI_2	Sample source:Tissue (PJI) | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_2.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-PJI_3	Sample source:Tissue (PJI) | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_3.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-PJI_4	Sample source:Tissue (PJI) | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_4.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-PJI_5	Sample source:Tissue (PJI) | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_5.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-Crain_1	Sample source:Galea | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_1.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-Crain_2	Sample source:Galea | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_2.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-Crain_3	Sample source:Galea | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_3.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-Crain_4	Sample source:Galea | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_4.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-Crain_5	Sample source:Galea | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_5.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-PJI_1	Sample source:Tissue (PJI) | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_1.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-PJI_2	Sample source:Tissue (PJI) | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_2.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-PJI_3	Sample source:Tissue (PJI) | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_3.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-PJI_4	Sample source:Tissue (PJI) | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_4.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-PJI_5	Sample source:Tissue (PJI) | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_5.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-Crain_1	Sample source:Galea | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_1.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-Crain_2	Sample source:Galea | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_2.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-Crain_3	Sample source:Galea | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_3.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-Crain_4	Sample source:Galea | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_4.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-Crain_5	Sample source:Galea | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_5.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-PJI_1	Sample source:Tissue (PJI) | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_1.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-PJI_2	Sample source:Tissue (PJI) | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_2.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-PJI_3	Sample source:Tissue (PJI) | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_3.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-PJI_4	Sample source:Tissue (PJI) | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_4.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-PJI_5	Sample source:Tissue (PJI) | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_5.mzXML
#COLLECTION
CO:COLLECTION_SUMMARY            	The intracellular metabolome of granulocytes isolated from the galea and PJI
CO:COLLECTION_SUMMARY            	tissue of mouse craniotomy and PJI models, respectively, were profiled by
CO:COLLECTION_SUMMARY            	LC-HRMS. Fifteen mice were infected for each biofilm model (craniotomy and PJI)
CO:COLLECTION_SUMMARY            	with single-cell suspensions prepared by pooling tissue homogenates from 3 mice
CO:COLLECTION_SUMMARY            	into one sample, resulting in 5 biological replicates. Samples were then
CO:COLLECTION_SUMMARY            	enriched for granulocytes using Ly6G magnetic beads (Miltenyi Biotec, Bergishch
CO:COLLECTION_SUMMARY            	Gladbach, Germany). Magnetic beads were utilized for purification instead of
CO:COLLECTION_SUMMARY            	FACS to limit alterations in cellular metabolism that may occur with extended
CO:COLLECTION_SUMMARY            	processing times. Samples were washed twice in 1X PBS, resuspended in 80% MetOH,
CO:COLLECTION_SUMMARY            	and frozen at -80°C for 15 min. Metabolomics was also performed on supernatants
CO:COLLECTION_SUMMARY            	prepared from brain, galea, and PJI tissue homogenates at days 3, 7, and 14
CO:COLLECTION_SUMMARY            	post-infection. Similar to granulocyte intracellular metabolite collection,
CO:COLLECTION_SUMMARY            	tissue supernatants were centrifuged at high speed to remove residual cellular
CO:COLLECTION_SUMMARY            	material, combined with 100% MetOH to yield an 80% MetOH solution, and frozen at
CO:COLLECTION_SUMMARY            	-80°C.
CO:SAMPLE_TYPE                   	Extracellular infection sites homogenate, intracellular granulocyte
#TREATMENT
TR:TREATMENT_SUMMARY             	All mice were group-housed at 21-23°C (22°C average) and 30-70% humidity (55%
TR:TREATMENT_SUMMARY             	average) under 12 h light/dark cycle with free access to food (2019S Teklad
TR:TREATMENT_SUMMARY             	Global 19% Protein Extruded Rodent Diet; Inotiv, West Lafayette, IN) and water.
TR:TREATMENT_SUMMARY             	S. aureus craniotomy infection was established in male and female 8- to
TR:TREATMENT_SUMMARY             	10-week-old C57BL/6J mice (RRID:IMSR_JAX:000664). Briefly, ketamine and xylazine
TR:TREATMENT_SUMMARY             	were administered to achieve a surgical plane of anesthesia before skin
TR:TREATMENT_SUMMARY             	disinfection with betadine. A midline incision of the scalp was made, whereupon
TR:TREATMENT_SUMMARY             	a bone flap was created using a high-speed pneumatic drill (Stryker Corporation,
TR:TREATMENT_SUMMARY             	Kalamazoo, MI). The bone flap was incubated with 0.5 mL of S. aureus USA300
TR:TREATMENT_SUMMARY             	LAC13C diluted to 10^6/mL in brain-heart infusion broth at 37°C for 5 min and
TR:TREATMENT_SUMMARY             	subsequently rinsed in PBS, blotted dry on a sterile field, and reimplanted.
TR:TREATMENT_SUMMARY             	This procedure reliably produces an infectious inoculum of 10^3 CFUs per bone
TR:TREATMENT_SUMMARY             	flap, which more accurately models surgical site infection that progresses to
TR:TREATMENT_SUMMARY             	biofilm formation. The scalp incision was closed using 6-0 nylon suture and
TR:TREATMENT_SUMMARY             	animals were continuously monitored until they regained ambulation, and daily
TR:TREATMENT_SUMMARY             	thereafter. S. aureus PJI was established in male and female 8- to 10-week-old
TR:TREATMENT_SUMMARY             	C57BL/6J mice. Mice were anesthetized with ketamine and xylazine prior to skin
TR:TREATMENT_SUMMARY             	disinfection with betadine and initial incision of the quadriceps. A secondary
TR:TREATMENT_SUMMARY             	incision was made to laterally displace the patellar tendon and a burr hole was
TR:TREATMENT_SUMMARY             	created in the distal end of the femur with a 26-gauge needle. An
TR:TREATMENT_SUMMARY             	orthopedic-grade nickel-titanium Kirschner wire (0.6 mm diameter; Custom Wire
TR:TREATMENT_SUMMARY             	Technologies, Port Washington, WI) was inserted through the burr hole into the
TR:TREATMENT_SUMMARY             	medullary canal and 10^3 CFUs of S. aureus USA300 LAC13C were inoculated at the
TR:TREATMENT_SUMMARY             	tip of the wire and the incision was closed with nylon suture. Mice received
TR:TREATMENT_SUMMARY             	buprenorphine SR to alleviate any post-surgical pain and were continuously
TR:TREATMENT_SUMMARY             	monitored until ambulatory and daily thereafter.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	A 13C15N-labeled canonical amino acid (13C15N-CAA) mix (Cambridge Isotope
SP:SAMPLEPREP_SUMMARY            	Laboratories, Andover, MA) was added to the extraction buffer comprised of 80%
SP:SAMPLEPREP_SUMMARY            	MeOH as an internal standard. Samples were then pelleted, and supernatants were
SP:SAMPLEPREP_SUMMARY            	dried by vacuum centrifuge for storage at -80°C. Cell pellets were stored for
SP:SAMPLEPREP_SUMMARY            	protein quantification to normalize metabolite peak areas. Prior to LC-HRMS,
SP:SAMPLEPREP_SUMMARY            	samples were reconstituted in 100 µL of 50% MetOH, pelleted, and the resulting
SP:SAMPLEPREP_SUMMARY            	supernatant was analyzed.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Metabolite separation was performed by liquid chromatography using a XBridge
CH:CHROMATOGRAPHY_SUMMARY        	Amide analytical column (150 × 2.1mm ID; 1.7µm particle size; Waters
CH:CHROMATOGRAPHY_SUMMARY        	Corporation, Milford, MA) and a binary solvent system infused at a flow rate of
CH:CHROMATOGRAPHY_SUMMARY        	0.3 ml/min. A guard XBridge Amide column (20 × 2.1mm ID; 3.5µm particle
CH:CHROMATOGRAPHY_SUMMARY        	size; Waters Corporation) was connected in front of the analytical column.
CH:CHROMATOGRAPHY_SUMMARY        	Mobile phase A was composed of ammonium acetate and ammonium hydroxide (10 mM
CH:CHROMATOGRAPHY_SUMMARY        	each) containing 5% acetonitrile in LC-MS grade water; mobile phase B was 100%
CH:CHROMATOGRAPHY_SUMMARY        	LC-MS grade acetonitrile. The pH of mobile phase A was adjusted to 8.0 using
CH:CHROMATOGRAPHY_SUMMARY        	glacial acetic acid. The UHPLC pumps were operated in gradient mode. The amide
CH:CHROMATOGRAPHY_SUMMARY        	column was maintained at 40°C, and the autosampler temperature was held at 5°C
CH:CHROMATOGRAPHY_SUMMARY        	throughout data acquisition. The injection volume for all samples was 5 µl with
CH:CHROMATOGRAPHY_SUMMARY        	the 13C15N-CAA mix as the internal standard.
