#METABOLOMICS WORKBENCH hormel101_20160720_143619_mwtab.txt DATATRACK_ID:683 STUDY_ID:ST000443 ANALYSIS_ID:AN000694 PROJECT_ID:PR000330 VERSION 1 CREATED_ON August 5, 2016, 10:26 am #PROJECT PR:PROJECT_TITLE Impact of Long-Term Poor and Good Glycemic Control on Metabolomics Alterations PR:PROJECT_TITLE in Type 1 Diabetic People. PR:PROJECT_TYPE Targeted GC-MS of plasma levels of tricarboxylic acid (TCA) cycle metabolites PR:PROJECT_SUMMARY The objective of the study was to determine whether T1D with good glycemic PR:PROJECT_SUMMARY control have persistent abnormalities of metabolites and pathways that exist in PR:PROJECT_SUMMARY T1D with poor glycemic control. PR:INSTITUTE Mayo Clinic PR:DEPARTMENT Endocrinology PR:LABORATORY Mayo Clinic Metabolomics Resource Core PR:LAST_NAME Nair PR:FIRST_NAME Sreekumaran PR:ADDRESS 200 First Street SW, Rochester, MN 55905 PR:EMAIL Nair.K@mayo.edu PR:PHONE 507-285-2415 #STUDY ST:STUDY_TITLE Quantitative measurements of TCA cycle metabolites in T1D poor control, good ST:STUDY_TITLE control, and controls. ST:STUDY_TYPE Quantitative measurements of plasma levels of tricarboxylic acid (TCA) cycle ST:STUDY_TYPE metabolites ST:STUDY_SUMMARY The objective of the study was to determine whether T1D with good glycemic ST:STUDY_SUMMARY control have persistent abnormalities of metabolites and pathways that exist in ST:STUDY_SUMMARY T1D with poor glycemic control. ST:INSTITUTE Mayo Clinic ST:DEPARTMENT Endocrinology ST:LABORATORY Mayo Clinic Metabolomics Resource Core ST:LAST_NAME Nair ST:FIRST_NAME Sreekumaran ST:ADDRESS 200 First Street SW, Rochester, MN 55905 ST:EMAIL Nair.K@mayo.edu ST:PHONE 507-285-2415 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS ms3332-1 sample1 treatment:T1D good glycemic control SUBJECT_SAMPLE_FACTORS ms3332-2 sample2 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-3 sample3 treatment:T1D good glycemic control SUBJECT_SAMPLE_FACTORS ms3332-4 sample4 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-5 sample5 treatment:T1D good glycemic control SUBJECT_SAMPLE_FACTORS ms3332-6 sample6 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-7 sample7 treatment:T1D good glycemic control SUBJECT_SAMPLE_FACTORS ms3332-8 sample8 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-9 sample9 treatment:T1D good glycemic control SUBJECT_SAMPLE_FACTORS ms3332-10 sample10 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-19 sample11 treatment:T1D good glycemic control SUBJECT_SAMPLE_FACTORS ms3332-20 sample12 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-21 sample13 treatment:T1D good glycemic control SUBJECT_SAMPLE_FACTORS ms3332-22 sample14 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-23 sample15 treatment:T1D good glycemic control SUBJECT_SAMPLE_FACTORS ms3332-20 sample16 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-11 sample17 treatment:T1D good glycemic control SUBJECT_SAMPLE_FACTORS ms3332-12 sample18 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-13 sample19 treatment:T1D good glycemic control SUBJECT_SAMPLE_FACTORS ms3332-14 sample20 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-15 sample21 treatment:T1D good glycemic control SUBJECT_SAMPLE_FACTORS ms3332-16 sample22 