#METABOLOMICS WORKBENCH ryant_20201130_125121 DATATRACK_ID:2336 STUDY_ID:ST001617 ANALYSIS_ID:AN002652 PROJECT_ID:000000 VERSION 1 CREATED_ON December 3, 2020, 11:16 am #PROJECT PR:PROJECT_TITLE Unique metabolomic profile of skeletal muscle in chronic limb threatening PR:PROJECT_TITLE ischemia PR:PROJECT_SUMMARY This project is focused on a cross-sectional analysis of non-PAD controls and PR:PROJECT_SUMMARY CLTI patients undergoing either a vascular intervention or undergoing limb PR:PROJECT_SUMMARY amputation was performed and involved a detailed assessment of the limb muscle PR:PROJECT_SUMMARY metabolome using HR-MAS and solution state NMR spectroscopy. It was hypothesized PR:PROJECT_SUMMARY that patients undergoing limb amputation would present with altered muscle PR:PROJECT_SUMMARY metaolite features compared with non-PAD controls. PR:INSTITUTE University of Florida PR:DEPARTMENT Applied Physiology and Kinesiology PR:LABORATORY Rm 42 and Rm 43 PR:LAST_NAME Ryan PR:FIRST_NAME Terence PR:ADDRESS 1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA PR:EMAIL ryant@ufl.edu PR:PHONE 352-294-1700 PR:FUNDING_SOURCE National Institutes of Health and the National Heart, Lung, and Blood, Institute PR:FUNDING_SOURCE numbers R01-HL149704 (to T.E.R.) and R01-HL148597 (to S.T.S.); as well as the PR:FUNDING_SOURCE American Heart Association grant number 18CDA34110044 (to T.E.R.). A portion of PR:FUNDING_SOURCE this work was supported by pilot funds (awarded to T.E.R.) from The University PR:FUNDING_SOURCE of Florida Claude D. Pepper Older Americans Independence Center P30AG028740. (if PR:FUNDING_SOURCE applicable) PR:PROJECT_COMMENTS PAD metabolomic study via NMR. A detailed assessment of the limb muscle PR:PROJECT_COMMENTS metabolome using semi-solid and solution state NMR spectroscopy. PR:PUBLICATIONS Journal of Clinical Medicine PR:CONTRIBUTORS Ram B. Khattri, Kyoungrae Kim, Trace Thome, Zachary R. Salyers, Kerri A. PR:CONTRIBUTORS O’Malley, Scott A. Berceli, Salvatore T. Scali, Terence E. Ryan* #STUDY ST:STUDY_TITLE Unique metabolomic profile of skeletal muscle in chronic limb threatening ST:STUDY_TITLE ischemia (HR-MAS study) ST:STUDY_SUMMARY This project is focused on a cross-sectional analysis of non-PAD controls and ST:STUDY_SUMMARY CLTI patients undergoing either a vascular intervention or undergoing limb ST:STUDY_SUMMARY amputation was performed and involved a detailed assessment of the limb muscle ST:STUDY_SUMMARY metabolome using HR-MAS and solution state NMR spectroscopy. It was hypothesized ST:STUDY_SUMMARY that patients undergoing limb amputation would present with altered muscle ST:STUDY_SUMMARY metaolite features compared with non-PAD controls. ST:INSTITUTE University of Florida ST:DEPARTMENT Applied Physiology and Kinesiology ST:LABORATORY Rm 42 and Rm 43 ST:LAST_NAME Ryan ST:FIRST_NAME Terence ST:ADDRESS 1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA ST:EMAIL ryant@ufl.edu ST:PHONE 352-294-1700 ST:NUM_GROUPS 3 ST:TOTAL_SUBJECTS 30 ST:NUM_MALES 25 ST:NUM_FEMALES 5 ST:STUDY_COMMENTS PAD metabolomic study via NMR. A detailed assessment of the limb muscle ST:STUDY_COMMENTS