#METABOLOMICS WORKBENCH xin_hu_emory_20210505_122815_mwtab.txt DATATRACK_ID:2620 STUDY_ID:ST001797 ANALYSIS_ID:AN002919 PROJECT_ID:000000 VERSION 1 CREATED_ON May 20, 2021, 8:11 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 ST:STUDY_TYPE Untargeted MS anlaysis ST:STUDY_SUMMARY We tested XLE quantification of chemicals in a non-fortified reference material: ST:STUDY_SUMMARY SRM-1957. Results show that XLE detected 29 out of 32 chemicals with certified ST:STUDY_SUMMARY or estimated reference values in the ng/kg range in SRM-1957; 16 out of 29 ST:STUDY_SUMMARY chemicals were quantified at >65% of the reference levels. 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: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 - N1957_1 type:SRM1957 RAW_FILE_NAME=N1957_1.mzXML SUBJECT_SAMPLE_FACTORS - N1957_2 type:SRM1957 RAW_FILE_NAME=N1957_2.mzXML SUBJECT_SAMPLE_FACTORS - N1957_3 type:SRM1957 RAW_FILE_NAME=N1957_3.mzXML SUBJECT_SAMPLE_FACTORS - N1957_4 type:SRM1957 RAW_FILE_NAME=N1957_4.mzXML SUBJECT_SAMPLE_FACTORS - N1957_5 type:SRM1957 RAW_FILE_NAME=N1957_5.mzXML SUBJECT_SAMPLE_FACTORS - N1957_6 type:SRM1957 RAW_FILE_NAME=N1957_6.mzXML SUBJECT_SAMPLE_FACTORS - N1957_7 type:SRM1957 RAW_FILE_NAME=N1957_7.mzXML SUBJECT_SAMPLE_FACTORS - N1957_8 type:SRM1957 RAW_FILE_NAME=N1957_8.mzXML SUBJECT_SAMPLE_FACTORS - N1957_9 type:SRM1957 RAW_FILE_NAME=N1957_9.mzXML #COLLECTION CO:COLLECTION_SUMMARY SRM1957 are non-fortified standard reference material of human serum developed CO:COLLECTION_SUMMARY for analysis organic contaminants. Aliquots were purchased from National CO:COLLECTION_SUMMARY Institute 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 N1957_1 N1957_2 N1957_3 N1957_4 N1957_5 N1957_6 N1957_7 N1957_8 N1957_9 Factors type:SRM1957 type:SRM1957 type:SRM1957 type:SRM1957 type:SRM1957 type:SRM1957 type:SRM1957 type:SRM1957 type:SRM1957 Hexachlorobenzene 7549 10654 8877 5247 6306 6919 9358 10507 10504 p,p'-DDE 83460 44940 64270 98420 22830 69710 42910 38050 34710 PBDE-100 1221 748 0 0 863 907 1090 1326 606 PBDE-17 0 0 0 0 0 0 0 899 920 PBDE-28_33 631 0 0 0 0 748 0 0 0 PBDE-47 34858 23841 14716 25148 26312 28651 16014 16696 12028 PBDE-66 0 0 0 0 0 0 663 756 0 PBDE-85 0 0 0 0 0 1767 0 0 0 PBDE-99 1159 2273 0 2974 0 0 0 1147 0 PCB-118 756 0 0 0 0 0 4004 1944 853 PCB-138 770 1184 2565 864 1025 1331 3750 1633 918 PCB-146 0 0 0 0 0 0 0 654 0 PCB-153 10475 8814 6700 0 3741 4351 4646 3199 3757 PCB-170 0 0 0 0 788 0 0 796 0 PCB-18 0 0 0 0 0 0 3650 3386 6344 PCB-180 5977 2989 2025 1658 815 1215 3952 1853 882 PCB-183 0 0 0 0 0 0 0 2958 0 PCB-187 0 0 0 0 0 0 0 3375 0 PCB-194 0 0 0 0 0 0 0 754 0 PCB-196+203 0 0 0 0 0 0 4737 2316 3796 PCB-199 0 0 0 0 0 0 0 636 0 PCB-206 0 0 0 0 0 0 0 0 988 PCB-28 0 0 0 0 0 0 4451 3501 4691 PCB-66 0 0 0 0 1972 0 979 683 787 PCB-74 0 0 0 0 599 0 4614 2755 3356 PCB-99 0 0 0 0 0 0 955 2623 757 trans-Nonachlor 5243 1294 1158 1145 2308 3249 3548 7753 5046 β-HCH 2509 2037 1686 0 2513 1176 0 2480 598 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name mz time Hexachlorobenzene 283.8095932 370 p,p'-DDE 245.999634 625 PBDE-100 403.7863985 988 PBDE-17 245.9674656 649 PBDE-28_33 245.9678244 678 PBDE-47 325.8759129 853 PBDE-66 325.8754356 883 PBDE-85 403.787037 1080 PBDE-99 403.7857562 1032 PCB-118 325.8795652 670 PCB-138 359.8409576 741 PCB-146 359.8408106 692 PCB-153 359.8407527 701 PCB-170 393.8021046 886 PCB-18 186.0230 402 PCB-180 393.8017648 843 PCB-183 393.8020289 770 PCB-187 393.8022476 762 PCB-194 427.762329 987 PCB-196+203 429.7597024 907 PCB-199 357.8249969 897 PCB-206 463.7204993 1040 PCB-28 255.96056 447 PCB-66 291.9188337 554 PCB-74 291.9188948 546 PCB-99 325.8797781 590 trans-Nonachlor 408.7833357 593 β-HCH 180.9372369 367 METABOLITES_END #END