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Waters ACQUITY UPLC BEH Amide (150 x 2.1mm,1.7um)
CH:SOLVENT_A                     	ammonium acetate and ammonium hydroxide (10 mM each) containing 5% acetonitrile
CH:SOLVENT_A                     	in LC-MS grade water
CH:SOLVENT_B                     	acetonitrile
CH:FLOW_GRADIENT                 	B% at initial 85%
CH:FLOW_RATE                     	0.3ml/min
CH:COLUMN_TEMPERATURE            	40
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Orbitrap Exploris 480
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	A HRMS Orbitrap (Exploris 480; Thermo Fisher Scientific) was operated in
MS:MS_COMMENTS                   	polarity switching mode and was used for untargeted metabolomics in a
MS:MS_COMMENTS                   	data-dependent MS/MS acquisition mode (DDA). Electrospray ionization (ESI)
MS:MS_COMMENTS                   	parameters were optimized as follows: electrospray ion voltage of -2700V and
MS:MS_COMMENTS                   	3500V in negative and positive mode respectively, ion transfer tube temperature
MS:MS_COMMENTS                   	was maintained at 400°C, and m/z scan range was 70-1050 Da. Orbitrap resolution
MS:MS_COMMENTS                   	for precursor ion scans were maintained at 240,000 for intracellular and 120,000
MS:MS_COMMENTS                   	for extracellular whereas fragment ion scans were maintained at 120,000 and
MS:MS_COMMENTS                   	60,000, for intracellular and extracellular respectively. Normalized collision
MS:MS_COMMENTS                   	energies at 30, 50, and 150% were used for fragmentation and data was acquired
MS:MS_COMMENTS                   	in profile mode.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Relative intensities
MS_METABOLITE_DATA_START
Samples	d3-Crain_1	d3-Crain_2	d3-Crain_3	d3-Crain_4	d3-Crain_5	d3-PJI_1	d3-PJI_2	d3-PJI_3	d3-PJI_4	d3-PJI_5	d7-Crain_1	d7-Crain_2	d7-Crain_3	d7-Crain_4	d7-Crain_5	d7-PJI_1	d7-PJI_2	d7-PJI_3	d7-PJI_4	d7-PJI_5	d14-Crain_1	d14-Crain_2	d14-Crain_3	d14-Crain_4	d14-Crain_5	d14-PJI_1	d14-PJI_2	d14-PJI_3	d14-PJI_4	d14-PJI_5
Factors	Sample source:Galea | Experimental Variable:d3	Sample source:Galea | Experimental Variable:d3	Sample source:Galea | Experimental Variable:d3	Sample source:Galea | Experimental Variable:d3	Sample source:Galea | Experimental Variable:d3	Sample source:Tissue (PJI) | Experimental Variable:d3	Sample source:Tissue (PJI) | Experimental Variable:d3	Sample source:Tissue (PJI) | Experimental Variable:d3	Sample source:Tissue (PJI) | Experimental Variable:d3	Sample source:Tissue (PJI) | Experimental Variable:d3	Sample source:Galea | Experimental Variable:d7	Sample source:Galea | Experimental Variable:d7	Sample source:Galea | Experimental Variable:d7	Sample source:Galea | Experimental Variable:d7	Sample source:Galea | Experimental Variable:d7	Sample source:Tissue (PJI) | Experimental Variable:d7	Sample source:Tissue (PJI) | Experimental Variable:d7	Sample source:Tissue (PJI) | Experimental Variable:d7	Sample source:Tissue (PJI) | Experimental Variable:d7	Sample source:Tissue (PJI) | Experimental Variable:d7	Sample source:Galea | Experimental Variable:d14	Sample source:Galea | Experimental Variable:d14	Sample source:Galea | Experimental Variable:d14	Sample source:Galea | Experimental Variable:d14	Sample source:Galea | Experimental Variable:d14	Sample source:Tissue (PJI) | Experimental Variable:d14	Sample source:Tissue (PJI) | Experimental Variable:d14	Sample source:Tissue (PJI) | Experimental Variable:d14	Sample source:Tissue (PJI) | Experimental Variable:d14	Sample source:Tissue (PJI) | Experimental Variable:d14
(-)-Chloramphenicol	1.07E+04	2.71E+04	1.12E+04	3.83E+03	9.52E+03	1.77E+04	1.38E+04	1.22E+04	1.12E+04	3.28E+03	4.19E+03	4.37E+04	3.51E+03	6.20E+03	4.73E+03	4.34E+03	1.52E+04	6.74E+03	2.38E+03	2.69E+04	5.41E+05	5.08E+04	1.68E+05	2.40E+06	4.94E+05	3.31E+03	3.01E+03	1.37E+04	5.44E+03	7.70E+03
(-)-pinellic acid	5.61E+04	3.23E+04	3.18E+04	1.49E+04	2.92E+04	7.40E+04	4.23E+04	6.06E+04	5.98E+04	2.50E+04	1.64E+04	5.77E+04	8.23E+03	3.63E+04	4.40E+04	3.65E+04	5.91E+04	3.80E+04	4.76E+04	1.60E+05	2.13E+05	3.58E+04	6.10E+04	6.09E+04	1.03E+05	3.47E+04	2.84E+04	8.89E+04	5.66E+04	1.40E+04
12-HSA	1.34E+05	8.90E+04	8.74E+04	5.38E+04	7.82E+04	1.96E+05	1.11E+05	1.35E+05	1.42E+05	7.52E+04	4.88E+04	1.42E+05	4.05E+04	1.84E+05	1.34E+05	1.04E+05	1.24E+05	1.35E+05	1.16E+05	3.92E+05	5.56E+05	1.37E+05	1.68E+05	2.10E+05	2.87E+05	1.25E+05	7.72E+04	1.57E+05	1.23E+05	5.01E+04
12-Hydroxylauric acid	5.03E+03	3.88E+03	4.16E+03	3.05E+03	3.88E+03	6.78E+03	3.73E+03	5.18E+03	4.92E+03	3.91E+03	2.83E+03	6.84E+03	2.30E+03	5.98E+03	5.13E+03	3.24E+03	4.90E+03	5.79E+03	5.57E+03	1.21E+04	2.66E+04	4.79E+03	7.74E+03	1.47E+04	9.10E+03	3.28E+03	3.15E+03	4.57E+03	5.26E+03	1.75E+03
13S-hydroxyoctadecadienoic acid	1.81E+04	1.25E+04	1.64E+04	1.24E+05	1.76E+04	2.43E+04	1.61E+04	1.70E+04	2.44E+04	1.16E+04	6.94E+03	1.63E+04	9.62E+03	1.33E+04	1.95E+04	1.20E+04	1.75E+04	1.32E+04	1.59E+04	3.68E+04	8.47E+04	1.74E+04	2.08E+04	3.98E+04	4.83E+04	1.54E+04	8.16E+03	2.24E+04	1.65E+04	4.23E+03
16-Heptadecyne-1,2,4-triol	3.07E+04	2.12E+04	1.73E+04	1.12E+04	2.01E+04	7.32E+04	7.89E+04	3.66E+04	3.64E+04	1.78E+04	1.13E+04	1.18E+05	8.00E+03	1.76E+04	2.20E+04	2.32E+04	4.78E+04	2.60E+04	2.83E+04	6.09E+04	2.14E+05	3.89E+04	3.05E+04	1.42E+05	5.99E+04	2.82E+04	1.30E+04	2.14E+04	3.38E+04	1.77E+04
1-Acetylcyclohexyl acetate	1.74E+04	1.28E+04	1.04E+04	8.25E+03	1.24E+04	2.48E+04	1.76E+04	1.93E+04	1.85E+04	1.64E+04	5.86E+03	2.20E+04	4.50E+03	1.49E+04	1.43E+04	1.01E+04	1.28E+04	1.48E+04	1.65E+04	3.76E+04	6.70E+04	1.49E+04	1.44E+04	2.45E+04	6.30E+04	1.32E+04	8.78E+03	3.19E+04	1.27E+04	6.05E+03
1-nonanoic acid	1.66E+04	1.51E+04	6.24E+03	1.89E+04	7.52E+03	1.14E+04	1.35E+04	2.05E+04	6.39E+03	6.59E+03	4.32E+03	9.19E+03	3.14E+03	1.10E+04	1.40E+04	4.56E+03	7.19E+03	1.37E+04	5.73E+03	1.57E+04	7.66E+04	5.89E+03	7.78E+03	1.09E+04	4.81E+04	1.58E+04	3.13E+03	1.45E+04	6.51E+03	2.32E+03
1-stearoyl-2-arachidonoyl-sn-glycero-3-phosphoethanolamine	4.83E+03	4.03E+03	2.09E+03	1.05E+03	2.30E+03	9.58E+03	6.28E+03	5.13E+03	6.02E+03	3.66E+03	7.56E+02	8.62E+03	5.06E+02	2.55E+03	1.89E+03	3.14E+03	9.13E+03	5.71E+03	4.12E+03	4.49E+03	1.06E+04	6.71E+03	5.24E+03	6.12E+02	2.00E+04	3.19E+03	2.86E+03	9.19E+03	4.71E+03	2.10E+03
2-Dodecylbenzenesulfonic acid	1.82E+05	1.44E+05	6.22E+04	6.38E+04	7.14E+04	1.29E+05	9.61E+04	1.92E+05	1.09E+05	3.72E+04	2.64E+04	5.19E+04	2.08E+04	1.15E+05	9.61E+04	5.99E+04	4.32E+04	7.99E+04	6.17E+04	1.91E+05	2.19E+05	4.78E+04	7.57E+04	1.04E+05	5.54E+05	5.15E+04	2.85E+04	1.66E+05	5.96E+04	1.22E+04
2-Hydroxy-4-oxohenicosyl acetate	1.90E+04	1.34E+04	1.11E+04	5.00E+03	8.90E+03	3.46E+04	2.75E+04	2.54E+04	2.27E+04	1.02E+04	5.93E+03	2.16E+04	4.99E+03	9.82E+03	1.78E+04	1.38E+04	1.83E+04	1.53E+04	2.16E+04	5.72E+04	1.03E+05	1.77E+04	2.25E+04	2.76E+04	3.36E+04	1.57E+04	8.11E+03	2.25E+04	2.15E+04	7.00E+03
3-Hydroxy-3-(2,3,4-trimethoxyphenyl)propanoic acid	2.71E+04	1.43E+04	9.70E+03	2.19E+03	1.14E+04	1.19E+04	9.69E+03	1.20E+04	1.12E+04	4.89E+03	3.34E+03	1.25E+04	1.94E+03	4.99E+03	5.65E+03	2.66E+03	2.04E+04	2.06E+04	9.22E+03	1.66E+04	3.91E+04	7.11E+03	1.82E+04	3.00E+04	1.51E+04	2.40E+03	1.02E+04	5.26E+03	1.93E+04	3.95E+03
3-Hydroxysebacic acid	2.74E+04	5.49E+03	1.37E+04	1.90E+03	1.64E+04	5.19E+04	1.09E+04	8.98E+03	3.28E+04	1.67E+04	2.47E+03	1.21E+04	2.95E+03	8.51E+03	5.63E+03	2.02E+04	2.00E+04	1.15E+04	7.64E+03	2.18E+04	1.11E+05	7.99E+03	2.76E+04	1.55E+04	2.55E+04	7.28E+03	2.75E+03	1.42E+04	1.21E+04	3.54E+03
3-oxolauric acid	6.18E+04	4.64E+04	3.52E+04	1.78E+04	4.19E+04	1.47E+05	9.75E+04	6.61E+04	7.02E+04	3.66E+04	2.00E+04	1.04E+05	1.84E+04	3.34E+04	4.08E+04	4.41E+04	7.95E+04	5.21E+04	6.65E+04	1.22E+05	2.90E+05	4.32E+04	1.35E+05	6.29E+04	2.54E+04	7.57E+04	4.71E+04	5.12E+04	1.38E+05	1.22E+04
