#METABOLOMICS WORKBENCH yli_20221114_095317 DATATRACK_ID:3570 STUDY_ID:ST002351 ANALYSIS_ID:AN003837 VERSION 1 CREATED_ON 09-06-2023 #PROJECT PR:PROJECT_TITLE Characterizing the intrauterine environment via untargeted metabolomics PR:PROJECT_TITLE profiling of maternal blood collected during pregnancy (Atlanta ECHO Cohort) PR:PROJECT_TYPE C18 Reversed-Phase Broad Spectrum Metabolomics PR:PROJECT_SUMMARY This project is funded by the NIH Environmental influences on Child Health PR:PROJECT_SUMMARY Outcomes (ECHO) program, and uses data from five cohorts: Healthy Start, Project PR:PROJECT_SUMMARY Viva, Atlanta ECHO, Paneth Cohort, and PETALS. This project will generate new PR:PROJECT_SUMMARY untargeted metabolomics profiling data at two time-points in pregnancy for all PR:PROJECT_SUMMARY women in order to identify trimester-specific metabolomics signatures and PR:PROJECT_SUMMARY longitudinal metabolomics trajectories associated with offspring birth size; PR:PROJECT_SUMMARY identify the contribution of maternal sociodemographic and perinatal PR:PROJECT_SUMMARY characteristics to the metabolomics signatures. PR:INSTITUTE NC HHEAR Hub PR:DEPARTMENT Untargeted Analysis PR:LABORATORY Sumner Lab PR:LAST_NAME Li PR:FIRST_NAME Yuanyuan PR:ADDRESS 500 Laureate Way, Kannapolis, NC 28081 PR:EMAIL yuanyli4@unc.edu PR:PHONE 9843770693 PR:FUNDING_SOURCE This research was supported by the Environmental influences on Child Health PR:FUNDING_SOURCE Outcomes (ECHO) program, Office of The Director, National Institutes of Health. PR:FUNDING_SOURCE The content is solely the responsibility of the authors and does not necessarily PR:FUNDING_SOURCE represent the official views of the National Institutes of Health. The Atlanta PR:FUNDING_SOURCE ECHO Cohort is supported by the following ECHO Program Collaborators: ECHO PR:FUNDING_SOURCE Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: PR:FUNDING_SOURCE Smith PB, Newby KL, Benjamin DK; U2C OD023375; ECHO Data Analysis Center: Johns PR:FUNDING_SOURCE Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: PR:FUNDING_SOURCE Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier D; PR:FUNDING_SOURCE U24 OD023382; North Carolina Human Health Exposure Analysis Resource Hub: PR:FUNDING_SOURCE Research Triangle Institute: Fennell T, University of North Carolina at Chapel PR:FUNDING_SOURCE Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857; PR:FUNDING_SOURCE Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., PR:FUNDING_SOURCE Rockville, Maryland: O’Brien B; U24 ES026539 PR:DOI http://dx.doi.org/10.21228/M8HD93 #STUDY ST:STUDY_TITLE Characterizing the intrauterine environment via untargeted metabolomics ST:STUDY_TITLE profiling of maternal blood collected during pregnancy (Atlanta ECHO Cohort) ST:STUDY_TYPE Prospective Cohort Study ST:STUDY_SUMMARY The Atlanta ECHO Cohort is comprised of African American mother-child dyads who ST:STUDY_SUMMARY reside in the greater metropolitan Atlanta area. Enrollment begins during ST:STUDY_SUMMARY pregnancy from prenatal care clinics affiliated with two metro hospital systems. ST:STUDY_SUMMARY During pregnancy, data and biospecimens are obtained during an initial study ST:STUDY_SUMMARY visit (between 8-14 weeks gestation) and during a second study visit (between ST:STUDY_SUMMARY 24-30 weeks gestation). In the perinatal period, we collect the residual newborn ST:STUDY_SUMMARY blood spot (obtained from the neonate during the delivery hospitalization for ST:STUDY_SUMMARY state metabolic screening). Details of pregnancy complications, birth, and ST:STUDY_SUMMARY neonatal outcomes are ascertained via medical record abstraction. Children are ST:STUDY_SUMMARY then followed annually via in-person clinical assessments and maternal ST:STUDY_SUMMARY questionnaire data. This research was supported by the Environmental influences ST:STUDY_SUMMARY on Child Health Outcomes (ECHO) program, Office of The Director, National ST:STUDY_SUMMARY Institutes