metabolome using semi-solid and solution state NMR spectroscopy. ST:PUBLICATIONS Journal of Clinical Medicine #SUBJECT SU:SUBJECT_TYPE Human SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:AGE_OR_AGE_RANGE 55-82 years SU:GENDER Male and female SU:HUMAN_RACE unknown SU:HUMAN_ETHNICITY unknown SU:HUMAN_MEDICATIONS Asprin, ACE inhibitor, statins, cilostazol SU:HUMAN_SMOKING_STATUS Controls = 4, CLTI Pre-surgery = 7, and CLTI Amputation = 9 #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 - CLTI Amputation1B Group:CLTI_Amputation RAW_FILE_NAME=CLTI Amputation1B.raw SUBJECT_SAMPLE_FACTORS - CLTI Amputation2B Group:CLTI_Amputation RAW_FILE_NAME=CLTI Amputation2B.raw SUBJECT_SAMPLE_FACTORS - CLTI Amputation3B Group:CLTI_Amputation RAW_FILE_NAME=CLTI Amputation3B.raw SUBJECT_SAMPLE_FACTORS - CLTI Amputation4B Group:CLTI_Amputation RAW_FILE_NAME=CLTI Amputation4B.raw SUBJECT_SAMPLE_FACTORS - CLTI Amputation6B Group:CLTI_Amputation RAW_FILE_NAME=CLTI Amputation6B.raw SUBJECT_SAMPLE_FACTORS - CLTI Amputation7B Group:CLTI_Amputation RAW_FILE_NAME=CLTI Amputation7B.raw SUBJECT_SAMPLE_FACTORS - CLTI Amputation8B Group:CLTI_Amputation RAW_FILE_NAME=CLTI Amputation8B.raw SUBJECT_SAMPLE_FACTORS - CLTI Amputation9B Group:CLTI_Amputation RAW_FILE_NAME=CLTI Amputation9B.raw SUBJECT_SAMPLE_FACTORS - CLTI Amputation10B Group:CLTI_Amputation RAW_FILE_NAME=CLTI Amputation10B.raw SUBJECT_SAMPLE_FACTORS - CLTI Pre-surgery1B Group:CLTI_Pre_surgery RAW_FILE_NAME=CLTI Pre-surgery1B.raw SUBJECT_SAMPLE_FACTORS - CLTI Pre-surgery2B Group:CLTI_Pre_surgery RAW_FILE_NAME=CLTI Pre-surgery2B.raw SUBJECT_SAMPLE_FACTORS - CLTI Pre-surgery5B Group:CLTI_Pre_surgery RAW_FILE_NAME=CLTI Pre-surgery5B.raw SUBJECT_SAMPLE_FACTORS - CLTI Pre-surgery6B Group:CLTI_Pre_surgery RAW_FILE_NAME=CLTI Pre-surgery6B.raw SUBJECT_SAMPLE_FACTORS - CLTI Pre-surgery7B Group:CLTI_Pre_surgery RAW_FILE_NAME=CLTI Pre-surgery7B.raw SUBJECT_SAMPLE_FACTORS - CLTI Pre-surgery9B Group:CLTI_Pre_surgery RAW_FILE_NAME=CLTI Pre-surgery9B.raw SUBJECT_SAMPLE_FACTORS - Control1B Group:Control RAW_FILE_NAME=Control1B.raw SUBJECT_SAMPLE_FACTORS - Control2B Group:Control RAW_FILE_NAME=Control2B.raw SUBJECT_SAMPLE_FACTORS - Control3B Group:Control RAW_FILE_NAME=Control3B.raw SUBJECT_SAMPLE_FACTORS - Control4B Group:Control RAW_FILE_NAME=Control4B.raw SUBJECT_SAMPLE_FACTORS - Control5B Group:Control RAW_FILE_NAME=Control5B.raw SUBJECT_SAMPLE_FACTORS - Control6B Group:Control RAW_FILE_NAME=Control6B.raw SUBJECT_SAMPLE_FACTORS - Control7B Group:Control RAW_FILE_NAME=Control7B.raw SUBJECT_SAMPLE_FACTORS - Control8B Group:Control RAW_FILE_NAME=Control8B.raw SUBJECT_SAMPLE_FACTORS - Control9B Group:Control RAW_FILE_NAME=Control9B.raw SUBJECT_SAMPLE_FACTORS - Control10B Group:Control RAW_FILE_NAME=Control10B.raw #COLLECTION CO:COLLECTION_SUMMARY Gastrocnemius muscle specimens were obtained from ten older adult non-PAD CO:COLLECTION_SUMMARY controls (Control), ten patients with critical limb ischemia (CLI) undergoing CO:COLLECTION_SUMMARY surgical intervention (CLTI Pre-surgery), and ten CLI patients undergoing limb CO:COLLECTION_SUMMARY amputation (CLTI