#METABOLOMICS WORKBENCH tcavallo_20161230_102843 DATATRACK_ID:811 STUDY_ID:ST000532 ANALYSIS_ID:AN000811 PROJECT_ID:PR000391 VERSION 1 CREATED_ON January 24, 2017, 1:01 pm #PROJECT PR:PROJECT_TITLE CHEAR Plasma Reference Material ProficiencyTest GC-MS PR:PROJECT_TYPE CHEAR-UT-1 PR:PROJECT_SUMMARY CHEAR Reference Material Plasma was provided by Emory University. The material PR:PROJECT_SUMMARY was prepared and analyzed using the GC-TOF MS workflow of the Eastern Regional PR:PROJECT_SUMMARY Metabolomics Resource Core (protocols available in metabolomics workbench). PR:PROJECT_SUMMARY Seven samples were injected of the sample reference material prepared in PR:PROJECT_SUMMARY replicate. PR:INSTITUTE RTI International PR:DEPARTMENT Discovery-Sciences-Technology (DST) PR:LABORATORY RTI CHEAR Analytical Hub - Untargeted Analysis Resource Core (UARC) PR:LAST_NAME Fennell PR:FIRST_NAME Timothy PR:ADDRESS 3040 E Cornwallis Road, Research Triangle Park, NC 27709 PR:EMAIL fennell@rti.org PR:PHONE 919-485-2781 #STUDY ST:STUDY_TITLE CHEAR Plasma Reference Material Plasma GC-MS ST:STUDY_TYPE Metabolomics ST:STUDY_SUMMARY CHEAR Reference Material Plasma was provided by Emory University. The material ST:STUDY_SUMMARY was prepared and analyzed using the GC-TOF MS workflow of the Eastern Regional ST:STUDY_SUMMARY Metabolomics Resource Core (protocols available in metabolomics workbench). ST:STUDY_SUMMARY Seven samples were injected of the sample reference material prepared in ST:STUDY_SUMMARY replicate. ST:INSTITUTE RTI International ST:DEPARTMENT Discovery-Sciences-Technology (DST) ST:LABORATORY RTI CHEAR Analytical Hub - Untargeted Analysis Resource Core ST:LAST_NAME Fennell ST:FIRST_NAME Timothy ST:ADDRESS 3040 E Cornwallis Road, Research Triangle Park, NC 27709 ST:EMAIL fennell@rti.org ST:PHONE 919-485-2781 #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 - CHE000054 0202a SAMPLE TYPE:Replicate CHEAR ID=CHE000054; Aliquot ID=202a; BINBASE ID=161012atbcsa07; SPECIES=human; MATRIX=PLASMA SUBJECT_SAMPLE_FACTORS - CHE000054 0203a SAMPLE TYPE:Replicate CHEAR ID=CHE000054; Aliquot ID=203a; BINBASE ID=161012atbcsa08; SPECIES=human; MATRIX=PLASMA SUBJECT_SAMPLE_FACTORS - CHE000054 0204a SAMPLE TYPE:Replicate CHEAR ID=CHE000054; Aliquot ID=204a; BINBASE ID=161012atbcsa09; SPECIES=human; MATRIX=PLASMA SUBJECT_SAMPLE_FACTORS - CHE000054 0205a SAMPLE TYPE:Replicate CHEAR ID=CHE000054; Aliquot ID=205a; BINBASE ID=161012atbcsa10; SPECIES=human; MATRIX=PLASMA SUBJECT_SAMPLE_FACTORS - CHE000054 0206a SAMPLE TYPE:Replicate CHEAR ID=CHE000054; Aliquot ID=206a; BINBASE ID=161012atbcsa11; SPECIES=human; MATRIX=PLASMA SUBJECT_SAMPLE_FACTORS - CHE000054 0207a SAMPLE TYPE:Replicate CHEAR ID=CHE000054; Aliquot ID=207a; BINBASE ID=161012atbcsa12; SPECIES=human; MATRIX=PLASMA SUBJECT_SAMPLE_FACTORS - CHE000054 0208a SAMPLE TYPE:Replicate CHEAR ID=CHE000054; Aliquot ID=208a; BINBASE ID=161012atbcsa13; SPECIES=human; MATRIX=PLASMA #COLLECTION CO:COLLECTION_SUMMARY CHEAR Reference Material Plasma was provided by Emory University. #TREATMENT TR:TREATMENT_SUMMARY No treatment. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY liquots of 30µL of plasma were mixed with 1000µL of (3:3:2) ACN:IPA:H2O by SP:SAMPLEPREP_SUMMARY vortex mixer then centrifuged. Supernatant recovered. Archived 450µL of SP:SAMPLEPREP_SUMMARY supernatant. Continued prep of remaining 450µL supernatant. Evaporated to SP:SAMPLEPREP_SUMMARY dryness w/speed vac. Samples resuspended in 450µL of 1:1 ACN:H2O. Evaporated to SP:SAMPLEPREP_SUMMARY dryness w/speed vac. Formation of methoximes by adding 10µL of 40mg/mL MeOx in SP:SAMPLEPREP_SUMMARY pyridine to each sample. Placed onto Thermomixer for 90 min at 30⁰C. Then FAME SP:SAMPLEPREP_SUMMARY retention index markers and MSTFA added to each sample for derivatization step. SP:SAMPLEPREP_SUMMARY Samples returned to Thermomixer for 45 min at 70⁰C. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Leco Pegasus 4D GC CH:COLUMN_NAME Restek Rtx-5Sil MS (30 x 0.25mm, 0.25um) CH:FLOW_GRADIENT Constant Flow CH:FLOW_RATE 1mL/min CH:COLUMN_PRESSURE 10.4 psi CH:INJECTION_TEMPERATURE 250 degrees C CH:RETENTION_INDEX FAME retention index markers C8-C30 CH:SAMPLE_INJECTION 0.5uL CH:ANALYTICAL_TIME 20 minutes CH:TRANSFERLINE_TEMPERATURE 280 degrees C #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Leco Pegasus 4D GCxGC TOF MS:INSTRUMENT_TYPE GC x GC-TOF MS:MS_TYPE EI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS intensity MS_METABOLITE_DATA_START Samples CHE000054 0206a CHE000054 0205a CHE000054 0208a CHE000054 0207a CHE000054 0202a CHE000054 0203a CHE000054 0204a Factors SAMPLE TYPE:Replicate SAMPLE TYPE:Replicate SAMPLE TYPE:Replicate SAMPLE TYPE:Replicate SAMPLE TYPE:Replicate SAMPLE TYPE:Replicate SAMPLE TYPE:Replicate 1,5-anhydroglucitol 95943 106819 128037 91880 89315 115481 113011 1-monopalmitin 1149 969 1218 765 879 1093 1150 1-monostearin 775 718 845 577 596 731 750 2-hydroxybutanoic acid 86104 71415 81628 58359 61420 76076 83160 3-(4-hydroxyphenyl)propionic acid 1893 2370 2065 1897 1635 1777 1647 3,4-dihydroxycinnamic acid 112 115 131 66 123 69 124 4-hydroxypyridine 10314 9009 10755 6913 8382 8804 9694 5-aminovaleric acid lactame 1224 1862 2361 1509 1851 2537 2680 alanine 913993 1108382 1312082 919770 934814 1181244 1295958 alpha-ketoglutarate 4554 6252 7530 5416 5323 6898 6205 arabitol 1471 1323 1069 796 1153 811 1211 arachidiic acid 24328 22962 29116 18667 22560 23102 25879 arachidonic acid 1564 1386 2072 1665 1680 1915 2105 asparagine dehydrated 1739 4130 3334 2837 2897 2297 2722 behenic acid 1092 1094 732 929 1049 1136 1203 benzoic acid 102771 94923 109969 76141 83617 103014 109037 beta alanine 1494 2476 3004 1986 2075 2937 2646 beta-hydroxybutyric acid 198166 162801 175270 133079 130704 190322 206656 C08 FAME ISTD 670243 518543 543189 426416 492571 542657 615521 C09 FAME ISTD 641560 553230 599344 451928 514527 582181 627942 C10 FAME ISTD 632771 645770 696433 565077 609611 675506 676494 C12 FAME ISTD 476495 580399 643802 459929 546824 660901 639279 C14 FAME ISTD 402363 432847 470282 371335 422768 462441 459131 C16 FAME ISTD 549135 536048 593403 443881 511789 572774 577540 C18 FAME ISTD 196152 172794 196762 150454 167454 184769 190402 C20 FAME ISTD 192736 173487 194482 150719 170767 183048 187390 C22 FAME ISTD 169385 145332 158596 125627 143657 155954 154612 C24 FAME ISTD 138792 120599 132015 107958 120588 134043 128791 C26 FAME ISTD 104873 97209 100233 81132 92576 105092 95884 C28 FAME ISTD 83865 79919 81866 66778 78757 86961 75961 C30 FAME ISTD 50402 50974 50124 40645 48633 56068 48479 capric acid 1758 2176 2318 1576 2128 2552 2121 caprylic acid 5962 4856 6258 4206 5021 5608 5372 citric acid 1392895 1560030 1823976 1361979 1316162 1656863 1636420 creatine major dehydrated TMS3 28919 47389 54082 26518 30745 44395 45082 cyanoalanine 146 163 134 91 138 63 154 