#METABOLOMICS WORKBENCH sdasari_20150618_9777721_mwtab.txt DATATRACK_ID:328 STUDY_ID:ST000221 ANALYSIS_ID:AN000328 PROJECT_ID:PR000181 VERSION 1 CREATED_ON 2016-09-17 #PROJECT PR:PROJECT_TITLE Identification of altered metabolites in normal plasma cells, slow progression PR:PROJECT_TITLE myeloma and fast progression multiple meyloma PR:PROJECT_TYPE Untargeted LC-MS Metabolomics PR:PROJECT_SUMMARY This experiment is comparing the metabolite profiles between normal plasma PR:PROJECT_SUMMARY and clonal plasma cells from multiple myeloma patients. PR:INSTITUTE Mayo Clinic PR:DEPARTMENT Neurology PR:LAST_NAME Gonsalves PR:FIRST_NAME Wilson PR:ADDRESS 200 First Street SW, Rochester, MN 55905 PR:EMAIL Dasari.Surendra@mayo.edu PR:PHONE 507-284-0513 #STUDY ST:STUDY_TITLE Normal plasma cells,Low proliferation multiple myeloma and High proliferation ST:STUDY_TITLE myeloma cells ST:STUDY_TYPE differential metabolomics ST:STUDY_SUMMARY CD138 sorted bone marrow plasma cell were obtained from a normal patient, a ST:STUDY_SUMMARY myelow with slow proliferation and a multiple myelow with rapid proliferation. ST:INSTITUTE Mayo Clinic ST:DEPARTMENT Hematology ST:LAST_NAME Gonsalves ST:FIRST_NAME William ST:ADDRESS - ST:EMAIL Dasari.Surendra@mayo.edu ST:PHONE - ST:NUM_GROUPS 1 ST:TOTAL_SUBJECTS 1 #SUBJECT SU:SUBJECT_TYPE Human 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 - BM049394SC004 Cognitive Status:fast MM SUBJECT_SAMPLE_FACTORS - BM049429SC001 Cognitive Status:normal SUBJECT_SAMPLE_FACTORS - BM049411SC003 Cognitive Status:slow MM #COLLECTION CO:COLLECTION_SUMMARY Bone marrow cells were collected after fasting. Collected cells were CD138 CO:COLLECTION_SUMMARY and frozen in -80C until analysis. CO:SAMPLE_TYPE Bone marrow cells CO:COLLECTION_LOCATION Illiac crest #TREATMENT TR:TREATMENT_SUMMARY - #SAMPLEPREP SP:SAMPLEPREP_SUMMARY 1) Incubated cell pellets on ice for 15 min 2) Added PBS to cells and vortex SP:SAMPLEPREP_SUMMARY Sonicate for 1 min in ice bath 4) Added MeOH+ACN mixture to cells and vortex SP:SAMPLEPREP_SUMMARY Sonicated for 1 min in ice bath 6) Incubated on ice for 30 min + spin for 20 SP:SAMPLEPREP_SUMMARY at 18K RPM 7) Combined 110 ul from each sample into a pool sample (330 ul SP:SAMPLEPREP_SUMMARY Split pool into two fractions of 150 ul each (hilic & C18) 9) Split remaning SP:SAMPLEPREP_SUMMARY into 150 ul x 2 (hilic & C18) #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY C18 CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME high-strength silica 2.1 150 mm, 1.8 m; Waters CH:CHROMATOGRAPHY_COMMENTS Metabolite separation was achieved using an Acquity UPLC system (Waters, CH:CHROMATOGRAPHY_COMMENTS MA) with both hydrophilic interaction chromatography (HILIC) (ethylene-bridged CH:CHROMATOGRAPHY_COMMENTS 2.1150 mm, 1.7 mm; Waters) and reversed-phase liquid chromatography C18 (RPLC) CH:CHROMATOGRAPHY_COMMENTS silica 2.1150 mm, 1.8 m; Waters). For each column, the run time was 20 min at a CH:CHROMATOGRAPHY_COMMENTS rate of 400 L/min. Reverse-phase chromatography was performed using 99% solvent CH:CHROMATOGRAPHY_COMMENTS (5 mmol/L NH4 acetate, 0.1% formic acid, and 1% acetonitrile) to 100% solvent B CH:CHROMATOGRAPHY_COMMENTS acetonitrile with 