#METABOLOMICS WORKBENCH schrodm_20230117_073337 DATATRACK_ID:3692 STUDY_ID:ST002443 ANALYSIS_ID:AN003980 PROJECT_ID:PR001574 VERSION 1 CREATED_ON January 17, 2023, 8:06 am #PROJECT PR:PROJECT_TITLE Characterizing the intrauterine environment via untargeted metabolomics PR:PROJECT_TITLE profiling of maternal blood collected during pregnancy (ARCH 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, ARCH 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 Nutrition Research Institute, UNC-CH, 500 Laureate Way, Kannapolis, NC 28081 PR:EMAIL yuanyli4@unc.edu PR:PHONE 984-377-0693 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 ARCH ECHO PR:FUNDING_SOURCE 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 #STUDY ST:STUDY_TITLE Characterizing the intrauterine environment via untargeted metabolomics ST:STUDY_TITLE profiling of maternal blood collected during pregnancy (ARCH Cohort) ST:STUDY_TYPE Prospective Cohort Study ST:STUDY_SUMMARY "The ARCH Cohort is a pregnancy cohort of approximately 1,000 women recruited at ST:STUDY_SUMMARY the first prenatal visit largely in Lansing, MI between 2008 and 2016. Blood was ST:STUDY_SUMMARY collected when possible at the onset of prenatal care and at the time of the ST:STUDY_SUMMARY glucose tolerance test (late 2nd, early 3rd trimester). Please contact Jean ST:STUDY_SUMMARY Kerver at kerverje@msu.edu for questions related to the subject characteristics ST:STUDY_SUMMARY and outcomes. This research was supported by the Environmental influences on ST:STUDY_SUMMARY Child Health Outcomes (ECHO) OIF 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. ARCH is an ECHO cohort which is supported by the following ECHO ST:STUDY_SUMMARY Program Collaborators: ECHO Coordinating Center: Duke Clinical Research ST:STUDY_SUMMARY Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ST:STUDY_SUMMARY ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public ST:STUDY_SUMMARY Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, ST:STUDY_SUMMARY North 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 Michigan State University ST:DEPARTMENT Department of Epidemiology and Biostatistics ST:LAST_NAME Kerver ST:FIRST_NAME Jean ST:ADDRESS 909 Wilson Road, East Lansing, MI, 48824 ST:EMAIL kerverje@msu.edu ST:PHONE 517-290-5062 ST:TOTAL_SUBJECTS 240 ST:STUDY_COMMENTS HHEAR Project EM19-0009, ECHO Project EC0374 #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 - PZPa_HHEAR_1 Sample Type:HHEAR Pool HHEAR-ECHO ID=N/A; 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RAW_FILE_NAME=PZPa_S_196 SUBJECT_SAMPLE_FACTORS - PZPa_S_197 Sample Type:Study Sample HHEAR-ECHO ID=EM-8PWW8-P-01-12902; RAW_FILE_NAME=PZPa_S_197 SUBJECT_SAMPLE_FACTORS - PZPa_S_198 Sample Type:Study Sample HHEAR-ECHO ID=EM-Q9DB9-P-01-12902; RAW_FILE_NAME=PZPa_S_198 SUBJECT_SAMPLE_FACTORS - PZPa_S_199 Sample Type:Study