#METABOLOMICS WORKBENCH wpathmasiri_20240216_114625 DATATRACK_ID:4651 STUDY_ID:ST003110 ANALYSIS_ID:AN005093 VERSION 1 CREATED_ON 03-06-2024 #PROJECT #STUDY #SUBJECT #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data #COLLECTION #TREATMENT #SAMPLEPREP #CHROMATOGRAPHY #ANALYSIS #END #METABOLOMICS WORKBENCH wpathmasiri_20240216_114625 DATATRACK_ID:4651 STUDY_ID:ST003110 ANALYSIS_ID:AN005093 VERSION 1 CREATED_ON 03-06-2024 #PROJECT PR:PROJECT_TITLE Untargeted Metabolomics Reveals Signatures of a Healthy Lifestyle PR:PROJECT_TYPE C18 Reversed-Phase Broad Spectrum Metabolomics PR:PROJECT_SUMMARY This cross-sectional study investigated differences in the plasma metabolome in PR:PROJECT_SUMMARY two groups of adults that were of similar age but varied markedly in body PR:PROJECT_SUMMARY composition and dietary and physical activity patterns. Study participants PR:PROJECT_SUMMARY included 52 adults in the lifestyle group (LIFE) (28 males, 24 females) and 52 PR:PROJECT_SUMMARY in the control group (CON) (27 males, 25 females). The results using an PR:PROJECT_SUMMARY extensive untargeted UPLC-HRMS analysis with 10,535 metabolite peaks identified PR:PROJECT_SUMMARY 83 metabolites and 16 metabolic pathways that differentiated LIFE and CON PR:PROJECT_SUMMARY groups. A novel metabolite signature of positive lifestyle habits emerged from PR:PROJECT_SUMMARY this analysis highlighted by lower plasma levels of numerous bile acids, an PR:PROJECT_SUMMARY amino acid profile characterized by higher histidine and lower glutamic acid, PR:PROJECT_SUMMARY glutamine, β-alanine, phenylalanine, tyrosine, and proline, an elevated vitamin PR:PROJECT_SUMMARY D status, higher levels of beneficial fatty acids and gut microbiome catabolism PR:PROJECT_SUMMARY metabolites from plant substrates, and reduced levels of N-glycan degradation PR:PROJECT_SUMMARY metabolites and environmental contaminants. This study established that the PR:PROJECT_SUMMARY plasma metabolome is strongly associated with body composition and lifestyle PR:PROJECT_SUMMARY habits. The robust lifestyle metabolite signature identified in this study is PR:PROJECT_SUMMARY consistent with an improved life expectancy and a reduced risk for chronic PR:PROJECT_SUMMARY disease. PR:INSTITUTE Metabolomics and Exposome Laboratory at UNC Nutrition Research Institute PR:DEPARTMENT Untargeted Analysis PR:LABORATORY Sumner Lab PR:LAST_NAME Rushing PR:FIRST_NAME Blake PR:ADDRESS Nutrition Research Institute , University of North Carolina at Chapel Hill, 500 PR:ADDRESS Laureate Way Kannapolis, NC 28081 PR:EMAIL blake_rushing@unc.edu PR:PHONE (704) 250-5000 PR:DOI http://dx.doi.org/10.21228/M80B0B #STUDY ST:STUDY_TITLE Influence of Lifestyle Factors on Epigenetic and Proteomic Biomarkers: A ST:STUDY_TITLE Cross-Sectional Comparison ST:STUDY_TYPE Cross-sectional Study ST:STUDY_SUMMARY This cross-sectional study investigated differences in the plasma metabolome in ST:STUDY_SUMMARY two groups of adults that were of similar age but varied markedly in body ST:STUDY_SUMMARY composition and dietary and physical activity patterns. Study participants ST:STUDY_SUMMARY included 52 adults in the lifestyle group (LIFE) (28 males, 24 females) and 52 ST:STUDY_SUMMARY in the control group (CON) (27 males, 25 females). The results using an ST:STUDY_SUMMARY extensive untargeted UPLC-HRMS analysis with 10,535 metabolite peaks identified ST:STUDY_SUMMARY 83 metabolites and 16 metabolic pathways that differentiated LIFE and CON ST:STUDY_SUMMARY groups. A novel metabolite signature of positive lifestyle habits emerged from ST:STUDY_SUMMARY this analysis highlighted by lower plasma levels of numerous bile acids, an ST:STUDY_SUMMARY amino acid profile characterized by higher histidine and lower glutamic acid, ST:STUDY_SUMMARY glutamine, β-alanine, phenylalanine, tyrosine, and proline, an elevated vitamin ST:STUDY_SUMMARY D status, higher levels of beneficial fatty acids and gut microbiome catabolism ST:STUDY_SUMMARY metabolites from plant substrates, and reduced levels of N-glycan degradation ST:STUDY_SUMMARY metabolites and environmental contaminants. This study established that the ST:STUDY_SUMMARY plasma metabolome is strongly associated with body composition and lifestyle ST:STUDY_SUMMARY habits. The robust lifestyle metabolite signature identified in this study is ST:STUDY_SUMMARY consistent with an improved life expectancy and a reduced risk for chronic ST:STUDY_SUMMARY disease. ST:INSTITUTE Appalachian State University ST:DEPARTMENT Department of Biology ST:LABORATORY Human Performance Laboratory