#METABOLOMICS WORKBENCH jburgess_20160305_080332 STUDY_ID:ST000359 ANALYSIS_ID:AN000589 PROJECT_ID:PR000285 VERSION 1 CREATED_ON March 10, 2016, 8:03 pm #PROJECT PR:PROJECT_TITLE Metabolic Profiling of Anxiety Prone HSV-Latently Infected Obese Mice PR:PROJECT_TYPE Broad Spectrum LCMS PR:PROJECT_SUMMARY The biological factors that lead children from low socioeconomic backgrounds to PR:PROJECT_SUMMARY be at greater risk for the development of anxiety and learning problems are not PR:PROJECT_SUMMARY well understood. While it is clear that there are genetic components to the risk PR:PROJECT_SUMMARY of developing mental health disorders, a role for environmental factors in PR:PROJECT_SUMMARY inducing these problems has been suggested. Many of these factors that affect PR:PROJECT_SUMMARY the brain likely involve exposures that occur early in life. Two such factors PR:PROJECT_SUMMARY are the higher rates of herpes simplex virus (HSV)-1 seropositivity and PR:PROJECT_SUMMARY prevalence of obesity among these children. HSV-1 infection has been associated PR:PROJECT_SUMMARY with impaired cognition during childhood and mental health problems in PR:PROJECT_SUMMARY adulthood. Additionally, obese adults are shown to have higher HSV-1 titers. PR:PROJECT_SUMMARY Similarly, other studies have correlated a diet high in saturated fat and PR:PROJECT_SUMMARY obesity with increased risk of mood disorders and anxiety. A mouse model of PR:PROJECT_SUMMARY obese HSV-1 latent infection was developed. Broad spectrum metabolomics analysis PR:PROJECT_SUMMARY was performed to better understand the metabolomic profile of hippocampus and to PR:PROJECT_SUMMARY compare this metabolomics profile with that of the hypothalamus, microglia, and PR:PROJECT_SUMMARY peripheral blood mononuclear cells. Brain tissue samples for metabolomics PR:PROJECT_SUMMARY experiments were generated in the following manner: 3-week old mice were placed PR:PROJECT_SUMMARY on a 10% low fat (LF) diet and acclimated for one week prior to intranasal HSV-1 PR:PROJECT_SUMMARY infection or mock infection with PBS. Fourteen days post-infection mice were PR:PROJECT_SUMMARY randomized to either a 45% high fat diet (HF) or remained on the LF diet. Eight PR:PROJECT_SUMMARY weeks post-diet transition, mice were euthanized and brain tissue samples were PR:PROJECT_SUMMARY collected and processed for metabolomics. PR:INSTITUTE University of North Carolina , Chapel Hill PR:DEPARTMENT Gillings School of Public Health PR:LABORATORY Department of Nutrition PR:LAST_NAME Sheridan PR:FIRST_NAME Patricia PR:ADDRESS 2002 Hooker Research Center, CB#7461, Chapel Hill NC, 27599 PR:EMAIL patricia_sheridan@med.unc.edu PR:PHONE 919-843-6434 PR:FUNDING_SOURCE NIH, Common Fund, Pilot and Feasibility #STUDY ST:STUDY_TITLE Broad Spectrum MS analysis of mouse microglia cells from Anxiety Prone ST:STUDY_TITLE HSV-Latently Infected Obese Mice ST:STUDY_TYPE Broad Spectrum LCMS ST:STUDY_SUMMARY A mouse model of obese HSV-1 latent infection was developed. Broad spectrum ST:STUDY_SUMMARY metabolomics analysis was performed to better understand the metabolomic profile ST:STUDY_SUMMARY of microglia cells and to compare this metabolomics profile with that of the ST:STUDY_SUMMARY hippocampus, hypothalamus, and peripheral blood mononuclear cells. Microglia ST:STUDY_SUMMARY cell samples for metabolomics experiments were generated in the following ST:STUDY_SUMMARY manner: 3-week old mice were placed on a 10% low fat (LF) diet and acclimated ST:STUDY_SUMMARY for one week prior to