3-Oxotetradecanoic acid	4.44E+04	3.23E+04	3.04E+04	1.54E+04	2.73E+04	7.02E+04	4.33E+04	3.93E+04	4.96E+04	2.58E+04	1.20E+04	5.91E+04	1.10E+04	1.47E+04	2.78E+04	3.23E+04	8.18E+04	3.92E+04	4.20E+04	9.65E+04	2.10E+05	4.56E+04	5.95E+04	6.90E+04	3.55E+04	4.07E+04	1.92E+04	3.79E+04	5.13E+04	1.90E+04
4-(1,2-Dihydroxy-2-propanyl)-1-methyl-1,2-cyclohexanediol	9.93E+03	4.32E+03	1.06E+04	2.84E+03	9.79E+03	1.14E+04	1.02E+04	1.23E+04	1.09E+04	4.44E+03	2.66E+03	1.48E+04	2.48E+03	6.00E+03	7.28E+03	3.03E+03	8.50E+03	4.78E+03	5.76E+03	2.33E+04	2.76E+04	1.40E+04	7.66E+03	2.54E+04	2.35E+04	6.65E+03	4.91E+03	9.39E+03	3.54E+03	4.35E+03
4-Undecylbenzenesulfonic acid	2.47E+05	1.89E+05	7.77E+04	7.50E+04	8.35E+04	1.41E+05	1.33E+05	2.30E+05	1.18E+05	4.45E+04	2.48E+04	6.16E+04	2.19E+04	1.02E+05	7.34E+04	4.50E+04	5.93E+04	8.38E+04	6.87E+04	2.18E+05	2.85E+05	5.29E+04	8.37E+04	8.90E+04	5.95E+05	5.56E+04	4.02E+04	1.99E+05	6.88E+04	3.81E+04
5-Hydroxydecanoic acid	2.70E+03	3.95E+03	3.36E+03	9.70E+02	2.31E+03	2.21E+04	1.32E+04	3.14E+03	5.42E+03	1.80E+03	2.39E+03	8.14E+03	2.10E+03	5.33E+03	3.74E+03	3.64E+03	9.89E+03	4.22E+03	4.35E+03	7.34E+03	6.68E+04	4.16E+03	5.72E+03	5.05E+03	6.50E+03	6.82E+03	1.45E+03	1.02E+04	7.82E+03	1.27E+03
5-hydroxyisouric acid	8.70E+03	9.20E+03	4.77E+04	3.25E+04	1.20E+04	1.72E+04	3.37E+03	2.84E+03	8.48E+03	6.37E+03	2.20E+03	2.03E+03	4.64E+03	4.70E+03	5.19E+03	3.19E+03	2.68E+03	2.47E+03	3.70E+03	3.52E+03	3.68E+03	2.41E+03	2.87E+03	1.65E+03	1.04E+03	5.29E+03	1.35E+03	2.98E+03	3.54E+03	6.94E+02
9-Oxononanoic acid	9.44E+04	6.32E+04	5.93E+04	1.45E+05	6.66E+04	9.45E+04	8.77E+04	9.25E+04	9.61E+04	4.00E+04	2.34E+04	1.08E+05	2.07E+04	5.23E+04	5.46E+04	4.41E+04	6.23E+04	5.58E+04	5.64E+04	1.52E+05	2.74E+05	8.07E+04	9.69E+04	1.32E+05	3.25E+05	5.39E+04	2.85E+04	1.80E+05	5.27E+04	3.29E+04
Arachidic acid	1.99E+04	1.09E+04	9.65E+03	6.80E+03	1.41E+04	3.19E+04	2.02E+04	1.12E+04	1.70E+04	7.36E+03	7.98E+03	6.07E+04	3.59E+03	1.87E+04	1.36E+04	1.13E+04	2.93E+04	1.09E+04	1.53E+04	3.92E+04	3.37E+05	4.08E+04	3.22E+04	1.58E+05	8.38E+04	1.58E+04	4.05E+03	1.62E+04	9.72E+03	6.41E+03
Azelaic acid	5.16E+04	8.23E+04	9.71E+03	6.74E+03	7.39E+03	1.05E+05	4.81E+04	4.83E+04	4.29E+04	1.83E+04	1.18E+04	3.31E+04	1.29E+04	3.60E+04	3.85E+04	2.61E+04	2.85E+04	3.89E+04	2.56E+04	8.14E+04	1.90E+05	3.92E+04	6.51E+04	5.03E+04	9.89E+04	3.87E+04	1.67E+04	3.98E+04	4.07E+04	9.42E+03
Benzoic acid	8.83E+04	7.33E+04	7.07E+04	4.08E+04	5.77E+04	9.11E+04	4.97E+04	8.52E+04	8.94E+04	6.76E+04	9.03E+04	1.50E+05	2.95E+04	7.53E+04	1.12E+05	7.69E+04	7.43E+04	1.10E+05	1.12E+05	2.75E+05	3.08E+05	5.96E+04	7.54E+04	1.24E+05	1.49E+05	6.92E+04	3.97E+04	1.90E+05	6.60E+04	1.03E+04
Biacetyl	2.30E+04	2.83E+04	4.85E+04	3.23E+04	4.92E+04	5.63E+04	4.84E+04	4.01E+04	7.32E+04	2.99E+04	2.97E+04	4.62E+04	2.45E+04	4.41E+04	3.11E+04	2.88E+04	4.93E+04	5.11E+04	5.31E+04	8.08E+04	5.17E+04	3.38E+04	4.54E+04	1.13E+04	2.89E+04	6.97E+04	3.03E+04	6.06E+04	6.93E+04	1.02E+04
Citric acid	3.22E+05	1.82E+05	1.84E+05	9.21E+04	1.30E+05	2.48E+05	1.42E+05	1.59E+05	1.32E+05	7.64E+04	1.38E+05	1.52E+05	1.68E+04	1.79E+05	7.07E+04	4.32E+04	5.11E+04	6.92E+04	6.48E+04	1.49E+05	1.94E+05	4.62E+04	5.30E+04	1.12E+05	9.46E+04	3.94E+04	2.24E+04	6.81E+04	4.81E+04	1.37E+04
CMPF	7.03E+03	4.53E+03	4.36E+03	2.04E+03	4.81E+03	9.41E+03	8.40E+03	7.61E+03	6.47E+03	4.99E+03	1.93E+03	6.30E+03	2.06E+03	3.24E+03	3.38E+03	4.25E+03	4.53E+03	3.84E+03	4.03E+03	1.44E+04	1.35E+04	5.64E+03	4.46E+03	1.05E+04	1.17E+04	4.89E+03	3.21E+03	7.63E+03	4.73E+03	2.85E+03
Cyclopentadecanolide	1.08E+04	8.48E+03	6.24E+03	7.22E+03	8.48E+03	3.09E+03	4.80E+03	9.00E+03	8.76E+03	5.12E+03	4.02E+03	4.58E+03	3.57E+03	9.86E+03	8.21E+03	7.73E+03	4.62E+03	8.14E+03	8.41E+03	1.02E+04	3.18E+04	3.27E+03	1.28E+04	1.84E+04	4.66E+04	1.50E+04	3.28E+03	1.83E+04	8.90E+03	2.67E+03
Dimethyl (3-oxocyclohexyl)malonate	2.54E+04	8.77E+03	1.48E+04	9.77E+03	1.41E+04	3.50E+04	2.35E+04	3.08E+04	2.91E+04	8.23E+03	8.39E+03	2.66E+04	6.99E+03	1.74E+04	1.96E+04	1.50E+04	1.86E+04	1.34E+04	2.04E+04	3.37E+04	4.08E+04	2.00E+04	1.70E+04	1.96E+04	5.39E+04	1.22E+04	1.16E+04	4.04E+04	2.44E+04	8.68E+03
DL-Lactic Acid	3.80E+05	7.11E+05	2.63E+05	2.31E+05	2.04E+05	1.69E+06	4.19E+05	5.23E+05	1.37E+06	1.02E+06	3.05E+05	4.47E+05	6.08E+05	5.56E+05	6.27E+05	6.19E+05	8.63E+05	2.56E+05	8.14E+05	5.17E+05	1.42E+06	4.62E+05	6.77E+05	2.37E+05	9.55E+05	8.33E+05	3.31E+05	5.88E+05	1.92E+06	2.27E+05
Epomediol	2.08E+04	1.96E+04	1.84E+04	1.04E+04	2.11E+04	5.00E+04	2.30E+04	2.73E+04	2.81E+04	1.67E+04	8.70E+03	4.67E+04	6.87E+03	3.80E+04	1.53E+04	1.48E+04	2.58E+04	1.58E+04	2.61E+04	5.77E+04	1.05E+05	4.02E+04	3.90E+04	6.50E+04	8.20E+04	1.35E+04	1.20E+04	7.23E+04	2.34E+04	1.49E+04
frescolat ML	4.39E+04	3.12E+04	3.02E+04	1.34E+04	2.90E+04	5.16E+04	3.13E+04	4.75E+04	4.84E+04	2.74E+04	1.74E+04	3.23E+04	1.59E+04	1.60E+04	3.63E+04	3.99E+04	2.83E+04	5.13E+04	4.77E+04	1.25E+05	1.10E+05	1.78E+04	5.63E+04	2.93E+04	4.31E+04	5.22E+04	3.88E+04	4.76E+04	3.56E+04	7.75E+03
Glycolic acid	4.63E+04	4.92E+04	6.96E+04	4.07E+04	9.65E+04	4.13E+04	4.07E+04	4.93E+04	5.33E+04	2.85E+04	2.22E+04	5.85E+04	1.81E+04	3.35E+04	6.18E+04	3.11E+04	2.09E+04	3.54E+04	6.16E+04	1.58E+05	2.77E+05	5.35E+04	7.28E+04	4.05E+05	2.76E+05	3.41E+04	2.83E+04	3.60E+04	6.36E+04	1.74E+04
Itaconic acid	1.17E+05	1.36E+05	2.57E+05	1.67E+05	2.37E+05	2.74E+05	2.35E+05	1.90E+05	3.67E+05	1.41E+05	1.47E+05	2.33E+05	1.28E+05	2.17E+05	1.59E+05	1.42E+05	2.47E+05	2.45E+05	2.57E+05	3.64E+05	2.59E+05	1.66E+05	2.28E+05	3.75E+04	1.21E+05	3.33E+05	1.48E+05	2.78E+05	3.32E+05	5.04E+04
Juniperic acid	6.41E+04	4.89E+04	3.74E+04	2.50E+04	3.90E+04	1.30E+05	7.71E+04	6.52E+04	6.99E+04	3.07E+04	2.05E+04	1.16E+05	1.48E+04	3.43E+04	4.41E+04	3.50E+04	9.19E+04	4.75E+04	5.72E+04	1.29E+05	4.29E+05	8.01E+04	6.26E+04	1.60E+05	9.32E+04	4.44E+04	2.64E+04	6.99E+04	5.40E+04	2.96E+04
Linoelaidic Acid	1.68E+05	1.15E+05	6.73E+05	1.09E+06	5.89E+05	1.41E+05	9.42E+04	1.74E+05	2.11E+05	1.06E+05	1.06E+05	1.90E+05	1.22E+05	5.54E+05	5.14E+05	2.39E+05	1.20E+05	1.40E+05	1.62E+05	2.36E+05	8.53E+05	2.09E+05	5.53E+05	1.25E+06	2.61E+06	4.07E+05	8.01E+04	1.58E+05	2.49E+05	2.82E+04
L-Pyroglutamic acid	2.62E+04	6.59E+04	5.10E+04	8.22E+04	2.80E+04	4.10E+04	1.60E+04	1.62E+04	1.54E+04	6.90E+03	7.00E+03	1.42E+04	9.50E+03	1.06E+04	1.27E+04	8.66E+03	2.81E+03	3.95E+03	1.30E+04	1.12E+04	6.45E+04	1.48E+04	2.08E+04	1.04E+05	3.35E+04	2.45E+04	2.03E+03	7.93E+03	4.29E+04	2.55E+03
L-α-PALMITIN	4.60E+04	2.57E+04	3.00E+04	1.09E+04	2.64E+04	6.51E+04	3.90E+04	4.63E+04	4.34E+04	2.14E+04	1.81E+04	6.51E+04	1.34E+04	4.57E+04	4.74E+04	2.86E+04	3.44E+04	4.78E+04	3.99E+04	9.12E+04	2.52E+05	6.15E+04	6.56E+04	1.34E+05	1.35E+05	2.78E+04	2.05E+04	4.22E+04	3.67E+04	1.37E+04
Methyl laurate	1.46E+04	1.42E+04	1.65E+04	1.11E+04	1.80E+04	2.25E+04	2.35E+04	1.55E+04	3.11E+04	1.61E+04	9.91E+03	2.13E+04	6.31E+03	3.56E+04	1.00E+04	9.16E+03	2.48E+04	2.72E+04	1.20E+04	3.24E+04	6.41E+04	2.20E+04	1.39E+04	4.05E+04	1.94E+05	1.11E+04	1.04E+04	3.20E+04	1.03E+04	4.81E+03
MHPG	2.31E+04	7.40E+03	1.32E+04	2.17E+03	1.17E+04	3.17E+04	1.36E+04	8.75E+03	1.22E+04	7.86E+03	2.79E+03	2.29E+04	4.40E+03	1.65E+04	1.23E+04	3.79E+03	3.00E+04	1.62E+04	1.11E+04	2.84E+04	4.79E+04	1.59E+04	1.30E+04	1.37E+04	3.99E+04	3.02E+03	7.24E+03	1.09E+04	8.92E+03	7.78E+03
myristic acid	6.62E+05	5.79E+05	3.65E+05	2.91E+05	4.10E+05	1.01E+06	5.45E+05	6.76E+05	7.90E+05	4.06E+05	9.25E+04	3.31E+05	6.77E+04	1.05E+05	1.52E+05	1.96E+05	2.70E+05	2.03E+05	2.76E+05	4.48E+05	1.11E+06	1.83E+05	3.25E+05	4.29E+05	8.97E+05	2.71E+05	1.04E+05	3.21E+05	3.05E+05	7.03E+04
N-Acetyl-L-aspartic acid	7.15E+03	2.63E+04	1.74E+04	1.33E+04	3.35E+04	1.06E+04	8.55E+03	1.59E+04	1.66E+04	5.08E+03	1.98E+04	2.43E+04	2.87E+04	3.91E+04	1.02E+04	1.57E+04	3.41E+04	1.57E+04	1.18E+04	2.71E+04	9.23E+04	3.39E+04	4.54E+04	5.66E+03	3.17E+04	6.70E+03	1.44E+04	2.05E+04	4.43E+04	4.31E+03
Oleic acid	1.42E+05	8.99E+04	4.67E+05	5.58E+05	4.19E+05	1.40E+05	1.09E+05	1.46E+05	1.48E+05	7.02E+04	7.22E+04	1.78E+05	7.10E+04	3.18E+05	3.39E+05	1.37E+05	1.11E+05	9.56E+04	1.10E+05	2.02E+05	1.14E+06	3.65E+05	4.53E+05	1.69E+06	1.78E+06	1.80E+05	5.85E+04	1.56E+05	1.39E+05	4.04E+04