0.1% formic acid). The gradient was 0 min, 0% B; 1 min, 0% B; CH:CHROMATOGRAPHY_COMMENTS min, 5% B; 13.0 min, 100% B; 16 min, 100% B; 16.5 min, 0% B; and 20 min, 0% B. CH:CHROMATOGRAPHY_COMMENTS hydrophilic interaction chromatography gradient was as follows: 0 min, 100% B; CH:CHROMATOGRAPHY_COMMENTS min, 100% B; 5 min, 90% B; 13.0 min, 0% B; 16 min, 0% B; 16.5 min, 100% B; and CH:CHROMATOGRAPHY_COMMENTS min, 100% B. The injection volume of each sample was 5 L and column was CH:CHROMATOGRAPHY_COMMENTS at 50C. #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6220 TOF MS:INSTRUMENT_TYPE TOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS C18-NESI (-ve ESI) A 6220 ToF-MS (Agilent Technologies) was operated in both MS:MS_COMMENTS and negative electrospray ionization (ESI) modes using a scan range of 50?1,200 MS:MS_COMMENTS The mass accuracy and mass resolution were 5 parts per million (ppm) and 20,000 MS:MS_COMMENTS respectively. The instrument settings were as follows: nebulizer gas MS:MS_COMMENTS 325°C, capillary voltage 3.5 kV, capillary temperature 300°C, fragmentor MS:MS_COMMENTS 150 V, skimmer voltage 58 V, octapole voltage 250 V, cycle time 0.5 s, and run MS:MS_COMMENTS 15.0 min. MS:MS_COMMENTS Negative C18 #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak area MS_METABOLITE_DATA_START Samples BM049394SC004 BM049429SC001 BM049411SC003 Factors Cognitive Status:fast MM Cognitive Status:normal Cognitive Status:slow MM 1-(2E_6E-phytadienyl)-2-(2E_6E-phytadienyl)-sn-glycero-3-phosphocholine 697.2111 271.7892 185.1338 Cyclopassifloside VII 675.4060 0.0000 6.1144 Dihydrozeatin-9-N-glucoside-O-glucoside 530.6018 570.8102 84.1046 Evasterioside E 2442.3706 31.4246 50.3428 Gingerglycolipid B 796.9873 0.0000 0.0000 Megalomicin B 4115.6411 6612.9672 484.0743 Neocasomorphin 4225.2320 113.8715 152.4182 PC(16:0/5:0(COOH)) 351.7045 2.9158 10.8483 Phenformin 449.1076 134.6065 29.2300 PS(20:3(8Z_11Z_14Z)/0:0) 592.0977 319.2372 39.3507 PS(20:3(8Z_11Z_14Z)/0:0)_1 366.1264 516.8746 222.8173 PS(21:0/0:0) 1077.0463 40.5113 7.2310 PS(21:0/0:0)_1 1363.4347 368.3055 342.1764 PS(22:1(11Z)/0:0) 931.5299 0.0000 10.0384 Vignatic acid A 387.8382 147.6978 170.1651 197.060476_8.162866667 290.3808725 216.2473498 100.590187 169.0884532_3.39205 115.7672996 113.8517466 118.1733398 528.3080347_11.63933333 1957.174722 1305.618093 820.8563253 164.0719899_3.39205 142.3563031 140.8409122 28.07427199 540.3293522_11.37383333 1487.010201 1300.474876 107.6009788 478.2915402_11.55636667 997.7163207 785.9975171 390.8010268 524.2766613_10.80133333 5519.785989 4724.469984 951.258738 476.2763965_10.84281667 1223.507098 1228.159801 393.1616229 208.0976367_10.41966667 1991.077405 1871.061481 1038.460147 672.1366554_10.39478333 37.41891479 102.0421152 84.79765933 500.2762336_10.85111667 4367.954156 3800.574804 2013.756183 210.0781561_9.689533333 96.73576265 115.657688 357.1609939 526.2922932_11.08343333 3479.717582 2816.894495 966.8831079 619.2876334_10.00481667 876.0341288 774.3645706 335.9127706 564.3291788_10.9009 1563.024558 1588.273851 79.66470403 599.3181467_11.64763333 739.8728767 545.7978954 182.0172049 791.5826922_13.7053 0 12.25319744 0 877.6634991_13.7053 1793.602184 1483.25882 1050.413274 