intranasal HSV-1 infection or mock infection with PBS. ST:STUDY_SUMMARY Fourteen days post-infection mice were randomized to either a 45% high fat diet ST:STUDY_SUMMARY (HF) or remained on the LF diet. Eight weeks post-diet transition, mice were ST:STUDY_SUMMARY euthanized and microglia cell samples were collected and processed for ST:STUDY_SUMMARY metabolomics. ST:INSTITUTE RTI ST:DEPARTMENT Discovery Sciences ST:LABORATORY System and Translational Sciences ST:LAST_NAME Sumner ST:FIRST_NAME Susan ST:ADDRESS 3040 Cornwallis Rd, RTP,NC ST:EMAIL ssumner@rti.org ST:PHONE 919-541-7479 ST:NUM_GROUPS 4 ST:TOTAL_SUBJECTS 31 ST:NUM_MALES 31 #SUBJECT SU:SUBJECT_TYPE mouse SU:SUBJECT_SPECIES Mus musculus SU:GENOTYPE_STRAIN C57Bl/6 SU:AGE_OR_AGE_RANGE 5.5 mo SU:WEIGHT_OR_WEIGHT_RANGE 29.1 g-49.2 g SU:GENDER male SU:ANIMAL_ANIMAL_SUPPLIER JAX SU:ANIMAL_HOUSING 4/cage SU:ANIMAL_FEED Purina chow #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS 432 Micro432 HSV-1:no | Diet at Week 2:LF Age=4.5 mo; Weight (g) =29.6 SUBJECT_SAMPLE_FACTORS 419 Micro419 HSV-1:no | Diet at Week 2:HF Age=4.5 mo; Weight (g) =33.2 SUBJECT_SAMPLE_FACTORS 418 Micro418 HSV-1:no | Diet at Week 2:HF Age=4.5 mo; Weight (g) =30.8 SUBJECT_SAMPLE_FACTORS 425 Micro425 HSV-1:yes | Diet at Week 2:HF Age=4.5 mo; Weight (g) =33.2 SUBJECT_SAMPLE_FACTORS 422 Micro422 HSV-1:no | Diet at Week 2:LF Age=4.5 mo; Weight (g) =30.3 SUBJECT_SAMPLE_FACTORS 430 Micro430 HSV-1:yes | Diet at Week 2:LF Age=4.5 mo; Weight (g) =25.2 SUBJECT_SAMPLE_FACTORS 441 Micro441 HSV-1:yes | Diet at Week 2:HF Age=4.5 mo; Weight (g) =38.6 SUBJECT_SAMPLE_FACTORS 433 Micro433 HSV-1:no | Diet at Week 2:LF Age=4.5 mo; Weight (g) =26.4 SUBJECT_SAMPLE_FACTORS 439 Micro439 HSV-1:yes | Diet at Week 2:HF Age=4.5 mo; Weight (g) =33.8 SUBJECT_SAMPLE_FACTORS 421 Micro421 HSV-1:no | Diet at Week 2:LF Age=4.5 mo; Weight (g) =30.8 SUBJECT_SAMPLE_FACTORS 435 Micro435 HSV-1:yes | Diet at Week 2:LF Age=4.5 mo; Weight (g) =24.9 SUBJECT_SAMPLE_FACTORS 424 Micro424 HSV-1:no | Diet at Week 2:LF Age=4.5 mo; Weight (g) =29.8 SUBJECT_SAMPLE_FACTORS 417 Micro417 HSV-1:no | Diet at Week 2:HF Age=4.5 mo; Weight (g) =32.6 SUBJECT_SAMPLE_FACTORS 446 Micro446 HSV-1:No | Diet at Week 2:HF Age=4.5 mo; Weight (g) =39 SUBJECT_SAMPLE_FACTORS 444 Micro444 HSV-1:No | Diet at Week 2:HF Age=4.5 mo; Weight (g) =38.1 SUBJECT_SAMPLE_FACTORS 436 Micro436 HSV-1:yes | Diet at Week 2:LF Age=4.5 mo; Weight (g) =29.8 SUBJECT_SAMPLE_FACTORS 445 Micro445 HSV-1:No | Diet at Week 2:HF Age=4.5 mo; Weight (g) =37.7 SUBJECT_SAMPLE_FACTORS 438 Micro438 HSV-1:yes | Diet at Week 2:LF Age=4.5 mo; Weight (g) =29.9 SUBJECT_SAMPLE_FACTORS 423 Micro423 HSV-1:no | Diet at Week 2:LF Age=4.5 mo; Weight (g) =30.7 SUBJECT_SAMPLE_FACTORS 429 Micro429 HSV-1:yes | Diet at Week 2:LF Age=4.5 mo; Weight (g) =28.6 SUBJECT_SAMPLE_FACTORS 427 Micro427 HSV-1:yes | Diet at Week 2:HF Age=4.5 mo; Weight (g) =31.4 SUBJECT_SAMPLE_FACTORS 434 Micro434 HSV-1:no | Diet at Week 2:LF Age=4.5 mo; Weight (g) =26.2 SUBJECT_SAMPLE_FACTORS 420 Micro420 HSV-1:no | Diet at Week 2:HF Age=4.5 mo; Weight (g) =33.8 SUBJECT_SAMPLE_FACTORS 440 Micro440 HSV-1:yes | Diet at Week 2:HF Age=4.5 mo; Weight (g) =38.3 SUBJECT_SAMPLE_FACTORS 426 Micro426 HSV-1:yes | Diet at Week 2:HF Age=4.5 mo; Weight (g) =30.9 SUBJECT_SAMPLE_FACTORS 431 Micro431 HSV-1:no | Diet at Week 2:LF Age=4.5 mo; Weight (g) =32.7 SUBJECT_SAMPLE_FACTORS 442 Micro442 HSV-1:yes | Diet at Week 2:HF Age=4.5 mo; Weight (g) =33.4 SUBJECT_SAMPLE_FACTORS 443 Micro443 HSV-1:No | Diet at Week 