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-24 sample23 treatment:T1D good glycemic control SUBJECT_SAMPLE_FACTORS ms3332-25 sample24 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-26 sample25 treatment:T1D good glycemic control SUBJECT_SAMPLE_FACTORS ms3332-16 sample26 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-17 sample27 treatment:T1D good glycemic control SUBJECT_SAMPLE_FACTORS ms3332-18 sample28 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-27 sample29 treatment:T1D good glycemic control SUBJECT_SAMPLE_FACTORS ms3332-28 sample30 treatment:ND SUBJECT_SAMPLE_FACTORS ms3333-25 sample31 treatment:ND SUBJECT_SAMPLE_FACTORS ms3333-14 sample32 treatment:ND SUBJECT_SAMPLE_FACTORS ms3333-27 sample33 treatment:ND SUBJECT_SAMPLE_FACTORS ms3333-4 sample34 treatment:ND SUBJECT_SAMPLE_FACTORS ms3333-6 sample35 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-36 sample36 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-50 sample37 treatment:ND SUBJECT_SAMPLE_FACTORS ms3333-20 sample38 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-28 sample39 treatment:ND SUBJECT_SAMPLE_FACTORS ms3333-8 sample40 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-14 sample41 treatment:ND SUBJECT_SAMPLE_FACTORS ms3333-10 sample42 treatment:ND SUBJECT_SAMPLE_FACTORS ms3333-12 sample43 treatment:ND SUBJECT_SAMPLE_FACTORS ms3332-54 sample44 treatment:ND SUBJECT_SAMPLE_FACTORS ms3333-1 sample45 treatment:T1D poor glycemic control SUBJECT_SAMPLE_FACTORS ms3333-21 sample46 treatment:T1D poor glycemic control SUBJECT_SAMPLE_FACTORS ms3333-7 sample47 treatment:T1D poor glycemic control SUBJECT_SAMPLE_FACTORS ms3333-22 sample48 treatment:T1D poor glycemic control SUBJECT_SAMPLE_FACTORS ms3333-9 sample49 treatment:T1D poor glycemic control SUBJECT_SAMPLE_FACTORS ms3333-11 sample50 treatment:T1D poor glycemic control SUBJECT_SAMPLE_FACTORS ms3333-23 sample51 treatment:T1D poor glycemic control SUBJECT_SAMPLE_FACTORS ms3333-13 sample52 treatment:T1D poor glycemic control SUBJECT_SAMPLE_FACTORS ms3333-15 sample53 treatment:T1D poor glycemic control SUBJECT_SAMPLE_FACTORS ms3333-3 sample54 treatment:T1D poor glycemic control SUBJECT_SAMPLE_FACTORS ms3333-5 sample55 treatment:T1D poor glycemic control SUBJECT_SAMPLE_FACTORS ms3333-16 sample56 treatment:T1D poor glycemic control SUBJECT_SAMPLE_FACTORS ms3333-18 sample57 treatment:T1D poor glycemic control SUBJECT_SAMPLE_FACTORS ms3333-19 sample58 treatment:T1D poor glycemic control #COLLECTION CO:COLLECTION_SUMMARY At 5:00 AM after an overnight fast, baseline blood samples were collected from CO:COLLECTION_SUMMARY study participants. Plasma samples were stored at 80°C until analysis. CO:SAMPLE_TYPE Blood. Plasma was isolated for MS analysis. #TREATMENT TR:TREATMENT_SUMMARY Participants were admitted to the Clinical Research Unit at St Mary’s Hospital TR:TREATMENT_SUMMARY (Rochester, Minnesota) the evening before the study and spent overnight in the TR:TREATMENT_SUMMARY Clinical Research Unit. The participants were given a standard meal on the TR:TREATMENT_SUMMARY evening of the admission after which they fasted overnight. Participants with TR:TREATMENT_SUMMARY T1D were treated with insulin as per their usual individual programs. At 5:00 AM TR:TREATMENT_SUMMARY after an overnight fast, baseline blood samples were collected from study TR:TREATMENT_SUMMARY participants. Plasma samples were stored at 80°C until analysis. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Plasma quality-control samples used in the study were prepared from pooled SP:SAMPLEPREP_SUMMARY plasma spiked with a selection of metabolites to mimic elevated levels of SP:SAMPLEPREP_SUMMARY metabolites during I− (insulin withdrawn) condition. Plasma was spiked with a SP:SAMPLEPREP_SUMMARY standard mixture (3:1 ratio of plasma to spiking solution) containing 100 μg/mL SP:SAMPLEPREP_SUMMARY niacin, hypoxanthine, leucine, isoleucine, phenylalanine, tryptophan, citric SP:SAMPLEPREP_SUMMARY acid, glucose, hippuric acid, and taurocholic acid dissolved in 1:1 SP:SAMPLEPREP_SUMMARY acetonitrile/water. All plasma samples (200 μL) were thawed on ice at 4°C SP:SAMPLEPREP_SUMMARY followed by deproteinization with methanol (1:4 ratio of plasma to methanol) and SP:SAMPLEPREP_SUMMARY vortexed for 10 s, followed by incubation at −20°C for 2 h. The samples were SP:SAMPLEPREP_SUMMARY then centrifuged at 15,871g for 30 min at 4°C. The supernatants were SP:SAMPLEPREP_SUMMARY lyophilized (Savant, Holbrook, NY) and stored at −20°C prior to analysis. The SP:SAMPLEPREP_SUMMARY samples were reconstituted in 50% H2O/acetonitrile and passed through a Microcon SP:SAMPLEPREP_SUMMARY YM3 filter (Millipore Corporation). The supernatants were transferred to SP:SAMPLEPREP_SUMMARY analytical vials, stored in the autosampler at 4°C, and analyzed within 48 h of SP:SAMPLEPREP_SUMMARY reconstitution in buffer. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY The derivatized extracts were analyzed with an Agilent 6890 gas chromatograph CH:CHROMATOGRAPHY_SUMMARY coupled with an Agilent 5973 mass selective detector. The 1-μL aliquots of the CH:CHROMATOGRAPHY_SUMMARY extracts were injected into a DB5-MS capillary column (30 m × 250 μm i.d., CH:CHROMATOGRAPHY_SUMMARY 0.25-μm film thickness; J&W Scientific, Folson, CA) using PTV injection CH:CHROMATOGRAPHY_SUMMARY (Gerstel CIS4 injector) in the splitless mode. The temperature of the PTV was 70 CH:CHROMATOGRAPHY_SUMMARY °C during injection, and 0.6 min after injection, the temperature was raised to CH:CHROMATOGRAPHY_SUMMARY 300 °C at a rate of 2 °C/s and held at 300 °C for 20 min. The initial GC oven CH:CHROMATOGRAPHY_SUMMARY temperature was 70 °C, 5 min after injection the GC oven temperature was CH:CHROMATOGRAPHY_SUMMARY increased with 5 °C/min to 320 °C and held for 5 min at 320 °C. Helium was CH:CHROMATOGRAPHY_SUMMARY used as a carrier gas and pressure programmed such that the helium flow was kept CH:CHROMATOGRAPHY_SUMMARY constant at a flow rate of 1.7 mL/min. Detection was achieved using MS detection CH:CHROMATOGRAPHY_SUMMARY in electron impact mode and full scan monitoring mode (m/z 15−800). The CH:CHROMATOGRAPHY_SUMMARY temperature of the ion source was set at 250 °C and that of the quadrupole at CH:CHROMATOGRAPHY_SUMMARY 200 °C. CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent 6890 CH:COLUMN_NAME None #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Agilent 5973 MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS micromolar MS_METABOLITE_DATA_START Samples