Amputation). Five pre-surgery patients underwent bypass CO:COLLECTION_SUMMARY interventions and five underwent endovascular procedures. Muscle specimens were CO:COLLECTION_SUMMARY collected within the confines of the operating rooms (CLI patients) or via CO:COLLECTION_SUMMARY percutaneous muscle biopsy using sterile procedures. A portion of the muscle was CO:COLLECTION_SUMMARY quickly trimmed of fat/connective tissue and snap frozen in liquid nitrogen for CO:COLLECTION_SUMMARY metabolomics analysis. CO:SAMPLE_TYPE Muscle CO:COLLECTION_METHOD Muscle specimens were collected within the confines of the operating rooms (CLI CO:COLLECTION_METHOD patients) or via percutaneous muscle biopsy using sterile procedures. A portion CO:COLLECTION_METHOD of the muscle was quickly trimmed of fat/connective tissue and snap frozen in CO:COLLECTION_METHOD liquid nitrogen for metabolomics analysis. CO:STORAGE_CONDITIONS -80℃ CO:STORAGE_VIALS cryovials #TREATMENT TR:TREATMENT_SUMMARY This was a prospective cohort study that examined the metabolomic profile of TR:TREATMENT_SUMMARY skeletal muscle from CLTI patients undergoing surgerical intervention or TR:TREATMENT_SUMMARY amputation, as well as a cohort of non-PAD controls. CLTI patients exhibited TR:TREATMENT_SUMMARY severe symptomology (Rutherford Classification 3-6), with high incidence of TR:TREATMENT_SUMMARY common PAD risk factors including hypertension, hyperlipidemia, coronary artery TR:TREATMENT_SUMMARY disease, and diabetes.Ten patients with critical limb ischemia (CLI) undergoing TR:TREATMENT_SUMMARY surgical intervention (CLTI Pre-surgery), and ten CLI patients undergoing limb TR:TREATMENT_SUMMARY amputation (CLTI Amputation). Five pre-surgery patients underwent bypass TR:TREATMENT_SUMMARY interventions and five underwent endovascular procedures. Muscle specimens were TR:TREATMENT_SUMMARY collected within the confines of the operating rooms (CLI patients) or via TR:TREATMENT_SUMMARY percutaneous muscle biopsy using sterile procedures. TR:HUMAN_FASTING non-fasted TR:HUMAN_ENDP_CLINICAL_SIGNS N/A #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Semi-solid HR-MAS spectra were collected on intact human gastrocnemius muscle SP:SAMPLEPREP_SUMMARY specimens. SP:PROCESSING_METHOD None SP:PROCESSING_STORAGE_CONDITIONS -80℃ SP:EXTRACT_STORAGE -80℃ SP:SAMPLE_RESUSPENSION None SP:SAMPLE_SPIKING None #ANALYSIS AN:DATA_FORMAT fid, 1r #NMR NM:INSTRUMENT_NAME Bruker Avance 800 MHz with HR-MAS console NM:INSTRUMENT_TYPE FT-NMR NM:NMR_EXPERIMENT_TYPE 1D-1H NM:FIELD_FREQUENCY_LOCK Deuterium NM:STANDARD_CONCENTRATION None NM:SPECTROMETER_FREQUENCY 800 MHz NM:NMR_PROBE HR-MAS probe NM:NMR_SOLVENT For HR-MAS experiments, deuterium oxide was used. NM:NMR_TUBE_SIZE 3.2 mm inside diameter plastic insert was used for HR-MAS experiments. NM:SHIMMING_METHOD Topshim NM:PULSE_SEQUENCE noesypr1d NM:WATER_SUPPRESSION presat NM:PULSE_WIDTH 90-degree NM:RECEIVER_GAIN 28.5 NM:OFFSET_FREQUENCY 4.96 Hz NM:CHEMICAL_SHIFT_REF_CPD None NM:TEMPERATURE 4 oC NM:NUMBER_OF_SCANS 256 NM:DUMMY_SCANS 8 NM:ACQUISITION_TIME 2.0447 s NM:RELAXATION_DELAY 2 s NM:SPECTRAL_WIDTH 8012.8 Hz