#METABOLOMICS WORKBENCH wilk0211_20190227_114504 DATATRACK_ID:1654 STUDY_ID:ST001148 ANALYSIS_ID:AN001894 PROJECT_ID:PR000767 VERSION 1 CREATED_ON March 12, 2019, 1:01 pm #PROJECT PR:PROJECT_TITLE Effect of harvesting technique and storage on the metabolic profile of PR:PROJECT_TITLE patient-derived skin fibroblasts PR:PROJECT_TYPE MS targeted metabolomics PR:PROJECT_SUMMARY Using human skin fibroblasts, we measured the effect of different cell PR:PROJECT_SUMMARY harvesting techniques (trypsinization, scraping, and methanol fixation) as well PR:PROJECT_SUMMARY as storage time on the metabolic profiles of organic acids, amino acids, and PR:PROJECT_SUMMARY acylcarnitines. PR:INSTITUTE Mayo Clinic - Rochester PR:DEPARTMENT Neurology PR:LAST_NAME Wilkins PR:FIRST_NAME Jordan PR:ADDRESS 200 First St SW PR:EMAIL wilkins.jordan@mayo.edu PR:PHONE 5072933857 #STUDY ST:STUDY_TITLE Effect of cell harvesting technique and storage on metabolic profiles in human ST:STUDY_TITLE skin fibroblasts ST:STUDY_TYPE Method ST:STUDY_SUMMARY Human skin fibroblasts were cultured in MEM media supplemented with 15% FBS. ST:STUDY_SUMMARY Cells were harvested using (i) trypsinization, (ii) scraping, (iii) methanol ST:STUDY_SUMMARY fixation and scraping, (iV) methanol fixation, scraping, and drying. Targeted ST:STUDY_SUMMARY metabolomics analysis of organic acids, amino acids, and acycarnitines was ST:STUDY_SUMMARY conducted at Mayo Clinic Metabolomics Core Facility. ST:INSTITUTE Mayo Clinic ST:DEPARTMENT Neurology ST:LAST_NAME Wilkins ST:FIRST_NAME Jordan ST:ADDRESS 200 First St SW ST:EMAIL wilkins.jordan@mayo.edu ST:PHONE 5072933857 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:AGE_OR_AGE_RANGE 21 SU:GENDER Male SU:CELL_BIOSOURCE_OR_SUPPLIER Mayo Clinic SU:CELL_STRAIN_DETAILS 10268 SU:CELL_PASSAGE_NUMBER 10 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - 10268_1 Harvest_technique:Trypsin | Storage_time:48_hours SUBJECT_SAMPLE_FACTORS - 10268_2 Harvest_technique:Trypsin | Storage_time:48_hours SUBJECT_SAMPLE_FACTORS - 10268_3 Harvest_technique:Trypsin | Storage_time:48_hours SUBJECT_SAMPLE_FACTORS - 10268_4 Harvest_technique:Scraping | Storage_time:48_hours SUBJECT_SAMPLE_FACTORS - 10268_5 Harvest_technique:Scraping | Storage_time:48_hours SUBJECT_SAMPLE_FACTORS - 10268_6 Harvest_technique:Scraping | Storage_time:48_hours SUBJECT_SAMPLE_FACTORS - 10268_7 Harvest_technique:Methanol | Storage_time:48_hours SUBJECT_SAMPLE_FACTORS - 10268_8 Harvest_technique:Methanol | Storage_time:48_hours SUBJECT_SAMPLE_FACTORS - 10268_9 Harvest_technique:Methanol | Storage_time:48_hours SUBJECT_SAMPLE_FACTORS - 10268_10 Harvest_technique:Methanol_dry | Storage_time:48_hours SUBJECT_SAMPLE_FACTORS - 10268_11 Harvest_technique:Methanol_dry | Storage_time:48_hours SUBJECT_SAMPLE_FACTORS - 10268_12 Harvest_technique:Methanol_dry | Storage_time:48_hours SUBJECT_SAMPLE_FACTORS - 10268_13 Harvest_technique:Trypsin | Storage_time:2_weeks SUBJECT_SAMPLE_FACTORS - 10268_14 Harvest_technique:Trypsin | Storage_time:2_weeks SUBJECT_SAMPLE_FACTORS - 10268_15 Harvest_technique:Trypsin | Storage_time:2_weeks SUBJECT_SAMPLE_FACTORS - 10268_16 Harvest_technique:Scraping | Storage_time:2_weeks SUBJECT_SAMPLE_FACTORS - 10268_17 Harvest_technique:Scraping | Storage_time:2_weeks SUBJECT_SAMPLE_FACTORS - 10268_18 Harvest_technique:Scraping | Storage_time:2_weeks SUBJECT_SAMPLE_FACTORS - 10268_19 Harvest_technique:Methanol | Storage_time:2_weeks SUBJECT_SAMPLE_FACTORS - 10268_20 Harvest_technique:Methanol | Storage_time:2_weeks SUBJECT_SAMPLE_FACTORS - 10268_21 Harvest_technique:Methanol | Storage_time:2_weeks SUBJECT_SAMPLE_FACTORS - 10268_22 Harvest_technique:Methanol_dry | Storage_time:2_weeks SUBJECT_SAMPLE_FACTORS - 10268_23 Harvest_technique:Methanol_dry | Storage_time:2_weeks SUBJECT_SAMPLE_FACTORS - 10268_24 Harvest_technique:Methanol_dry | Storage_time:2_weeks SUBJECT_SAMPLE_FACTORS - 10268_25 Harvest_technique:Trypsin | Storage_time:4_weeks SUBJECT_SAMPLE_FACTORS - 10268_26 Harvest_technique:Trypsin | Storage_time:4_weeks SUBJECT_SAMPLE_FACTORS - 10268_27 Harvest_technique:Trypsin | Storage_time:4_weeks SUBJECT_SAMPLE_FACTORS - 10268_28 Harvest_technique:Scraping | Storage_time:4_weeks SUBJECT_SAMPLE_FACTORS - 10268_29 Harvest_technique:Scraping | Storage_time:4_weeks SUBJECT_SAMPLE_FACTORS - 10268_30 Harvest_technique:Scraping | Storage_time:4_weeks SUBJECT_SAMPLE_FACTORS - 10268_31 Harvest_technique:Methanol | Storage_time:4_weeks SUBJECT_SAMPLE_FACTORS - 10268_32 Harvest_technique:Methanol | Storage_time:4_weeks SUBJECT_SAMPLE_FACTORS - 10268_33 Harvest_technique:Methanol | Storage_time:4_weeks SUBJECT_SAMPLE_FACTORS - 10268_34 Harvest_technique:Methanol_dry | Storage_time:4_weeks SUBJECT_SAMPLE_FACTORS - 10268_35 Harvest_technique:Methanol_dry | Storage_time:4_weeks SUBJECT_SAMPLE_FACTORS - 10268_36 Harvest_technique:Methanol_dry | Storage_time:4_weeks #COLLECTION CO:COLLECTION_SUMMARY Prior to harvest, media was removed and cells were rinsed with PBS. For CO:COLLECTION_SUMMARY trypsinization, cells were dissociated by incubation with 0.05% trypsin at 37ºC CO:COLLECTION_SUMMARY for 5 minutes. Dissociated cells were resuspended in MEM (containing 15% FBS) to CO:COLLECTION_SUMMARY neutralize trypsin activity. Cells were centrifuged at 1000 x g for 5 min at CO:COLLECTION_SUMMARY 4ºC. Pelleted cells were resuspended in PBS and centrifuged at 1000 x g for 5 CO:COLLECTION_SUMMARY min at 4°C. The supernatant was aspirated, cell pellets were flash frozen in CO:COLLECTION_SUMMARY liquid nitrogen, and frozen pellets were stored at -80ºC. For scraping, cells CO:COLLECTION_SUMMARY were detached in 1 mL PBS using a rubber scraper. Detached cells were CO:COLLECTION_SUMMARY centrifuged at 1000 x g for 5 min at 4°C, and PBS was aspirated. Cell pellets CO:COLLECTION_SUMMARY were flash frozen in liquid nitrogen and stored at -80ºC. For methanol CO:COLLECTION_SUMMARY fixation, 1 mL 80% methanol (pre-chilled on dry ice) was added to cells. Tissue CO:COLLECTION_SUMMARY