sample1 sample2 sample3 sample4 sample5 sample6 sample7 sample8 sample9 sample10 sample11 sample12 sample13 sample14 sample15 sample16 sample17 sample18 sample19 sample20 sample21 sample22 sample23 sample24 sample25 sample26 sample27 sample28 sample29 sample30 sample31 sample32 sample33 sample34 sample35 sample36 sample37 sample38 sample39 sample40 sample41 sample42 sample43 sample44 sample45 sample46 sample47 sample48 sample49 sample50 sample51 sample52 sample53 sample54 sample55 sample56 sample57 sample58 Factors treatment:T1D good glycemic control treatment:ND treatment:T1D good glycemic control treatment:ND treatment:T1D good glycemic control treatment:ND treatment:T1D good glycemic control treatment:ND treatment:T1D good glycemic control treatment:ND treatment:T1D good glycemic control treatment:ND treatment:T1D good glycemic control treatment:ND treatment:T1D good glycemic control treatment:ND treatment:T1D good glycemic control treatment:ND treatment:T1D good glycemic control treatment:ND treatment:T1D good glycemic control treatment:ND treatment:T1D good glycemic control treatment:ND treatment:T1D good glycemic control treatment:ND treatment:T1D good glycemic control treatment:ND treatment:T1D good glycemic control treatment:ND treatment:ND treatment:ND treatment:ND treatment:ND treatment:ND treatment:ND treatment:ND treatment:ND treatment:ND treatment:ND treatment:ND treatment:ND treatment:ND treatment:ND treatment:T1D poor glycemic control treatment:T1D poor glycemic control treatment:T1D poor glycemic control treatment:T1D poor glycemic control treatment:T1D poor glycemic control treatment:T1D poor glycemic control treatment:T1D poor glycemic control treatment:T1D poor glycemic control treatment:T1D poor glycemic control treatment:T1D poor glycemic control treatment:T1D poor glycemic control treatment:T1D poor glycemic control treatment:T1D poor glycemic control treatment:T1D poor glycemic control Citrate 129.409 151.45 132.09 160.582 139.822 240.655 150.787 161.508 206.101 111.822 151.149 161.742 121.657 128.015 212.104 161.439 140.06 155.595 154.462 137.393 97.643 154.683 156.613 146.307 159.329 153.502 128.017 158.248 169.83 121.615 165.4 134.54 101.11 182.76 146.54 151.19 135.915 120.29 121.44 160.7 138.44 151 143.61 176.203 140.49 146.08 145.09 146.63 143.91 106.71 137.517 139.24 120.96 125.09 171.23 130.4 111.603 106.19 cis.Aconitate 7.016 6.715 8.198 6.811 8.733 8.616 8.088 7.312 10.19 6.864 9.68 11.902 11.789 8.937 11.614 12.388 9.276 9.012 10.06 8.776 9.095 9.349 10.314 9.595 9.435 9.371 9.847 8.88 12.047 10.757 NA 9.71 8.35 8.23 8.49 7.71 3.824 8.37 7.61 9.43 8.84 9.69 8.93 4.636 8.26 8.81 7.22 9.26 9.95 9.46 4.447 8.01 9.4 8.89 11.61 6.42 4.853 8.53 Isocitrate 5.174 4.823 3.193 4.693 3.529 8.331 5.241 6.378 6.651 5.752 2.368 5.993 1.16 5.479 9.255 6.631 6.755 6.608 5.65 7.009 NA 3.999 5.607 2.916 4.518 6.191 4.886 5.016 NA 5.537 5.37 5.39 3.81 5.85 6.1 4.66 5.568 3.71 6.2 6.81 6.75 6.08 5.79 6.512 4.26 5.89 6.07 4.53 1.98 5.48 5.69 7.85 1.56 2.28 4.65 1.51 3.653 2.56 Ketoglutarate 17.043 7.759 9.32 8.77 12.235 14.028 10.882 13.581 13.709 7.695 12.939 15.085 9.809 12.485 11.337 15.079 8.577 12.148 12.049 10.911 10.316 14.67 21.656 10.782 11.113 14.517 11.444 13.338 12.055 9.916 