NM:NUM_DATA_POINTS_ACQUIRED 16384 points NM:REAL_DATA_POINTS 65536 NM:LINE_BROADENING 0.5 Hz NM:ZERO_FILLING 65,536 points NM:APODIZATION Exponential NM:BASELINE_CORRECTION_METHOD Spline NM:CHEMICAL_SHIFT_REF_STD Alanine doublet at 1.46 ppm #NMR_METABOLITE_DATA NMR_METABOLITE_DATA:UNITS A.U. NMR_METABOLITE_DATA_START Samples CLTI Amputation1B CLTI Amputation2B CLTI Amputation3B CLTI Amputation4B CLTI Amputation6B CLTI Amputation7B CLTI Amputation8B CLTI Amputation9B CLTI Amputation10B CLTI Pre-surgery1B CLTI Pre-surgery2B CLTI Pre-surgery5B CLTI Pre-surgery6B CLTI Pre-surgery7B CLTI Pre-surgery9B Control1B Control2B Control3B Control4B Control5B Control6B Control7B Control8B Control9B Control10B Factors Group:CLTI_Amputation Group:CLTI_Amputation Group:CLTI_Amputation Group:CLTI_Amputation Group:CLTI_Amputation Group:CLTI_Amputation Group:CLTI_Amputation Group:CLTI_Amputation Group:CLTI_Amputation Group:CLTI_Pre_surgery Group:CLTI_Pre_surgery Group:CLTI_Pre_surgery Group:CLTI_Pre_surgery Group:CLTI_Pre_surgery Group:CLTI_Pre_surgery Group:Control Group:Control Group:Control Group:Control Group:Control Group:Control Group:Control Group:Control Group:Control Group:Control Caprate 1551.435009 19272.77922 19765.96697 20886.53302 26545.51978 21320.25958 21728.43875 9761.603044 10487.68556 20321.59869 9923.434213 11824.88871 17680.5786 1169.686947 5850.051607 8631.300572 16351.01911 18203.3365 7063.66292 26200.27514 12003.13494 6607.499913 14691.30089 13485.21917 7605.969989 CH3-lipid 224125.3283 1005208.575 937843.0114 551717.8773 1543304.908 547227.193 671286.7011 307424.9219 1628391.18 1156141.183 749907.3305 443455.381 1350368.217 380583.1517 510605.3794 732525.8771 199979.2402 516007.2441 333131.3161 625320.0765 662878.0236 172953.286 482728.1686 756392.1776 280960.6525 2-Aminobutyrate 20638.51377 45894.56372 40513.76504 26734.77663 67556.75625 27502.66641 31220.13401 16473.00353 54517.54998 37451.30314 26449.62752 19678.15971 47451.98572 20703.60896 26393.14217 36061.65118 15620.67776 26038.25866 14574.54804 30781.53386 33029.38286 10404.9706 28441.6048 27595.55734 11253.49135 (CH2)nlipids+lactate 1124010.746 5233779.925 5106118.908 2698652.314 7963165.474 2813313.212 3549584.876 1466423.887 8289366.699 5894748.264 3671004.287 2241356.882 7452814.529 1950661.143 2428250.189 3835762.003 894146.2406 2654209.539 1826801.741 3129758.225 3456292.832 865590.3316 2620332.513 3767006.24 1364114.407 Lactate 322054.4574 1018574.867 780749.1139 545692.989 1545031.184 550106.6358 610979.6827 363065.8989 1843197.069 1293599.569 849505.714 458905.5639 1039675.331 374885.5746 614218.6117 634955.9737 150920.5631 427114.7525 266877.0917 603205.4978 670195.0738 131006.9156 389642.2845 818616.5765 276171.0707 Alanine 11106.83277 7766.663642 6006.755553 1964.852258 4724.001129 5740.832585 7207.774649 2814.444653 15453.3969 1113.82846 1676.116284 2442.282093 12517.7873 10450.89259 7657.988005 9991.766718 82.30495909 2488.091613 6837.054769 1427.97793 11304.39549 2519.425124 8851.586181 2298.096929 1024.384924 (CH2-CH2-CO-) lipids 102781.908 589303.2171 554757.491 296348.7106 916817.5135 297426.3882 366754.1258 168485.5872 