#METABOLOMICS WORKBENCH ryant_20220217_085911 DATATRACK_ID:3095 STUDY_ID:ST002188 ANALYSIS_ID:AN003582 PROJECT_ID:PR001395 VERSION 1 CREATED_ON June 6, 2022, 9:07 pm #PROJECT PR:PROJECT_TITLE Metabolomic profiling reveals muscle metabolic changes following iliac PR:PROJECT_TITLE arteriovenous fistula creation in mice: part 1 PR:PROJECT_TYPE Study of the skeletal muscle metabolome in mice with iliac arteriovenous fistula PR:PROJECT_TYPE via 1H NMR PR:PROJECT_SUMMARY In the present study, we hypothesize that the creation of an iliac AVF would PR:PROJECT_SUMMARY result in significant alterations to the limb muscle metabolome. Recently, our PR:PROJECT_SUMMARY group developed a new murine model to address the pathophysiology of PR:PROJECT_SUMMARY access-related hand dysfunction (ARHD) in mice, where AVF creation is performed PR:PROJECT_SUMMARY in the iliac artery/vein. Because of the anatomical location of the AVF PR:PROJECT_SUMMARY creation, this model produces clinically relevant changes in the mouse hindlimb PR:PROJECT_SUMMARY including hemodynamic alterations, muscle weakness, and mitochondrial function PR:PROJECT_SUMMARY impairment. 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 (NIH) grant R01HL148597 (STS). Additional salary PR:FUNDING_SOURCE support was provided by NIH grants R01DK119274 (SAB) and R01HL149704 (TER), and PR:FUNDING_SOURCE a postdoctoral fellowship (POST903198) from the American Heart Association (KK). PR:PROJECT_COMMENTS Metabolomic study via NMR PR:PUBLICATIONS Frontiers PR:CONTRIBUTORS Ram B. Khattri, Kyoungrae Kim, Erik M. Anderson, Brian Fazzone, Kenneth C PR:CONTRIBUTORS Harland, Qiongyao Hu, Victoria R. Palzkill, Tomas A. Cort, Kerri A. O’Malle, PR:CONTRIBUTORS Scott A. Berceli, Terence E. Ryan, Salvatore T. Scali #STUDY ST:STUDY_TITLE Metabolomic profiling reveals muscle metabolic changes following iliac ST:STUDY_TITLE arteriovenous fistula creation in mice ST:STUDY_TYPE Study of the skeletal muscle metabolome in mice with iliac arteriovenous fistula ST:STUDY_TYPE via 1H NMR ST:STUDY_SUMMARY In the present study, we hypothesize that the creation of an iliac AVF would ST:STUDY_SUMMARY result in significant alterations to the limb muscle metabolome. Recently, our ST:STUDY_SUMMARY group developed a new murine model to address the pathophysiology of ST:STUDY_SUMMARY access-related hand dysfunction (ARHD) in mice, where AVF creation is performed ST:STUDY_SUMMARY in the iliac artery/vein. Because of the anatomical location of the AVF ST:STUDY_SUMMARY creation, this model produces clinically relevant changes in the mouse hindlimb ST:STUDY_SUMMARY including hemodynamic alterations, muscle weakness, and mitochondrial function ST:STUDY_SUMMARY impairment. 