cyclohexylamine 1tms 2231 3748 2987 3465 3359 3031 2990 cysteine 3132 4543 5592 3959 4235 4983 4818 cystine 8160 8488 11737 7596 7784 9127 9124 enolpyruvate nist 4767 3844 4378 3078 3273 4295 4277 fructose 94769 99019 121753 86573 84634 108365 107091 fructose 2 60010 59809 71042 56466 55429 66387 66780 gluconic acid 670 725 861 614 661 811 809 glucose 1034983 1220184 1244169 1218788 763729 1233787 741347 glucose 2 728057 743028 769952 665858 648025 784255 790991 glucose-6-phosphate minor 27 31 32 60 32 25 22 glutamine 3TMS major 173610 188384 227459 161326 167896 202142 201623 glutamine dehydrated 2028 3474 4126 2832 2953 3853 3226 glutaric acid 267 437 393 279 95 421 275 glyceric acid 8777 10157 11821 8223 8793 11445 10435 glycerol 150898 146747 163642 117856 132189 158732 149596 glycine 621689 612173 763880 518767 530515 685646 698555 glycolic acid 5827 4682 4897 3819 3876 4831 5103 heptadecanoic acid 11169 10732 13346 6253 10729 11207 11754 hexitol 680 649 813 559 571 766 698 hexose pertms nist 18725 196477 48407 31059 68206 126135 48830 histidine 2 75028 82251 99321 71237 71547 87466 88659 hydroxylamine 6747 5036 5834 4950 4847 5274 5867 isoleucine 142618 167821 202343 139714 141689 180126 185463 isothreonic acid 2 2193 2888 3354 2321 2455 3263 3053 itaconic acid 6463 8734 9912 8273 8544 9423 8178 lactic acid 4889466 4849083 5153098 4306216 4368688 5020300 5084584 lauric acid 1969 2076 2095 1719 2084 2352 1743 leucine 318862 371293 471751 294768 300716 399388 418409 linoleic acid 8799 8932 10155 7647 7896 9428 9715 lysine 165049 170543 212498 146112 147015 185510 189238 maleimide 21748 23041 18661 15001 23798 17325 13422 malic acid 322 1483 1871 1271 1389 1823 1695 mannose 22707 24032 28222 21044 20297 24878 25378 methionine (likely) 18909 27311 30773 22756 23273 29249 27474 methylmaleic acid 184 542 620 583 146 526 493 methyltetrahydrophenanthrenone2 nist 4613 38276 40084 32811 58982 53357 38662 myo-inositol 43469 43436 53322 37493 37699 47479 49419 myristic acid 3806 4244 5040 3650 3894 4630 4412 N-acetyl-L-aspartic acid 1 146 129 123 117 176 136 120 N-acetyl-L-aspartic acid TMS3 228 236 294 233 229 294 258 n-methylalanine 40246 43571 52442 37176 37460 47000 51924 octadecanol 872 868 942 830 1159 1089 912 oleic acid 5987 7331 12885 7060 7349 5831 13835 oxalic acid 204042 145885 159362 130874 150418 191796 138608 oxoproline 177257 226670 267266 186280 193227 239492 212102 palmitic acid 558072 497709 280741 481881 482316 510499 512535 palmitoleic acid 8057 8924 12460 7974 8435 6989 10158 pelargonic acid 10547 12665 15208 9239 12292 12847 12999 phenylalanine 43143 58492 62569 47233 45514 57392 52625 phosphate 1930794 1896506 2147755 1722829 1608746 1928547 1925409 phthalate ester 70437 62710 100006 61869 88937 95209 66096 phthalic acid 18154 22760 26486 18888 19662 23495 23690 phthalic acid mono-2-ethylhexylester 137898 136091 167137 112897 115480 147720 152825 pipecolinic acid 5836 8237 9432 6660 7078 7238 8599 proline 507623 645491 772769 550085 544828 688643 700255 pseudo-uridine 2989 3035 3527 2353 2540 3445 3215 putrescine 221 172 258 119 272 242 204 ribitol (likely) 3087 3493 4047 2986 3015 3729 3765 salicylic acid 1923 2562 3073 2106 2073 2840 2759 serine 79062 118315 138858 90945 104043 125342 120776 serine minor 13685 2984 3937 4949 2030 2632 3994 stearic acid 2807621 2717142 3295336 2216516 2611267 2575436 2900733 succinic acid 47304 44171 54578 37003 41122 46013 