995695.0952 689080.5147 454444.9664 254101.5919 818373.8693 249035.7067 313397.2266 461352.2831 86726.19952 282232.027 188588.9968 340148.3758 405806.6234 87977.36211 279727.0278 445432.3615 152819.1402 (CH=CH-CH2-CH2) lipids 191173.1486 882466.7402 812810.8706 460510.4251 1522895.203 461719.2533 580796.4152 270461.8235 1565308.842 1083614.964 770822.0649 382667.2242 1193929.974 343876.1457 518986.0222 725698.2539 172937.2663 474514.2474 316846.3877 532082.7176 588329.4084 150379.3165 431651.309 715487.8546 258776.8772 Glutamine 16878.33137 15191.4304 11016.05033 8859.548684 14105.43814 18990.97623 11286.8191 7265.737983 24605.53281 8965.523716 6033.6823 3856.102619 13102.50017 5275.191684 6500.640413 19337.75126 12865.46104 9041.348125 18309.66414 8596.810231 15936.41499 8726.436345 21039.30098 6096.090031 2606.9394 O-Cresol? 148.5219721 7903.277893 6940.005401 4373.243989 7865.724835 5604.82066 4834.093293 2238.403894 8044.985796 8819.89171 4586.801217 3627.231506 17614.33138 1935.420863 3197.510664 10215.83147 3316.05438 47099.92581 5110.167607 6256.159197 9697.94099 15360.71746 7365.807457 4973.489239 2165.826314 (CH2-CH2-CO) lipids 111074.7813 578029.2466 547193.8805 300443.9491 932621.358 298796.6832 379453.2326 168402.9916 986845.7387 697180.3928 465052.9561 255032.8914 788451.5748 227151.2475 305553.2157 452982.0439 91999.17213 292226.5546 188241.8724 342319.9968 393864.4676 89468.03323 272903.872 452970.1777 156829.8055 Pyruvate/3-Hydroxyisovalerate 524.4770364 1094.55023 668.6957272 70.56481829 1173.934386 2289.2859 953.9883554 334.3944975 696.8668686 2116.677169 1596.504298 1097.404356 1473.028555 891.9808046 3190.231448 1753.259418 499.4055339 264.4912098 870.4533626 209.9789587 2210.617708 757.8967149 1206.266225 1948.968586 650.9048653 Lipid Y 47952.31847 172656.591 143500.0541 63055.0987 430561.0537 112914.092 225204.4138 62859.11468 316688.0195 419718.4567 400738.6902 361377.6319 487216.7313 371729.2607 431031.0951 354841.9327 98487.28616 228308.363 214253.0842 171262.6206 325632.2786 175649.3748 215137.4488 378321.915 356669.0149 Creatine 79817.50487 35071.40255 25634.25389 20026.65831 38928.00511 61448.46295 54956.74036 21389.56204 48744.72064 29422.1114 31901.94227 41029.41501 31166.15775 40168.90765 54032.98325 44743.00267 28289.95059 42317.48954 60379.00109 38401.73787 49518.96231 46032.17839 48661.31536 33340.62004 39187.42223 Thymol/Methylmalonate? 24675.18706 13864.64654 6255.719749 11307.80969 23012.65363 26190.86472 25881.42106 9480.738951 30268.53174 129746.8512 163104.9172 159386.9715 155041.2577 172331.5419 179521.1517 128874.0837 33442.81283 84791.11598 26489.47902 39588.32356 55409.34151 92249.78157 35691.48346 152293.3421 177805.4421 Taurine 42722.21532 32422.76572 26774.84045 16481.16177 53731.3792 81993.2223 79715.27816 26014.63397 58514.68827 52036.67891 60620.85687 68304.0865 64340.30679 76273.93735 82373.85857 66806.10457 39041.80734 47628.94012 82301.10297 33633.3019 78970.00667 57068.29107 68720.70874 63115.86249 74138.49188 Unknown3.32to3.36ppm 522.4633205 952.1887746 4618.711396 1829.028317 9224.384209 304.7836103 1563.185511 53.7600067 11007.33954 