2:HF Age=4.5 mo; Weight (g) =37.8 SUBJECT_SAMPLE_FACTORS 437 Micro437 HSV-1:yes | Diet at Week 2:LF Age=4.5 mo; Weight (g) =27.6 SUBJECT_SAMPLE_FACTORS 428 Micro428 HSV-1:yes | Diet at Week 2:LF Age=4.5 mo; Weight (g) =28.9 SUBJECT_SAMPLE_FACTORS - Total_Pool_01 HSV-1:- | Diet at Week 2:- Age=-; Weight (g) =- SUBJECT_SAMPLE_FACTORS - Total_Pool_02 HSV-1:- | Diet at Week 2:- Age=-; Weight (g) =- SUBJECT_SAMPLE_FACTORS - Total_Pool_03 HSV-1:- | Diet at Week 2:- Age=-; Weight (g) =- SUBJECT_SAMPLE_FACTORS - Total_Pool_04 HSV-1:- | Diet at Week 2:- Age=-; Weight (g) =- SUBJECT_SAMPLE_FACTORS - Total_Pool_05 HSV-1:- | Diet at Week 2:- Age=-; Weight (g) =- #COLLECTION CO:COLLECTION_SUMMARY For extraction, 500 µL of an ice-cold solution of 90:10 Methanol:Chloroform CO:COLLECTION_SUMMARY (v/v) ¬was added to the tubes containing the thawed cell pellets. MagNA lyser CO:COLLECTION_SUMMARY ceramic beads were added to the tubes, and a MagNA lyser was used to beat the CO:COLLECTION_SUMMARY samples for two 30 seconds pulses at 2,000 rpm placing samples on cold block for CO:COLLECTION_SUMMARY 5 min in between pulses. Sonication for 5 min was performed to the beaten cell CO:COLLECTION_SUMMARY samples, followed by centrifugation at room temperature and at 16,000rcf for 4 CO:COLLECTION_SUMMARY min. A 350 µL aliquot of each experimental cell sample homogenized supernatant CO:COLLECTION_SUMMARY was transferred to a labeled 2.0 mL Lo-Bind Eppendorf tube CO:SAMPLE_TYPE brain homogenate #TREATMENT TR:TREATMENT_SUMMARY 3-week old mice were placed on a 10% low fat (LF) diet and acclimated for one TR:TREATMENT_SUMMARY week prior to intranasal HSV-1 infection or mock infection with PBS. Fourteen TR:TREATMENT_SUMMARY days post-infection mice were randomized to either a 45% high fat diet (HF) or TR:TREATMENT_SUMMARY remained on the LF diet. Eight weeks post-diet transition, mice were euthanized TR:TREATMENT_SUMMARY and brain tissue samples were collected and processed for metabolomics. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY For extraction, 500 µL of an ice-cold solution of 90:10 Methanol:Chloroform SP:SAMPLEPREP_SUMMARY (v/v) ¬was added to the tubes containing the thawed cell pellets. MagNA lyser SP:SAMPLEPREP_SUMMARY ceramic beads were added to the tubes, and a MagNA lyser was used to beat the SP:SAMPLEPREP_SUMMARY samples for two 30 seconds pulses at 2,000 rpm placing samples on cold block for SP:SAMPLEPREP_SUMMARY 5 min in between pulses. Sonication for 5 min was performed to the beaten cell SP:SAMPLEPREP_SUMMARY samples, followed by centrifugation at room temperature and at 16,000rcf for 4 SP:SAMPLEPREP_SUMMARY min. A 350 µL aliquot of each experimental cell sample homogenized supernatant SP:SAMPLEPREP_SUMMARY was transferred to a labeled 2.0 mL Lo-Bind Eppendorf tube Analytical pooled SP:SAMPLEPREP_SUMMARY samples were created by combining a 125 µL aliquot from each experimental SP:SAMPLEPREP_SUMMARY sample in a labeled 20 mL glass vial. The QC pooled sample was vortexed for 30 SP:SAMPLEPREP_SUMMARY sec and 350 µL aliquots were transferred to 5 labeled 2.0 mL Lo-Bind eppendorf SP:SAMPLEPREP_SUMMARY tubes. Next, the supernatants of the study and QC pooled samples were dried SP:SAMPLEPREP_SUMMARY overnight using a lyophilizer. The residue was reconstituted in 50 µL of SP:SAMPLEPREP_SUMMARY reconstitution buffer (95:5 Water:Methanol, v/v) and mixed on an analog vortex SP:SAMPLEPREP_SUMMARY mixer for 1 min with speed set at 10. Then, the samples were centrifuged at room SP:SAMPLEPREP_SUMMARY temperature for 4 min at 16,000 rcf, and the supernatants were transferred to SP:SAMPLEPREP_SUMMARY autosampler vials for data acquisition. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY UPLC-MS spectra were collected for all samples. UPLC was performed on a Waters CH:CHROMATOGRAPHY_SUMMARY Acquity UPLC with an Acquity HSS T3 column (2.1x 100mm x 1.8 µm) at 50 °C CH:CHROMATOGRAPHY_SUMMARY using the reversed-phase separation. Water with 0.1% formic acid (mobile phase CH:CHROMATOGRAPHY_SUMMARY A) and methanol with 0.1% formic acid (mobile phase B) were used as mobile CH:CHROMATOGRAPHY_SUMMARY phases and the metabolites were chromatographically separated using a gradient CH:CHROMATOGRAPHY_SUMMARY separation: (see the 3. Sheridan-Mice-Microglia Cells_RP-Metadata and Analytical CH:CHROMATOGRAPHY_SUMMARY Metadata.xlsx file for the flow gradient). Mass spectroscopy analysis was CH:CHROMATOGRAPHY_SUMMARY performed using a Synapt G2 Si ESI-Q-TOF using a 10 µL injection volume. CH:CHROMATOGRAPHY_SUMMARY UPLC-MS data were collected over 70-1000 m/z in both positive and negative CH:CHROMATOGRAPHY_SUMMARY modes. CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Synapt G2 Si ESI-Q-TOF CH:COLUMN_NAME Acquity HSS T3 CH:FLOW_RATE .4ml/min CH:METHODS_FILENAME RTI-RCMRC-RP CH:SOLVENT_A Water with 0.1% formic acid CH:SOLVENT_B Methanol with 0.1% formic acid CH:INJECTION_TEMPERATURE 8 C #ANALYSIS AN:ANALYSIS_TYPE MS AN:OPERATOR_NAME Suraj Dhungana AN:DETECTOR_TYPE TOF AN:SOFTWARE_VERSION Mass Lynx 4.1 AN:ACQUISITION_DATE 7/16/15 AN:ANALYSIS_PROTOCOL_FILE RTI-RCMRC-RP-NEG AN:DATA_FORMAT Continuum #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Waters Synapt G2 Si QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS intensity MS_METABOLITE_DATA_START Samples Total_Pool_01 Total_Pool_02 Total_Pool_03 Total_Pool_04 Total_Pool_05 Micro417 Micro418 Micro419 Micro420 Micro443 Micro444 Micro445 Micro446 Micro425 Micro426 Micro427 Micro439 Micro440 Micro441 Micro442 Micro421 Micro422 Micro423 Micro424 Micro431 Micro432 Micro433 Micro434 Micro428 Micro429 Micro430 Micro435 Micro436 Micro437 Micro438 Factors HSV-1:- | Diet at Week 2:- HSV-1:- | Diet at Week 2:- HSV-1:- | Diet at Week 2:- HSV-1:- | Diet at Week 2:- HSV-1:- | Diet at Week 2:- HSV-1:no | Diet at Week 2:HF HSV-1:no | Diet at Week 2:HF HSV-1:no | Diet at Week 2:HF HSV-1:no | Diet at Week 2:HF HSV-1:No | Diet at Week 2:HF HSV-1:No | Diet at Week 2:HF HSV-1:No | Diet at Week 2:HF HSV-1:No | Diet at Week 2:HF HSV-1:yes | Diet at Week 2:HF HSV-1:yes | Diet at Week 2:HF HSV-1:yes | Diet at Week 2:HF HSV-1:yes | Diet at Week 2:HF HSV-1:yes | Diet at Week 2:HF HSV-1:yes | Diet at Week 2:HF HSV-1:yes | Diet at Week 2:HF HSV-1:no | Diet at Week 2:LF HSV-1:no | Diet at Week 2:LF HSV-1:no | Diet at Week 2:LF HSV-1:no | Diet at Week 2:LF HSV-1:no | Diet at Week 2:LF HSV-1:no | Diet at Week 2:LF HSV-1:no | Diet at Week 2:LF HSV-1:no | Diet at Week 2:LF HSV-1:yes | Diet at Week 2:LF HSV-1:yes | Diet at Week 2:LF HSV-1:yes | Diet at Week 2:LF HSV-1:yes | Diet at Week 2:LF HSV-1:yes | Diet at Week 2:LF HSV-1:yes | Diet at Week 2:LF HSV-1:yes | Diet at Week 2:LF 15.84_279.2295m/z 156.4668474 201.0170729 215.2127156 168.7584637 209.711573 67.99917904 44.61093846 67.73882265 23.37455705 501.3475997 284.494715 122.9225047 142.9330742 183.3088073 291.888133 284.7729357 173.4851663 234.4204366 464.8972689 831.6771214 99.82936224 134.7940671 159.2813366 241.2558496 148.1826344 84.41099492 130.0394153 203.4693053 203.855605 277.9295955 