culture plates were rocked by hand for about 20 seconds to evenly distribute CO:COLLECTION_SUMMARY methanol solution. Cells were detached in 80% methanol using a rubber scraper. CO:COLLECTION_SUMMARY Suspensions were transferred to an Eppendorf tube and stored at -80ºC. For CO:COLLECTION_SUMMARY methanol fixation and drying, cells were harvested in 1 mL 80% methanol, and the CO:COLLECTION_SUMMARY supernatant was evaporated using a Speed Vacuum at room temperature. Dried CO:COLLECTION_SUMMARY samples were stored at -80ºC. Frozen samples were stored at -80ºC for 48 CO:COLLECTION_SUMMARY hours, two weeks, or four weeks. CO:SAMPLE_TYPE Cultured fibroblasts CO:COLLECTION_FREQUENCY 48 hours, 2 weeks, and 4 weeks CO:STORAGE_CONDITIONS -80℃ CO:STORAGE_VIALS 1.5 mL Eppendorf tubes #TREATMENT TR:TREATMENT_SUMMARY Collection method and storage time were variable as described in collection TR:TREATMENT_SUMMARY summary #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Targeted metabolomics analysis of amino acids, acylcarnitines, and metabolites SP:SAMPLEPREP_SUMMARY of the TCA cycle was conducted at the Mayo Clinic Metabolomics Center. Samples SP:SAMPLEPREP_SUMMARY harvested in methanol were dried using a Speed Vacuum. Cell pellets were SP:SAMPLEPREP_SUMMARY sonicated in 100 µL PBS and spiked with 15 - 25 µL of the respective internal SP:SAMPLEPREP_SUMMARY standards. Proteins were removed by adding 450 µL of cold 1:1 SP:SAMPLEPREP_SUMMARY methanol/acetonitrile solution with subsequent centrifugation for 15 minutes SP:SAMPLEPREP_SUMMARY (18,000 x g at 4ºC). Supernatants were transferred to a 1 mL dram and dried SP:SAMPLEPREP_SUMMARY under a nitrogen stream for approximately 30 minutes. Prior to detection, TCA SP:SAMPLEPREP_SUMMARY cycle metabolites were derivatized using ethoxyamine and then with MtBSTFA + 1% SP:SAMPLEPREP_SUMMARY tBDMCS (N-Methyl-N-(t-Butyldimethylsilyl)-Trifluoroacetamide + 1% SP:SAMPLEPREP_SUMMARY t-Butyldimethylchlorosilane). Amino acids were derivatized with SP:SAMPLEPREP_SUMMARY 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate using the Waters AccQ-Fluor SP:SAMPLEPREP_SUMMARY Reagent Kit (Cat. # WATO52880). Acylcarnitines were reconstituted in buffer SP:SAMPLEPREP_SUMMARY containing 99% MeOH, 1% H2O, 1 mM ammonium formate, and 0.1% formic acid. SP:SAMPLEPREP_SUMMARY Organic acids were detected with an Agilent 5977A gas chromatography/mass SP:SAMPLEPREP_SUMMARY spectrometry (GC/MS) under electron impact and single ion monitoring conditions. SP:SAMPLEPREP_SUMMARY Analytes were separated on an Agilent DB-5MS column (30 m x 0.25 mm x 0.25μm). SP:SAMPLEPREP_SUMMARY Sample injection volume was 1 μL performed in splitless mode. The inlet SP:SAMPLEPREP_SUMMARY temperature was maintained at 250°C. The carrier gas was helium set at a flow SP:SAMPLEPREP_SUMMARY rate of 0.9 ml/min. The initial oven temperature was 120°C set with the SP:SAMPLEPREP_SUMMARY following ramp rates: Ramp to 180°C at 25°C/min; Ramp to 270°C at 