6882.298237 1254.924488 2094.035073 2799.93388 5455.911086 3472.098999 6859.164632 130.4226851 34697.70034 3899.900976 1153.599774 7203.711365 15237.73686 4480.911276 1105.81205 7202.939733 Taurine.1 26522.75233 18257.31526 19130.13025 7462.355298 36291.20938 56745.72121 68273.35339 15696.94312 40206.67207 47132.82582 57561.71313 62900.00211 55446.4661 68509.88964 75936.96304 60648.54283 25978.4934 43334.58298 64484.83065 25296.0691 65219.52134 50838.28067 52625.36528 57787.36777 69131.45242 Unknown-3.77to3.87ppm 35343.06821 21875.63814 18112.98689 16387.69905 154315.6611 157527.6165 162898.6747 32395.36286 139169.3513 233571.4267 266446.1014 310875.1587 265843.3327 332968.5561 356948.7528 246963.254 76313.46051 174554.0739 207856.1739 95725.79434 255735.4851 165724.4982 178536.7845 262744.5938 317046.1133 Creatine.1 68718.16956 31710.32347 23093.58732 18734.80723 34124.60815 56538.26536 46720.05278 19358.3525 43049.00082 25042.4465 27444.2899 32483.50721 28927.91362 36689.32498 39388.69511 47480.44211 24694.40946 33066.7065 53956.08942 25333.59803 47539.34486 42414.67122 45718.42695 29751.15398 37044.21767 CH2OCOr(glyceryl) 44273.57708 183665.9875 171472.4046 100313.0217 292728.0132 101156.9097 123573.9652 59953.16684 300165.1004 206655.0054 141715.0812 81195.88711 233666.6834 77115.51352 98318.64448 147665.5139 32911.8777 113266.0126 60844.47568 110961.9542 129459.2525 44901.38028 95689.23759 140617.3055 55723.02814 Glucose 6594.27545 3603.384523 3406.102496 1192.828297 138.3370527 1807.646179 1469.640034 567.1224727 837.1665742 1763.725103 629.6230266 3446.766374 3085.322219 3938.894289 2010.878733 2938.080109 1615.037862 1644.561801 4141.494634 1157.277021 4771.894349 1363.426408 3219.330954 2472.783201 2763.780723 CH-glycerol 22116.45767 88425.27367 85233.34474 49702.31424 148299.8848 46762.73928 63054.21826 26681.22142 170023.8186 113529.1156 73559.98306 40781.15708 121593.2998 36705.15706 51254.99386 70411.06607 15470.93831 47011.03926 30564.06832 54973.36068 57396.62367 14012.66161 41866.74403 73636.86327 26700.51814 CH= lipids 122522.2735 559830.4267 520913.2365 275115.0868 1028634.381 270863.7033 354595.7864 166830.5982 960483.9892 686971.0335 504357.6808 227528.1483 740639.9245 212689.0271 330923.4576 468909.7315 93037.51448 284511.5731 180561.0741 316849.1981 365649.1561 93526.62466 254871.3869 456473.7475 160216.7482 Unknown-5.86to5.91ppm 188.4964123 2354.804515 2048.570735 202.9434725 1388.746315 161.8992454 190.2078599 76.8156845 2169.833304 295.9678654 717.8668228 146.9073283 1984.665169 101.2329937 155.6650121 194.5870128 62.26104607 130.5087794 218.1015496 214.0231948 128.3279514 79.21025776 297.7964307 211.2516099 97.62572665 NMR_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Caprate CH3-lipid 2-Aminobutyrate (CH2)nlipids+lactate Lactate Alanine (CH2-CH2-CO-) lipids (CH=CH-CH2-CH2) lipids Glutamine O-Cresol? (CH2-CH2-CO) lipids Pyruvate/3-Hydroxyisovalerate Lipid Y Creatine Thymol/Methylmalonate? Taurine Unknown3.32to3.36ppm Taurine.1 Unknown-3.77to3.87ppm Creatine.1 CH2OCOr(glyceryl) Glucose CH-glycerol CH= lipids Unknown-5.86to5.91ppm METABOLITES_END #END