of Health. The content is solely the responsibility of the authors ST:STUDY_SUMMARY and does not necessarily represent the official views of the National Institutes ST:STUDY_SUMMARY of Health. The Atlanta ECHO Cohort is supported by the following ECHO Program ST:STUDY_SUMMARY Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, ST:STUDY_SUMMARY Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data ST:STUDY_SUMMARY Analysis Center: Johns Hopkins University Bloomberg School of Public Health, ST:STUDY_SUMMARY Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North ST:STUDY_SUMMARY Carolina: Catellier D; U24 OD023382 North Carolina Human Health Exposure ST:STUDY_SUMMARY Analysis Resource Hub: Research Triangle Institute: Fennell T, University of ST:STUDY_SUMMARY North Carolina at Chapel Hill: Sumner S, University of North Carolina at ST:STUDY_SUMMARY Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource ST:STUDY_SUMMARY Coordinating Center: Westat, Inc., Rockville, Maryland: O’Brien B; U24 ST:STUDY_SUMMARY ES026539 ST:INSTITUTE Emory University ST:DEPARTMENT School of Medicine, Department of Gynecology & Obstetrics; School of Public ST:DEPARTMENT Health, Department of Environmental Health ST:LAST_NAME Dunlop ST:FIRST_NAME Anne ST:ADDRESS 101 Woodruff Circle, Rm 4303 Woodruff Memorial Building, Atlanta, GA 30322 ST:EMAIL amlang@emory.edu ST:PHONE (404) 712-8520 ST:SUBMIT_DATE 2022-11-14 #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 N/A PZDu_HHEAR_1 Sample Type:HHEAR Pool RAW_FILE_NAME=PZDu_HHEAR_1 SUBJECT_SAMPLE_FACTORS N/A PZDu_HHEAR_10 Sample Type:HHEAR Pool RAW_FILE_NAME=PZDu_HHEAR_10 SUBJECT_SAMPLE_FACTORS N/A PZDu_HHEAR_11 Sample Type:HHEAR Pool RAW_FILE_NAME=PZDu_HHEAR_11 SUBJECT_SAMPLE_FACTORS N/A PZDu_HHEAR_12 Sample Type:HHEAR Pool RAW_FILE_NAME=PZDu_HHEAR_12 SUBJECT_SAMPLE_FACTORS N/A PZDu_HHEAR_13 Sample Type:HHEAR Pool RAW_FILE_NAME=PZDu_HHEAR_13 SUBJECT_SAMPLE_FACTORS N/A 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EM-JNHU3-S-01-10901 PZDu_S_562 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_562 SUBJECT_SAMPLE_FACTORS EM-9KKX7-S-01-10901 PZDu_S_563 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_563 SUBJECT_SAMPLE_FACTORS EM-XHY56-S-01-10901 PZDu_S_564 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_564 SUBJECT_SAMPLE_FACTORS EM-DY9C0-S-01-10901 PZDu_S_565 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_565 SUBJECT_SAMPLE_FACTORS EM-9DE48-S-01-10901 PZDu_S_566 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_566 SUBJECT_SAMPLE_FACTORS EM-8VUE6-S-01-10901 PZDu_S_567 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_567 SUBJECT_SAMPLE_FACTORS EM-ALZC9-S-01-10901 PZDu_S_568 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_568 SUBJECT_SAMPLE_FACTORS EM-E4MR8-S-01-10901 PZDu_S_569 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_569 SUBJECT_SAMPLE_FACTORS EM-H5L56-S-01-10901 PZDu_S_57 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_57 SUBJECT_SAMPLE_FACTORS EM-P51P6-S-01-10901 PZDu_S_570 Sample Type:Study Sample 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Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_65 SUBJECT_SAMPLE_FACTORS EM-6QL53-S-01-10901 PZDu_S_66 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_66 SUBJECT_SAMPLE_FACTORS EM-G3NH9-S-01-10901 PZDu_S_67 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_67 SUBJECT_SAMPLE_FACTORS EM-R6201-S-01-10901 PZDu_S_68 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_68 SUBJECT_SAMPLE_FACTORS EM-QYTC0-S-01-10901 PZDu_S_69 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_69 SUBJECT_SAMPLE_FACTORS EM-3ZF29-S-01-10901 PZDu_S_7 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_7 SUBJECT_SAMPLE_FACTORS EM-CZMU9-S-01-10901 PZDu_S_70 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_70 SUBJECT_SAMPLE_FACTORS EM-5K4A5-S-01-10901 PZDu_S_71 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_71 SUBJECT_SAMPLE_FACTORS EM-VL308-S-01-10901 