6°C/min; SP:SAMPLEPREP_SUMMARY Ramp to 325°C at 30°C/min. The transfer line temperature was 280°C. SP:SAMPLEPREP_SUMMARY Concentrations of lactic acid, fumaric acid, succinic acid, oxaloacetic acid, SP:SAMPLEPREP_SUMMARY alpha-ketoglutaric acid, malic acid, 2-hydroxyglutaric acid, cis-aconitic acid, SP:SAMPLEPREP_SUMMARY citric acid, and isocitric acid were measured against a calibration curve. Amino SP:SAMPLEPREP_SUMMARY acids were analyzed using Thermo TSQ Quantum Ultra mass spectrometer (West Palm SP:SAMPLEPREP_SUMMARY Beach, FL) coupled with a Waters ACQUITY ultra performance liquid chromatography SP:SAMPLEPREP_SUMMARY BEH C18 column (2.1 mm x 150 mm x. 1.7 μm). Data acquisition was performed SP:SAMPLEPREP_SUMMARY using selected reaction monitoring (SRM) and positive electrospray ionization SP:SAMPLEPREP_SUMMARY (ESI). Injection volume was 1 μL. The column flow rate was 400 μL/min with an SP:SAMPLEPREP_SUMMARY isothermal set at 43°C. Mobile phase A was 1% acetonitrile in 0.1% formic acid, SP:SAMPLEPREP_SUMMARY and mobile phase B was 100% acetonitrile. The mass spectrometer was operated SP:SAMPLEPREP_SUMMARY with 4000 capillary voltage, 50 sheath gas, 20 auxiliary gas, and 15 sweep gas. SP:SAMPLEPREP_SUMMARY The capillary temperature was 2700°C. Collision gas was 1.5 Torr and collision SP:SAMPLEPREP_SUMMARY energy was 25 V. The tube lens was kept at 90 V. The concentration of amino SP:SAMPLEPREP_SUMMARY acids was calculated against a calibration curve. Acylcarnitines were analyzed SP:SAMPLEPREP_SUMMARY using a Waters ACQUITY UPLC system (Milford, MA) coupled with a Thermo TSQ SP:SAMPLEPREP_SUMMARY Quantiva tandem mass spectrometer (West Palm Beach, FL) in SRM and positive SP:SAMPLEPREP_SUMMARY (H)ESI mode. Analytes were separated on a Waters ACQUITY UPLC BEH C8 column (2.1 SP:SAMPLEPREP_SUMMARY mm x 150 mm x. 1.7 μm) with an isothermal temperature of 43°C. The mass SP:SAMPLEPREP_SUMMARY spectrometer capillary voltage was set to 4000 with a sheath gas 30, auxiliary SP:SAMPLEPREP_SUMMARY gas 5, and sweep gas 2. The ion transfer tube was maintained at 300°C with the SP:SAMPLEPREP_SUMMARY vaporizer at 40°C, collision gas at 1.5 Torr, and collision energy at 12 V. SP:SAMPLEPREP_SUMMARY Concentrations of carnitine, acetylcarnitine, propionylcarnitine, SP:SAMPLEPREP_SUMMARY butyrylcarnitine, isovalerylcarnitine, octanoylcarnitine, lauroylcarnitine, SP:SAMPLEPREP_SUMMARY myristoylcarnitine, palmitoylcarnitine, oleoylcarnitine, and stearoylcarnitine SP:SAMPLEPREP_SUMMARY were measured against a calibration curve. Data sets collected by GC/MS were SP:SAMPLEPREP_SUMMARY analyzed using Mass Hunter GC/MS Quantitation software version B.07 (Agilent). SP:SAMPLEPREP_SUMMARY Analysis of LC/MS data sets was performed using Xcalibur Quant browser (Thermo SP:SAMPLEPREP_SUMMARY Scientific). #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Amino acids were analyzed using Thermo TSQ Quantum Ultra mass spectrometer (West CH:CHROMATOGRAPHY_SUMMARY Palm Beach, FL) coupled with a