ST:LAST_NAME Nieman ST:FIRST_NAME David ST:ADDRESS North Carolina Research Campus, 600 Laureate Way, Kannapolis, NC 28081 ST:EMAIL niemandc@appstate.edu ST:PHONE (828) 773-0056 ST:SUBMIT_DATE 2024-02-16 #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 1 S_1 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=1; RAW_FILE_NAME=S_1.raw SUBJECT_SAMPLE_FACTORS 10 S_10 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=9; RAW_FILE_NAME=S_10.raw SUBJECT_SAMPLE_FACTORS 100 S_100 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=121; RAW_FILE_NAME=S_100.raw SUBJECT_SAMPLE_FACTORS 101 S_101 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=122; RAW_FILE_NAME=S_101.raw SUBJECT_SAMPLE_FACTORS 102 S_102 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=123; RAW_FILE_NAME=S_102.raw SUBJECT_SAMPLE_FACTORS 107 S_107 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=131; RAW_FILE_NAME=S_107.raw SUBJECT_SAMPLE_FACTORS 108 S_108 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=132; RAW_FILE_NAME=S_108.raw SUBJECT_SAMPLE_FACTORS 11 S_11 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=10; RAW_FILE_NAME=S_11.raw SUBJECT_SAMPLE_FACTORS 15 S_15 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=17; RAW_FILE_NAME=S_15.raw SUBJECT_SAMPLE_FACTORS 17 S_17 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=18; RAW_FILE_NAME=S_17.raw SUBJECT_SAMPLE_FACTORS 18 S_18 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=19; RAW_FILE_NAME=S_18.raw SUBJECT_SAMPLE_FACTORS 19 S_19 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=20; RAW_FILE_NAME=S_19.raw SUBJECT_SAMPLE_FACTORS 2 S_2 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=2; RAW_FILE_NAME=S_2.raw SUBJECT_SAMPLE_FACTORS 22 S_22 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=22; RAW_FILE_NAME=S_22.raw SUBJECT_SAMPLE_FACTORS 27 S_27 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=30; RAW_FILE_NAME=S_27.raw SUBJECT_SAMPLE_FACTORS 28 S_28 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=31; RAW_FILE_NAME=S_28.raw SUBJECT_SAMPLE_FACTORS 3 S_3 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=3; RAW_FILE_NAME=S_3.raw SUBJECT_SAMPLE_FACTORS 30 S_30 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=33; RAW_FILE_NAME=S_30.raw SUBJECT_SAMPLE_FACTORS 32 S_32 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=35; RAW_FILE_NAME=S_32.raw SUBJECT_SAMPLE_FACTORS 33 S_33 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=36; RAW_FILE_NAME=S_33.raw SUBJECT_SAMPLE_FACTORS 35 S_35 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=41; RAW_FILE_NAME=S_35.raw SUBJECT_SAMPLE_FACTORS 37 S_37 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=43; RAW_FILE_NAME=S_37.raw SUBJECT_SAMPLE_FACTORS 40 S_40 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=46; RAW_FILE_NAME=S_40.raw SUBJECT_SAMPLE_FACTORS 43 S_43 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=49; RAW_FILE_NAME=S_43.raw SUBJECT_SAMPLE_FACTORS 47 S_47 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=55; RAW_FILE_NAME=S_47.raw SUBJECT_SAMPLE_FACTORS 49 S_49 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=57; RAW_FILE_NAME=S_49.raw SUBJECT_SAMPLE_FACTORS 50 S_50 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=58; RAW_FILE_NAME=S_50.raw SUBJECT_SAMPLE_FACTORS 52 S_52 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=60; RAW_FILE_NAME=S_52.raw SUBJECT_SAMPLE_FACTORS 54 S_54 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=62; RAW_FILE_NAME=S_54.raw SUBJECT_SAMPLE_FACTORS 56 S_56 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=67; RAW_FILE_NAME=S_56.raw SUBJECT_SAMPLE_FACTORS 58 S_58 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=68; RAW_FILE_NAME=S_58.raw SUBJECT_SAMPLE_FACTORS 60 S_60 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=70; RAW_FILE_NAME=S_60.raw SUBJECT_SAMPLE_FACTORS 63 S_63 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=73; RAW_FILE_NAME=S_63.raw SUBJECT_SAMPLE_FACTORS 64 S_64 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=74; RAW_FILE_NAME=S_64.raw SUBJECT_SAMPLE_FACTORS 65 S_65 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=75; RAW_FILE_NAME=S_65.raw SUBJECT_SAMPLE_FACTORS 69 S_69 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=82; RAW_FILE_NAME=S_69.raw SUBJECT_SAMPLE_FACTORS 73 S_73 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=86; RAW_FILE_NAME=S_73.raw SUBJECT_SAMPLE_FACTORS 76 S_76 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=92; RAW_FILE_NAME=S_76.raw SUBJECT_SAMPLE_FACTORS 77 S_77 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=93; RAW_FILE_NAME=S_77.raw