#METABOLOMICS WORKBENCH xin_hu_emory_20210505_133124_mwtab.txt DATATRACK_ID:2622 STUDY_ID:ST001798 ANALYSIS_ID:AN002920 PROJECT_ID:000000 VERSION 1 CREATED_ON May 20, 2021, 8:49 pm #PROJECT PR:PROJECT_TYPE Targeted GC-MS quantitative analysis PR:PROJECT_SUMMARY Complementing the genome with an understanding of the human exposome is an PR:PROJECT_SUMMARY important challenge for contemporary science and technology. Tens of thousands PR:PROJECT_SUMMARY of chemicals are used in commerce, yet cost for targeted environmental chemical PR:PROJECT_SUMMARY analysis limits surveillance to a few hundred known hazards. To overcome PR:PROJECT_SUMMARY limitations which prevent scaling to thousands of chemicals, we developed a PR:PROJECT_SUMMARY single-step express liquid extraction (XLE), gas chromatography high-resolution PR:PROJECT_SUMMARY mass spectrometry (GC-HRMS) analysis and computational pipeline to PR:PROJECT_SUMMARY operationalize the human exposome. We show that the workflow supports PR:PROJECT_SUMMARY quantification of environmental chemicals in human plasma (200 µL) and tissue PR:PROJECT_SUMMARY (≤ 100 mg) samples. The method also provides high resolution, sensitivity and PR:PROJECT_SUMMARY selectivity for exposome epidemiology of mass spectral features without a priori PR:PROJECT_SUMMARY knowledge of chemical identity. The simplicity of the method can facilitate PR:PROJECT_SUMMARY harmonization of environmental biomonitoring between laboratories and enable PR:PROJECT_SUMMARY population level human exposome research with limited sample volume. PR:INSTITUTE Emory University PR:DEPARTMENT Medicine/Pulmonary PR:LABORATORY Dean Jones PR:FIRST_NAME Xin PR:ADDRESS Emory University Whitehead building (Rm 225), 615 Michael Street, Atlanta, PR:ADDRESS Georgia, 30322, USA PR:EMAIL xin.hu2@emory.edu PR:PHONE 4047275091 PR:FUNDING_SOURCE This study was supported by the NIEHS, U2C ES030163 (DPJ), U2C ES030859 (DIW) PR:FUNDING_SOURCE and P30 ES019776 (CJM), NIDDK RC2 DK118619 (KNL), NHLBI R01 HL086773 (DPJ), US PR:FUNDING_SOURCE Department of Defense W81XWH2010103 (DPJ), and the Chris M. Carlos and Catharine PR:FUNDING_SOURCE Nicole Jockisch Carlos Endowment Fund in Primary Sclerosing Cholangitis (PSC) PR:FUNDING_SOURCE (KNL). PR:PROJECT_TITLE A scalable workflow for the human exposome PR:LAST_NAME Hu PR:CONTRIBUTORS Xin Hu, Douglas I. Walker, Yongliang Liang, M. Ryan Smith, Michael L. Orr, Brian PR:CONTRIBUTORS D. Juran, Chunyu Ma, Karan Uppal, Michael Koval, Greg S. Martin, David C. PR:CONTRIBUTORS Neujahr, Carmen J. Marsit, Young-Mi Go, Kurt Pennell, Gary W. Miller, PR:CONTRIBUTORS Konstantinos N. Lazaridis, Dean P. Jones #STUDY ST:STUDY_TITLE SRM1957 validation using GC-XLE (2) ST:STUDY_SUMMARY We evaluated quantification using XLE by testing 68 different chemicals (PCB, ST:STUDY_SUMMARY PBDEs, chlorinated pesticides) in SRM-1958 using external calibration curves ST:STUDY_SUMMARY (0.05 to 2 ng/mL) and comparing measured values to the reference concentrations ST:STUDY_SUMMARY reported for SRM. We identified all 40 PCBs that are reported with a reference ST:STUDY_SUMMARY mass fraction (including certified values and non-certified estimates) in the ST:STUDY_SUMMARY range of 46.6 to 490 ng/kg in SRM-1958 certificate of analysis (issue date: 11 ST:STUDY_SUMMARY October 2018). Quantification without adjustment for recovery was reproducible ST:STUDY_SUMMARY with 29 PCB qualifications at >70% and 35 PCBs at >65% of the