#METABOLOMICS WORKBENCH jburgess_20160305_080332 STUDY_ID:ST000359 ANALYSIS_ID:AN000588 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-POS 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 POSITIVE #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 0.68_164.9286m/z 83.57667369 86.84512809 69.5503799 85.80979573 76.10521175 72.85011421 110.1430056 59.00927231 100.2405694 64.94955693 53.38492823 63.63039841 90.27919526 46.37790945 47.37924965 63.41982207 73.46627888 61.09710178 38.25603362 45.56003572 78.06640205 104.0914711 85.87611447 79.17116122 62.5184824 117.3772775 93.77969248 88.24023597 99.88325628 66.77417848 102.575476 77.12107393 99.17927533 59.1765395 84.89864548 8.52_366.0414m/z 24.80252504 27.94906315 28.04768175 26.5334161 21.17966855 30.85893619 46.95227808 16.67395667 48.29335321 4.09365845 23.57983064 14.52930368 40.19099242 0 0.000212658 13.55613659 17.44536192 16.27600233 6.141837002 0.479153135 19.37400586 33.70347718 46.4375888 38.78275369 6.107291647 47.20053829 36.20048849 16.96342778 29.90738658 20.8279141 47.32046337 19.03271311 43.50305305 6.352464927 46.10314834 14.54_581.3291m/z 171.8897029 157.0474751 177.3178379 131.6323173 160.5332035 148.4026654 92.00617077 92.34660404 95.09178862 113.0369278 78.10956122 103.2497141 50.83731856 311.3887163 603.102336 129.069382 94.17770433 645.1739184 357.7596875 231.7300002 104.7439068 179.4935256 103.9756622 76.48199866 202.1158474 146.2991253 86.97516438 43.73147275 82.3352293 71.53577519 182.2258184 282.9838808 142.0286814 124.4909599 83.65406319 4.81_617.4030m/z 32.2930755 36.9976643 31.93279816 35.47092061 33.01383971 25.75883171 55.51499277 14.43642135 54.98898405 20.41149985 6.80932466 11.36707205 49.299479 6.117726306 7.079195287 27.36803572 25.23931897 15.35164815 19.62857208 10.12129479 36.97492932 44.58136405 66.6432483 52.85724236 24.32968438 61.37670859 51.83160211 39.57323851 62.46402141 31.78932438 46.76352095 32.80639227 57.41870038 16.86739551 27.37687107 3.72_528.3165m/z 138.3437487 147.2818518 127.4730823 140.3835572 145.1841722 83.00141005 164.0653241 75.97548824 181.3811421 93.92019106 64.1831793 88.01801835 176.1198868 55.20051651 63.51463204 77.89291795 111.0556371 93.54283184 76.12589966 64.2912882 131.4045814 182.3323669 165.3598467 160.6409827 87.67565737 211.1322729 227.0520418 132.1919274 200.5373527 134.4601485 181.8116958 94.93467531 182.3717927 72.38705917 101.90177 5.01_320.1935m/z 107.6242767 116.6279319 94.19371855 101.2661415 108.2808333 86.87789147 117.0499182 62.22174684 123.9212211 69.64717407 55.04871784 83.93691628 138.6708402 58.19089092 67.44069455 80.93379559 95.30835142 92.77513998 62.12786802 66.45672947 111.2591533 123.408596 150.47533 115.614223 90.01719067 133.7495448 136.1990288 107.3860432 167.1083884 107.8575353 119.6529827 107.1602594 129.5191054 85.36332341 86.55739476 4.47_300.6934m/z 28.54045546 30.02687395 34.40321528 33.72609422 26.61177088 20.76436013 52.21702948 16.54665822 38.54936853 20.42524477 8.648046041 17.04662465 45.12223331 11.42139446 4.888001725 17.8669102 28.06710769 9.164758623 13.92516358 11.03332276 23.29371039 37.94159116 