808.5741681_13.7053 2434.817379 2661.237157 149.3483007 671.266041_12.83411667 233.4288807 135.1654062 186.7515464 441.2029728_13.1328 4117.55128 2320.181377 625.1442895 891.5784301_13.7053 130.8259508 39.02924838 0 122.0345828_18.57571667 23.62778643 18.43657827 37.09574827 826.56188_19.38881667 24111.83766 9105.584096 25338.67403 885.5492237_15.87913333 62493.27595 23944.60833 39485.55544 859.5707156_13.7053 622.9691362 247.845041 0 906.6184419_15.55555 692.6487387 370.8073786 2580.147767 642.3609948_11.90483333 282.4222284 221.4331021 0 720.4086886_11.97121667 1063.071896 1000.554625 0 710.3860773_11.89655 427.3623895 336.4351876 0 685.3575101_11.77208333 298.5545083 249.7623378 0 694.392325_11.81356667 1501.922017 1373.450155 0 638.3659774_12.06248333 2363.713247 1795.215873 0 698.2860766_12.83411667 741.9280382 621.8919469 585.9389256 681.2951164_12.83411667 640.5255205 513.4134858 443.6861704 694.4286295_12.66816667 886.8874226 558.131082 0 706.3565028_12.06248333 445.6185401 275.1364179 0 664.385352_12.21183333 2826.111591 2066.027279 0 153.0657071_3.39205 149.2114854 137.9730817 152.8471916 263.8533923_0.936116667 213.7861688 205.7392239 547.9957829 265.8536298_0.944416667 350.3228216 412.2122202 586.3505222 197.8077571_0.944416667 8857.121751 8769.231044 20973.74897 372.9995855_0.927833333 58.1502597 70.3437953 214.7779116 102.0562098_0.927833333 130.409268 143.0128712 26.81625366 313.064623_0.936116667 0 0 494.3990426 195.8107828_0.944416667 7516.960081 7416.00988 18074.99117 209.8340127_0.944416667 263.527773 236.6222511 338.2610154 202.019732_0.944416667 409.2970379 404.0372274 170.9271951 159.0088498_0.952716667 1091.790112 1004.314682 432.4199762 199.8047957_0.944416667 4998.279324 5082.053231 11859.66607 343.9941745_0.944416667 190.3026318 175.1648206 5755.395907 291.0824524_0.944416667 39.16175408 23.60779498 4000.264788 325.8342982_0.819966667 50.39384455 73.97993111 153.3734849 180.9009543_0.828266667 13.44781955 65.4595661 87.10863548 232.9257215_0.894633333 1297.246292 990.3854711 403.1086334 138.965913_0.795066667 240.1924942 217.2263258 108.626374 206.9725179_0.819966667 13278.46194 12657.95247 5279.702643 162.9830515_0.819966667 362.9284716 364.7586555 163.993866 172.9559813_0.894633333 867.617099 791.1811051 264.4387272 162.9078616_0.919533333 172.4406895 131.4096919 533.0255204 315.0406028_0.927833333 2048.740296 2206.410626 6032.333545 146.0459054_0.927833333 885.1147207 917.8332456 316.3129757 346.8768225_0.894633333 1815.758966 1579.61618 440.8592475 124.0083607_0.902933333 261.9829082 290.4823216 178.9500771 225.9878652_0.919533333 152.3557155 88.49009991 33.46453832 135.031384_1.367566667 244.0848789 266.0609455 15.35792382 237.0064876_1.367566667 131.8502414 128.0529063 189.0258562 400.9430015_1.367566667 80.08986195 55.3973856 202.8039081 187.0419351_1.367566667 538.830094 541.6695904 1388.143824 89.02470786_1.367566667 224.8996863 213.1537496 427.4292348 178.920231_1.367566667 338.4977588 318.2621652 674.0129052 186.045487_1.367566667 88.85294839 118.1264243 317.7018552 441.1139865_3.39205 557.8363528 574.9351713 546.0774094 482.0120373_3.39205 304.2538891 371.3081572 413.2508441 363.1722382_3.39205 143.2210579 190.8499143 147.9306156 