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 4 ST:TOTAL_SUBJECTS 34 ST:NUM_MALES All ST:STUDY_COMMENTS Metabolomic study via NMR ST:PUBLICATIONS Frontiers #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:GENOTYPE_STRAIN C57BL/6J SU:AGE_OR_AGE_RANGE 13 weeks SU:WEIGHT_OR_WEIGHT_RANGE (28.4±1.1 g (control SHAM mice) vs 26.4±2.1 g (control AVF mice) vs 23.6±2.1 SU:WEIGHT_OR_WEIGHT_RANGE (CKD SHAM mice) vs 24.7±1.3 (CKD AVF mice), N=8-10/group). SU:GENDER Male SU:ANIMAL_ANIMAL_SUPPLIER Jackson Labs (Stock # 000664) SU:ANIMAL_HOUSING Housed in a temperature of 22 oC SU:ANIMAL_LIGHT_CYCLE 12-hour light/12-hour dark SU:ANIMAL_FEED Ad libitum (Casein control diet vs. adenine-supplemented diet to induce CKD) SU:ANIMAL_WATER free access to food and water (3-5 animals per cage). SU:ANIMAL_INCLUSION_CRITERIA (3-5 animals per cage). #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 - Sample name Factor1***:Group RAW_FILE_NAME=raw data file names SUBJECT_SAMPLE_FACTORS - Terence_MS18_Quad_Org Factor1***:CKD_AVF RAW_FILE_NAME=Terence_MS18_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS26_Quad_Org Factor1***:CKD_AVF RAW_FILE_NAME=Terence_MS26_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS27_Quad_Org Factor1***:CKD_AVF RAW_FILE_NAME=Terence_MS27_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS28_Quad_Org Factor1***:CKD_AVF RAW_FILE_NAME=Terence_MS28_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS29_Quad_Org Factor1***:CKD_AVF RAW_FILE_NAME=Terence_MS29_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS30_Quad_Org Factor1***:CKD_AVF RAW_FILE_NAME=Terence_MS30_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS36_Quad_Org Factor1***:CKD_AVF RAW_FILE_NAME=Terence_MS36_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS37_Quad_Org Factor1***:CKD_AVF RAW_FILE_NAME=Terence_MS37_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS39_Quad_Org Factor1***:CKD_AVF RAW_FILE_NAME=Terence_MS39_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS40_Quad_Org Factor1***:CKD_AVF RAW_FILE_NAME=Terence_MS40_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS6_Quad_Org Factor1***:CKD_SHAM RAW_FILE_NAME=Terence_MS6_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS7_Quad_Org Factor1***:CKD_SHAM RAW_FILE_NAME=Terence_MS7_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS8_Quad_Org Factor1***:CKD_SHAM RAW_FILE_NAME=Terence_MS8_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS9_Quad_Org Factor1***:CKD_SHAM RAW_FILE_NAME=Terence_MS9_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS10_Quad_Org Factor1***:CKD_SHAM RAW_FILE_NAME=Terence_MS10_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS16_Quad_Org Factor1***:CKD_SHAM RAW_FILE_NAME=Terence_MS16_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS17_Quad_Org Factor1***:CKD_SHAM RAW_FILE_NAME=Terence_MS17_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS20_Quad_Org Factor1***:CKD_SHAM RAW_FILE_NAME=Terence_MS20_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS14_Quad_Org Factor1***:CON_AVF RAW_FILE_NAME=Terence_MS14_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS22_Quad_Org Factor1***:CON_AVF RAW_FILE_NAME=Terence_MS22_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS23_Quad_Org Factor1***:CON_AVF RAW_FILE_NAME=Terence_MS23_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS24_Quad_Org Factor1***:CON_AVF RAW_FILE_NAME=Terence_MS24_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS25_Quad_Org Factor1***:CON_AVF RAW_FILE_NAME=Terence_MS25_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS32_Quad_Org Factor1***:CON_AVF RAW_FILE_NAME=Terence_MS32_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS33_Quad_Org