Palmitic Acid	3.95E+06	2.06E+06	1.98E+06	1.75E+06	2.40E+06	2.60E+06	1.41E+06	3.59E+06	3.59E+06	1.42E+06	1.64E+06	2.80E+06	1.08E+06	5.96E+06	3.90E+06	2.74E+06	1.74E+06	3.60E+06	3.02E+06	8.78E+06	8.08E+06	1.25E+06	3.91E+06	3.07E+06	1.12E+07	2.26E+06	1.55E+06	4.24E+06	2.21E+06	4.48E+05
p-Cresylsulfate	1.01E+04	9.91E+03	6.17E+04	2.50E+04	5.56E+04	1.36E+04	1.33E+04	1.02E+04	1.08E+04	5.37E+03	7.62E+03	3.34E+04	6.89E+03	1.25E+04	2.81E+04	1.18E+04	1.37E+04	3.00E+03	5.13E+03	8.59E+03	1.16E+05	3.49E+04	3.73E+04	2.29E+05	8.44E+04	4.34E+03	1.44E+03	4.77E+03	8.06E+03	3.36E+03
pentadecanoic acid	6.15E+04	5.09E+04	4.85E+04	4.65E+04	6.22E+04	5.72E+04	3.65E+04	6.21E+04	6.94E+04	4.01E+04	2.48E+04	4.43E+04	2.07E+04	7.60E+04	7.67E+04	5.59E+04	3.73E+04	6.60E+04	6.15E+04	1.49E+05	1.64E+05	3.64E+04	7.58E+04	8.78E+04	3.35E+05	6.23E+04	3.36E+04	9.13E+04	5.62E+04	1.37E+04
Propyl (9E)-12-hydroxy-9-octadecenoate	2.99E+04	1.85E+04	1.52E+04	8.56E+03	1.05E+04	2.86E+04	2.21E+04	2.38E+04	2.25E+04	1.27E+04	7.49E+03	2.75E+04	7.41E+03	2.76E+04	2.06E+04	2.25E+04	1.85E+04	1.93E+04	2.18E+04	6.85E+04	9.23E+04	2.88E+04	2.67E+04	3.52E+04	3.21E+04	2.01E+04	1.69E+04	2.12E+04	2.71E+04	1.12E+04
RG1300000	2.26E+05	1.67E+05	1.45E+05	1.42E+05	1.40E+05	4.22E+05	2.33E+05	2.34E+05	2.52E+05	1.14E+05	7.62E+04	2.94E+05	5.82E+04	1.05E+05	1.53E+05	1.67E+05	2.66E+05	1.93E+05	2.32E+05	4.82E+05	1.29E+06	2.22E+05	2.68E+05	4.01E+05	4.98E+05	2.03E+05	9.87E+04	2.23E+05	2.45E+05	8.16E+04
RH3422500	4.66E+03	2.50E+03	2.96E+03	2.79E+03	2.21E+03	5.18E+03	4.16E+03	3.30E+03	5.36E+03	1.46E+03	1.45E+03	3.72E+03	1.70E+03	4.76E+03	5.22E+03	2.56E+03	2.16E+03	2.51E+03	3.10E+03	7.21E+03	9.96E+03	3.16E+03	3.83E+03	2.54E+03	1.83E+04	3.80E+03	1.21E+03	4.00E+03	3.59E+03	1.32E+03
Stearic acid	1.29E+06	7.01E+05	7.79E+05	4.61E+05	1.06E+06	1.58E+06	1.13E+06	1.16E+06	1.17E+06	4.39E+05	6.21E+05	2.37E+06	3.48E+05	2.22E+06	1.59E+06	8.16E+05	1.25E+06	1.12E+06	1.00E+06	3.61E+06	1.01E+07	2.36E+06	2.21E+06	5.13E+06	5.92E+06	7.53E+05	5.67E+05	1.60E+06	9.44E+05	4.02E+05
Succinic acid	5.62E+04	6.13E+04	5.45E+04	1.74E+04	2.31E+04	5.72E+04	4.56E+04	4.39E+04	5.05E+04	4.74E+04	5.01E+04	6.95E+04	3.81E+04	2.03E+04	5.12E+04	6.60E+04	8.01E+04	5.85E+04	8.22E+04	1.10E+05	1.48E+05	4.17E+04	6.54E+04	8.01E+04	9.29E+04	1.29E+05	5.22E+04	2.44E+04	7.76E+04	2.61E+04
Thapsic acid	2.98E+04	2.20E+04	1.29E+04	7.17E+03	1.60E+04	7.16E+04	5.00E+04	3.19E+04	3.08E+04	1.89E+04	9.37E+03	5.58E+04	6.92E+03	2.88E+04	2.18E+04	1.94E+04	7.21E+04	2.80E+04	3.07E+04	7.01E+04	1.93E+05	3.56E+04	6.05E+04	4.71E+04	1.63E+04	2.61E+04	1.49E+04	7.29E+04	5.23E+04	1.40E+04
Traumatic Acid	1.21E+05	8.61E+04	8.30E+04	4.25E+04	5.63E+04	1.35E+05	9.03E+04	1.40E+05	1.34E+05	6.32E+04	4.10E+04	1.43E+05	2.90E+04	9.22E+04	1.10E+05	6.15E+04	1.21E+05	1.17E+05	1.32E+05	1.67E+05	5.41E+05	1.26E+05	1.45E+05	2.28E+05	2.72E+05	9.75E+04	4.28E+04	1.49E+05	1.15E+05	4.24E+04
Undecanedioic acid	1.37E+04	1.10E+04	3.51E+03	2.01E+03	4.42E+03	4.67E+04	8.19E+03	8.45E+03	1.18E+04	1.46E+04	2.57E+03	1.71E+04	4.98E+03	5.03E+03	5.40E+03	5.52E+03	9.51E+03	1.05E+04	1.40E+04	5.88E+04	8.06E+04	7.50E+03	3.35E+04	8.19E+03	3.41E+04	2.72E+04	1.09E+04	8.36E+03	1.03E+04	2.93E+03
Valproic acid	6.04E+04	5.57E+04	7.02E+05	2.43E+05	5.94E+05	3.09E+05	2.78E+05	8.60E+04	1.12E+05	4.86E+04	4.59E+04	3.63E+05	3.77E+04	1.98E+05	2.55E+05	9.66E+04	2.78E+05	2.60E+04	6.00E+04	4.78E+04	1.97E+06	1.85E+05	4.13E+05	2.75E+06	9.65E+05	4.24E+04	2.10E+04	4.47E+04	1.33E+05	1.96E+04
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Human Metabolome Database
(-)-Chloramphenicol	HMDB0014589
(-)-pinellic acid	HMDB0004708
12-HSA	HMDB0061706
12-Hydroxylauric acid	HMDB0002059
13S-hydroxyoctadecadienoic acid	HMDB0004667
16-Heptadecyne-1,2,4-triol	HMDB0035473
1-Acetylcyclohexyl acetate	HMDB0039710
1-nonanoic acid	HMDB0000847
1-stearoyl-2-arachidonoyl-sn-glycero-3-phosphoethanolamine	HMDB0009003
2-Dodecylbenzenesulfonic acid	HMDB0031031
2-Hydroxy-4-oxohenicosyl acetate	HMDB0036344
3-Hydroxy-3-(2,3,4-trimethoxyphenyl)propanoic acid	HMDB0142079
3-Hydroxysebacic acid	HMDB0000350
3-oxolauric acid	HMDB0010727
3-Oxotetradecanoic acid	HMDB0010730
4-(1,2-Dihydroxy-2-propanyl)-1-methyl-1,2-cyclohexanediol	HMDB0039974
4-Undecylbenzenesulfonic acid	HMDB0032549
5-Hydroxydecanoic acid	HMDB0040329
5-hydroxyisouric acid	HMDB0030097
9-Oxononanoic acid	HMDB0094711
Arachidic acid	HMDB0002212
Azelaic acid	HMDB0000784
Benzoic acid	HMDB0001870
Biacetyl	HMDB0003407
Citric acid	HMDB0000094
CMPF	HMDB0061112
Cyclopentadecanolide	HMDB0034455
Dimethyl (3-oxocyclohexyl)malonate	HMDB0032496
DL-Lactic Acid	HMDB0144295
Epomediol	HMDB0041376
frescolat ML	HMDB0037212
Glycolic acid	HMDB0000115
Itaconic acid	HMDB0002092
Juniperic acid	HMDB0006294
Linoelaidic Acid	HMDB0006270
L-Pyroglutamic acid	HMDB0000267
L-α-PALMITIN	HMDB0005356
Methyl laurate	HMDB0031018
MHPG	HMDB0001490
myristic acid	HMDB0000806
N-Acetyl-L-aspartic acid	HMDB0000812
Oleic acid	HMDB0000207
Palmitic Acid	HMDB0000220
p-Cresylsulfate	HMDB0011635
pentadecanoic acid	HMDB0000826
Propyl (9E)-12-hydroxy-9-octadecenoate	HMDB0032476
RG1300000	HMDB0247617
RH3422500	HMDB0034844
Stearic acid	HMDB0000827
Succinic acid	HMDB0000254
Thapsic acid	HMDB0000672
Traumatic Acid	HMDB0000933
Undecanedioic acid	HMDB0000888
Valproic acid	HMDB0001877
METABOLITES_END
#END


#METABOLOMICS WORKBENCH dhananjay_shinde5_20240914_131224 DATATRACK_ID:5191 STUDY_ID:ST003482 ANALYSIS_ID:AN005717 PROJECT_ID:PR002139
VERSION             	1
CREATED_ON             	September 14, 2024, 3:09 pm
#PROJECT
PR:PROJECT_TITLE                 	Tissue niche influences immune and metabolic profiles to Staphylococcus aureus
PR:PROJECT_TITLE                 	biofilm infection
PR:PROJECT_TYPE                  	Intracellular data
PR:PROJECT_SUMMARY               	Infection is a devastating post-surgical complication, often requiring
PR:PROJECT_SUMMARY               	additional procedures and prolonged antibiotic therapy. This is especially
PR:PROJECT_SUMMARY               	relevant for craniotomy and prosthetic joint infections (PJI), both of which are
PR:PROJECT_SUMMARY               	characterized by biofilm formation on the bone or implant surface, respectively,
PR:PROJECT_SUMMARY               	with S. aureus representing a primary cause. The local tissue microenvironment
PR:PROJECT_SUMMARY               	likely has profound effects on immune attributes that can influence treatment
PR:PROJECT_SUMMARY               	efficacy, which becomes critical to consider when developing therapeutics for
PR:PROJECT_SUMMARY               	biofilm infections. However, the extent to which distinct tissue niches
PR:PROJECT_SUMMARY               	influence immune function during biofilm development remains relatively unknown.
PR:PROJECT_SUMMARY               	To address this, we compare the metabolomic, transcriptomic, and functional
PR:PROJECT_SUMMARY               	attributes of leukocytes in mouse models of S. aureus craniotomy and PJI
PR:PROJECT_SUMMARY               	complemented with patient samples from both infection modalities, which reveals
PR:PROJECT_SUMMARY               	profound tissue niche-dependent differences in nucleic acid, amino acid, and
PR:PROJECT_SUMMARY               	lipid metabolism with links to immune modulation. These signatures are both
PR:PROJECT_SUMMARY               	spatially and temporally distinct, differing not only between infection sites
PR:PROJECT_SUMMARY               	but evolving over time within a single model. Collectively, this demonstrates
PR:PROJECT_SUMMARY               	that biofilms elicit unique immune and metabolic responses that are heavily
PR:PROJECT_SUMMARY               	influenced by the local tissue microenvironment, which will likely have
PR:PROJECT_SUMMARY               	important implications when designing therapeutic approaches targeting these
PR:PROJECT_SUMMARY               	infections.