reference levels. ST:STUDY_SUMMARY Eleven out of 13 PBDE/PBBs and all 17 organochlorine pesticides were ST:STUDY_SUMMARY identifiable and reproducibly quantified in this experiment. Therefore, XLE ST:STUDY_SUMMARY provides sufficient recovery to support accurate absolute quantification of a ST:STUDY_SUMMARY broad range of environmental chemicals. Overall, XLE supported measurement of 68 ST:STUDY_SUMMARY out of the 70 chemicals that are in the ng/kg range in SRM-1958. ST:INSTITUTE Emory University ST:DEPARTMENT Medicine/Pulmonary ST:LABORATORY Dean Jones ST:LAST_NAME Hu ST:FIRST_NAME Xin ST:ADDRESS Emory University Whitehead building (Rm 225), 615 Michael Street ST:EMAIL xin.hu2@emory.edu ST:STUDY_TYPE Untargeted MS anlaysis ST:PHONE 4047275091 #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]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - N1958_1 type:SRM1958 RAW_FILE_NAME=N1958_1.mzXML SUBJECT_SAMPLE_FACTORS - N1958_2 type:SRM1958 RAW_FILE_NAME=N1958_2.mzXML SUBJECT_SAMPLE_FACTORS - N1958_3 type:SRM1958 RAW_FILE_NAME=N1958_3.mzXML SUBJECT_SAMPLE_FACTORS - N1958_4 type:SRM1958 RAW_FILE_NAME=N1958_4.mzXML SUBJECT_SAMPLE_FACTORS - N1958_5 type:SRM1958 RAW_FILE_NAME=N1958_5.mzXML SUBJECT_SAMPLE_FACTORS - N1958_6 type:SRM1958 RAW_FILE_NAME=N1958_6.mzXML SUBJECT_SAMPLE_FACTORS - N1958_7 type:SRM1958 RAW_FILE_NAME=N1958_7.mzXML SUBJECT_SAMPLE_FACTORS - N1958_8 type:SRM1958 RAW_FILE_NAME=N1958_8.mzXML SUBJECT_SAMPLE_FACTORS - N1958_9 type:SRM1958 RAW_FILE_NAME=N1958_9.mzXML #COLLECTION CO:COLLECTION_SUMMARY SRM1958 are fortified standard reference material of human serum developed for CO:COLLECTION_SUMMARY analysis organic contaminants. Aliquots were purchased from National Institute CO:COLLECTION_SUMMARY of Standards & Technology (NIST). CO:SAMPLE_TYPE Blood (serum) #TREATMENT TR:TREATMENT_SUMMARY 50 µL formic acid (Emprove® Essential DAC, Sigma-Aldrich) was added to 200 µL TR:TREATMENT_SUMMARY SRM aliquots and immediately followed by addition of 200 µL hexane – ethyl TR:TREATMENT_SUMMARY acetate (2:1 v/v, ≥99% pure, Sigma-Aldrich) containing the internal standards TR:TREATMENT_SUMMARY (final concentration: 1 ng/mL). The sample mixture was shaken vigorously on ice TR:TREATMENT_SUMMARY using multi-tube vortexer (VWR VX-2500) for 1 h and centrifuged at 1000 g, 4 °C TR:TREATMENT_SUMMARY for 10 min. The sample mixture was chilled during entire extraction procedure. TR:TREATMENT_SUMMARY The organic supernatant was transferred to a new tube with 25 mg MgSO4 TR:TREATMENT_SUMMARY (≥99.99% pure, Sigma-Aldrich) and vortexed vigorously to remove water. After TR:TREATMENT_SUMMARY 10 min centrifugation at 1000 g, 80 µL of the final supernatant was spiked with TR:TREATMENT_SUMMARY instrumental internal standards (final concentration: 1 ng/mL) for analysis. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Same as treatment #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Samples were analyzed with three injections using GC-HRMS with a Thermo CH:CHROMATOGRAPHY_SUMMARY Scientific Q Exactive GC hybrid quadrupole Orbitrap mass spectrometer with 2 µL CH:CHROMATOGRAPHY_SUMMARY per injection. A capillary DB-5MS column (15 m × 0.25 mm × 0.25 µm film CH:CHROMATOGRAPHY_SUMMARY thickness) was used with the following temperature program: hold 75 °C for 1 CH:CHROMATOGRAPHY_SUMMARY min, 25 °C/min to 180 °C, 6 °C/min to 250 °C, 20 °C/min to 350 °C and hold CH:CHROMATOGRAPHY_SUMMARY for 5 min. The flow rate of the helium