Factor1***:CON_AVF RAW_FILE_NAME=Terence_MS33_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS35_Quad_Org Factor1***:CON_AVF RAW_FILE_NAME=Terence_MS35_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS1_Quad_Org Factor1***:CON_SHAM RAW_FILE_NAME=Terence_MS1_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS2_Quad_Org Factor1***:CON_SHAM RAW_FILE_NAME=Terence_MS2_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS3_Quad_Org Factor1***:CON_SHAM RAW_FILE_NAME=Terence_MS3_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS4_Quad_Org Factor1***:CON_SHAM RAW_FILE_NAME=Terence_MS4_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS5_Quad_Org Factor1***:CON_SHAM RAW_FILE_NAME=Terence_MS5_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS11_Quad_Org Factor1***:CON_SHAM RAW_FILE_NAME=Terence_MS11_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS12_Quad_Org Factor1***:CON_SHAM RAW_FILE_NAME=Terence_MS12_Quad_Org.raw SUBJECT_SAMPLE_FACTORS - Terence_MS15_Quad_Org Factor1***:CON_SHAM RAW_FILE_NAME=Terence_MS15_Quad_Org.raw #COLLECTION CO:COLLECTION_SUMMARY Two weeks after surgery, mice were anesthetized using isoflurane and the left CO:COLLECTION_SUMMARY quadriceps muscle was rapidly dissected and immediately frozen in liquid CO:COLLECTION_SUMMARY nitrogen and stored at -80oC for metabolite extraction. Euthanasia was carried CO:COLLECTION_SUMMARY out by thoracotomy followed by cervical dislocation. CO:SAMPLE_TYPE Muscle CO:COLLECTION_METHOD While under isoflurane anesthesia, tissues were rapidly dissected and snap CO:COLLECTION_METHOD frozen in liquid nitrogen and stored at -80°C until metabolite extraction CO:COLLECTION_LOCATION University of Florida, Applied Physiology and Kinesiology, 1864 stadium RD, CO:COLLECTION_LOCATION Gainesville, FL 32611 CO:STORAGE_CONDITIONS -80℃ CO:STORAGE_VIALS cryovials #TREATMENT TR:TREATMENT_SUMMARY Briefly, mice were acclimated to a standard casein-based chow (Control Diet) for TR:TREATMENT_SUMMARY 7 days. Thereafter, the animals were randomly assign to either remain on Control TR:TREATMENT_SUMMARY Diet (non-CKD groups) or receive adenine-supplemented diet to induce CKD. TR:TREATMENT_SUMMARY Following three weeks of the casein/adenine diet, animals were randomly assigned TR:TREATMENT_SUMMARY to receive either an arteriovenous fistula surgery (AVF) or sham surgery (SHAM) TR:ANIMAL_VET_TREATMENTS none TR:ANIMAL_ANESTHESIA isoflurane TR:ANIMAL_FASTING non-fasted TR:ANIMAL_ENDP_EUTHANASIA Euthanasia was carried out by thoracotomy followed by cervical dislocation. TR:ANIMAL_ENDP_TISSUE_COLL_LIST Skeletal muscle (quadriceps) TR:ANIMAL_ENDP_TISSUE_PROC_METHOD Two weeks after surgery, mice were anesthetized using isoflurane and the left TR:ANIMAL_ENDP_TISSUE_PROC_METHOD quadriceps muscle was rapidly dissected and immediately frozen in liquid TR:ANIMAL_ENDP_TISSUE_PROC_METHOD nitrogen and stored at -80oC for metabolite extraction. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Weights of frozen quadriceps specimens were weighed using a microbalance SP:SAMPLEPREP_SUMMARY (Mettler-Toledo, Columbus, OH, USA). Next, a slightly modified FOLCH extraction SP:SAMPLEPREP_SUMMARY was performed to extract aqueous and lipid phase metabolites. The aqueous phase SP:SAMPLEPREP_SUMMARY was lyophilized overnight (Labconco Corporation, Kansas, MO, USA) and the lipid SP:SAMPLEPREP_SUMMARY phase was dried by passing inert nitrogen gas. The resulting aqueous and lipid SP:SAMPLEPREP_SUMMARY phase dry powders were stored at -80oC until analysis using nuclear magnetic SP:SAMPLEPREP_SUMMARY resonance (NMR). The dry powder of aqueous phase samples was dissolved in 50 µL SP:SAMPLEPREP_SUMMARY of phosphate buffer system (50 mM, pH 7.2) consisting of 0.5 mM D6-DSS, 2 mM SP:SAMPLEPREP_SUMMARY EDTA and 0.2% NaN2. Lipid phase dry powders were dissolved in 70 µL of CDCl3 SP:SAMPLEPREP_SUMMARY supplemented with 10 mM of pyrazine (as internal NMR standard). All samples were SP:SAMPLEPREP_SUMMARY loaded into 1.5 mm optical density (O.D.) NMR tubes. SP:PROCESSING_METHOD Lyophilization and Homogenization SP:PROCESSING_STORAGE_CONDITIONS -80℃ SP:EXTRACTION_METHOD Modified FOLCH extraction SP:EXTRACT_STORAGE -80℃ SP:SAMPLE_RESUSPENSION Deuterated chloroform (80 microliter) with 10 mM pyrazine was used to re-suspend SP:SAMPLE_RESUSPENSION organic phase samples. SP:SAMPLE_SPIKING 10 mM of pyrazine for organic phase samples. #ANALYSIS AN:DATA_FORMAT fid, 1r #NMR NM:INSTRUMENT_NAME Bruker Avance III Cryo800 MHz 54 mm NM:INSTRUMENT_TYPE FT-NMR NM:NMR_EXPERIMENT_TYPE 1D-1H NM:FIELD_FREQUENCY_LOCK Deuterated chloroform NM:STANDARD_CONCENTRATION 10mM pyrazine NM:SPECTROMETER_FREQUENCY 799.9 NM:NMR_PROBE CP TXI CryoProbe NM:NMR_SOLVENT Deuterated chloroform NM:NMR_TUBE_SIZE 1.5 mm O.D. NM:SHIMMING_METHOD Topshim NM:PULSE_SEQUENCE noesypr1d NM:WATER_SUPPRESSION none NM:PULSE_WIDTH 90-degree NM:RECEIVER_GAIN 228 NM:OFFSET_FREQUENCY None NM:CHEMICAL_SHIFT_REF_CPD CDCl3 at 7.26 ppm and pyrazine at 8.61 ppm NM:TEMPERATURE 25 o C NM:NUMBER_OF_SCANS 128 scans NM:DUMMY_SCANS 8 NM:ACQUISITION_TIME 4 s NM:RELAXATION_DELAY 1 s NM:SPECTRAL_WIDTH 9523.8 NM:NUM_DATA_POINTS_ACQUIRED 38094 NM:REAL_DATA_POINTS 65536 NM:LINE_BROADENING 0.22 Hz NM:ZERO_FILLING 65,536 points NM:APODIZATION Exponential NM:BASELINE_CORRECTION_METHOD Spline NM:CHEMICAL_SHIFT_REF_STD 7.26ppm for CDCl3 NM:BINNED_INCREMENT None NM:BINNED_DATA_NORMALIZATION_METHOD None NM:BINNED_DATA_CHEMICAL_SHIFT_RANGE None NM:BINNED_DATA_EXCLUDED_RANGE None #NMR_METABOLITE_DATA NMR_METABOLITE_DATA:UNITS A.U. NMR_METABOLITE_DATA_START Samples Terence_MS18_Quad_Org Terence_MS26_Quad_Org Terence_MS27_Quad_Org Terence_MS28_Quad_Org Terence_MS29_Quad_Org Terence_MS30_Quad_Org Terence_MS36_Quad_Org Terence_MS37_Quad_Org Terence_MS39_Quad_Org Terence_MS40_Quad_Org Terence_MS6_Quad_Org Terence_MS7_Quad_Org Terence_MS8_Quad_Org Terence_MS9_Quad_Org Terence_MS10_Quad_Org Terence_MS16_Quad_Org Terence_MS17_Quad_Org Terence_MS20_Quad_Org Terence_MS14_Quad_Org Terence_MS22_Quad_Org Terence_MS23_Quad_Org Terence_MS24_Quad_Org Terence_MS25_Quad_Org Terence_MS32_Quad_Org Terence_MS33_Quad_Org Terence_MS35_Quad_Org Terence_MS1_Quad_Org Terence_MS2_Quad_Org Terence_MS3_Quad_Org Terence_MS4_Quad_Org Terence_MS5_Quad_Org Terence_MS11_Quad_Org Terence_MS12_Quad_Org Terence_MS15_Quad_Org Factors