PR:INSTITUTE                     	UNMC
PR:DEPARTMENT                    	Department of pathology, microbiology and immunology
PR:LABORATORY                    	Pathology, Microbiology and Immunology
PR:LAST_NAME                     	Dhananjay
PR:FIRST_NAME                    	Shinde
PR:ADDRESS                       	DRC 2, 7066, UNMC, Emily st
PR:EMAIL                         	dhananjay.shinde@unmc.edu
PR:PHONE                         	4025597623
#STUDY
ST:STUDY_TITLE                   	Tissue niche influences immune and metabolic profiles to Staphylococcus aureus
ST:STUDY_TITLE                   	biofilm infection
ST:STUDY_SUMMARY                 	Infection is a devastating post-surgical complication, often requiring
ST:STUDY_SUMMARY                 	additional procedures and prolonged antibiotic therapy. This is especially
ST:STUDY_SUMMARY                 	relevant for craniotomy and prosthetic joint infections (PJI), both of which are
ST:STUDY_SUMMARY                 	characterized by biofilm formation on the bone or implant surface, respectively,
ST:STUDY_SUMMARY                 	with S. aureus representing a primary cause. The local tissue microenvironment
ST:STUDY_SUMMARY                 	likely has profound effects on immune attributes that can influence treatment
ST:STUDY_SUMMARY                 	efficacy, which becomes critical to consider when developing therapeutics for
ST:STUDY_SUMMARY                 	biofilm infections. However, the extent to which distinct tissue niches
ST:STUDY_SUMMARY                 	influence immune function during biofilm development remains relatively unknown.
ST:STUDY_SUMMARY                 	To address this, we compare the metabolomic, transcriptomic, and functional
ST:STUDY_SUMMARY                 	attributes of leukocytes in mouse models of S. aureus craniotomy and PJI
ST:STUDY_SUMMARY                 	complemented with patient samples from both infection modalities, which reveals
ST:STUDY_SUMMARY                 	profound tissue niche-dependent differences in nucleic acid, amino acid, and
ST:STUDY_SUMMARY                 	lipid metabolism with links to immune modulation. These signatures are both
ST:STUDY_SUMMARY                 	spatially and temporally distinct, differing not only between infection sites
ST:STUDY_SUMMARY                 	but evolving over time within a single model. Collectively, this demonstrates
ST:STUDY_SUMMARY                 	that biofilms elicit unique immune and metabolic responses that are heavily
ST:STUDY_SUMMARY                 	influenced by the local tissue microenvironment, which will likely have
ST:STUDY_SUMMARY                 	important implications when designing therapeutic approaches targeting these
ST:STUDY_SUMMARY                 	infections.
ST:INSTITUTE                     	UNMC
ST:DEPARTMENT                    	Department of pathology, microbiology and immunology
ST:LAST_NAME                     	Shinde
ST:FIRST_NAME                    	Dhananjay
ST:ADDRESS                       	DRC 2, 7066, UNMC, Emily st
ST:EMAIL                         	dhananjay.shinde@unmc.edu
ST:PHONE                         	4025597623
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:GENDER                        	Male and female
#FACTORS
#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           	-	d3-Crain_1	Sample source:Galea | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_1.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-Crain_2	Sample source:Galea | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_2.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-Crain_3	Sample source:Galea | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_3.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-Crain_4	Sample source:Galea | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_4.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-Crain_5	Sample source:Galea | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_5.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-PJI_1	Sample source:Tissue (PJI) | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_1.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-PJI_2	Sample source:Tissue (PJI) | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_2.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-PJI_3	Sample source:Tissue (PJI) | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_3.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-PJI_4	Sample source:Tissue (PJI) | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_4.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d3-PJI_5	Sample source:Tissue (PJI) | Experimental Variable:d3	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_5.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-Crain_1	Sample source:Galea | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_1.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-Crain_2	Sample source:Galea | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_2.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-Crain_3	Sample source:Galea | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_3.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-Crain_4	Sample source:Galea | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_4.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-Crain_5	Sample source:Galea | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_5.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-PJI_1	Sample source:Tissue (PJI) | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_1.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-PJI_2	Sample source:Tissue (PJI) | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_2.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-PJI_3	Sample source:Tissue (PJI) | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_3.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-PJI_4	Sample source:Tissue (PJI) | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_4.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d7-PJI_5	Sample source:Tissue (PJI) | Experimental Variable:d7	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_5.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-Crain_1	Sample source:Galea | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_1.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-Crain_2	Sample source:Galea | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_2.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-Crain_3	Sample source:Galea | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_3.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-Crain_4	Sample source:Galea | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_4.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-Crain_5	Sample source:Galea | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_5.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-PJI_1	Sample source:Tissue (PJI) | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_1.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-PJI_2	Sample source:Tissue (PJI) | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_2.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-PJI_3	Sample source:Tissue (PJI) | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_3.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-PJI_4	Sample source:Tissue (PJI) | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_4.mzXML
SUBJECT_SAMPLE_FACTORS           	-	d14-PJI_5	Sample source:Tissue (PJI) | Experimental Variable:d14	Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_5.mzXML
#COLLECTION
CO:COLLECTION_SUMMARY            	The intracellular metabolome of granulocytes isolated from the galea and PJI
CO:COLLECTION_SUMMARY            	tissue of mouse craniotomy and PJI models, respectively, were profiled by
CO:COLLECTION_SUMMARY            	LC-HRMS. Fifteen mice were infected for each biofilm model (craniotomy and PJI)
CO:COLLECTION_SUMMARY            	with single-cell suspensions prepared by pooling tissue homogenates from 3 mice
CO:COLLECTION_SUMMARY            	into one sample, resulting in 5 biological replicates. Samples were then
CO:COLLECTION_SUMMARY            	enriched for granulocytes using Ly6G magnetic beads (Miltenyi Biotec, Bergishch
CO:COLLECTION_SUMMARY            	Gladbach, Germany). Magnetic beads were utilized for purification instead of
CO:COLLECTION_SUMMARY            	FACS to limit alterations in cellular metabolism that may occur with extended
CO:COLLECTION_SUMMARY            	processing times. Samples were washed twice in 1X PBS, resuspended in 80% MetOH,
CO:COLLECTION_SUMMARY            	and frozen at -80°C for 15 min. Metabolomics was also performed on supernatants
CO:COLLECTION_SUMMARY            	prepared from brain, galea, and PJI tissue homogenates at days 3, 7, and 14
CO:COLLECTION_SUMMARY            	post-infection. Similar to granulocyte intracellular metabolite collection,
CO:COLLECTION_SUMMARY            	tissue supernatants were centrifuged at high speed to remove residual cellular
CO:COLLECTION_SUMMARY            	material, combined with 100% MetOH to yield an 80% MetOH solution, and frozen at
CO:COLLECTION_SUMMARY            	-80°C.
CO:SAMPLE_TYPE                   	Extracellular infection sites homogenate, intracellular granulocyte
#TREATMENT
TR:TREATMENT_SUMMARY             	All mice were group-housed at 21-23°C (22°C average) and 30-70% humidity (55%
TR:TREATMENT_SUMMARY             	average) under 12 h light/dark cycle with free access to food (2019S Teklad
TR:TREATMENT_SUMMARY             	Global 19% Protein Extruded Rodent Diet; Inotiv, West Lafayette, IN) and water.
TR:TREATMENT_SUMMARY             	S. aureus craniotomy infection was established in male and female 8- to
TR:TREATMENT_SUMMARY             	10-week-old C57BL/6J mice (RRID:IMSR_JAX:000664). Briefly, ketamine and xylazine
TR:TREATMENT_SUMMARY             	were administered to achieve a surgical plane of anesthesia before skin
TR:TREATMENT_SUMMARY             	disinfection with betadine. A midline incision of the scalp was made, whereupon
TR:TREATMENT_SUMMARY             	a bone flap was created using a high-speed pneumatic drill (Stryker Corporation,
TR:TREATMENT_SUMMARY             	Kalamazoo, MI). The bone flap was incubated with 0.5 mL of S. aureus USA300
TR:TREATMENT_SUMMARY             	LAC13C diluted to 10^6/mL in brain-heart infusion broth at 37°C for 5 min and
TR:TREATMENT_SUMMARY             	subsequently rinsed in PBS, blotted dry on a sterile field, and reimplanted.
TR:TREATMENT_SUMMARY             	This procedure reliably produces an infectious inoculum of 10^3 CFUs per bone
TR:TREATMENT_SUMMARY             	flap, which more accurately models surgical site infection that progresses to
TR:TREATMENT_SUMMARY             	biofilm formation. The scalp incision was closed using 6-0 nylon suture and
TR:TREATMENT_SUMMARY             	animals were continuously monitored until they regained ambulation, and daily
TR:TREATMENT_SUMMARY             	thereafter. S. aureus PJI was established in male and female 8- to 10-week-old
TR:TREATMENT_SUMMARY             	C57BL/6J mice. Mice were anesthetized with ketamine and xylazine prior to skin
TR:TREATMENT_SUMMARY             	disinfection with betadine and initial incision of the quadriceps. A secondary
TR:TREATMENT_SUMMARY             	incision was made to laterally displace the patellar tendon and a burr hole was
TR:TREATMENT_SUMMARY             	created in the distal end of the femur with a 26-gauge needle. An
TR:TREATMENT_SUMMARY             	orthopedic-grade nickel-titanium Kirschner wire (0.6 mm diameter; Custom Wire
TR:TREATMENT_SUMMARY             	Technologies, Port Washington, WI) was inserted through the burr hole into the
TR:TREATMENT_SUMMARY             	medullary canal and 10^3 CFUs of S. aureus USA300 LAC13C were inoculated at the
TR:TREATMENT_SUMMARY             	tip of the wire and the incision was closed with nylon suture. Mice received
TR:TREATMENT_SUMMARY             	buprenorphine SR to alleviate any post-surgical pain and were continuously
TR:TREATMENT_SUMMARY             	monitored until ambulatory and daily thereafter.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	A 13C15N-labeled canonical amino acid (13C15N-CAA) mix (Cambridge Isotope
SP:SAMPLEPREP_SUMMARY            	Laboratories, Andover, MA) was added to the extraction buffer comprised of 80%
SP:SAMPLEPREP_SUMMARY            	MeOH as an internal standard. Samples were then pelleted, and supernatants were
SP:SAMPLEPREP_SUMMARY            	dried by vacuum centrifuge for storage at -80°C. Cell pellets were stored for
SP:SAMPLEPREP_SUMMARY            	protein quantification to normalize metabolite peak areas. Prior to LC-HRMS,
SP:SAMPLEPREP_SUMMARY            	samples were reconstituted in 100 µL of 50% MetOH, pelleted, and the resulting
SP:SAMPLEPREP_SUMMARY            	supernatant was analyzed.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Metabolite separation was performed by liquid chromatography using a XBridge
CH:CHROMATOGRAPHY_SUMMARY        	Amide analytical column (150 × 2.1mm ID; 1.7µm particle size; Waters
CH:CHROMATOGRAPHY_SUMMARY        	Corporation, Milford, MA) and a binary solvent system infused at a flow rate of
CH:CHROMATOGRAPHY_SUMMARY        	0.3 ml/min. A guard XBridge Amide column (20 × 2.1mm ID; 3.5µm particle
CH:CHROMATOGRAPHY_SUMMARY        	size; Waters Corporation) was connected in front of the analytical column.
CH:CHROMATOGRAPHY_SUMMARY        	Mobile phase A was composed of ammonium acetate and ammonium hydroxide (10 mM
CH:CHROMATOGRAPHY_SUMMARY        	each) containing 5% acetonitrile in LC-MS grade water; mobile phase B was 100%
CH:CHROMATOGRAPHY_SUMMARY        	LC-MS grade acetonitrile. The pH of mobile phase A was adjusted to 8.0 using
CH:CHROMATOGRAPHY_SUMMARY        	glacial acetic acid. The UHPLC pumps were operated in gradient mode. The amide
CH:CHROMATOGRAPHY_SUMMARY        	column was maintained at 40°C, and the autosampler temperature was held at 5°C
CH:CHROMATOGRAPHY_SUMMARY        	throughout data acquisition. The injection volume for all samples was 5 µl with
CH:CHROMATOGRAPHY_SUMMARY        	the 13C15N-CAA mix as the internal standard.