196.6123212 355.6672709 463.8316415 237.713915 352.8856657 0.68_94.9229m/z 38.98851521 43.71150703 40.68729016 43.44157718 37.95582923 37.86626543 78.09759762 22.83536432 96.55468829 23.24590136 22.10039227 20.43100631 62.53303252 13.77825684 16.76639585 34.34867878 21.84570388 31.54643971 11.45650114 18.25312959 26.99234668 78.60612498 89.27499474 49.88177955 28.33762204 58.33447538 85.21807529 45.82429105 92.20082872 41.46025586 78.22742525 25.1365286 71.25448031 20.04005934 47.30511896 0.68_92.9258m/z 68.11501291 83.00174389 74.41014311 78.63794002 67.52999549 67.2686803 150.5385307 42.84630688 169.911505 40.05705522 36.19612639 35.87706866 112.8693716 22.64241429 30.77438899 54.604234 51.09495395 50.2978683 20.02765587 30.67794839 46.10702895 137.6971535 159.340569 93.93223539 40.13530689 108.7279851 155.1428193 82.15800858 162.5177644 71.29237834 145.0688169 49.03418828 129.7703223 35.16450717 93.51050245 0.68_272.7926m/z 27.40804427 27.70701306 25.40570052 26.02247381 21.57865567 25.95822008 62.16435018 24.57972834 72.80810129 14.1425325 13.44688845 9.064278886 48.84649474 7.481706038 9.251259652 20.60072064 27.03813442 13.34979437 2.503816323 11.67323259 15.66631473 61.55002603 58.61554502 27.25704923 17.79444627 47.61870971 64.63520261 32.24134903 63.90840263 27.76397495 52.02814998 18.16700714 47.92447589 9.578675235 48.17396976 0.65_294.8213m/z 39.2112649 48.03866581 53.33736938 38.29273148 40.54433879 43.01876353 78.81984828 26.70260997 74.17472695 32.74397917 20.60152602 34.89401628 49.9304898 14.01677397 19.29069894 43.77747628 40.4919024 34.5019848 8.011683507 20.96411068 31.54509969 63.62436518 62.2892625 30.45641264 33.15457432 57.20143818 61.44998768 44.53120178 69.96347708 44.39776013 63.25587016 41.39673581 58.50045586 29.05002776 36.66867243 2.52_134.0448m/z 57.50110411 76.87966382 62.78338306 67.29717662 55.44237294 30.64176056 17.58431377 29.27850973 68.56522782 37.09620357 79.58928103 41.56685088 139.6371273 15.03283971 20.12454165 30.26750464 37.59860413 36.70376992 14.93121424 32.61778739 75.99089062 68.01798006 91.34930008 597.3853237 35.90778004 34.4991618 60.07530978 42.5267868 41.7959574 8.457916753 59.40005676 35.85401935 27.17806639 42.6405379 50.51543291 0.68_270.7956m/z 171.5182439 215.3658577 183.2232421 187.5410472 161.4829668 165.7983183 500.6226503 91.27555692 522.620953 91.62945345 92.86639327 81.84938197 305.8460499 45.19595912 77.40344857 132.6658082 121.2529704 131.8687968 38.91356178 53.52293907 89.64431407 441.5196954 466.1271291 152.893841 98.07856808 375.8764377 472.4562878 190.8272922 432.8154558 160.0314189 467.2721051 128.5157743 394.391924 63.15074651 257.8855282 8.52_195.0420m/z 238.5807684 269.9313578 287.3593859 297.1559636 240.6609167 271.8001016 458.415108 194.3861635 558.7256906 174.5917176 209.7885542 226.7243932 326.3767991 103.816912 221.7215506 152.7558384 226.2509154 245.8044473 106.5888349 148.7126726 180.0484189 388.3464507 623.2723118 358.9441452 190.0024963 327.1065993 400.9184331 209.2420419 417.9062826 256.4135133 407.210632 222.2691275 490.1316325 133.2524232 419.0149453 9.27_353.0459m/z 47.51105286 58.06128342 52.18547336 53.16999188 48.839622 74.34614995 129.7217158 79.20124577 120.4601635 66.44935844 58.55948887 53.67155214 90.12136696 27.41255378 39.76376933 53.93836434 64.50051981 59.16926382 34.39471749 39.31988218 45.30256981 104.5556789 99.50287958 92.89055429 34.65042613 60.89368595 63.98327105 37.4343028 73.28825247 37.44187262 70.85883535 59.2378339 86.6527683 29.57109823 88.97298861 