Waters ACQUITY ultra performance liquid CH:CHROMATOGRAPHY_SUMMARY chromatography BEH C18 column (2.1 mm x 150 mm x. 1.7 μm). Data acquisition was CH:CHROMATOGRAPHY_SUMMARY performed using selected reaction monitoring (SRM) and positive electrospray CH:CHROMATOGRAPHY_SUMMARY ionization (ESI). Injection volume was 1 μL. The column flow rate was 400 CH:CHROMATOGRAPHY_SUMMARY μL/min with an isothermal set at 43°C. Mobile phase A was 1% acetonitrile in CH:CHROMATOGRAPHY_SUMMARY 0.1% formic acid, and mobile phase B was 100% acetonitrile. The mass CH:CHROMATOGRAPHY_SUMMARY spectrometer was operated with 4000 capillary voltage, 50 sheath gas, 20 CH:CHROMATOGRAPHY_SUMMARY auxiliary gas, and 15 sweep gas. The capillary temperature was 2700°C. CH:CHROMATOGRAPHY_SUMMARY Collision gas was 1.5 Torr and collision energy was 25 V. The tube lens was kept CH:CHROMATOGRAPHY_SUMMARY at 90 V. The concentration of amino acids was calculated against a calibration CH:CHROMATOGRAPHY_SUMMARY curve. CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Waters Acquity BEH C18 (150 x 2mm, 1.7um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Quantiva MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Xcalibur acquisition and Quantitation 4.0.27 (Thermo Electron) #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS micromol metabolite per gram protein MS_METABOLITE_DATA_START Samples 10268_1 10268_2 10268_3 10268_4 10268_5 10268_6 10268_7 10268_8 10268_9 10268_10 10268_11 10268_12 10268_13 10268_14 10268_15 10268_16 10268_17 10268_18 10268_19 10268_20 10268_21 10268_22 10268_23 10268_24 10268_25 10268_26 10268_27 10268_28 10268_29 10268_30 10268_31 10268_32 10268_33 10268_34 10268_35 10268_36 Factors Harvest_technique:Trypsin | Storage_time:48_hours Harvest_technique:Trypsin | Storage_time:48_hours Harvest_technique:Trypsin | Storage_time:48_hours Harvest_technique:Scraping | Storage_time:48_hours Harvest_technique:Scraping | Storage_time:48_hours Harvest_technique:Scraping | Storage_time:48_hours Harvest_technique:Methanol | Storage_time:48_hours Harvest_technique:Methanol | Storage_time:48_hours Harvest_technique:Methanol | Storage_time:48_hours Harvest_technique:Methanol_dry | Storage_time:48_hours Harvest_technique:Methanol_dry | Storage_time:48_hours Harvest_technique:Methanol_dry | Storage_time:48_hours Harvest_technique:Trypsin | Storage_time:2_weeks Harvest_technique:Trypsin | Storage_time:2_weeks Harvest_technique:Trypsin | Storage_time:2_weeks Harvest_technique:Scraping | Storage_time:2_weeks Harvest_technique:Scraping | Storage_time:2_weeks Harvest_technique:Scraping | Storage_time:2_weeks Harvest_technique:Methanol | Storage_time:2_weeks Harvest_technique:Methanol | Storage_time:2_weeks Harvest_technique:Methanol | Storage_time:2_weeks Harvest_technique:Methanol_dry | Storage_time:2_weeks Harvest_technique:Methanol_dry | Storage_time:2_weeks Harvest_technique:Methanol_dry | Storage_time:2_weeks Harvest_technique:Trypsin | Storage_time:4_weeks Harvest_technique:Trypsin | Storage_time:4_weeks Harvest_technique:Trypsin | Storage_time:4_weeks Harvest_technique:Scraping | Storage_time:4_weeks Harvest_technique:Scraping | Storage_time:4_weeks Harvest_technique:Scraping | Storage_time:4_weeks Harvest_technique:Methanol | Storage_time:4_weeks Harvest_technique:Methanol | Storage_time:4_weeks Harvest_technique:Methanol | Storage_time:4_weeks Harvest_technique:Methanol_dry | Storage_time:4_weeks Harvest_technique:Methanol_dry | Storage_time:4_weeks Harvest_technique:Methanol_dry | Storage_time:4_weeks Acetylcarnitine 0.110172375 0.072997782 0.074863233 0.010930405 0.011742433 0.011991277 0.144057023 0.128559713 0.149776582 0.119466938 0.13228137 0.145567593 0.11753547 0.112622434 0.129129802 0.049164582 0.069701877 0.063026604 0.142276272 0.168403398 0.162241609 0.209860878 0.18694457 0.154486329 0.085088383 0.078079001 0.090138078 0.038808539 0.04606206 0.039273604 0.127887025 0.120702553 0.133558514 0.154573777 0.126722286 0.123378801 Butyrylcarnitine 0.110180067 0.084762019 0.079683405 0.00739317 0.006880029 0.007139659 0.033643622 0.028694667 0.033986455 0.031528841 0.034312664 0.035937015 0.095989122 0.094199821 0.107961421 0.009967147 0.013895837 0.012727733 0.030241792 0.037332292 0.034684794 0.045215531 0.042741399 0.035003858 0.076660233 0.072822167 0.085304736 0.012497285 0.01492127 0.012338094 0.032858914 0.030125992 0.033715529 0.040820576 0.036128294 0.030810564 Carnitine 0.186670032 0.156674248 0.142517844 0.04464898 0.042800405 0.041829738 0.471197048 0.463418218 0.574029198 0.450561645 0.525280211 0.570237836 0.293835027 0.281426492 0.324786006 0.196103909 0.271490122 0.255926921 0.511843296 0.619307763 0.615737843 0.794317556 0.729775162 0.611911141 0.165819366 0.151797529 0.174472952 0.105127163 0.12972025 0.113673704 0.462021801 0.468410998 0.520233984 0.603801269 0.509815906 0.446871653 Isovalerylcarnitine 0.016735625 0.011303587 0.011033604 0.00688215 0.006479985 0.006746745 0.013067685 0.013441138 0.01510474 0.01249725 0.014484783 0.014880054 0.012830083 0.012488415 0.014493691 0.008970364 0.012650659 0.011520673 0.012513368 0.015755658 0.015386938 0.018312186 0.017435796 0.014520018 0.008439953 0.008187127 0.009560995 0.005310934 0.006457421 0.00535959 0.010918368 0.010494447 0.012574113 0.013115836 0.011745154 0.00972114 Lauroylcarnitine 0.000697532 0.000540155 0.000484311 0.000439643 0.000407184 0.000433901 0.000331339 0.000388237 0.00042098 0.000320819 0.000391315 0.00039277 0.000484086 0.0005121 0.000571498 0.000593955 0.000834631 0.000751339 0.000373982 0.000476052 0.000482352 0.000565133 0.000532492 0.00044929 0.000467737 0.000465846 0.000609396 0.000688193 0.000828517 0.000664843 0.000424983 0.000420172 0.000486288 0.000512719 0.000457035 0.000390632 Myristoylcarnitine 0.009131418 0.006999727 0.006896239 0.004676067 0.004327349 0.004520405 0.00362041 0.004230168 0.004735508 0.00348981 0.00418718 0.004304335 0.008112408 0.008266255 0.009387162 0.007631178 0.010761838 0.009731389 0.004725312 