#METABOLOMICS WORKBENCH sdasari_20150618_9777721_mwtab.txt DATATRACK_ID:328 STUDY_ID:ST000221 ANALYSIS_ID:AN000330 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 HILIC CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME ethylene-bridged hybrid 2.1 150 mm, 1.7 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 HILIC-PESI (-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 MS:MS_COMMENTS m/z. The mass accuracy and mass resolution were 5 parts per million (ppm) and MS:MS_COMMENTS ppm, 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 HILEC #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 2560.1308 608.4826 36.9886 1-(2-Hydroxyphenylamino)-1-deoxy-beta-D-gentiobioside 1_2-carbamate 188.1796 2577.8287 1415.6908 1-O-alpha-D-glucopyranosyl-(2-hexadecanoyloxy)-eicosan-1-ol 12447.7015 5481.9067 2137.1980 27-nor-campestan-3beta_4beta_5alpha_6alpha_7beta_8beta_14alpha_15alpha_24-nonol 0.0000 739.0876 32.6015 3-iodo-2E-acrylic acid 49.3657 37.9381 26.2703 3^-N-Acetyl-4^-O-(10_12-octadecadienoyl)fusarochromanone 0.0000 2353.6865 117.7451 5-Ethynyl-5^-(1-propynyl)-2_2^-bithiophene 474.2662 307.2734 47.4384 Apraclonidine 2865.5773 2875.8259 15695.6452 Bradykinin hydroxyproline 2477.7353 323.9787 1416.0494 Brassilexin 9.1521 8.1304 3.1737 Chloramphenicol 402.1937 533.1110 2220.0918 Cyclolinopeptide F 1617.7522 554.9108 310.6887 Cysteineglutathione disulfide 622.4092 622.6780 1661.5568 Cytosine 1486.9224 137.3011 66.1993 Dehydrocarpaine I 40.8894 2291.1114 276.0107 Ecabet 810.8448 1565.8013 707.9398 Echinasteroside C 15-O-sulfate 458.1482 3706.0297 438.3469 Ethacrynic acid 3282.8364 3708.0279 12481.8477 Evasterioside C 221.6601 333.4275 833.1497 Feruloylcholine 107.0646 2.0506 232.9266 Glycerol triundecanoate 641.8820 673.1351 2338.7989 Guanabenz 763.2557 348.7630 112.5720 Guanidinosuccinic acid 147.3207 76.5797 76.4623 Hydroxyzine 52.1081 63.4048 98.9577 Kuwanon Z 45.3157 74.9777 364.8663 Levamisole 237.1806 931.8171 386.7346 Megalomicin B 23685.8791 10542.0416 6034.6156 Methylarsonite 5107.8816 6076.9523 11097.7629 Methylpyrazine 119.1868 5126.9447 387.6763 N-Decanoylglycine 111.3985 54.7095 665.6268 N-Glycoloylganglioside GM2 131.3683 34.0436 39.5215 N-Methylcalystegine B2 705.4161 816.8209 1557.8372 N-Nitrosothialdine 50.2272 32.8095 9.1100 N-Nonanoylglycine 202.4402 165.9612 346.9480 N-Undecanoylglycine 47.3163 30.5140 1026.2850 N-Undecylbenzenesulfonic acid 120143.6139 248859.3362 136235.6798 Octafluorocyclobutane 10.5781 6.8475 4.3410 Pantothenic acid 867.3596 1905.2045 342.7124 PA(P-20:0/21:0) 131.8896 19.6035 20.8831 Peonidin 4573.9553 4970.1683 20232.4751 PG(O-20:0/22:6(4Z_7Z_10Z_13Z_16Z_19Z)) 1789.3925 209.2230 469.8369 Pimozide 3.1482 966.2784 94.1948 PS(20:3(8Z_11Z_14Z)/0:0) 437.6932 305.7747 151.9467 PS(P-20:0/17:2(9Z_12Z)) 72738.7903 25046.6272 24616.1018 Pyrazinamide 1972.7043 2080.8768 414.6604 S-(2_5-Dimethyl-3-furanyl) 2-furancarbothioate 2962.7934 2938.4099 880.8374 S-Acetyl dihydroasparagusic acid 1299.4914 1356.6196 2898.0821 (±)-Sulfobutanedioic acid 60.8053 102.8604 334.4585 Taurine 6975.6309 3368.2420 562.2565 Terlipressin 2361.6650 1025.1264 216.8232 Vignatic acid A 640.8294 64.8943 107.3718 Vinaginsenoside R3 876.7019 176.0777 284.1168 Xanthosine 1532.0708 43.1328 0.0000 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 1-(2-Hydroxyphenylamino)-1-deoxy-beta-D-gentiobioside 1,2-carbamate 480.1114611 0.827 Mayo_RI HMDB37551 MAYO_ID 1-O-alpha-D-glucopyranosyl-(2-hexadecanoyloxy)-eicosan-1-ol 751.5464339 4.618816667 Mayo_RI 