#METABOLOMICS WORKBENCH chichen@umn.edu_20160629_144517 DATATRACK_ID:656 STUDY_ID:ST000437 ANALYSIS_ID:AN000688 PROJECT_ID:PR000337 VERSION 1 CREATED_ON August 2, 2016, 12:40 pm #PROJECT PR:PROJECT_TITLE Ring trial in CHEAR untargeted resource PR:PROJECT_SUMMARY To conduct metabolomic studies comparing 20 males and 20 females as well as 10 PR:PROJECT_SUMMARY pool controls and 5 spike-in samples using the extraction methods, MS or NMR PR:PROJECT_SUMMARY platforms these laboratories deem best suited to fulfill the comparative aims of PR:PROJECT_SUMMARY the trial with respect to qualitative metabolome analysis, quantitative PR:PROJECT_SUMMARY differences between study groups, and finding, identifying and estimating PR:PROJECT_SUMMARY quantitative differences in the spiked-in samples. PR:INSTITUTE University of Minesota PR:DEPARTMENT Department of Food Science and Nutrition PR:LABORATORY Nutritional Metabolomics PR:LAST_NAME Chen PR:FIRST_NAME Chi PR:ADDRESS 1334 Eckles Ave W, St Paul, MN 55108 PR:EMAIL chichen@umn.edu PR:PHONE 612-624-7704 PR:FUNDING_SOURCE NIEHS #STUDY ST:STUDY_TITLE Metabolomic analysis of human plasma samples spiked with unknown chemicals ST:STUDY_TYPE Untargeted metabolomics for sample classification ST:STUDY_SUMMARY To conduct metabolomic studies comparing 20 males and 20 females as well as 10 ST:STUDY_SUMMARY pool controls and 5 spike-in samples using MS platform to perform both ST:STUDY_SUMMARY qualitative and quantitative metabolome analysis to classify the sample groups ST:STUDY_SUMMARY (male, female, pooled) and to identify the spiked-in compounds. ST:INSTITUTE University of Minesota ST:DEPARTMENT Department of Food Science and Nutrition ST:LABORATORY Nutritional Metabolomics ST:LAST_NAME Chen ST:FIRST_NAME Chi ST:ADDRESS 1334 Eckles Ave W, St Paul, MN 55108 ST:EMAIL chichen@umn.edu ST:PHONE 612-624-7704 ST:NUM_GROUPS 4 ST:TOTAL_SUBJECTS 55 ST:NUM_MALES 20 ST:NUM_FEMALES 20 #SUBJECT SU:SUBJECT_TYPE Human SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:GENDER male and female #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - S039 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - std3 Sample type:Spiked SUBJECT_SAMPLE_FACTORS - S048 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S036 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S034 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S049 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S050 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S029 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S010 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S045 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S024 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S042 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S046 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S007 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S040 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S022 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - Std1 Sample type:Spiked SUBJECT_SAMPLE_FACTORS - S017 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S019 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S018 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S006 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S026 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S025 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S032 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S033 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - std4 Sample type:Spiked SUBJECT_SAMPLE_FACTORS - S012 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S037 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S030 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S011 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S021 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S001 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S009 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S016 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S013 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S002 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S008 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - std5 Sample type:Spiked SUBJECT_SAMPLE_FACTORS - S031 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S020 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S027 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S028 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S005 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S043 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S047 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S003 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S004 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S044 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - Std2 Sample type:Spiked SUBJECT_SAMPLE_FACTORS - S023 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S038 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S035 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S014 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S041 Sample type:Unknown SUBJECT_SAMPLE_FACTORS - S015 Sample type:Unknown #COLLECTION CO:COLLECTION_SUMMARY Human Plasma collected and processed by BioreclamationIVT (Catalog No. HMPLLIHP) CO:COLLECTION_SUMMARY supplemented with blinded spiked-in standards as determined by PENN STATE in CO:COLLECTION_SUMMARY consultation with National Institute of Standards and Technology and the CO:COLLECTION_SUMMARY advisory board for the NIH Regional Comprehensive Metabolomics Resource Cores. CO:SAMPLE_TYPE Plasma #TREATMENT TR:TREATMENT_SUMMARY 10 pooled plasma samples prepared by combining equal amounts of aliquots of each TR:TREATMENT_SUMMARY male and each female sample. 5 plasma samples supplemented with blinded TR:TREATMENT_SUMMARY spiked-in standards. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Human plasma samples were thawed on ice. Aliquots of 50 uL plasma samples were SP:SAMPLEPREP_SUMMARY mixed with 500uL of 66% acetonitrile in water. The mixture was vortexed at SP:SAMPLEPREP_SUMMARY 3000rpm for 30s, and centrifuged at 21,000 × g at 4 oC for 10 min. An aliquot SP:SAMPLEPREP_SUMMARY of 200 uL supernatant was transferred to HPLC vials and analyzed by LC/MS. SP:PROCESSING_METHOD Extraction, vortex, centrifugation SP:PROCESSING_STORAGE_CONDITIONS On ice SP:EXTRACTION_METHOD 66% acetontrile in water SP:EXTRACT_STORAGE - 20 C #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Targeted method for sample classification CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Acquity BEH C8 column CH:FLOW_GRADIENT 0-0.5min-0.5%B, 0.5-4min gradient to 60%B, 4-8min gradient to 100%B, CH:FLOW_GRADIENT 8-9min-100%B, 9-9.2min gradient to 0.5%B, 9.2-10min-0.5%B CH:FLOW_RATE 0.5mL/min CH:COLUMN_TEMPERATURE 60 C CH:SOLVENT_A Water with 0.1% formic acid CH:SOLVENT_B Acetonitrile with 0.1% formic acid CH:SAMPLE_INJECTION 5 uL CH:WEAK_WASH_SOLVENT_NAME 50% methanol in water CH:WEAK_WASH_VOLUME 500 uL CH:STRONG_WASH_SOLVENT_NAME 50% isopropanol, 25% acetonitrile and 25% water CH:STRONG_WASH_VOLUME 500 uL CH:SAMPLE_LOOP_SIZE 20 uL #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Waters Xevo-G2-S MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MASS_ACCURACY <5 ppm #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS nM MS_METABOLITE_DATA_START Samples S039 std3 S048 S036 S034 S049 S050 S029 S010 S045 S024 S042 S046 S007 S040 S022 Std1 S017 S019 S018 S006 S026 S025 S032 S033 std4 S012 S037 S030 S011 S021 S001 S009 S016 S013 S002 S008 std5 S031 S020 S027 S028 S005 S043 S047 S003 S004 S044 Std2 S023 S038 S035 S014 S041 S015 Factors Sample type:Unknown Sample type:Spiked Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Spiked Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Spiked Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Spiked Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Spiked Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Sample type:Unknown Testosterone 8.9 6.5 8.4 21.4 N/D 7.4 N/D N/D 11.7 5.8 19.3 N/D 8.3 0.1 12.7 8.9 8.9 17.9 N/D 11.2 N/D 0.2 N/D N/D 9.3 2.5 1.2 N/D 9.2 N/D N/D N/D 12.8 N/D N/D 21 8.5 7.1 41.6 6.8 1.4 5.8 N/D 17.4 9.9 7.8 14.5 7.9 10.4 N/D 14.9 15.3 8.3 3.5 9.8 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z PubChem ID KEGG ID Testosterone 3.29 289.2162 8144176 C00535 METABOLITES_END #END