#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