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52.64148603 50.37263829 54.20022522 45.26152585 33.52175129 5.557212074 26.11003931 34.1869573 34.57024842 63.64841353 43.5402302 60.11704593 7.689661265 10.89989307 19.11617824 29.45013984 32.46992369 25.96045226 52.53020134 98.9898266 33.30125348 35.48447341 479.8414154 26.2359902 17.5868459 30.9495085 59.81461466 19.14538793 82.81795931 42.12016817 27.05601764 0 28.00179249 18.17820076 12.60_449.1476m/z 246.5532461 266.7545543 250.5157296 233.6499378 199.8658601 186.5890171 773.9203377 158.466654 104.099583 496.3062994 151.321669 249.6245552 1173.350588 121.7181453 206.7435457 199.253936 323.0152529 122.3177991 1345.821029 148.6489942 122.6198787 153.023933 118.1051083 68.55502441 185.2875442 125.0614149 212.8745598 163.4284729 520.4386421 116.8483048 194.6497474 144.8301236 341.2998903 175.3286777 349.0363029 8.52_443.0163m/z 641.089076 687.1383357 648.7995812 682.6088911 703.377237 752.5833272 1118.151669 822.8394668 1100.178626 683.5937206 650.7940132 731.7574354 682.100287 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195.2934453 103.7119094 140.6477759 211.1123943 123.482889 225.9144776 151.0388057 639.933766 377.8454252 513.1494214 405.653279 15.50_421.2239m/z 10332.06958 10596.95825 12694.61269 10771.50339 11298.663 1747.607595 1470.585811 22965.01434 4435.811154 484.0184441 1159.488783 5174.589904 11038.07541 60671.8041 66859.89461 19540.34124 2106.255436 1216.551655 2847.057608 7365.361292 10360.66541 1295.949892 2647.968156 3215.839583 49747.4363 27428.26509 13407.78166 1788.316128 6121.067197 6577.046135 41071.47264 8951.603463 1439.449431 2456.818041 3835.128088 13.89_269.2092m/z 178.6022627 170.230717 157.4895584 189.9732323 187.6193471 160.9129646 120.0926956 193.9647994 197.790819 196.3202213 151.9012514 176.2899816 131.1658587 230.4916041 258.5048763 184.3205 172.3326943 191.9848434 123.1144254 162.6683829 143.880218 117.3262082 194.4904467 147.5409065 168.5010763 156.0795155 166.1986488 160.8657045 137.5515097 149.8139356 194.6855549 301.9985358 142.4474079 208.6054347 179.893639 15.48_489.2111m/z 158.4157121 142.2540051 167.9500061 163.3856371 146.6966661 21.88852606 24.71728108 505.3939768 51.38920149 0 4.122355661 58.26416988 128.1595464 1666.363752 1460.466591 283.7372338 19.35838371 5.7540174 29.17119709 105.6719438 124.5330047 11.6928615 27.80134886 22.50617615 1114.067859 655.9801277 190.8546575 22.55718164 65.31547465 89.14857967 873.7830053 112.3258275 0 35.12416201 44.65944012 1.54_151.0236m/z 35.31695212 31.44684574 31.55846396 27.78315828 29.42554045 24.64624048 9.59499166 9.61562442 23.76642299 24.23501567 47.57651658 32.09768651 37.36839573 11.40671198 3.407483131 19.80123886 21.92350373 28.62052011 15.64792498 47.10663616 47.42534219 19.09921839 22.47233019 553.2306659 18.36881308 11.97373932 18.97976873 43.51478589 26.96298692 76.52029686 26.10704361 12.65776288 14.48236867 26.5360815 12.34389533 12.54_283.1077m/z 592.7919764 549.3841451 477.0311776 525.7258299 430.9467564 368.9065814 480.2354635 584.7395614 776.5459203 639.1786743 611.9879071 503.0302875 857.6922509 533.8120833 551.2106211 748.788347 689.082904 666.5519497 449.1839804 491.2599014 474.5669816 557.0162477 584.8541932 431.3936098 698.0081493 373.963638 506.2930943 363.0693639 678.1095917 454.161768 339.8417502 540.021819 585.2662414 754.9565935 671.5828801 13.96_437.2197m/z 49.59384864 45.74906907 39.6602861 38.83894381 36.85786001 7.690229488 4.801901452 92.14248195 12.89143408 0 0 10.08215781 0 570.8848445 226.585099 