Sample HHEAR-ECHO ID=EM-AV6J7-P-01-12902; RAW_FILE_NAME=PZPa_S_199 SUBJECT_SAMPLE_FACTORS - PZPa_S_200 Sample Type:Study Sample HHEAR-ECHO ID=EM-W4QB1-P-01-12902; RAW_FILE_NAME=PZPa_S_200 SUBJECT_SAMPLE_FACTORS - PZPa_S_201 Sample Type:Study Sample HHEAR-ECHO ID=EM-X9JR1-P-01-12902; RAW_FILE_NAME=PZPa_S_201 SUBJECT_SAMPLE_FACTORS - PZPa_S_202 Sample Type:Study Sample HHEAR-ECHO ID=EM-JD3D3-P-01-12902; RAW_FILE_NAME=PZPa_S_202 SUBJECT_SAMPLE_FACTORS - PZPa_S_203 Sample Type:Study Sample HHEAR-ECHO ID=EM-FAHU6-P-01-12902; RAW_FILE_NAME=PZPa_S_203 SUBJECT_SAMPLE_FACTORS - PZPa_S_204 Sample Type:Study Sample HHEAR-ECHO ID=EM-PTWK1-P-01-12902; RAW_FILE_NAME=PZPa_S_204 SUBJECT_SAMPLE_FACTORS - PZPa_S_205 Sample Type:Study Sample HHEAR-ECHO ID=EM-PDGH9-P-01-12902; RAW_FILE_NAME=PZPa_S_205 SUBJECT_SAMPLE_FACTORS - PZPa_S_206 Sample Type:Study Sample HHEAR-ECHO ID=EM-WS8N0-P-01-12902; RAW_FILE_NAME=PZPa_S_206 SUBJECT_SAMPLE_FACTORS - PZPa_S_207 Sample Type:Study Sample HHEAR-ECHO ID=EM-KAXW1-P-01-12902; RAW_FILE_NAME=PZPa_S_207 SUBJECT_SAMPLE_FACTORS - PZPa_S_208 Sample Type:Study Sample HHEAR-ECHO ID=EM-9Z579-P-01-12902; RAW_FILE_NAME=PZPa_S_208 SUBJECT_SAMPLE_FACTORS - PZPa_S_209 Sample Type:Study Sample HHEAR-ECHO ID=EM-83S19-P-01-12902; RAW_FILE_NAME=PZPa_S_209 SUBJECT_SAMPLE_FACTORS - PZPa_S_210 Sample Type:Study Sample HHEAR-ECHO ID=EM-8RQ96-P-01-12902; RAW_FILE_NAME=PZPa_S_210 SUBJECT_SAMPLE_FACTORS - PZPa_S_211 Sample Type:Study Sample HHEAR-ECHO ID=EM-5D3M5-P-01-12902; RAW_FILE_NAME=PZPa_S_211 SUBJECT_SAMPLE_FACTORS - PZPa_S_212 Sample Type:Study Sample HHEAR-ECHO ID=EM-G3VD9-P-01-12902; RAW_FILE_NAME=PZPa_S_212 SUBJECT_SAMPLE_FACTORS - PZPa_S_213 Sample Type:Study Sample HHEAR-ECHO ID=EM-9NF78-P-01-12902; RAW_FILE_NAME=PZPa_S_213 SUBJECT_SAMPLE_FACTORS - PZPa_S_214 Sample Type:Study Sample HHEAR-ECHO ID=EM-96SY3-P-01-12902; RAW_FILE_NAME=PZPa_S_214 SUBJECT_SAMPLE_FACTORS - PZPa_S_215 Sample Type:Study Sample HHEAR-ECHO ID=EM-XYPJ1-P-01-12902; RAW_FILE_NAME=PZPa_S_215 SUBJECT_SAMPLE_FACTORS - PZPa_S_216 Sample Type:Study Sample HHEAR-ECHO ID=EM-WSRN4-P-01-12902; RAW_FILE_NAME=PZPa_S_216 SUBJECT_SAMPLE_FACTORS - PZPa_S_217 Sample Type:Study Sample HHEAR-ECHO ID=EM-SZDC2-P-01-12902; RAW_FILE_NAME=PZPa_S_217 SUBJECT_SAMPLE_FACTORS - PZPa_S_218 Sample Type:Study Sample HHEAR-ECHO ID=EM-PN2S3-P-01-12902; RAW_FILE_NAME=PZPa_S_218 SUBJECT_SAMPLE_FACTORS - PZPa_S_219 Sample Type:Study Sample HHEAR-ECHO ID=EM-KNTF3-P-01-12902; RAW_FILE_NAME=PZPa_S_219 SUBJECT_SAMPLE_FACTORS - PZPa_S_220 Sample Type:Study Sample HHEAR-ECHO ID=EM-6UNH1-P-01-12902; RAW_FILE_NAME=PZPa_S_220 SUBJECT_SAMPLE_FACTORS - PZPa_S_221 Sample Type:Study Sample HHEAR-ECHO ID=EM-70GW5-P-01-12902; RAW_FILE_NAME=PZPa_S_221 SUBJECT_SAMPLE_FACTORS - PZPa_S_222 Sample Type:Study Sample HHEAR-ECHO ID=EM-Z9SW9-P-01-12902; RAW_FILE_NAME=PZPa_S_222 