8.65 8.25 11.11 11.4 9.77 13.78 9.407 12.68 9.93 9.19 10.91 9.69 9.96 10.064 10.56 12.71 10.59 8.67 13.38 11.06 9.434 13.44 11.26 9.7 13.28 12.43 7.275 8.7 succinate 4.163 2.579 2.817 2.607 3.253 4.951 3.258 3.821 3.809 2.495 3.749 4.896 4.6 3.405 4.523 4.874 2.616 3.459 3.437 2.896 2.739 5.719 4.683 3.454 4.082 5.88 4.388 5.7 5.643 3.055 4.59 2.43 5.14 4.08 2.82 5.26 3.773 4.12 NA 3.08 2.94 3.25 2.72 4.711 3.84 5.18 3.72 4.61 4.03 3.74 1.802 3.28 4.66 3.21 4.14 4.54 1.17 3.91 fummarate 1.927 0.997 1.553 1.046 1.445 2.322 1.728 1.753 1.834 1.174 2.099 2.54 1.837 1.738 2.468 2.561 1.47 1.737 1.762 2.033 1.394 2.908 3.674 1.954 2.231 2.98 2.133 3.127 2.214 2.172 1.77 1.21 2.09 2.21 1.2 1.78 1.518 2.78 1.49 2.04 2.11 1.63 1.72 2.429 1.32 2.15 1.49 2.04 2.18 2.04 1.288 1.81 1.8 1.38 1.81 1.94 0.929 1.59 Malate 4.83 2.246 4.431 2.865 3.739 7.132 4.769 4.479 4.598 2.52 3.831 4.183 3.627 3.35 6.496 4.154 4.136 3.927 4.839 4.02 2.469 5.412 6.337 3.521 4.308 5.365 4.661 5.004 4.339 3.083 4.39 2.09 4.56 4.68 2.55 3.84 2.683 6.23 3.16 4.42 3.98 3.67 3.84 8.92 3.61 4.79 4.2 3.45 4.74 3.51 2.603 4.43 4.51 3.5 5.56 3.54 2.74 2.41 Oxaloacetate 0.988 0.685 0.638 0.449 1.094 0.198 0.471 1.539 0.455 0.862 1.306 1.434 1.372 1.562 1.494 1.417 0.832 0.541 0.673 0.764 1.139 1.04 1.437 1.334 1.406 1.602 1.425 1.774 1.439 1.448 NA 0.63 1.27 2.19 0.74 1.28 0.357 1.42 1.45 1.31 0.79 2.22 0.98 0.655 0.78 1.41 1.54 1.26 0.39 1.38 1.033 0.73 1.38 0.85 0.87 1.51 0.988 1.26 Aspartate 0.426 0.882 2.113 0.291 1.509 0.24 0.832 2.505 0.157 1.076 1.518 1.743 2.61 0.924 0.969 1.718 2.751 0.731 2.614 1.378 1.893 2.393 1.21 1.313 1.546 2.34 2.617 1.173 0.648 1.544 NA 0.79 0.81 1.25 0.92 0.76 0.69 1.22 1.55 2.18 1.43 0.79 0.91 1.303 3.66 1.1 1.25 2.06 1.57 1.71 1.364 1.33 1.41 1.88 1.11 1 0.54 1.26 Glutamate 119.374 86.447 118.767 74.218 120.377 23.964 39.453 369.402 31.138 133.69 288.466 261.847 234.399 155.741 99.413 273.395 195.569 157.737 180.506 170.106 435.011 332.078 422.406 346.439 300.114 382.792 214.579 234.834 60.201 593.966 NA 212.32 58.94 76.25 124.62 85.99 56.348 133.94 402.74 341.29 180.65 156.21 247.71 127.927 273.56 123.93 305.62 257.66 182.52 263.24 241.339 135.19 166.74 273.12 131.69 71.24 98.966 280.48 Lacatate 80.199 59.895 27.472 73.667 44.132 70.29 50.013 85.155 56.179 64.61 172.298 133.209 72.363 114.601 127.77 134.176 53.29 99.514 208.27 44.562 98.247 NA 83.018 121.839 71.672 NA 88.389 17.251 120.304 162.041 82.6 97.11 247.64 188.32 152.55 123.54 102.913 NA 171.37 63.66 47.18 52.41 61.51 212.425 49.99 188.48 42.37 62.51 90.09 126.8 77.364 123.04 133.02 47.79 156.93 103.32 80.37 51.88 Alanine 383.673 356.072 399.334 385.052 405.646 434.298 357.49 440.415 381.397 311.723 386.988 392.706 333.001 347.981 432.635 393.753 419.033 428.607 383.098 398.638 385.287 422.021 314.328 375.073 380.41 428.762 345.497 353.356 348.808 416.794 68.74 431.45 395.21 465.32 410.93 399.83 362.082 427.84 420.21 376.5 408.15 371.21 316.97 384.216 389.61 413.54 372.58 373.2 439.15 430.95 359.997 453.24 440.81 350.66 457.72 355.98 251.086 331.75 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Citrate cis.Aconitate Isocitrate Ketoglutarate succinate fummarate Malate Oxaloacetate Aspartate Glutamate Lacatate Alanine METABOLITES_END #END