55.59926903 0 0 7.493258847 0 25.81373829 0 0 0 134.5306988 163.8308303 67.86889763 0 19.88040505 33.4801742 642.0817771 24.56594702 0 0 0 10.79_309.0988m/z 50.34746982 49.30913909 54.12588068 44.01323198 47.42660764 9.616561307 9.339711484 82.46325843 21.38136939 0 0 4.305921789 13.70190438 522.6728374 284.9366131 59.38895519 0 0 11.57107963 14.7852133 40.65168269 0 5.639470679 3.865679484 159.386019 199.455982 46.65458807 0 28.81233129 24.58350312 549.6048337 24.6749424 0 4.119250314 14.6856084 11.52_315.1935m/z 17.61002734 20.49352427 18.9246611 18.06291414 16.41143067 0 0 18.43763219 9.233266157 36.25372209 116.3754654 91.27523429 83.56390037 0 0 0 12.57815994 23.46007354 21.95741685 19.21151936 27.19775124 5.050737443 36.76610462 9.800352446 13.26580858 1.460844101 0 19.85867222 0 8.462841211 0 42.08166944 5.672977309 52.30178187 17.95663434 14.16_295.2249m/z 82.38135628 74.95044272 77.12487114 80.02449656 103.9823497 22.58319165 6.063435054 13.75507483 16.06254006 190.1266485 153.8829955 125.9797629 86.95662326 163.475844 47.2899628 81.42938623 20.99699329 45.35441555 40.62736784 96.62440593 15.04323096 33.84378149 24.41954853 7.969068023 190.3615177 48.47148431 145.4856719 140.6167464 118.2504913 77.39981551 702.2000113 26.08990584 20.80544658 86.57125174 107.5412869 9.92_309.0985m/z 45.52029253 47.52585392 46.64820762 42.03267355 43.19420096 10.20626012 9.687262178 72.04853708 32.09982406 0 0 9.793807435 15.90861464 476.3246087 250.6294321 57.19144051 0.241998582 0 15.70946685 7.217141152 29.09201586 0 11.40475507 7.534437227 139.4625309 135.9173409 47.82722492 4.017333984 28.6227906 25.95925421 605.1371905 27.48499565 0 0 10.87339245 14.20_293.2081m/z 37.56886053 35.18327974 33.53791155 30.54571816 31.96312954 7.637648469 7.6543801 22.96813657 14.41142118 54.50454862 43.82301017 57.25280091 31.01576308 50.47032743 29.20765216 36.39102978 8.678359808 13.31581798 15.66870752 15.75403062 4.05688686 8.931559125 13.39772384 7.688159109 91.07949587 27.80127434 50.48946766 66.5231989 59.23237661 39.74407288 189.8380323 9.353701729 14.74754661 15.66849179 36.43996649 14.06_271.2249m/z 142.3671636 129.9346899 112.1955477 131.0517096 145.4962112 110.2015704 110.8964742 154.6877054 176.9928498 149.3553336 117.2638223 117.4829242 110.2309211 181.22246 184.737736 141.619534 132.856346 130.4770765 108.7934406 110.8610273 109.779351 116.1240906 142.0282916 118.0764333 160.2975639 111.6465182 135.8060213 113.0808946 123.0940159 96.26946283 151.4499727 181.407796 119.9984433 205.8250128 131.8608489 1.08_346.0544m/z 632.7326012 641.6750681 576.7370429 517.3886663 686.8556945 63.66023845 861.4604843 1583.753338 99.27110128 1075.975563 1403.625345 2486.255309 1808.739866 1184.449581 325.8917221 467.9850869 1765.6224 1241.02501 2234.063162 1132.822956 269.8038088 518.7720777 1196.048648 979.896281 127.9792659 885.781657 301.0744153 113.4950828 168.5845042 0 806.208385 1831.209673 2525.11628 1778.676479 1807.655088 15.84_531.2690m/z 298.5113812 305.5086102 353.935204 339.7914604 276.2781323 96.96442807 168.2274073 357.6918952 180.8092544 646.9918922 728.2929816 983.2599863 1365.010027 180.5208678 155.7631912 199.3723275 458.0409192 719.5228232 399.0714409 472.5623842 258.219196 241.3413247 576.3213913 495.6675201 221.7493629 147.8729046 134.8489298 290.8123266 107.9237642 206.2910671 196.1163821 696.6214085 646.3194768 847.621326 571.8209544 1.30_135.0287m/z 54.91893266 57.92294368 54.19720214 58.42476646 46.57220099 35.66515109 24.18983544 33.81601257 43.80536736 67.59649909 97.51262381 