SUBJECT_SAMPLE_FACTORS - PZPa_S_223 Sample Type:Study Sample HHEAR-ECHO ID=EM-QF0W4-P-01-12902; RAW_FILE_NAME=PZPa_S_223 SUBJECT_SAMPLE_FACTORS - PZPa_S_224 Sample Type:Study Sample HHEAR-ECHO ID=EM-HH2P2-P-01-12902; RAW_FILE_NAME=PZPa_S_224 SUBJECT_SAMPLE_FACTORS - PZPa_S_225 Sample Type:Study Sample HHEAR-ECHO ID=EM-FL031-P-01-12902; RAW_FILE_NAME=PZPa_S_225 SUBJECT_SAMPLE_FACTORS - PZPa_S_226 Sample Type:Study Sample HHEAR-ECHO ID=EM-WZ488-P-01-12902; RAW_FILE_NAME=PZPa_S_226 SUBJECT_SAMPLE_FACTORS - PZPa_S_227 Sample Type:Study Sample HHEAR-ECHO ID=EM-X4NB3-P-01-12902; RAW_FILE_NAME=PZPa_S_227 SUBJECT_SAMPLE_FACTORS - PZPa_S_228 Sample Type:Study Sample HHEAR-ECHO ID=EM-G5XS9-P-01-12902; RAW_FILE_NAME=PZPa_S_228 SUBJECT_SAMPLE_FACTORS - PZPa_S_229 Sample Type:Study Sample HHEAR-ECHO ID=EM-48355-P-01-12902; RAW_FILE_NAME=PZPa_S_229 SUBJECT_SAMPLE_FACTORS - PZPa_S_230 Sample Type:Study Sample HHEAR-ECHO ID=EM-6S3D0-P-01-12902; RAW_FILE_NAME=PZPa_S_230 SUBJECT_SAMPLE_FACTORS - PZPa_S_231 Sample Type:Study Sample HHEAR-ECHO ID=EM-GUE13-P-01-12902; RAW_FILE_NAME=PZPa_S_231 SUBJECT_SAMPLE_FACTORS - PZPa_S_232 Sample Type:Study Sample HHEAR-ECHO ID=EM-MDUB1-P-01-12902; RAW_FILE_NAME=PZPa_S_232 SUBJECT_SAMPLE_FACTORS - PZPa_S_233 Sample Type:Study Sample HHEAR-ECHO ID=EM-GQ567-P-01-12902; RAW_FILE_NAME=PZPa_S_233 SUBJECT_SAMPLE_FACTORS - PZPa_S_234 Sample Type:Study Sample HHEAR-ECHO ID=EM-QW7W0-P-01-12902; RAW_FILE_NAME=PZPa_S_234 SUBJECT_SAMPLE_FACTORS - PZPa_S_235 Sample Type:Study Sample HHEAR-ECHO ID=EM-7TBD2-P-01-12902; RAW_FILE_NAME=PZPa_S_235 SUBJECT_SAMPLE_FACTORS - PZPa_S_236 Sample Type:Study Sample HHEAR-ECHO ID=EM-SVQC8-P-01-12902; RAW_FILE_NAME=PZPa_S_236 SUBJECT_SAMPLE_FACTORS - PZPa_S_237 Sample Type:Study Sample HHEAR-ECHO ID=EM-R8CB1-P-01-12902; RAW_FILE_NAME=PZPa_S_237 SUBJECT_SAMPLE_FACTORS - PZPa_S_238 Sample Type:Study Sample HHEAR-ECHO ID=EM-LNNQ4-P-01-12902; RAW_FILE_NAME=PZPa_S_238 SUBJECT_SAMPLE_FACTORS - PZPa_S_239 Sample Type:Study Sample HHEAR-ECHO ID=EM-V2J93-P-01-12902; RAW_FILE_NAME=PZPa_S_239 SUBJECT_SAMPLE_FACTORS - PZPa_S_240 Sample Type:Study Sample HHEAR-ECHO ID=EM-MZ4F7-P-01-12902; RAW_FILE_NAME=PZPa_S_240 SUBJECT_SAMPLE_FACTORS - PZPa_SP_1 Sample Type:Study Pool HHEAR-ECHO ID=EM-JTYQ2-QCSP-01-12902; RAW_FILE_NAME=PZPa_SP_1 SUBJECT_SAMPLE_FACTORS - PZPa_SP_2 Sample Type:Study Pool HHEAR-ECHO ID=EM-F4HY6-QCSP-02-12902; RAW_FILE_NAME=PZPa_SP_2 SUBJECT_SAMPLE_FACTORS - PZPa_SP_3 Sample Type:Study Pool HHEAR-ECHO ID=EM-3VW62-QCSP-03-12902; RAW_FILE_NAME=PZPa_SP_3 SUBJECT_SAMPLE_FACTORS - PZPa_SP_4 Sample Type:Study Pool HHEAR-ECHO ID=EM-3BKQ9-QCSP-04-12902; RAW_FILE_NAME=PZPa_SP_4 SUBJECT_SAMPLE_FACTORS - PZPa_SP_5 Sample Type:Study Pool HHEAR-ECHO ID=EM-30UJ3-QCSP-05-12902; RAW_FILE_NAME=PZPa_SP_5 SUBJECT_SAMPLE_FACTORS - PZPa_SP_6 Sample Type:Study Pool HHEAR-ECHO