48.37041571 34.60585458 12.53390697 47.92372935 49.39182574 37.7051314 53.75754879 32.56595153 42.82342442 26.85500227 41.80836212 18.11596602 17.90357882 8.52_234.1025n 560.7749947 532.4417267 529.5058895 579.7312923 724.5477614 543.0059548 646.951463 361.2557065 701.3923549 427.6568202 445.3313288 381.5181527 554.4872872 329.0713303 353.4679519 400.342801 425.1267461 486.9577752 294.9736647 352.5265342 407.7826105 597.9051019 727.3234973 531.2262717 414.8740797 616.4151538 600.5560389 497.3088608 719.444671 523.9282682 707.1031743 453.2522803 641.8983483 378.0049959 716.7830327 4.87_360.2184m/z 73.29730059 71.99290013 68.76334465 67.34438444 77.1484664 74.23614713 36.89584772 56.4976923 58.94959907 64.11194119 60.02536319 69.8526443 64.64744758 70.33913523 95.94615043 87.10787795 78.7565273 88.87202288 72.74605084 85.42553713 60.53993261 55.08513465 59.17776911 51.05630093 86.79248842 51.0550312 50.31534969 54.96552169 50.55441312 62.24641587 52.00784281 88.45897142 63.19126889 75.51972462 78.72788229 15.01_572.3576n 7877.102303 6576.723831 7652.212571 7432.461978 7254.905257 6841.135392 6212.462352 9054.897821 7716.429508 11951.75402 10416.8354 10197.31047 5347.571155 12976.62626 18058.81024 9376.960025 9145.494406 11811.83818 10418.03259 12176.30336 6878.066127 3879.300718 5324.693314 5488.341706 12718.854 6732.856446 5155.659929 4577.092206 4278.849722 6705.919123 5465.682313 9716.801168 4354.046081 13457.95984 7233.027248 15.01_616.3807n 7431.409407 5327.512081 6084.019024 6033.070006 5941.781553 5790.559338 6124.216555 9088.64285 6094.578918 9539.436652 8281.396442 8413.136177 4278.57746 12559.57082 15537.094 7239.366522 8119.817717 9378.843956 9870.09181 9788.54257 6139.503548 3861.369977 4064.766934 4675.082412 10290.44556 6789.911195 4617.442779 3572.053721 3280.441578 5163.678132 5070.645321 8697.794392 3485.091868 10377.58732 5699.282618 4.93_555.3640m/z 33.72082933 34.234402 34.43378849 34.40211057 34.03668255 18.60857658 50.36930084 4.231095284 50.38966037 7.750929334 0 3.233793389 38.58464015 0 0 18.79107741 32.06113748 0 0 0 22.53571518 45.6334389 67.12710375 23.70276668 3.744171157 62.32006406 68.32420977 36.94166903 68.4276185 45.40287222 49.4936827 5.316638863 36.65798843 0 9.101713643 9.21_261.0065m/z 1389.491348 1197.506945 1494.990076 1366.339528 1212.57401 825.7148119 929.8348153 680.7260766 1868.40807 719.5739187 323.2813927 769.7299011 592.3914707 477.1813539 539.2612327 954.6013949 1017.708249 1090.924209 540.6846223 415.5010632 3361.967083 2109.047493 2550.373679 1204.182468 648.8627 609.1659432 1610.880392 2582.32495 2027.266886 2262.831226 1313.596743 1251.76395 1568.949153 844.4875354 841.6048487 4.43_616.3946n 232.1298682 259.7624316 268.0976916 301.767462 296.2626652 179.3167658 354.1803977 135.4152534 388.7532287 135.5138344 83.25390284 135.8767706 432.0088056 71.91972749 106.4094542 179.7262257 223.3676254 129.7862561 132.9928093 97.85292783 245.7920277 343.257329 396.7563374 326.0982194 173.7754881 413.1134046 396.1701183 255.8523047 485.1185223 274.1532827 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617.4029689 4.808633333 3.72_528.3165m/z 