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Waters ACQUITY UPLC BEH Amide (150 x 2.1mm,1.7um)
CH:SOLVENT_A                     	ammonium acetate and ammonium hydroxide (10 mM each) containing 5% acetonitrile
CH:SOLVENT_A                     	in LC-MS grade water
CH:SOLVENT_B                     	acetonitrile
CH:FLOW_GRADIENT                 	B% at initial 85%
CH:FLOW_RATE                     	0.3ml/min
CH:COLUMN_TEMPERATURE            	40
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Orbitrap Exploris 480
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	A HRMS Orbitrap (Exploris 480; Thermo Fisher Scientific) was operated in
MS:MS_COMMENTS                   	polarity switching mode and was used for untargeted metabolomics in a
MS:MS_COMMENTS                   	data-dependent MS/MS acquisition mode (DDA). Electrospray ionization (ESI)
MS:MS_COMMENTS                   	parameters were optimized as follows: electrospray ion voltage of -2700V and
MS:MS_COMMENTS                   	3500V in negative and positive mode respectively, ion transfer tube temperature
MS:MS_COMMENTS                   	was maintained at 400°C, and m/z scan range was 70-1050 Da. Orbitrap resolution
MS:MS_COMMENTS                   	for precursor ion scans were maintained at 240,000 for intracellular and 120,000
MS:MS_COMMENTS                   	for extracellular whereas fragment ion scans were maintained at 120,000 and
MS:MS_COMMENTS                   	60,000, for intracellular and extracellular respectively. Normalized collision
MS:MS_COMMENTS                   	energies at 30, 50, and 150% were used for fragmentation and data was acquired
MS:MS_COMMENTS                   	in profile mode.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Relative intensities
MS_METABOLITE_DATA_START
Samples	d3-Crain_1	d3-Crain_2	d3-Crain_3	d3-Crain_4	d3-Crain_5	d3-PJI_1	d3-PJI_2	d3-PJI_3	d3-PJI_4	d3-PJI_5	d7-Crain_1	d7-Crain_2	d7-Crain_3	d7-Crain_4	d7-Crain_5	d7-PJI_1	d7-PJI_2	d7-PJI_3	d7-PJI_4	d7-PJI_5	d14-Crain_1	d14-Crain_2	d14-Crain_3	d14-Crain_4	d14-Crain_5	d14-PJI_1	d14-PJI_2	d14-PJI_3	d14-PJI_4	d14-PJI_5
Factors	Sample source:Galea | Experimental Variable:d3	Sample source:Galea | Experimental Variable:d3	Sample source:Galea | Experimental Variable:d3	Sample source:Galea | Experimental Variable:d3	Sample source:Galea | Experimental Variable:d3	Sample source:Tissue (PJI) | Experimental Variable:d3	Sample source:Tissue (PJI) | Experimental Variable:d3	Sample source:Tissue (PJI) | Experimental Variable:d3	Sample source:Tissue (PJI) | Experimental Variable:d3	Sample source:Tissue (PJI) | Experimental Variable:d3	Sample source:Galea | Experimental Variable:d7	Sample source:Galea | Experimental Variable:d7	Sample source:Galea | Experimental Variable:d7	Sample source:Galea | Experimental Variable:d7	Sample source:Galea | Experimental Variable:d7	Sample source:Tissue (PJI) | Experimental Variable:d7	Sample source:Tissue (PJI) | Experimental Variable:d7	Sample source:Tissue (PJI) | Experimental Variable:d7	Sample source:Tissue (PJI) | Experimental Variable:d7	Sample source:Tissue (PJI) | Experimental Variable:d7	Sample source:Galea | Experimental Variable:d14	Sample source:Galea | Experimental Variable:d14	Sample source:Galea | Experimental Variable:d14	Sample source:Galea | Experimental Variable:d14	Sample source:Galea | Experimental Variable:d14	Sample source:Tissue (PJI) | Experimental Variable:d14	Sample source:Tissue (PJI) | Experimental Variable:d14	Sample source:Tissue (PJI) | Experimental Variable:d14	Sample source:Tissue (PJI) | Experimental Variable:d14	Sample source:Tissue (PJI) | Experimental Variable:d14
(-)-Chloramphenicol	1.07E+04	2.71E+04	1.12E+04	3.83E+03	9.52E+03	1.77E+04	1.38E+04	1.22E+04	1.12E+04	3.28E+03	4.19E+03	4.37E+04	3.51E+03	6.20E+03	4.73E+03	4.34E+03	1.52E+04	6.74E+03	2.38E+03	2.69E+04	5.41E+05	5.08E+04	1.68E+05	2.40E+06	4.94E+05	3.31E+03	3.01E+03	1.37E+04	5.44E+03	7.70E+03
(-)-pinellic acid	5.61E+04	3.23E+04	3.18E+04	1.49E+04	2.92E+04	7.40E+04	4.23E+04	6.06E+04	5.98E+04	2.50E+04	1.64E+04	5.77E+04	8.23E+03	3.63E+04	4.40E+04	3.65E+04	5.91E+04	3.80E+04	4.76E+04	1.60E+05	2.13E+05	3.58E+04	6.10E+04	6.09E+04	1.03E+05	3.47E+04	2.84E+04	8.89E+04	5.66E+04	1.40E+04
12-HSA	1.34E+05	8.90E+04	8.74E+04	5.38E+04	7.82E+04	1.96E+05	1.11E+05	1.35E+05	1.42E+05	7.52E+04	4.88E+04	1.42E+05	4.05E+04	1.84E+05	1.34E+05	1.04E+05	1.24E+05	1.35E+05	1.16E+05	3.92E+05	5.56E+05	1.37E+05	1.68E+05	2.10E+05	2.87E+05	1.25E+05	7.72E+04	1.57E+05	1.23E+05	5.01E+04
12-Hydroxylauric acid	5.03E+03	3.88E+03	4.16E+03	3.05E+03	3.88E+03	6.78E+03	3.73E+03	5.18E+03	4.92E+03	3.91E+03	2.83E+03	6.84E+03	2.30E+03	5.98E+03	5.13E+03	3.24E+03	4.90E+03	5.79E+03	5.57E+03	1.21E+04	2.66E+04	4.79E+03	7.74E+03	1.47E+04	9.10E+03	3.28E+03	3.15E+03	4.57E+03	5.26E+03	1.75E+03
13S-hydroxyoctadecadienoic acid	1.81E+04	1.25E+04	1.64E+04	1.24E+05	1.76E+04	2.43E+04	1.61E+04	1.70E+04	2.44E+04	1.16E+04	6.94E+03	1.63E+04	9.62E+03	1.33E+04	1.95E+04	1.20E+04	1.75E+04	1.32E+04	1.59E+04	3.68E+04	8.47E+04	1.74E+04	2.08E+04	3.98E+04	4.83E+04	1.54E+04	8.16E+03	2.24E+04	1.65E+04	4.23E+03
16-Heptadecyne-1,2,4-triol	3.07E+04	2.12E+04	1.73E+04	1.12E+04	2.01E+04	7.32E+04	7.89E+04	3.66E+04	3.64E+04	1.78E+04	1.13E+04	1.18E+05	8.00E+03	1.76E+04	2.20E+04	2.32E+04	4.78E+04	2.60E+04	2.83E+04	6.09E+04	2.14E+05	3.89E+04	3.05E+04	1.42E+05	5.99E+04	2.82E+04	1.30E+04	2.14E+04	3.38E+04	1.77E+04
1-Acetylcyclohexyl acetate	1.74E+04	1.28E+04	1.04E+04	8.25E+03	1.24E+04	2.48E+04	1.76E+04	1.93E+04	1.85E+04	1.64E+04	5.86E+03	2.20E+04	4.50E+03	1.49E+04	1.43E+04	1.01E+04	1.28E+04	1.48E+04	1.65E+04	3.76E+04	6.70E+04	1.49E+04	1.44E+04	2.45E+04	6.30E+04	1.32E+04	8.78E+03	3.19E+04	1.27E+04	6.05E+03
1-nonanoic acid	1.66E+04	1.51E+04	6.24E+03	1.89E+04	7.52E+03	1.14E+04	1.35E+04	2.05E+04	6.39E+03	6.59E+03	4.32E+03	9.19E+03	3.14E+03	1.10E+04	1.40E+04	4.56E+03	7.19E+03	1.37E+04	5.73E+03	1.57E+04	7.66E+04	5.89E+03	7.78E+03	1.09E+04	4.81E+04	1.58E+04	3.13E+03	1.45E+04	6.51E+03	2.32E+03
1-stearoyl-2-arachidonoyl-sn-glycero-3-phosphoethanolamine	4.83E+03	4.03E+03	2.09E+03	1.05E+03	2.30E+03	9.58E+03	6.28E+03	5.13E+03	6.02E+03	3.66E+03	7.56E+02	8.62E+03	5.06E+02	2.55E+03	1.89E+03	3.14E+03	9.13E+03	5.71E+03	4.12E+03	4.49E+03	1.06E+04	6.71E+03	5.24E+03	6.12E+02	2.00E+04	3.19E+03	2.86E+03	9.19E+03	4.71E+03	2.10E+03
2-Dodecylbenzenesulfonic acid	1.82E+05	1.44E+05	6.22E+04	6.38E+04	7.14E+04	1.29E+05	9.61E+04	1.92E+05	1.09E+05	3.72E+04	2.64E+04	5.19E+04	2.08E+04	1.15E+05	9.61E+04	5.99E+04	4.32E+04	7.99E+04	6.17E+04	1.91E+05	2.19E+05	4.78E+04	7.57E+04	1.04E+05	5.54E+05	5.15E+04	2.85E+04	1.66E+05	5.96E+04	1.22E+04
2-Hydroxy-4-oxohenicosyl acetate	1.90E+04	1.34E+04	1.11E+04	5.00E+03	8.90E+03	3.46E+04	2.75E+04	2.54E+04	2.27E+04	1.02E+04	5.93E+03	2.16E+04	4.99E+03	9.82E+03	1.78E+04	1.38E+04	1.83E+04	1.53E+04	2.16E+04	5.72E+04	1.03E+05	1.77E+04	2.25E+04	2.76E+04	3.36E+04	1.57E+04	8.11E+03	2.25E+04	2.15E+04	7.00E+03