0.005911859 0.006090703 0.006988843 0.006476702 0.005536664 0.006595807 0.006620058 0.008167734 0.006749124 0.008099454 0.006543878 0.004147624 0.004154663 0.004952303 0.005079728 0.004502517 0.003828361 Octanoylcarnitine 0.007510205 0.005010712 0.004877716 0.005235566 0.004824507 0.005086324 0.003194812 0.00366703 0.003957707 0.003008161 0.00351406 0.003501893 0.005824569 0.006340563 0.007263617 0.008323529 0.011702281 0.010570713 0.00439871 0.005388781 0.005424402 0.00632594 0.005736082 0.004803252 0.004359619 0.004361054 0.005765839 0.006869837 0.00821049 0.006606764 0.003558722 0.003451449 0.00396786 0.004334092 0.003699018 0.003329744 Oleoylcarnitine 0.029822359 0.021110079 0.024241005 0.003693737 0.003572986 0.003371654 0.004212298 0.004697709 0.005462279 0.00397657 0.004738494 0.004785608 0.031602924 0.029034857 0.03471379 0.0042756 0.006174481 0.005666828 0.00445644 0.00606846 0.00664044 0.006868129 0.006812513 0.006036605 0.034117401 0.034496394 0.037119021 0.002672968 0.003296459 0.002834816 0.003841101 0.004416714 0.005345821 0.004351539 0.004565758 0.003548493 Palmitoylcarnitine 0.063818029 0.058080837 0.057757181 0.01034148 0.009543483 0.009860392 0.008563433 0.010735518 0.013140006 0.008384938 0.010908219 0.012297648 0.065203999 0.062145354 0.073047764 0.016735771 0.023683666 0.021453997 0.011168565 0.01479833 0.016183326 0.015995703 0.01633591 0.014946924 0.053633274 0.052075895 0.059438498 0.015378696 0.018591491 0.015234439 0.010202437 0.011555268 0.014418887 0.012457589 0.012393617 0.009308694 Propionylcarnitine 0.095468345 0.070621587 0.061250398 0.011051006 0.011413256 0.011544206 0.061599316 0.061331497 0.08614032 0.059976211 0.075655079 0.090529071 0.077645724 0.076707229 0.086827473 0.013758597 0.019914306 0.018191549 0.054214985 0.069778342 0.081951987 0.095553495 0.105079598 0.088780388 0.047023651 0.047793009 0.054679417 0.011214126 0.013518578 0.01112826 0.055740543 0.060537653 0.076845291 0.076320139 0.075513681 0.064203189 Stearoylcarnitine 0.00483388 0.004475497 0.004312497 0.001908353 0.001566481 0.001667907 0.001663425 0.001736676 0.001899997 0.00206063 0.002301541 0.002060134 0.008910198 0.008939096 0.010666249 0.006892827 0.009683906 0.0090716 0.004489888 0.00542522 0.005698899 0.007674003 0.007441679 0.006402503 0.00719342 0.006369769 0.007630332 0.004801928 0.006136839 0.005246079 0.00319283 0.003144278 0.003830271 0.004879673 0.00410635 0.003697223 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name rtimes (mins) Transition HMDB Acetylcarnitine 1.1 204.1>85.0 HMDB00201 Butyrylcarnitine 3 232.1>85.0 HMDB02013 Carnitine 1 162.1>85.0 HMDB00062 Isovalerylcarnitine 3.71 246.2>85.0 HMDB00688 Lauroylcarnitine 7.27 344.3>85.0 HMDB02250 Myristoylcarnitine 7.72 372.4>85.0 HMDB05066 Octanoylcarnitine 5.85 288.2>85.0 HMDB00791 Oleoylcarnitine 8.29 426.4>85.0 HMDB05065 Palmitoylcarnitine 8.15 400.4>85.0 HMDB00222 Propionylcarnitine 2.35 218.1>85.0 HMDB00824 Stearoylcarnitine 8.6 428.4>85.0 HMDB00848 METABOLITES_END #END