PZDu_S_72 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_72 SUBJECT_SAMPLE_FACTORS EM-K71T8-S-01-10901 PZDu_S_73 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_73 SUBJECT_SAMPLE_FACTORS EM-C1AN6-S-01-10901 PZDu_S_74 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_74 SUBJECT_SAMPLE_FACTORS EM-S0B98-S-01-10901 PZDu_S_75 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_75 SUBJECT_SAMPLE_FACTORS EM-HUM14-S-01-10901 PZDu_S_76 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_76 SUBJECT_SAMPLE_FACTORS EM-CMWB0-S-01-10901 PZDu_S_77 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_77 SUBJECT_SAMPLE_FACTORS EM-UBLP6-S-01-10901 PZDu_S_78 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_78 SUBJECT_SAMPLE_FACTORS EM-XKLH1-S-01-10901 PZDu_S_79 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_79 SUBJECT_SAMPLE_FACTORS EM-E9LA5-S-01-10901 PZDu_S_8 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_8 SUBJECT_SAMPLE_FACTORS EM-RSG71-S-01-10901 PZDu_S_80 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_80 SUBJECT_SAMPLE_FACTORS EM-ED9A9-S-01-10901 PZDu_S_81 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_81 SUBJECT_SAMPLE_FACTORS EM-D73T3-S-01-10901 PZDu_S_82 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_82 SUBJECT_SAMPLE_FACTORS EM-NJGM7-S-01-10901 PZDu_S_83 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_83 SUBJECT_SAMPLE_FACTORS EM-TQC78-S-01-10901 PZDu_S_84 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_84 SUBJECT_SAMPLE_FACTORS EM-8KTV4-S-01-10901 PZDu_S_85 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_85 SUBJECT_SAMPLE_FACTORS EM-Z1345-S-01-10901 PZDu_S_86 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_86 SUBJECT_SAMPLE_FACTORS EM-APZJ4-S-01-10901 PZDu_S_87 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_87 SUBJECT_SAMPLE_FACTORS EM-J17J8-S-01-10901 PZDu_S_88 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_88 SUBJECT_SAMPLE_FACTORS EM-ETW84-S-01-10901 PZDu_S_89 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_89 SUBJECT_SAMPLE_FACTORS EM-3CZQ2-S-01-10901 PZDu_S_9 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_9 SUBJECT_SAMPLE_FACTORS EM-KFFW8-S-01-10901 PZDu_S_90 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_90 SUBJECT_SAMPLE_FACTORS EM-45G28-S-01-10901 PZDu_S_91 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_91 SUBJECT_SAMPLE_FACTORS EM-323F2-S-01-10901 PZDu_S_92 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_92 SUBJECT_SAMPLE_FACTORS EM-TLF43-S-01-10901 PZDu_S_94 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_94 SUBJECT_SAMPLE_FACTORS EM-KNPW0-S-01-10901 PZDu_S_95 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_95 SUBJECT_SAMPLE_FACTORS EM-6PSQ7-S-01-10901 PZDu_S_96 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_96 SUBJECT_SAMPLE_FACTORS EM-74DH3-S-01-10901 PZDu_S_97 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_97 SUBJECT_SAMPLE_FACTORS EM-WX714-S-01-10901 PZDu_S_98 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_98 SUBJECT_SAMPLE_FACTORS EM-R60R9-S-01-10901 PZDu_S_99 Sample Type:Study Sample RAW_FILE_NAME=PZDu_S_99 #COLLECTION CO:COLLECTION_SUMMARY The Atlanta ECHO Cohort is comprised of African American mother-child dyads who CO:COLLECTION_SUMMARY reside in the greater metropolitan Atlanta area. Enrollment begins during CO:COLLECTION_SUMMARY pregnancy from prenatal care clinics affiliated with two metro hospital systems. CO:COLLECTION_SUMMARY During pregnancy, data and biospecimens are obtained during an initial study CO:COLLECTION_SUMMARY visit (between 8-14 weeks gestation) and during a second study visit (between CO:COLLECTION_SUMMARY 24-30 weeks gestation). In the perinatal period, we collect the residual newborn CO:COLLECTION_SUMMARY blood spot (obtained from the neonate during the delivery hospitalization for CO:COLLECTION_SUMMARY state metabolic screening). Details of pregnancy complications, birth, and