ID=EM-FAF66-QCSP-06-12902; RAW_FILE_NAME=PZPa_SP_6 SUBJECT_SAMPLE_FACTORS - PZPa_SP_7 Sample Type:Study Pool HHEAR-ECHO ID=EM-50SU9-QCSP-07-12902; RAW_FILE_NAME=PZPa_SP_7 SUBJECT_SAMPLE_FACTORS - PZPa_SP_8 Sample Type:Study Pool HHEAR-ECHO ID=EM-QCSY1-QCSP-08-12902; RAW_FILE_NAME=PZPa_SP_8 SUBJECT_SAMPLE_FACTORS - PZPa_SP_9 Sample Type:Study Pool HHEAR-ECHO ID=EM-87QN8-QCSP-09-12902; RAW_FILE_NAME=PZPa_SP_9 SUBJECT_SAMPLE_FACTORS - PZPa_SP_10 Sample Type:Study Pool HHEAR-ECHO ID=EM-VGGK0-QCSP-10-12902; RAW_FILE_NAME=PZPa_SP_10 SUBJECT_SAMPLE_FACTORS - PZPa_SP_11 Sample Type:Study Pool HHEAR-ECHO ID=EM-TQ200-QCSP-11-12902; RAW_FILE_NAME=PZPa_SP_11 SUBJECT_SAMPLE_FACTORS - PZPa_SP_12 Sample Type:Study Pool HHEAR-ECHO ID=EM-YKTQ2-QCSP-12-12902; RAW_FILE_NAME=PZPa_SP_12 SUBJECT_SAMPLE_FACTORS - PZPa_SP_13 Sample Type:Study Pool HHEAR-ECHO ID=EM-JAD23-QCSP-13-12902; RAW_FILE_NAME=PZPa_SP_13 SUBJECT_SAMPLE_FACTORS - PZPa_SP_14 Sample Type:Study Pool HHEAR-ECHO ID=EM-G0JL0-QCSP-14-12902; RAW_FILE_NAME=PZPa_SP_14 SUBJECT_SAMPLE_FACTORS - PZPa_SP_15 Sample Type:Study Pool HHEAR-ECHO ID=EM-37JL5-QCSP-15-12902; RAW_FILE_NAME=PZPa_SP_15 SUBJECT_SAMPLE_FACTORS - PZPa_SP_16 Sample Type:Study Pool HHEAR-ECHO ID=EM-7BH75-QCSP-16-12902; RAW_FILE_NAME=PZPa_SP_16 SUBJECT_SAMPLE_FACTORS - PZPa_SP_17 Sample Type:Study Pool HHEAR-ECHO ID=EM-QLU09-QCSP-17-12902; RAW_FILE_NAME=PZPa_SP_17 SUBJECT_SAMPLE_FACTORS - PZPa_SP_18 Sample Type:Study Pool HHEAR-ECHO ID=EM-SJR46-QCSP-18-12902; RAW_FILE_NAME=PZPa_SP_18 SUBJECT_SAMPLE_FACTORS - PZPa_SP_19 Sample Type:Study Pool HHEAR-ECHO ID=EM-AB5M6-QCSP-19-12902; RAW_FILE_NAME=PZPa_SP_19 SUBJECT_SAMPLE_FACTORS - PZPa_SP_20 Sample Type:Study Pool HHEAR-ECHO ID=EM-A55Y7-QCSP-20-12902; RAW_FILE_NAME=PZPa_SP_20 SUBJECT_SAMPLE_FACTORS - PZPa_SP_21 Sample Type:Study Pool HHEAR-ECHO ID=EM-52431-QCSP-21-12902; RAW_FILE_NAME=PZPa_SP_21 SUBJECT_SAMPLE_FACTORS - PZPa_SP_22 Sample Type:Study Pool HHEAR-ECHO ID=EM-PAG24-QCSP-22-12902; RAW_FILE_NAME=PZPa_SP_22 SUBJECT_SAMPLE_FACTORS - PZPa_SP_23 Sample Type:Study Pool HHEAR-ECHO ID=EM-YSPQ8-QCSP-23-12902; RAW_FILE_NAME=PZPa_SP_23 SUBJECT_SAMPLE_FACTORS - PZPa_SP_24 Sample Type:Study Pool HHEAR-ECHO ID=EM-G04E7-QCSP-24-12902; RAW_FILE_NAME=PZPa_SP_24 #COLLECTION CO:COLLECTION_SUMMARY The ARCH Cohort is a pregnancy cohort of approximately 1,000 women recruited at CO:COLLECTION_SUMMARY the first prenatal visit largely in Lansing, MI between 2008 and 2016. Blood was CO:COLLECTION_SUMMARY collected when possible at the onset of prenatal care and at the time of the CO:COLLECTION_SUMMARY glucose tolerance test (late 2nd, early 3rd trimester). Please contact Jean CO:COLLECTION_SUMMARY Kerver at kerverje@msu.edu for questions related to the subject characteristics CO:COLLECTION_SUMMARY and outcomes. CO:SAMPLE_TYPE Blood (plasma) CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY N/A #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Study samples and study pools (50 µL aliquots) were shipped from Michigan State SP:SAMPLEPREP_SUMMARY University to the NC HHEAR Hub on dry ice. HHEAR reference plasma (50 µL each) SP:SAMPLEPREP_SUMMARY and NIST plasma (1950) reference material (50 µL each) were provided by the NC SP:SAMPLEPREP_SUMMARY HHEAR Hub. LC-MS grade water (50 µL) was used as blank. All samples were thawed SP:SAMPLEPREP_SUMMARY at 4°C overnight before the preparation. Samples, including study samples, SP:SAMPLEPREP_SUMMARY study pool samples, HHEAR reference plasma, NIST reference plasma, and blanks SP:SAMPLEPREP_SUMMARY were mixed with 400 µL methanol containing 500 ng/mL tryptophan-d5 as internal SP:SAMPLEPREP_SUMMARY standard and vortexed by a multiple tube vortex mixer for 2 min at 5000 rpm at SP:SAMPLEPREP_SUMMARY room temperature. All samples were centrifuged at 16,000 rcf for 10 min at 4°C. SP:SAMPLEPREP_SUMMARY The supernatant (350 µL) was transferred into a pre-labeled 2.0 mL Lo-bind SP:SAMPLEPREP_SUMMARY Eppendorf tube, dried by a SpeedVac overnight, and stored at -80° C. For SP:SAMPLEPREP_SUMMARY immediate analysis, 100 µL of water-methanol solution (95:5, v/v) was used to SP:SAMPLEPREP_SUMMARY reconstitute the dried extracts. Samples were thoroughly mixed on a multiple SP:SAMPLEPREP_SUMMARY tube vortex mixer for 10 min at 5000 rpm at room temperature and then SP:SAMPLEPREP_SUMMARY centrifuged at 4°C for 10 min at 16,000 rcf. The supernatant was transferred to SP:SAMPLEPREP_SUMMARY pre-labeled autosampler vials for data acquisition by LC-MS. SP:PROCESSING_STORAGE_CONDITIONS On ice SP:EXTRACTION_METHOD Vortex with methanol containing 500ng/ml tryptophan-d5 as internal standard SP:EXTRACT_STORAGE -80℃ SP:SAMPLE_RESUSPENSION Water-Methanol (95:5, v/v) SP:SAMPLE_SPIKING Tryptophan-d5 stock solution at 500 ng/mL #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) CH:SOLVENT_A 100% water; 0.1% formic acid CH:SOLVENT_B 100% methanol; 0.1% formic acid 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:COLUMN_TEMPERATURE 50 CH:COLUMN_PRESSURE 6000-10000 psi CH:INJECTION_TEMPERATURE 8 CH:INTERNAL_STANDARD Tryptophan-d5 CH:ANALYTICAL_TIME 22 min CH:WEAK_WASH_SOLVENT_NAME 10% methanol/90% water; 0.1% formic acid CH:STRONG_WASH_SOLVENT_NAME 75% 2-Propanol/25% Water; 0.1% formic acid #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive HF-X Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Xcalibur 4.1.31.9 for data acquisition; Progenesis QI 2.4 for data preprocessing MS:CAPILLARY_TEMPERATURE 320 °C MS:CAPILLARY_VOLTAGE 3.5 kV MS:COLLISION_ENERGY 20-45, ramp MS:COLLISION_GAS N2 MS:DRY_GAS_TEMP 400°C MS:FRAGMENTATION_METHOD CID MS:DESOLVATION_GAS_FLOW 55 MS:MS_RESULTS_FILE ST002443_AN003980_Results.txt UNITS:Peak area Has m/z:Yes Has RT:Yes RT units:Minutes #END