3-Hydroxy-3-(2,3,4-trimethoxyphenyl)propanoic acid	2.71E+04	1.43E+04	9.70E+03	2.19E+03	1.14E+04	1.19E+04	9.69E+03	1.20E+04	1.12E+04	4.89E+03	3.34E+03	1.25E+04	1.94E+03	4.99E+03	5.65E+03	2.66E+03	2.04E+04	2.06E+04	9.22E+03	1.66E+04	3.91E+04	7.11E+03	1.82E+04	3.00E+04	1.51E+04	2.40E+03	1.02E+04	5.26E+03	1.93E+04	3.95E+03
3-Hydroxysebacic acid	2.74E+04	5.49E+03	1.37E+04	1.90E+03	1.64E+04	5.19E+04	1.09E+04	8.98E+03	3.28E+04	1.67E+04	2.47E+03	1.21E+04	2.95E+03	8.51E+03	5.63E+03	2.02E+04	2.00E+04	1.15E+04	7.64E+03	2.18E+04	1.11E+05	7.99E+03	2.76E+04	1.55E+04	2.55E+04	7.28E+03	2.75E+03	1.42E+04	1.21E+04	3.54E+03
3-oxolauric acid	6.18E+04	4.64E+04	3.52E+04	1.78E+04	4.19E+04	1.47E+05	9.75E+04	6.61E+04	7.02E+04	3.66E+04	2.00E+04	1.04E+05	1.84E+04	3.34E+04	4.08E+04	4.41E+04	7.95E+04	5.21E+04	6.65E+04	1.22E+05	2.90E+05	4.32E+04	1.35E+05	6.29E+04	2.54E+04	7.57E+04	4.71E+04	5.12E+04	1.38E+05	1.22E+04
3-Oxotetradecanoic acid	4.44E+04	3.23E+04	3.04E+04	1.54E+04	2.73E+04	7.02E+04	4.33E+04	3.93E+04	4.96E+04	2.58E+04	1.20E+04	5.91E+04	1.10E+04	1.47E+04	2.78E+04	3.23E+04	8.18E+04	3.92E+04	4.20E+04	9.65E+04	2.10E+05	4.56E+04	5.95E+04	6.90E+04	3.55E+04	4.07E+04	1.92E+04	3.79E+04	5.13E+04	1.90E+04
4-(1,2-Dihydroxy-2-propanyl)-1-methyl-1,2-cyclohexanediol	9.93E+03	4.32E+03	1.06E+04	2.84E+03	9.79E+03	1.14E+04	1.02E+04	1.23E+04	1.09E+04	4.44E+03	2.66E+03	1.48E+04	2.48E+03	6.00E+03	7.28E+03	3.03E+03	8.50E+03	4.78E+03	5.76E+03	2.33E+04	2.76E+04	1.40E+04	7.66E+03	2.54E+04	2.35E+04	6.65E+03	4.91E+03	9.39E+03	3.54E+03	4.35E+03
4-Undecylbenzenesulfonic acid	2.47E+05	1.89E+05	7.77E+04	7.50E+04	8.35E+04	1.41E+05	1.33E+05	2.30E+05	1.18E+05	4.45E+04	2.48E+04	6.16E+04	2.19E+04	1.02E+05	7.34E+04	4.50E+04	5.93E+04	8.38E+04	6.87E+04	2.18E+05	2.85E+05	5.29E+04	8.37E+04	8.90E+04	5.95E+05	5.56E+04	4.02E+04	1.99E+05	6.88E+04	3.81E+04
5-Hydroxydecanoic acid	2.70E+03	3.95E+03	3.36E+03	9.70E+02	2.31E+03	2.21E+04	1.32E+04	3.14E+03	5.42E+03	1.80E+03	2.39E+03	8.14E+03	2.10E+03	5.33E+03	3.74E+03	3.64E+03	9.89E+03	4.22E+03	4.35E+03	7.34E+03	6.68E+04	4.16E+03	5.72E+03	5.05E+03	6.50E+03	6.82E+03	1.45E+03	1.02E+04	7.82E+03	1.27E+03
5-hydroxyisouric acid	8.70E+03	9.20E+03	4.77E+04	3.25E+04	1.20E+04	1.72E+04	3.37E+03	2.84E+03	8.48E+03	6.37E+03	2.20E+03	2.03E+03	4.64E+03	4.70E+03	5.19E+03	3.19E+03	2.68E+03	2.47E+03	3.70E+03	3.52E+03	3.68E+03	2.41E+03	2.87E+03	1.65E+03	1.04E+03	5.29E+03	1.35E+03	2.98E+03	3.54E+03	6.94E+02
9-Oxononanoic acid	9.44E+04	6.32E+04	5.93E+04	1.45E+05	6.66E+04	9.45E+04	8.77E+04	9.25E+04	9.61E+04	4.00E+04	2.34E+04	1.08E+05	2.07E+04	5.23E+04	5.46E+04	4.41E+04	6.23E+04	5.58E+04	5.64E+04	1.52E+05	2.74E+05	8.07E+04	9.69E+04	1.32E+05	3.25E+05	5.39E+04	2.85E+04	1.80E+05	5.27E+04	3.29E+04
Arachidic acid	1.99E+04	1.09E+04	9.65E+03	6.80E+03	1.41E+04	3.19E+04	2.02E+04	1.12E+04	1.70E+04	7.36E+03	7.98E+03	6.07E+04	3.59E+03	1.87E+04	1.36E+04	1.13E+04	2.93E+04	1.09E+04	1.53E+04	3.92E+04	3.37E+05	4.08E+04	3.22E+04	1.58E+05	8.38E+04	1.58E+04	4.05E+03	1.62E+04	9.72E+03	6.41E+03
Azelaic acid	5.16E+04	8.23E+04	9.71E+03	6.74E+03	7.39E+03	1.05E+05	4.81E+04	4.83E+04	4.29E+04	1.83E+04	1.18E+04	3.31E+04	1.29E+04	3.60E+04	3.85E+04	2.61E+04	2.85E+04	3.89E+04	2.56E+04	8.14E+04	1.90E+05	3.92E+04	6.51E+04	5.03E+04	9.89E+04	3.87E+04	1.67E+04	3.98E+04	4.07E+04	9.42E+03
Benzoic acid	8.83E+04	7.33E+04	7.07E+04	4.08E+04	5.77E+04	9.11E+04	4.97E+04	8.52E+04	8.94E+04	6.76E+04	9.03E+04	1.50E+05	2.95E+04	7.53E+04	1.12E+05	7.69E+04	7.43E+04	1.10E+05	1.12E+05	2.75E+05	3.08E+05	5.96E+04	7.54E+04	1.24E+05	1.49E+05	6.92E+04	3.97E+04	1.90E+05	6.60E+04	1.03E+04
Biacetyl	2.30E+04	2.83E+04	4.85E+04	3.23E+04	4.92E+04	5.63E+04	4.84E+04	4.01E+04	7.32E+04	2.99E+04	2.97E+04	4.62E+04	2.45E+04	4.41E+04	3.11E+04	2.88E+04	4.93E+04	5.11E+04	5.31E+04	8.08E+04	5.17E+04	3.38E+04	4.54E+04	1.13E+04	2.89E+04	6.97E+04	3.03E+04	6.06E+04	6.93E+04	1.02E+04
Citric acid	3.22E+05	1.82E+05	1.84E+05	9.21E+04	1.30E+05	2.48E+05	1.42E+05	1.59E+05	1.32E+05	7.64E+04	1.38E+05	1.52E+05	1.68E+04	1.79E+05	7.07E+04	4.32E+04	5.11E+04	6.92E+04	6.48E+04	1.49E+05	1.94E+05	4.62E+04	5.30E+04	1.12E+05	9.46E+04	3.94E+04	2.24E+04	6.81E+04	4.81E+04	1.37E+04
CMPF	7.03E+03	4.53E+03	4.36E+03	2.04E+03	4.81E+03	9.41E+03	8.40E+03	7.61E+03	6.47E+03	4.99E+03	1.93E+03	6.30E+03	2.06E+03	3.24E+03	3.38E+03	4.25E+03	4.53E+03	3.84E+03	4.03E+03	1.44E+04	1.35E+04	5.64E+03	4.46E+03	1.05E+04	1.17E+04	4.89E+03	3.21E+03	7.63E+03	4.73E+03	2.85E+03
Cyclopentadecanolide	1.08E+04	8.48E+03	6.24E+03	7.22E+03	8.48E+03	3.09E+03	4.80E+03	9.00E+03	8.76E+03	5.12E+03	4.02E+03	4.58E+03	3.57E+03	9.86E+03	8.21E+03	7.73E+03	4.62E+03	8.14E+03	8.41E+03	1.02E+04	3.18E+04	3.27E+03	1.28E+04	1.84E+04	4.66E+04	1.50E+04	3.28E+03	1.83E+04	8.90E+03	2.67E+03
Dimethyl (3-oxocyclohexyl)malonate	2.54E+04	8.77E+03	1.48E+04	9.77E+03	1.41E+04	3.50E+04	2.35E+04	3.08E+04	2.91E+04	8.23E+03	8.39E+03	2.66E+04	6.99E+03	1.74E+04	1.96E+04	1.50E+04	1.86E+04	1.34E+04	2.04E+04	3.37E+04	4.08E+04	2.00E+04	1.70E+04	1.96E+04	5.39E+04	1.22E+04	1.16E+04	4.04E+04	2.44E+04	8.68E+03
DL-Lactic Acid	3.80E+05	7.11E+05	2.63E+05	2.31E+05	2.04E+05	1.69E+06	4.19E+05	5.23E+05	1.37E+06	1.02E+06	3.05E+05	4.47E+05	6.08E+05	5.56E+05	6.27E+05	6.19E+05	8.63E+05	2.56E+05	8.14E+05	5.17E+05	1.42E+06	4.62E+05	6.77E+05	2.37E+05	9.55E+05	8.33E+05	3.31E+05	5.88E+05	1.92E+06	2.27E+05
Epomediol	2.08E+04	1.96E+04	1.84E+04	1.04E+04	2.11E+04	5.00E+04	2.30E+04	2.73E+04	2.81E+04	1.67E+04	8.70E+03	4.67E+04	6.87E+03	3.80E+04	1.53E+04	1.48E+04	2.58E+04	1.58E+04	2.61E+04	5.77E+04	1.05E+05	4.02E+04	3.90E+04	6.50E+04	8.20E+04	1.35E+04	1.20E+04	7.23E+04	2.34E+04	1.49E+04
frescolat ML	4.39E+04	3.12E+04	3.02E+04	1.34E+04	2.90E+04	5.16E+04	3.13E+04	4.75E+04	4.84E+04	2.74E+04	1.74E+04	3.23E+04	1.59E+04	1.60E+04	3.63E+04	3.99E+04	2.83E+04	5.13E+04	4.77E+04	1.25E+05	1.10E+05	1.78E+04	5.63E+04	2.93E+04	4.31E+04	5.22E+04	3.88E+04	4.76E+04	3.56E+04	7.75E+03
Glycolic acid	4.63E+04	4.92E+04	6.96E+04	4.07E+04	9.65E+04	4.13E+04	4.07E+04	4.93E+04	5.33E+04	2.85E+04	2.22E+04	5.85E+04	1.81E+04	3.35E+04	6.18E+04	3.11E+04	2.09E+04	3.54E+04	6.16E+04	1.58E+05	2.77E+05	5.35E+04	7.28E+04	4.05E+05	2.76E+05	3.41E+04	2.83E+04	3.60E+04	6.36E+04	1.74E+04
Itaconic acid	1.17E+05	1.36E+05	2.57E+05	1.67E+05	2.37E+05	2.74E+05	2.35E+05	1.90E+05	3.67E+05	1.41E+05	1.47E+05	2.33E+05	1.28E+05	2.17E+05	1.59E+05	1.42E+05	2.47E+05	2.45E+05	2.57E+05	3.64E+05	2.59E+05	1.66E+05	2.28E+05	3.75E+04	1.21E+05	3.33E+05	1.48E+05	2.78E+05	3.32E+05	5.04E+04
Juniperic acid	6.41E+04	4.89E+04	3.74E+04	2.50E+04	3.90E+04	1.30E+05	7.71E+04	6.52E+04	6.99E+04	3.07E+04	2.05E+04	1.16E+05	1.48E+04	3.43E+04	4.41E+04	3.50E+04	9.19E+04	4.75E+04	5.72E+04	1.29E+05	4.29E+05	8.01E+04	6.26E+04	1.60E+05	9.32E+04	4.44E+04	2.64E+04	6.99E+04	5.40E+04	2.96E+04
Linoelaidic Acid	1.68E+05	1.15E+05	6.73E+05	1.09E+06	5.89E+05	1.41E+05	9.42E+04	1.74E+05	2.11E+05	1.06E+05	1.06E+05	1.90E+05	1.22E+05	5.54E+05	5.14E+05	2.39E+05	1.20E+05	1.40E+05	1.62E+05	2.36E+05	8.53E+05	2.09E+05	5.53E+05	1.25E+06	2.61E+06	4.07E+05	8.01E+04	1.58E+05	2.49E+05	2.82E+04
L-Pyroglutamic acid	2.62E+04	6.59E+04	5.10E+04	8.22E+04	2.80E+04	4.10E+04	1.60E+04	1.62E+04	1.54E+04	6.90E+03	7.00E+03	1.42E+04	9.50E+03	1.06E+04	1.27E+04	8.66E+03	2.81E+03	3.95E+03	1.30E+04	1.12E+04	6.45E+04	1.48E+04	2.08E+04	1.04E+05	3.35E+04	2.45E+04	2.03E+03	7.93E+03	4.29E+04	2.55E+03