91.75546897 96.72279348 29.4602707 28.80571583 48.12706578 57.32527792 54.72011706 36.67962293 88.94760068 74.94634436 33.95730101 69.4818741 381.0320882 47.58342713 24.54728343 38.46219278 93.35208797 47.74663485 136.1441973 58.48651358 39.47543599 42.21654294 76.03871293 36.63896672 14.42_319.2252m/z 514.3970033 498.3763505 436.8529052 430.5078751 401.6174724 2378.738992 83.78446395 215.8648738 229.2719 594.4827253 452.9001791 495.6909664 582.3999278 867.181554 266.0481923 306.5976745 225.6284745 303.9854721 207.4589772 885.6430074 517.1417118 860.0458954 715.3665486 1933.09034 198.6983527 93.06510325 104.2432909 793.2720947 514.8234792 463.8997573 522.2994308 332.9606117 155.3388831 273.810629 299.5656714 12.43_270.2046m/z 36.24418816 42.37595886 42.48583102 40.37095791 31.39709924 21.81984117 97.09736711 161.6049129 22.69849175 15.86850528 12.66271766 34.80074593 18.11702508 18.28398421 36.59940159 22.29566712 16.89233816 34.54899587 79.43295757 28.29670149 10.32545 259.6821291 15.26834849 15.02503042 50.29896237 27.82939456 37.99954326 44.65565516 109.839128 45.90064777 22.56609058 21.76364099 16.0071486 27.80002224 27.05223312 11.00_242.1731m/z 3773.352703 4057.40298 3881.407783 4112.718223 3194.970284 2377.821343 9544.113528 11124.73215 3192.487537 2060.309405 1119.741997 4891.446802 2253.801947 2060.326269 4005.661793 3498.199879 2246.58833 4675.432581 7670.413401 2806.963013 914.7379911 12562.95876 1913.091265 1591.532081 5649.026868 2962.00569 3520.146117 4843.336168 8445.595838 5413.572532 2291.971167 2601.989692 1641.400436 3587.305849 2314.369169 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name m/z Retention time (min) 15.84_279.2295m/z 279.2295382 15.83861667 0.68_94.9229m/z 94.92290065 0.675166667 0.68_92.9258m/z 92.92575233 0.675166667 0.68_272.7926m/z 272.7926439 0.675166667 0.65_294.8213m/z 294.8213464 0.6547 2.52_134.0448m/z 134.0447785 2.5167 0.68_270.7956m/z 270.7955603 0.675166667 8.52_195.0420m/z 195.0419523 8.519616667 9.27_353.0459m/z 353.0459197 9.269883333 8.42_371.0571m/z 371.0570783 8.417216667 6.15_298.9974m/z 298.9974417 6.1458 8.52_353.0467m/z 353.0466866 8.519616667 0.68_292.8250m/z 292.8250306 0.675166667 2.58_282.0823m/z 282.0823231 2.578133333 10.79_281.1395m/z 281.1394938 10.79093333 2.60_267.0717m/z 267.071741 2.5986 8.52_375.0289m/z 375.0288573 8.519616667 1.02_174.0383m/z 174.0383247 1.016133333 8.52_421.0340m/z 421.0339679 8.519616667 8.52_271.0735m/z 271.0735266 8.519616667 9.13_351.0304m/z 351.0303836 9.12655 8.52_504.9870m/z 504.987038 8.519616667 14.91_100.9308m/z 100.9308067 14.91141667 2.60_335.0596m/z 335.0596065 2.5986 12.60_449.1476m/z 449.1476176 12.59865 8.52_443.0163m/z 443.0163421 8.519616667 8.52_436.9992m/z 436.9991621 8.519616667 1.30_182.0103m/z 182.0103271 1.3028 15.50_421.2239m/z 421.2238866 15.49788333 13.89_269.2092m/z 269.2091992 13.89493333 15.48_489.2111m/z 489.211088 15.4774 1.54_151.0236m/z 151.0236427 1.541166667 12.54_283.1077m/z 283.1076502 12.53721667 13.96_437.2197m/z 437.2196641 13.95636667 10.79_309.0988m/z 309.0988397 10.79093333 11.52_315.1935m/z 315.1934912 11.52073333 14.16_295.2249m/z 295.2248672 14.16113333 9.92_309.0985m/z 309.0985011 9.917783333 14.20_293.2081m/z 293.2080636 14.2021 14.06_271.2249m/z 271.2249336 14.05875 1.08_346.0544m/z 346.0544199 1.077566667 15.84_531.2690m/z 531.2690288 15.83861667 1.30_135.0287m/z 135.0287161 1.3028 14.42_319.2252m/z 319.225212 14.41996667 12.43_270.2046m/z 270.2045657 12.43483333 11.00_242.1731m/z 242.1730509 10.9957 METABOLITES_END #END