114.0260437_1.375866667 40.27027887 44.37094046 107.10304 170.0916122_3.39205 5047.493306 5141.941106 4865.826319 431.0853914_3.39205 1261.993721 1193.175147 1385.02007 232.0123074_0.952716667 915.0323472 853.1343133 368.602419 130.0173513_0.952716667 271.1314197 249.8465622 118.1825625 246.02688_0.952716667 409.9921381 311.4540906 162.4610923 233.008737_0.952716667 1512.246576 1370.874935 606.4311615 248.0237148_0.952716667 1556.841404 1488.285244 693.075849 274.0210645_0.952716667 873.8666534 839.8307275 389.7845772 197.8825131_0.952716667 0 0 232.7433043 226.993515_1.110366667 568.9949311 580.0317966 218.2565232 347.0393316_1.185033333 1310.991133 1270.634231 37.50590128 346.0547803_1.2929 1223.109686 1107.325229 355.9744265 558.0635203_0.9942 588.280334 582.6259494 109.7723703 114.056106_1.027383333 194.251182 152.8318773 12.70109283 323.0283525_1.043983333 422.9841912 415.8731424 78.2717081 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name moverz_quant ri ri_type pubchem_id inchi_key kegg_id other_id other_id_type 1-(2E,6E-phytadienyl)-2-(2E,6E-phytadienyl)-sn-glycero-3-phosphocholine 844.6341267 13.697 Mayo_RI 74380415 MAYO_ID Cyclopassifloside VII 751.3672973 12.07908333 Mayo_RI HMDB36300 MAYO_ID Dihydrozeatin-9-N-glucoside-O-glucoside 566.207692 3.39205 Mayo_RI HMDB12212 MAYO_ID Evasterioside E 753.382268 11.85506667 Mayo_RI 123069331 MAYO_ID Gingerglycolipid B 713.3509372 11.76378333 Mayo_RI HMDB41094 MAYO_ID Megalomicin B 953.5372987 15.86253333 Mayo_RI 74380507 MAYO_ID Neocasomorphin 697.3557538 12.0293 Mayo_RI HMDB60164 MAYO_ID PC(16:0/5:0(COOH)) 608.3550871 11.7389 Mayo_RI 135642572 MAYO_ID Phenformin 242.0821654 10.13756667 Mayo_RI HMDB15050 MAYO_ID PS(20:3(8Z,11Z,14Z)/0:0) 568.2638888 10.85111667 Mayo_RI 123063849 MAYO_ID PS(20:3(8Z,11Z,14Z)/0:0)_2 528.2724244 11.33235 Mayo_RI 123063849 MAYO_ID PS(21:0/0:0) 588.3290566 10.9092 Mayo_RI 123063853 MAYO_ID PS(21:0/0:0)_2 566.3437098 11.62273333 Mayo_RI 123063853 MAYO_ID PS(22:1(11Z)/0:0) 578.3449667 12.73455 Mayo_RI 123063850 MAYO_ID Vignatic acid A 552.2730195 11.25766667 Mayo_RI HMDB33599 MAYO_ID 197.060476_8.162866667 197.060476 8.162866667 169.0884532_3.39205 169.0884532 3.39205 528.3080347_11.63933333 528.3080347 11.63933333 164.0719899_3.39205 164.0719899 3.39205 540.3293522_11.37383333 540.3293522 11.37383333 478.2915402_11.55636667 478.2915402 11.55636667 524.2766613_10.80133333 524.2766613 10.80133333 476.2763965_10.84281667 476.2763965 10.84281667 208.0976367_10.41966667 208.0976367 10.41966667 672.1366554_10.39478333 672.1366554 10.39478333 500.2762336_10.85111667 500.2762336 10.85111667 210.0781561_9.689533333 210.0781561 9.689533333 526.2922932_11.08343333 526.2922932 11.08343333 619.2876334_10.00481667 619.2876334 10.00481667 564.3291788_10.9009 564.3291788 10.9009 599.3181467_11.64763333 599.3181467 11.64763333 791.5826922_13.7053 791.5826922 13.7053 877.6634991_13.7053 877.6634991 13.7053 808.5741681_13.7053 808.5741681 13.7053 671.266041_12.83411667 671.266041 12.83411667 441.2029728_13.1328 441.2029728 13.1328 891.5784301_13.7053 891.5784301 13.7053 122.0345828_18.57571667 122.0345828 18.57571667 826.56188_19.38881667 826.56188 19.38881667 885.5492237_15.87913333 885.5492237 