L-α-PALMITIN	4.60E+04	2.57E+04	3.00E+04	1.09E+04	2.64E+04	6.51E+04	3.90E+04	4.63E+04	4.34E+04	2.14E+04	1.81E+04	6.51E+04	1.34E+04	4.57E+04	4.74E+04	2.86E+04	3.44E+04	4.78E+04	3.99E+04	9.12E+04	2.52E+05	6.15E+04	6.56E+04	1.34E+05	1.35E+05	2.78E+04	2.05E+04	4.22E+04	3.67E+04	1.37E+04
Methyl laurate	1.46E+04	1.42E+04	1.65E+04	1.11E+04	1.80E+04	2.25E+04	2.35E+04	1.55E+04	3.11E+04	1.61E+04	9.91E+03	2.13E+04	6.31E+03	3.56E+04	1.00E+04	9.16E+03	2.48E+04	2.72E+04	1.20E+04	3.24E+04	6.41E+04	2.20E+04	1.39E+04	4.05E+04	1.94E+05	1.11E+04	1.04E+04	3.20E+04	1.03E+04	4.81E+03
MHPG	2.31E+04	7.40E+03	1.32E+04	2.17E+03	1.17E+04	3.17E+04	1.36E+04	8.75E+03	1.22E+04	7.86E+03	2.79E+03	2.29E+04	4.40E+03	1.65E+04	1.23E+04	3.79E+03	3.00E+04	1.62E+04	1.11E+04	2.84E+04	4.79E+04	1.59E+04	1.30E+04	1.37E+04	3.99E+04	3.02E+03	7.24E+03	1.09E+04	8.92E+03	7.78E+03
myristic acid	6.62E+05	5.79E+05	3.65E+05	2.91E+05	4.10E+05	1.01E+06	5.45E+05	6.76E+05	7.90E+05	4.06E+05	9.25E+04	3.31E+05	6.77E+04	1.05E+05	1.52E+05	1.96E+05	2.70E+05	2.03E+05	2.76E+05	4.48E+05	1.11E+06	1.83E+05	3.25E+05	4.29E+05	8.97E+05	2.71E+05	1.04E+05	3.21E+05	3.05E+05	7.03E+04
N-Acetyl-L-aspartic acid	7.15E+03	2.63E+04	1.74E+04	1.33E+04	3.35E+04	1.06E+04	8.55E+03	1.59E+04	1.66E+04	5.08E+03	1.98E+04	2.43E+04	2.87E+04	3.91E+04	1.02E+04	1.57E+04	3.41E+04	1.57E+04	1.18E+04	2.71E+04	9.23E+04	3.39E+04	4.54E+04	5.66E+03	3.17E+04	6.70E+03	1.44E+04	2.05E+04	4.43E+04	4.31E+03
Oleic acid	1.42E+05	8.99E+04	4.67E+05	5.58E+05	4.19E+05	1.40E+05	1.09E+05	1.46E+05	1.48E+05	7.02E+04	7.22E+04	1.78E+05	7.10E+04	3.18E+05	3.39E+05	1.37E+05	1.11E+05	9.56E+04	1.10E+05	2.02E+05	1.14E+06	3.65E+05	4.53E+05	1.69E+06	1.78E+06	1.80E+05	5.85E+04	1.56E+05	1.39E+05	4.04E+04
Palmitic Acid	3.95E+06	2.06E+06	1.98E+06	1.75E+06	2.40E+06	2.60E+06	1.41E+06	3.59E+06	3.59E+06	1.42E+06	1.64E+06	2.80E+06	1.08E+06	5.96E+06	3.90E+06	2.74E+06	1.74E+06	3.60E+06	3.02E+06	8.78E+06	8.08E+06	1.25E+06	3.91E+06	3.07E+06	1.12E+07	2.26E+06	1.55E+06	4.24E+06	2.21E+06	4.48E+05
p-Cresylsulfate	1.01E+04	9.91E+03	6.17E+04	2.50E+04	5.56E+04	1.36E+04	1.33E+04	1.02E+04	1.08E+04	5.37E+03	7.62E+03	3.34E+04	6.89E+03	1.25E+04	2.81E+04	1.18E+04	1.37E+04	3.00E+03	5.13E+03	8.59E+03	1.16E+05	3.49E+04	3.73E+04	2.29E+05	8.44E+04	4.34E+03	1.44E+03	4.77E+03	8.06E+03	3.36E+03
pentadecanoic acid	6.15E+04	5.09E+04	4.85E+04	4.65E+04	6.22E+04	5.72E+04	3.65E+04	6.21E+04	6.94E+04	4.01E+04	2.48E+04	4.43E+04	2.07E+04	7.60E+04	7.67E+04	5.59E+04	3.73E+04	6.60E+04	6.15E+04	1.49E+05	1.64E+05	3.64E+04	7.58E+04	8.78E+04	3.35E+05	6.23E+04	3.36E+04	9.13E+04	5.62E+04	1.37E+04
Propyl (9E)-12-hydroxy-9-octadecenoate	2.99E+04	1.85E+04	1.52E+04	8.56E+03	1.05E+04	2.86E+04	2.21E+04	2.38E+04	2.25E+04	1.27E+04	7.49E+03	2.75E+04	7.41E+03	2.76E+04	2.06E+04	2.25E+04	1.85E+04	1.93E+04	2.18E+04	6.85E+04	9.23E+04	2.88E+04	2.67E+04	3.52E+04	3.21E+04	2.01E+04	1.69E+04	2.12E+04	2.71E+04	1.12E+04
RG1300000	2.26E+05	1.67E+05	1.45E+05	1.42E+05	1.40E+05	4.22E+05	2.33E+05	2.34E+05	2.52E+05	1.14E+05	7.62E+04	2.94E+05	5.82E+04	1.05E+05	1.53E+05	1.67E+05	2.66E+05	1.93E+05	2.32E+05	4.82E+05	1.29E+06	2.22E+05	2.68E+05	4.01E+05	4.98E+05	2.03E+05	9.87E+04	2.23E+05	2.45E+05	8.16E+04
RH3422500	4.66E+03	2.50E+03	2.96E+03	2.79E+03	2.21E+03	5.18E+03	4.16E+03	3.30E+03	5.36E+03	1.46E+03	1.45E+03	3.72E+03	1.70E+03	4.76E+03	5.22E+03	2.56E+03	2.16E+03	2.51E+03	3.10E+03	7.21E+03	9.96E+03	3.16E+03	3.83E+03	2.54E+03	1.83E+04	3.80E+03	1.21E+03	4.00E+03	3.59E+03	1.32E+03
Stearic acid	1.29E+06	7.01E+05	7.79E+05	4.61E+05	1.06E+06	1.58E+06	1.13E+06	1.16E+06	1.17E+06	4.39E+05	6.21E+05	2.37E+06	3.48E+05	2.22E+06	1.59E+06	8.16E+05	1.25E+06	1.12E+06	1.00E+06	3.61E+06	1.01E+07	2.36E+06	2.21E+06	5.13E+06	5.92E+06	7.53E+05	5.67E+05	1.60E+06	9.44E+05	4.02E+05
Succinic acid	5.62E+04	6.13E+04	5.45E+04	1.74E+04	2.31E+04	5.72E+04	4.56E+04	4.39E+04	5.05E+04	4.74E+04	5.01E+04	6.95E+04	3.81E+04	2.03E+04	5.12E+04	6.60E+04	8.01E+04	5.85E+04	8.22E+04	1.10E+05	1.48E+05	4.17E+04	6.54E+04	8.01E+04	9.29E+04	1.29E+05	5.22E+04	2.44E+04	7.76E+04	2.61E+04
Thapsic acid	2.98E+04	2.20E+04	1.29E+04	7.17E+03	1.60E+04	7.16E+04	5.00E+04	3.19E+04	3.08E+04	1.89E+04	9.37E+03	5.58E+04	6.92E+03	2.88E+04	2.18E+04	1.94E+04	7.21E+04	2.80E+04	3.07E+04	7.01E+04	1.93E+05	3.56E+04	6.05E+04	4.71E+04	1.63E+04	2.61E+04	1.49E+04	7.29E+04	5.23E+04	1.40E+04
Traumatic Acid	1.21E+05	8.61E+04	8.30E+04	4.25E+04	5.63E+04	1.35E+05	9.03E+04	1.40E+05	1.34E+05	6.32E+04	4.10E+04	1.43E+05	2.90E+04	9.22E+04	1.10E+05	6.15E+04	1.21E+05	1.17E+05	1.32E+05	1.67E+05	5.41E+05	1.26E+05	1.45E+05	2.28E+05	2.72E+05	9.75E+04	4.28E+04	1.49E+05	1.15E+05	4.24E+04
Undecanedioic acid	1.37E+04	1.10E+04	3.51E+03	2.01E+03	4.42E+03	4.67E+04	8.19E+03	8.45E+03	1.18E+04	1.46E+04	2.57E+03	1.71E+04	4.98E+03	5.03E+03	5.40E+03	5.52E+03	9.51E+03	1.05E+04	1.40E+04	5.88E+04	8.06E+04	7.50E+03	3.35E+04	8.19E+03	3.41E+04	2.72E+04	1.09E+04	8.36E+03	1.03E+04	2.93E+03
Valproic acid	6.04E+04	5.57E+04	7.02E+05	2.43E+05	5.94E+05	3.09E+05	2.78E+05	8.60E+04	1.12E+05	4.86E+04	4.59E+04	3.63E+05	3.77E+04	1.98E+05	2.55E+05	9.66E+04	2.78E+05	2.60E+04	6.00E+04	4.78E+04	1.97E+06	1.85E+05	4.13E+05	2.75E+06	9.65E+05	4.24E+04	2.10E+04	4.47E+04	1.33E+05	1.96E+04
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Human Metabolome Database
(-)-Chloramphenicol	HMDB0014589
(-)-pinellic acid	HMDB0004708
12-HSA	HMDB0061706
12-Hydroxylauric acid	HMDB0002059
13S-hydroxyoctadecadienoic acid	HMDB0004667
16-Heptadecyne-1,2,4-triol	HMDB0035473
1-Acetylcyclohexyl acetate	HMDB0039710
1-nonanoic acid	HMDB0000847
1-stearoyl-2-arachidonoyl-sn-glycero-3-phosphoethanolamine	HMDB0009003
2-Dodecylbenzenesulfonic acid	HMDB0031031
2-Hydroxy-4-oxohenicosyl acetate	HMDB0036344
3-Hydroxy-3-(2,3,4-trimethoxyphenyl)propanoic acid	HMDB0142079
3-Hydroxysebacic acid	HMDB0000350
3-oxolauric acid	HMDB0010727
3-Oxotetradecanoic acid	HMDB0010730
4-(1,2-Dihydroxy-2-propanyl)-1-methyl-1,2-cyclohexanediol	HMDB0039974
4-Undecylbenzenesulfonic acid	HMDB0032549
5-Hydroxydecanoic acid	HMDB0040329
5-hydroxyisouric acid	HMDB0030097
9-Oxononanoic acid	HMDB0094711
Arachidic acid	HMDB0002212
Azelaic acid	HMDB0000784
Benzoic acid	HMDB0001870
Biacetyl	HMDB0003407
Citric acid	HMDB0000094
CMPF	HMDB0061112
Cyclopentadecanolide	HMDB0034455
Dimethyl (3-oxocyclohexyl)malonate	HMDB0032496
DL-Lactic Acid	HMDB0144295
Epomediol	HMDB0041376
frescolat ML	HMDB0037212
Glycolic acid	HMDB0000115
Itaconic acid	HMDB0002092
Juniperic acid	HMDB0006294
Linoelaidic Acid	HMDB0006270
L-Pyroglutamic acid	HMDB0000267
L-α-PALMITIN	HMDB0005356
Methyl laurate	HMDB0031018
MHPG	HMDB0001490
myristic acid	HMDB0000806
N-Acetyl-L-aspartic acid	HMDB0000812
Oleic acid	HMDB0000207
Palmitic Acid	HMDB0000220
p-Cresylsulfate	HMDB0011635
pentadecanoic acid	HMDB0000826
Propyl (9E)-12-hydroxy-9-octadecenoate	HMDB0032476
RG1300000	HMDB0247617
RH3422500	HMDB0034844
Stearic acid	HMDB0000827
Succinic acid	HMDB0000254
Thapsic acid	HMDB0000672
Traumatic Acid	HMDB0000933
Undecanedioic acid	HMDB0000888
Valproic acid	HMDB0001877
METABOLITES_END
#END