CO:COLLECTION_SUMMARY neonatal outcomes are ascertained via medical record abstraction. Children are CO:COLLECTION_SUMMARY then followed annually via in-person clinical assessments and maternal CO:COLLECTION_SUMMARY questionnaire data. CO:SAMPLE_TYPE Blood (serum) CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY N/A #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Serum samples (100 µL per tube) were shipped from Emory University to the NC SP:SAMPLEPREP_SUMMARY HHEAR Hub on dry ice. The NC HHEAR Hub thawed and transferred 50 µL of the SP:SAMPLEPREP_SUMMARY study samples to a new set of tubes and used them for the analysis. An SP:SAMPLEPREP_SUMMARY additional 10 µL was taken from the original study sample and transferred to SP:SAMPLEPREP_SUMMARY another tube to make the total study pool for this project, and then distributed SP:SAMPLEPREP_SUMMARY with 50 µL per aliquot, used as Quality control study pools (QC study pools) SP:SAMPLEPREP_SUMMARY throughout the whole analysis. All sample aliquots (50 µL each) and QC study SP:SAMPLEPREP_SUMMARY pools (50 µL each) were stored at -80° C until the day for sample preparation. SP:SAMPLEPREP_SUMMARY HHEAR reference plasma (50 µL each) and NIST serum (909c) reference material SP:SAMPLEPREP_SUMMARY (50 µL each) were provided by the NC HHEAR Hub. The LC-MS grade water (50 µL) SP:SAMPLEPREP_SUMMARY was used as blank. Sample preparation was conducted by batch, and all samples SP:SAMPLEPREP_SUMMARY (except blank) were thawed at 4°C overnight before the preparation. Samples, SP:SAMPLEPREP_SUMMARY including study samples, study pool samples, HHEAR reference plasma, NIST SP:SAMPLEPREP_SUMMARY reference serum, and blanks, were mixed with 400 µL methanol containing 500 SP:SAMPLEPREP_SUMMARY ng/mL tryptophan-d5 as internal standard and vortexed by a multiple tube vortex SP:SAMPLEPREP_SUMMARY mixer for 2 min at 5000 rpm at room temperature. All samples were centrifuged at SP:SAMPLEPREP_SUMMARY 16,000 rcf for 10 min at 4°C. The supernatant (350 µL) was transferred into a SP:SAMPLEPREP_SUMMARY pre-labeled 2.0 mL Lo-bind Eppendorf tube, dried by a SpeedVac overnight, and SP:SAMPLEPREP_SUMMARY stored at -80° C. For immediate analysis, 100 µL of water-methanol solution SP:SAMPLEPREP_SUMMARY (95:5, v/v) was used to reconstitute the dried extracts. Samples were thoroughly SP:SAMPLEPREP_SUMMARY mixed on a multiple tube vortex mixer for 10 min at 5000 rpm at room temperature SP:SAMPLEPREP_SUMMARY and then centrifuged at 4°C for 10 min at 16,000 rcf. The supernatant was SP:SAMPLEPREP_SUMMARY transferred to pre-labeled autosampler vials for data acquisition by LC-MS. SP:PROCESSING_STORAGE_CONDITIONS On ice SP:EXTRACT_STORAGE Described in summary #CHROMATOGRAPHY CH:INSTRUMENT_NAME Thermo Scientific™ Vanquish™ UPHPLC CH:COLUMN_NAME Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,3um) CH:COLUMN_PRESSURE 6000-10000 psi CH:COLUMN_TEMPERATURE 50 CH:FLOW_GRADIENT Time(min) Flow Rate %A %B Curve 1. 0 0.4 99.0 1.0 5 2. 1.00 0.4 99.0 1.0 5 3. CH:FLOW_GRADIENT 16.00 0.4 1.0 99.0 5 4. 19.00 0.4 1.0 99.0 5 5. 19.50 0.4 99.0 1.0 5 6. 22.00 CH:FLOW_GRADIENT 0.4 99.0 1.0 5 CH:FLOW_RATE 0.4 ml/min CH:INJECTION_TEMPERATURE 8 CH:SOLVENT_A 100% water; 0.1% formic acid CH:SOLVENT_B 100% methanol; 0.1% formic acid CH:RANDOMIZATION_ORDER Randomized CH:CHROMATOGRAPHY_TYPE Reversed phase #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive HF-X Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:MS_COMMENTS Instrument: Thermo Q Exactive HFx Software: Xcalibur 4.1.31.9 for data MS:MS_COMMENTS acquisition; Progenesis QI 2.4 for data preprocessing MS:ION_MODE POSITIVE MS:CAPILLARY_TEMPERATURE 320 °C MS:CAPILLARY_VOLTAGE 3.5 KV MS:COLLISION_ENERGY 20-45, ramp MS:COLLISION_GAS N2 MS:DRY_GAS_FLOW 55 MS:DRY_GAS_TEMP 400°C MS:FRAGMENTATION_METHOD CID MS:DESOLVATION_GAS_FLOW 55 MS:MS_RESULTS_FILE ST002351_AN003837_Results.txt UNITS:Normalized intensity Has m/z:Yes Has RT:Yes RT units:Minutes #END