46866 taurine 944 1673 2071 1242 1492 2463 1961 threonine 62059 89454 105473 68683 77428 95919 88900 tryptophan 120393 116044 143916 97460 106714 124641 130133 tyrosine 129207 137129 166385 118713 114058 144406 146225 urea 427496 473035 532939 436040 403333 494065 422370 uric acid 74624 90784 103886 72623 73023 91486 95167 valine 486332 518363 631325 429007 437027 559777 597034 dioctylphtalate 2499007 2215785 2614192 1765408 1767307 2317016 2393843 polysiloxane 381 374 871 327 267 458 94 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Retention Index Moverz Quant Local ID PubChem CID KEGG ID mass spectrum quantmass 1,5-anhydroglucitol 635618 217 1-monopalmitin 902507 371 1-monostearin 960911 399 2-hydroxybutanoic acid 259826 131 3-(4-hydroxyphenyl)propionic acid 598786 179 3,4-dihydroxycinnamic acid 748751 219 4-hydroxypyridine 282530 152 5-aminovaleric acid lactame 274276 156 alanine 244959 116 alpha-ketoglutarate 508594 198 arabitol 569134 217 arachidiic acid 856790 117 arachidonic acid 833385 150 asparagine dehydrated 476109 100 behenic acid 920744 117 benzoic acid 338781 179 beta alanine 437797 248 beta-hydroxybutyric acid 280289 147 C08 FAME ISTD 262320 87 C09 FAME ISTD 323120 87 C10 FAME ISTD 381020 87 C12 FAME ISTD 487220 87 C14 FAME ISTD 582620 87 C16 FAME ISTD 668720 87 C18 FAME ISTD 747420 87 C20 FAME ISTD 819620 87 C22 FAME ISTD 886620 87 C24 FAME ISTD 948820 87 C26 FAME ISTD 1006900 87 C28 FAME ISTD 1061700 87 C30 FAME ISTD 1113100 87 capric acid 452601 229 caprylic acid 344270 201 citric acid 619803 273 creatine major dehydrated TMS3 503755 115 cyanoalanine 404025 141 cyclohexylamine 1tms 227703 128 cysteine 500494 220 cystine 805427 218 enolpyruvate nist 235895 147 fructose 640102 217 fructose 2 645861 217 gluconic acid 694524 333 glucose 651462 319 glucose 2 659971 319 glucose-6-phosphate minor 818222 387 glutamine 3TMS major 601223 156 glutamine dehydrated 519659 227 glutaric acid 422187 158 glyceric acid 379286 189 glycerol 346514 205 glycine 370374 174 glycolic acid 229350 177 heptadecanoic acid 751505 117 hexitol 704774 318 hexose pertms nist 682302 191 histidine 2 664188 154 hydroxylamine 256159 146 isoleucine 360577 158 isothreonic acid 2 492628 292 itaconic acid 387493 147 lactic acid 219176 117 lauric acid 548658 257 leucine 347456 158 linoleic acid 777348 337 lysine 665216 156 maleimide 244991 154 malic acid 464809 233 mannose 646323 319 methionine (likely) 483951 176 methylmaleic acid 419346 184 methyltetrahydrophenanthrenone2 nist 591810 197 myo-inositol 730658 217 myristic acid 634958 285 N-acetyl-L-aspartic acid 1 548361 158 N-acetyl-L-aspartic acid TMS3 552267 274 n-methylalanine 288139 130 octadecanol 755925 327 oleic acid 778813 159 oxalic acid 260471 147 oxoproline 486729 156 palmitic acid 714747 313 palmitoleic acid 706886 129 pelargonic acid 399814 215 phenylalanine 536679 218 phosphate 347293 299 phthalate ester 644918 149 phthalic acid 567608 147 phthalic acid mono-2-ethylhexylester 741335 223 pipecolinic acid 403737 156 proline 365741 142 pseudo-uridine 815915 217 putrescine 588605 174 ribitol (likely) 575025 217 salicylic acid 481513 267 serine 396979 204 serine minor 339553 116 stearic acid 788356 117 succinic acid 371358 147 taurine 556874 326 threonine 411756 218 tryptophan 780426 202 tyrosine 672235 218 urea 329250 100 uric acid 731820 441 valine 315026 144 dioctylphtalate 891860 149 polysiloxane 772952 429 METABOLITES_END #END