Factor1***:CKD_AVF Factor1***:CKD_AVF Factor1***:CKD_AVF Factor1***:CKD_AVF Factor1***:CKD_AVF Factor1***:CKD_AVF Factor1***:CKD_AVF Factor1***:CKD_AVF Factor1***:CKD_AVF Factor1***:CKD_AVF Factor1***:CKD_SHAM Factor1***:CKD_SHAM Factor1***:CKD_SHAM Factor1***:CKD_SHAM Factor1***:CKD_SHAM Factor1***:CKD_SHAM Factor1***:CKD_SHAM Factor1***:CKD_SHAM Factor1***:CON_AVF Factor1***:CON_AVF Factor1***:CON_AVF Factor1***:CON_AVF Factor1***:CON_AVF Factor1***:CON_AVF Factor1***:CON_AVF Factor1***:CON_AVF Factor1***:CON_SHAM Factor1***:CON_SHAM Factor1***:CON_SHAM Factor1***:CON_SHAM Factor1***:CON_SHAM Factor1***:CON_SHAM Factor1***:CON_SHAM Factor1***:CON_SHAM C18-CH3 chol+V26C1C1:W25 70.655 27.752 27.048 44.38 47.43 28.337 23.29 72.172 32.084 66.42 25.232 25.967 25.427 25.043 24.077 25.408 24.809 18.766 28.188 29.088 42.952 45.541 46.372 30.963 77.07 71.476 23.017 21.865 24.685 24.524 23.878 24.87 17.8 26.693 CH3-protons 691.372 542.156 529.016 711.428 723.311 558.769 444.949 778.385 646.073 767.537 534.006 560.07 539.011 581.321 509.561 494.932 499.877 353.489 617.729 596.191 760.731 730.727 777.242 596.775 891.093 814.731 488.463 432.123 523.476 498.733 518.475 513.221 419.103 645.773 Unknown triplet 47.82 75.074 67.171 52.913 59.529 63.571 47.483 47.845 87.211 52.24 75.041 82.798 83.264 81.629 72.646 77.75 80.451 55.44 77.28 69.167 59.774 55.632 58.911 61.744 58.088 48.565 75.66 58.773 87.584 77.393 86.444 84.933 58.483 70.04 C19-CH3 Chol 78.603 31.821 30.808 49.717 51.737 32.289 26.034 80.11 37.522 73.017 29.471 30.227 29.436 29.603 27.176 29.267 28.894 22.459 32.59 32.717 47.49 49.111 50.294 35.813 84.989 80.154 26.171 24.64 28.081 27.101 27.76 28.292 19.524 32.397 CH2n chol 1564.606 1581.85 1515.987 1831.23 1885.353 1638.207 1288.852 1962.366 1906.215 1782.603 1563.808 1671.069 1611.632 1673.785 1596.773 1470.487 1504.582 997.995 1776.557 1723.287 1911.15 1869.333 2028.682 1641.192 2054.032 1795.856 1511.123 1285.858 1605.03 1532.11 1591.346 1570.263 1340.238 1745.25 CH2n aliphatic chains 1782.641 1543.131 1589.467 2017.276 2064.158 1550.354 1286.89 2044.771 1857.252 2101.739 1559.24 1576.19 1533.595 1730.72 1362.829 1373.817 1460.445 981.043 1811.215 1694.042 2288.391 2144.381 2243.717 1742.721 2477.924 2254.487 1310.006 1216.18 1511.576 1380.294 1464.627 1415.052 1153.951 1988.173 Cholesterol2 6.093 3.945 5.21 3.389 2.961 4.424 3.142 4.161 5.485 7.337 4.422 4.875 6.439 5.634 3.955 4.973 6.289 3.157 3.832 5.767 3.431 2.122 4.995 4.197 8.394 3.514 0.873 3.458 4.145 3.161 3.165 2.043 1.393 7.973 CH2-CH2-COO-beta 304.403 283.645 285.927 333.161 336.014 285.267 228.577 343.515 341.476 342.368 281.785 294.298 291.415 314.454 254.473 260.166 277.643 186.711 321.716 311.742 368.244 344.038 371.829 303.254 406.411 365.969 234.654 222.758 269.913 249.387 264.909 256.591 203.829 370.374 CH2-CH=CH-CH2-alpha 367.882 328.123 338.329 423.01 446.086 330.223 264.698 432.222 392.24 422.919 322.128 326.333 318.887 342.987 283.129 293.73 304.24 200.336 380.363 353.959 469.301 441.216 485.356 370.84 508.935 450.237 270.312 252.098 304.787 290.86 292.192 294.983 240.578 400.996 CH2COO-alpha 259.806 253.099 259.097 307.984 316.092 259.809 214.246 306.305 302.567 