528.3164616 3.7176 5.01_320.1935m/z 320.1934515 5.0134 4.47_300.6934m/z 300.6934139 4.4679 8.52_234.1025n 273.0871767 8.519333333 4.87_360.2184m/z 360.2183986 4.870066667 15.01_572.3576n 595.3517172 15.01345 15.01_616.3807n 639.3762346 15.01345 4.93_555.3640m/z 555.3639907 4.9315 9.21_261.0065m/z 261.0064702 9.208183333 4.43_616.3946n 599.3913106 4.426933333 5.07_363.7291n 364.7364118 5.074833333 3.33_394.2574n 395.2647013 3.32855 5.83_527.2899m/z 527.2898723 5.825116667 5.40_418.2367m/z 418.2367141 5.3951 4.43_505.3257n 506.3329727 4.426933333 3.06_394.2573n 417.2473662 3.062333333 14.99_528.3338n 551.3269592 14.99298333 14.54_256.1254n 239.1221577 14.54248333 4.73_309.2028m/z 309.2028018 4.726716667 7.37_425.7559n 464.7571492 7.366633333 5.70_373.7297m/z 373.72965 5.70225 5.52_442.7651m/z 442.76506 5.51795 14.81_371.1858m/z 371.1857696 14.80868333 15.86_335.2341m/z 335.2340915 15.85875 8.34_191.1011m/z 191.1011437 8.335033333 4.39_534.3640m/z 534.3640083 4.385983333 15.84_441.2811m/z 441.2811072 15.83828333 2.93_284.1950m/z 284.195017 2.926366667 4.47_320.1937m/z 320.1936938 4.4679 14.97_507.3014m/z 507.3013919 14.9725 5.01_309.2021m/z 309.2021364 5.0134 7.37_474.7783n 513.7851219 7.366633333 7.14_892.5136m/z 892.513614 7.141383333 4.99_448.2411m/z 448.2411061 4.992916667 2.80_266.1853m/z 266.185321 2.803516667 5.70_390.2439m/z 390.24386 5.70225 14.93_463.2774m/z 463.2773779 14.93155 8.44_539.6551m/z 539.6550653 8.437416667 7.81_531.8007m/z 531.8007036 7.809916667 3.72_506.3339m/z 506.3339214 3.7176 4.77_291.1906m/z 291.1905784 4.767683333 8.52_374.5325m/z 374.5324832 8.519333333 7.10_419.7533m/z 419.7533483 7.100433333 5.12_617.4007m/z 617.4007487 5.115783333 15.98_393.2558m/z 393.2558113 15.98161667 5.24_364.7364m/z 364.7363901 5.23865 15.76_281.2552n 304.2449952 15.76373333 2.54_267.0965n 268.1037291 2.5373 7.95_224.1229m/z 224.1228738 7.953266667 3.74_377.2550m/z 377.2549787 3.738083333 7.35_421.2391m/z 421.2390501 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457.749348 7.1209 8.64_778.7161m/z 778.7160506 8.642183333 4.49_309.2027m/z 309.2027187 4.488366667 6.76_781.4458m/z 781.4457695 6.759683333 6.06_438.7704m/z 438.7704371 6.063466667 6.72_684.4039m/z 684.4039052 6.718733333 7.81_520.7982m/z 520.7982259 7.809916667 5.83_420.7714m/z 420.7714427 5.825116667 5.70_408.2427n 431.2617875 5.70225 7.14_474.2832m/z 474.2832402 7.141383333 6.45_553.8336m/z 553.8335667 6.452533333 8.54_222.0287n 206.0099693 8.5398 5.66_396.7501m/z 396.7501 5.6613 14.85_375.2312m/z 375.2312491 14.84963333 5.85_453.2748m/z 453.2748025 5.845583333 7.14_430.7492m/z 430.7492376 7.141383333 5.24_376.2293m/z 376.229319 5.23865 7.47_533.9953m/z 533.9952703 7.469016667 6.31_504.8043m/z 504.8043284 6.309183333 5.58_388.7470n 389.7435703 5.579383333 5.85_453.7826m/z 453.7825644 5.845583333 7.06_428.2223n 451.2602929 7.059466667 5.99_497.7987m/z 497.7986961 5.988933333 4.79_599.3917m/z 599.3917273 4.78815 8.52_354.0528n 355.0600881 8.519333333 6.01_438.2691m/z 438.269058 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566.2983739 14.87011667 9.04_695.7207m/z 695.7206503 9.044366667 METABOLITES_END #END