74380338 MAYO_ID 27-nor-campestan-3beta,4beta,5alpha,6alpha,7beta,8beta,14alpha,15alpha,24-nonol 497.3122625 0.934866667 Mayo_RI 123069005 MAYO_ID 3-iodo-2E-acrylic acid 178.9008032 3.440633333 Mayo_RI 135636028 MAYO_ID 3'-N-Acetyl-4'-O-(10,12-octadecadienoyl)fusarochromanone 617.3570086 1.316533333 Mayo_RI HMDB38566 MAYO_ID 5-Ethynyl-5'-(1-propynyl)-2,2'-bithiophene 208.9878063 5.440216667 Mayo_RI HMDB29735 MAYO_ID Apraclonidine 225.0096876 1.200366667 Mayo_RI HMDB15099 MAYO_ID Bradykinin hydroxyproline 1056.542056 5.324066667 Mayo_RI HMDB11728 MAYO_ID Brassilexin 173.0170487 1.9388 Mayo_RI HMDB39638 MAYO_ID Chloramphenicol 302.99395 1.167183333 Mayo_RI HMDB14589 MAYO_ID Cyclolinopeptide F 1104.467773 4.618816667 Mayo_RI HMDB36479 MAYO_ID Cysteineglutathione disulfide 463.0380319 1.142283333 Mayo_RI HMDB00656 MAYO_ID Cytosine 92.02550766 2.79345 Mayo_RI HMDB00630 MAYO_ID Dehydrocarpaine I 513.3091738 1.001233333 Mayo_RI HMDB30271 MAYO_ID Ecabet 401.0710189 0.98465 Mayo_RI HMDB15613 MAYO_ID Echinasteroside C 15-O-sulfate 749.3439874 1.034433333 Mayo_RI 123069339 MAYO_ID Ethacrynic acid 282.9945271 1.0925 Mayo_RI HMDB15039 MAYO_ID Evasterioside C 669.2955181 1.1174 Mayo_RI 123069329 MAYO_ID Feruloylcholine 298.187789 0.83895 Mayo_RI HMDB32800 MAYO_ID Glycerol triundecanoate 297.2425538 1.1174 Mayo_RI HMDB31089 MAYO_ID Guanabenz 250.9861991 2.785166667 Mayo_RI HMDB14767 MAYO_ID Guanidinosuccinic acid 196.0333306 5.257683333 Mayo_RI HMDB03157 MAYO_ID Hydroxyzine 355.1580143 1.366316667 Mayo_RI HMDB14697 MAYO_ID Kuwanon Z 629.1209025 1.1008 Mayo_RI HMDB33297 MAYO_ID Levamisole 203.0648751 1.158883333 Mayo_RI HMDB14986 MAYO_ID Megalomicin B 953.5372987 15.86253333 Mayo_RI 74380507 MAYO_ID Methylarsonite 144.9247767 3.66465 Mayo_RI HMDB12259 MAYO_ID Methylpyrazine 93.04574935 1.739683333 Mayo_RI HMDB33112 MAYO_ID N-Decanoylglycine 230.1750977 1.376433333 Mayo_RI HMDB13267 MAYO_ID N-Glycoloylganglioside GM2 778.5968936 6.095683333 Mayo_RI HMDB00770 MAYO_ID N-Methylcalystegine B2 210.0756116 1.150583333 Mayo_RI HMDB36604 MAYO_ID N-Nitrosothialdine 173.0212289 2.776866667 Mayo_RI HMDB31186 MAYO_ID N-Nonanoylglycine 214.1445967 1.4078 Mayo_RI HMDB13279 MAYO_ID N-Undecanoylglycine 242.176019 1.53225 Mayo_RI HMDB13286 MAYO_ID N-Undecylbenzenesulfonic acid 311.1678292 0.98465 Mayo_RI HMDB32549 MAYO_ID Octafluorocyclobutane 180.9700161 12.50101667 Mayo_RI HMDB31292 MAYO_ID Pantothenic acid 218.1034301 2.478166667 Mayo_RI HMDB00210 MAYO_ID PA(P-20:0/21:0) 779.5904126 6.162066667 Mayo_RI 123067316 MAYO_ID Peonidin 357.0130761 1.1008 Mayo_RI HMDB05797 MAYO_ID PG(O-20:0/22:6(4Z,7Z,10Z,13Z,16Z,19Z)) 871.5585259 5.564683333 Mayo_RI 123064904 MAYO_ID Pimozide 482.2025697 0.901666667 Mayo_RI HMDB15232 MAYO_ID PS(20:3(8Z,11Z,14Z)/0:0) 528.2724244 11.33235 Mayo_RI 123063849 MAYO_ID PS(P-20:0/17:2(9Z,12Z)) 766.5377656 4.6603 Mayo_RI 123063807 MAYO_ID Pyrazinamide 158.0126289 2.0882 Mayo_RI HMDB14483 MAYO_ID S-(2,5-Dimethyl-3-furanyl) 2-furancarbothioate 203.0168937 2.079916667 Mayo_RI HMDB39585 MAYO_ID S-Acetyl dihydroasparagusic acid 230.9559836 13.51326667 Mayo_RI HMDB35717 MAYO_ID (±)-Sulfobutanedioic acid 218.9575142 4.552433333 Mayo_RI HMDB40585 MAYO_ID Taurine 169.9860205 0.897033333 Mayo_RI HMDB00251 MAYO_ID Terlipressin 1225.487344 4.51095 Mayo_RI HMDB15569 MAYO_ID Vignatic acid A 552.2730195 11.25766667 Mayo_RI HMDB33599 MAYO_ID Vinaginsenoside R3 951.5252795 4.49435 Mayo_RI HMDB40493 MAYO_ID Xanthosine 283.0680545 2.992583333 Mayo_RI HMDB00299 MAYO_ID METABOLITES_END #END