carrier gas was 1 mL/min. Ion source and CH:CHROMATOGRAPHY_SUMMARY transfer line temperatures were 250°C and 280°C, respectively. Data were CH:CHROMATOGRAPHY_SUMMARY collected from 3 to 24.37 min with positive electron ionization (EI) mode (+70 CH:CHROMATOGRAPHY_SUMMARY eV), scanning from m/z 85.0000 to 850.0000 with a resolution of 60,000. CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Thermo Trace 1310 CH:COLUMN_NAME Agilent DB5-MS (15m x 0.25mm, 0.25um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE EI MS:ION_MODE POSITIVE MS:MS_COMMENTS Data were collected from 3 to 24.37 min with positive electron ionization (EI) MS:MS_COMMENTS mode (+70 eV), scanning from m/z 85.0000 to 850.0000 with a resolution of MS:MS_COMMENTS 60,000. Raw data were examined by checking signal-to-noise ratio, peak shape and MS:MS_COMMENTS spectral information for surrogate and internal standards using a 5 ppm m/z MS:MS_COMMENTS tolerance and 30 s retention time window in xCalibur Qualbrowser software. Data MS:MS_COMMENTS extraction was performed by XCMS to generate about 40,000 chemical features MS:MS_COMMENTS identified by spectral m/z and retention time. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS raw intensity MS_METABOLITE_DATA_START Samples N1958_1 N1958_2 N1958_3 N1958_4 N1958_5 N1958_6 N1958_7 N1958_8 N1958_9 Factors type:SRM1958 type:SRM1958 type:SRM1958 type:SRM1958 type:SRM1958 type:SRM1958 type:SRM1958 type:SRM1958 type:SRM1958 2,4'-DDD 88811 56590 35075 38218 19660 31623 23852 27328 32822 2,4'-DDE 127671 75990 45810 107158 64703 92661 75222 94010 78968 2,4'-DDT + 4,4'-DDD 15479 24506 12664 21020 32478 42528 47300 29390 25242 4,4'-DDE 191287 130782 149026 126541 128191 94669 155038 39516 84577 4,4'-DDT 8729 3330 2861 8344 6567 5302 4975 4314 5114 cis-Chlordane 18671 16853 16749 17320 16847 15482 18173 15767 19694 cis-Nonachlor 25695 14078 6112 13695 6177 12333 2297 11153 10512 Hexachlorobenzene 100319 99773 81618 74409 56963 69226 60612 64532 65151 Mirex 45160 32440 30600 33160 28900 25390 28290 30650 28240 Octachlorostyrene 11990 5268 4381 5690 7895 2149 7770 6229 7519 Oxychlordan 5920 6950 4177 3291 3547 3864 3170 1021 2548 PBB-153 11561 2538 5573 1642 1819 1122 3437 3393 1303 PBDE-100 30143 9526 11902 8517 17254 15381 14458 5981 14657 PBDE-153 30414 11186 12172 14943 12613 7114 4556 11455 13516 PBDE-154 27018 13913 12551 11560 9559 6247 11574 7411 5017 PBDE-17 40748 30041 25562 25963 30043 24885 21331 26604 17486 PBDE-183 1510 1165 2128 868 1258 1244 0 0 0 PBDE-28_33 44566 31396 10389 23890 27444 22578 25382 21616 23965 PBDE-47 41322 31880 43975 31082 21933 25791 18614 36909 29900 PBDE-66 19622 9730 6561 11208 6463 9623 8986 11142 7204 PBDE-85 19628 18996 12731 6159 8645 9761 14067 11508 15059 PBDE-99 28549 13646 18922 15303 7228 16257 11841 17210 15310 PCB-101 63963 53361 40000 38743 36159 31585 21960 38314 21425 PCB-105 79335 62774 52477 36433 38751 27813 32991 26697 22413 PCB-110 67885 41375 55604 47750 27172 27692 25539 31817 41333 PCB-114 3920 1960 1220 1325 0 687 0 447 423 PCB-118 91754 69751 61234 20675 20694 34258 30580 43668 34725 PCB-123 2099 3513 1002 1186 1426 1003 8473 553 441 PCB-128 45160 30900 28600 25310 18700 22890 24670 25880 21840 PCB-138+158 43267 77675 68116 33958 35533 49076 47740 37770 30812 PCB-146 35159 28819 30345 18794 22659 23530 10590 24048 22273 PCB-149 47909 35968 29202 25651 22144 23703 23477 18013 25739 PCB-151 42119 25808 22899 21123 19838 23260 22999 