15.87913333 859.5707156_13.7053 859.5707156 13.7053 906.6184419_15.55555 906.6184419 15.55555 642.3609948_11.90483333 642.3609948 11.90483333 720.4086886_11.97121667 720.4086886 11.97121667 710.3860773_11.89655 710.3860773 11.89655 685.3575101_11.77208333 685.3575101 11.77208333 694.392325_11.81356667 694.392325 11.81356667 638.3659774_12.06248333 638.3659774 12.06248333 698.2860766_12.83411667 698.2860766 12.83411667 681.2951164_12.83411667 681.2951164 12.83411667 694.4286295_12.66816667 694.4286295 12.66816667 706.3565028_12.06248333 706.3565028 12.06248333 664.385352_12.21183333 664.385352 12.21183333 153.0657071_3.39205 153.0657071 3.39205 263.8533923_0.936116667 263.8533923 0.936116667 265.8536298_0.944416667 265.8536298 0.944416667 197.8077571_0.944416667 197.8077571 0.944416667 372.9995855_0.927833333 372.9995855 0.927833333 102.0562098_0.927833333 102.0562098 0.927833333 313.064623_0.936116667 313.064623 0.936116667 195.8107828_0.944416667 195.8107828 0.944416667 209.8340127_0.944416667 209.8340127 0.944416667 202.019732_0.944416667 202.019732 0.944416667 159.0088498_0.952716667 159.0088498 0.952716667 199.8047957_0.944416667 199.8047957 0.944416667 343.9941745_0.944416667 343.9941745 0.944416667 291.0824524_0.944416667 291.0824524 0.944416667 325.8342982_0.819966667 325.8342982 0.819966667 180.9009543_0.828266667 180.9009543 0.828266667 232.9257215_0.894633333 232.9257215 0.894633333 138.965913_0.795066667 138.965913 0.795066667 206.9725179_0.819966667 206.9725179 0.819966667 162.9830515_0.819966667 162.9830515 0.819966667 172.9559813_0.894633333 172.9559813 0.894633333 162.9078616_0.919533333 162.9078616 0.919533333 315.0406028_0.927833333 315.0406028 0.927833333 146.0459054_0.927833333 146.0459054 0.927833333 346.8768225_0.894633333 346.8768225 0.894633333 124.0083607_0.902933333 124.0083607 0.902933333 225.9878652_0.919533333 225.9878652 0.919533333 135.031384_1.367566667 135.031384 1.367566667 237.0064876_1.367566667 237.0064876 1.367566667 400.9430015_1.367566667 400.9430015 1.367566667 187.0419351_1.367566667 187.0419351 1.367566667 89.02470786_1.367566667 89.02470786 1.367566667 178.920231_1.367566667 178.920231 1.367566667 186.045487_1.367566667 186.045487 1.367566667 441.1139865_3.39205 441.1139865 3.39205 482.0120373_3.39205 482.0120373 3.39205 363.1722382_3.39205 363.1722382 3.39205 114.0260437_1.375866667 114.0260437 1.375866667 170.0916122_3.39205 170.0916122 3.39205 431.0853914_3.39205 431.0853914 3.39205 232.0123074_0.952716667 232.0123074 0.952716667 130.0173513_0.952716667 130.0173513 0.952716667 246.02688_0.952716667 246.02688 0.952716667 233.008737_0.952716667 233.008737 0.952716667 248.0237148_0.952716667 248.0237148 0.952716667 274.0210645_0.952716667 274.0210645 0.952716667 197.8825131_0.952716667 197.8825131 0.952716667 226.993515_1.110366667 226.993515 1.110366667 347.0393316_1.185033333 347.0393316 1.185033333 346.0547803_1.2929 346.0547803 1.2929 558.0635203_0.9942 558.0635203 0.9942 114.056106_1.027383333 114.056106 1.027383333 323.0283525_1.043983333 323.0283525 1.043983333 1-(2E_6E-phytadienyl)-2-(2E_6E-phytadienyl)-sn-glycero-3-phosphocholine PS(20:3(8Z_11Z_14Z)/0:0) PS(20:3(8Z_11Z_14Z)/0:0)_1 PS(21:0/0:0)_1 METABOLITES_END #END