290.157 245.021 257.79 255.572 269.348 236.258 225.758 245.564 159.1 286.761 270.172 335.126 313.594 346.496 277.609 346.65 316.079 214.151 207.774 245.852 235.05 237.053 232.71 197.003 315.567 CH-CH2-CH=CH 260.089 421.392 404.766 409.118 396.197 439.408 302.859 305.085 515.694 317.249 426.946 474.88 474.251 457.874 464.247 433.241 445.757 288.517 469.834 447.438 425.059 374.912 466.109 411.871 376.158 250.229 426.963 353.089 483.579 463.532 470.212 462.938 365.807 372.556 N+CH33 290.602 377.11 376.386 395.683 431.415 384.813 371.402 346.969 426 327.774 367.71 389.653 387.283 379.97 379.428 351.756 367.06 254.952 396.836 383.829 418.687 377.84 479.084 368.557 366.897 342.778 339.941 345.516 372.931 383.694 367.85 358.979 340.011 375.678 Cholesterol3 176.249 23.677 28.865 30.184 29.496 26.062 18.015 69.495 33.374 71.925 36 33.377 35.951 38.699 23.965 33.39 71.33 73.49 35.08 27.213 32.069 34.518 28.236 178.727 72.578 72.398 26.208 24.145 28.47 23.046 32.443 28.568 15.108 139.923 3CH2 glycerophospholipids 159.703 390.705 299.169 521.087 323.473 354.212 423.084 256.175 737.816 272.034 730.579 426.256 525.06 459.44 328.038 525.033 230.624 142.813 481.928 401.955 461.532 488.404 300.495 199.278 324.327 141.221 463.648 299.565 431.398 376.185 434.754 472.286 283.451 174.167 CH2_TG1 80.742 191.836 265.475 163.196 367.929 105.731 91.465 108.482 174.529 115.361 203.523 253.803 192.363 169.573 195.274 206.53 103.18 64.007 197.763 389.637 242.58 158.593 508.118 95.809 135.575 99.394 256.763 95.593 147.68 145.921 135.275 155.663 153.085 109.781 CH2_TG2 103.596 142.689 172.35 138.291 175.207 116.452 107.064 129.631 151.589 131.148 151.458 172.66 153.245 151.889 143.835 147.78 118.207 77.828 161.58 184.118 169.01 142.677 246.452 113.234 147.385 125.369 158.058 105.854 132.362 128.93 127.403 130.778 119.292 138.053 1CH phospholipids+TG 61.344 94.668 111.991 97.022 111.716 85.102 68.66 75.602 103.694 81.995 98.745 110.027 100.203 96.253 92.999 95.831 81.336 52.531 105.559 112.585 111.284 93.4 139.386 81.768 88.542 55.38 100.056 69.76 93.547 88.351 88.787 87.458 74.907 70.232 CH phospholipid 58.588 77.897 87.112 88.629 93.522 82.132 67.261 74.864 92.572 72.712 78.673 88.096 86.723 86.733 82.438 77.335 80.645 50.472 89.631 83.075 94.214 85.861 104.952 83.439 83.628 54.324 74.39 68.32 82.922 85.615 74.479 76.459 68.638 71.64 CH TG 6.267 3.157 5.119 3.606 2.619 2.396 1.739 5.628 5.543 7.087 6.41 6.308 5.901 8.837 3.889 4.81 4.634 2.422 7.198 3.871 5.737 5.556 2.483 4.941 9.876 17.41 4.094 2.661 4.18 4.358 3.868 3.197 2.122 13.844 CH=CH 493.792 662.154 644.024 696.462 708.134 663.782 491.166 575.366 787.899 599.451 664.536 719.98 711.923 714.538 672.533 653.552 662.933 436.193 744.946 693.399 740.627 677.652 797.293 666.744 698.554 541.781 643 534.038 717.566 681.181 686.89 687.097 547.395 626.471 NMR_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name C18-CH3 chol+V26C1C1:W25 CH3-protons Unknown triplet C19-CH3 Chol CH2n chol CH2n aliphatic chains Cholesterol2 CH2-CH2-COO-beta CH2-CH=CH-CH2-alpha CH2COO-alpha CH-CH2-CH=CH N+CH33 Cholesterol3 3CH2 glycerophospholipids CH2_TG1 CH2_TG2 1CH phospholipids+TG CH phospholipid CH TG CH=CH METABOLITES_END #END