18244 17838 PCB-153 74714 43699 42394 40415 12756 41104 26024 20090 36624 PCB-156 70690 50064 29735 25939 27319 15807 37062 26226 34614 PCB-157 64880 48211 41238 43422 38457 32517 29738 26105 39404 PCB-167 47831 58219 38682 52479 32881 21284 35888 45624 27234 PCB-170 29418 22739 22472 23809 5560 19340 21125 18782 18310 PCB-172 28360 24680 18760 20260 21240 15480 17690 16690 17960 PCB-177 20760 18063 16701 17186 13975 12609 13128 13326 11403 PCB-178 29213 18745 15092 16854 13811 14322 16125 15766 14491 PCB-18 98824 92940 76466 72806 63290 66702 69496 62738 64853 PCB-180 40294 31567 19041 27903 22978 17898 22345 21436 22203 PCB-183 26238 15742 26782 26129 28368 4563 26319 13792 12358 PCB-187 35510 25197 22436 23984 18916 15713 19714 19994 16668 PCB-189 50414 37038 13847 30366 22425 27807 19807 17303 28541 PCB-194 31137 5788 3607 14653 13493 14949 8580 8232 12282 PCB-195 29486 13720 15244 15489 12973 13321 12390 12781 9312 PCB-196 25352 42871 39748 21103 30756 36935 26643 44364 37024 PCB-199 34126 16188 13781 15233 5563 13014 14165 14244 8015 PCB-206 29126 15524 14753 12434 5937 13236 8912 14591 8442 PCB-209 0 0 40840 22630 19660 27795 12004 9164 6477 PCB-28 114153 41345 88015 85940 78404 73475 78950 32187 72692 PCB-44 57064 40182 31570 38066 25520 25062 40547 37095 34533 PCB-49 60741 37603 45559 68873 21922 36019 41009 22542 33339 PCB-66 71878 101072 74794 68255 68142 59887 51750 46383 64133 PCB-74 83004 7767 80777 66180 46589 43546 48860 61286 38918 PCB-87 26569 20524 17296 18059 13490 10922 15053 12572 11452 PCB-99 60036 46468 38775 44701 26154 36570 31471 36600 33623 trans-Chlordane 26339 14854 18388 16252 2805 7747 9886 16067 7133 trans-Nonachlor 41899 36813 7616 10082 14798 17061 25415 17018 16743 α-BHC 20265 13510 15854 18134 15148 15226 13215 15470 14106 β-BHC 23619 23190 6065 16484 16457 17088 17133 17658 13139 γ-BHC 24305 19877 16721 23915 22214 20162 20500 17219 9144 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name mz time 2,4'-DDD 235.00766 621 2,4'-DDE 245.99993 567 2,4'-DDT + 4,4'-DDD 235.00768 672 4,4'-DDE 245.99997 614 4,4'-DDT 235.00767 725 cis-Chlordane 372.82566 560 cis-Nonachlor 408.78402 664 Hexachlorobenzene 283.80985 363 Mirex 271.80986 879 Octachlorostyrene 379.74413 522 Oxychlordan 114.99452 532 PBB-153 467.69946 1101 PBDE-100 403.78711 972 PBDE-153 483.69509 1135 PBDE-154 483.69525 1100 PBDE-17 245.96764 637 PBDE-183 561.6064 1217 PBDE-28_33 245.96769 663 PBDE-47 325.87603 838 PBDE-66 325.8616 872 PBDE-85 403.78678 1077 PBDE-99 403.78653 1018 PCB-101 325.88 573 PCB-105 325.87967 693 PCB-110 325.87998 618 PCB-114 325.87998 649 PCB-118 325.88009 657 PCB-123 325.88009 669 PCB-128 359.8409 763 PCB-138+158 359.84131 727 PCB-146 359.84106 679 PCB-149 359.84143 651 PCB-151 359.84124 636 PCB-153 359.84133 687 PCB-156 359.84084 804 PCB-157 359.84105 811 PCB-167 359.84082 772 PCB-170 393.80254 873 PCB-172 393.80256 817 PCB-177 393.8025 790 PCB-178 393.80212 737 PCB-18 186.02291 395 PCB-180 393.80247 829 PCB-183 393.80164 758 PCB-187 393.80207 748 PCB-189 393.80183 921 PCB-194 427.76332 973 PCB-195 429.76035 937 PCB-196 429.75964 894 PCB-199 357.82573 883 PCB-206 463.72094 1027 PCB-209 497.68295 1065 PCB-28 255.96097 438 PCB-44 219.98414 492 PCB-49 291.91916 473 PCB-66 291.91907 535 PCB-74 291.91903 544 PCB-87 253.94524 617 PCB-99 325.88001 578 trans-Chlordane 372.82561 578 trans-Nonachlor 408.78376 583 α-BHC 182.93435 360 β-BHC 182.93434 378 γ-BHC 182.93435 385 METABOLITES_END #END