SUBJECT_SAMPLE_FACTORS 78 S_78 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=94; RAW_FILE_NAME=S_78.raw SUBJECT_SAMPLE_FACTORS 79 S_79 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=95; RAW_FILE_NAME=S_79.raw SUBJECT_SAMPLE_FACTORS 80 S_80 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=96; RAW_FILE_NAME=S_80.raw SUBJECT_SAMPLE_FACTORS 81 S_81 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=97; RAW_FILE_NAME=S_81.raw SUBJECT_SAMPLE_FACTORS 82 S_82 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=98; RAW_FILE_NAME=S_82.raw SUBJECT_SAMPLE_FACTORS 84 S_84 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=100; RAW_FILE_NAME=S_84.raw SUBJECT_SAMPLE_FACTORS 86 S_86 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=105; RAW_FILE_NAME=S_86.raw SUBJECT_SAMPLE_FACTORS 87 S_87 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=106; RAW_FILE_NAME=S_87.raw SUBJECT_SAMPLE_FACTORS 89 S_89 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=108; RAW_FILE_NAME=S_89.raw SUBJECT_SAMPLE_FACTORS 90 S_90 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=109; RAW_FILE_NAME=S_90.raw SUBJECT_SAMPLE_FACTORS 93 S_93 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=112; RAW_FILE_NAME=S_93.raw SUBJECT_SAMPLE_FACTORS 94 S_94 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=113; RAW_FILE_NAME=S_94.raw SUBJECT_SAMPLE_FACTORS 97 S_97 Sample source:Blood | Phenotype:Control | Sample type:Study sample Analysis Order=118; RAW_FILE_NAME=S_97.raw SUBJECT_SAMPLE_FACTORS 103 S_103 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=124; RAW_FILE_NAME=S_103.raw SUBJECT_SAMPLE_FACTORS 104 S_104 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=125; RAW_FILE_NAME=S_104.raw SUBJECT_SAMPLE_FACTORS 105 S_105 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=126; RAW_FILE_NAME=S_105.raw SUBJECT_SAMPLE_FACTORS 106 S_106 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=127; RAW_FILE_NAME=S_106.raw SUBJECT_SAMPLE_FACTORS 109 S_109 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=133; RAW_FILE_NAME=S_109.raw SUBJECT_SAMPLE_FACTORS 110 S_110 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=134; RAW_FILE_NAME=S_110.raw SUBJECT_SAMPLE_FACTORS 12 S_12 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=14; RAW_FILE_NAME=S_12.raw SUBJECT_SAMPLE_FACTORS 13 S_13 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=15; RAW_FILE_NAME=S_13.raw SUBJECT_SAMPLE_FACTORS 14 S_14 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=16; RAW_FILE_NAME=S_14.raw SUBJECT_SAMPLE_FACTORS 21 S_21 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=21; RAW_FILE_NAME=S_21.raw SUBJECT_SAMPLE_FACTORS 23 S_23 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=23; RAW_FILE_NAME=S_23.raw SUBJECT_SAMPLE_FACTORS 24 S_24 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=27; RAW_FILE_NAME=S_24.raw SUBJECT_SAMPLE_FACTORS 25 S_25 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=28; RAW_FILE_NAME=S_25.raw SUBJECT_SAMPLE_FACTORS 26 S_26 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=29; RAW_FILE_NAME=S_26.raw SUBJECT_SAMPLE_FACTORS 29 S_29 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=32; RAW_FILE_NAME=S_29.raw SUBJECT_SAMPLE_FACTORS 31 S_31 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=34; RAW_FILE_NAME=S_31.raw SUBJECT_SAMPLE_FACTORS 34 S_34 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=40; RAW_FILE_NAME=S_34.raw SUBJECT_SAMPLE_FACTORS 36 S_36 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=42; RAW_FILE_NAME=S_36.raw SUBJECT_SAMPLE_FACTORS 38 S_38 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=44; RAW_FILE_NAME=S_38.raw SUBJECT_SAMPLE_FACTORS 39 S_39 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=45; RAW_FILE_NAME=S_39.raw SUBJECT_SAMPLE_FACTORS 41 S_41 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=47; RAW_FILE_NAME=S_41.raw SUBJECT_SAMPLE_FACTORS 42 S_42 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=48; RAW_FILE_NAME=S_42.raw SUBJECT_SAMPLE_FACTORS 44 S_44 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=53; RAW_FILE_NAME=S_44.raw SUBJECT_SAMPLE_FACTORS 46 S_46 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=54; RAW_FILE_NAME=S_46.raw SUBJECT_SAMPLE_FACTORS 48 S_48 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=56; RAW_FILE_NAME=S_48.raw SUBJECT_SAMPLE_FACTORS 5 S_5 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=4; RAW_FILE_NAME=S_5.raw SUBJECT_SAMPLE_FACTORS 51 S_51 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=59; RAW_FILE_NAME=S_51.raw SUBJECT_SAMPLE_FACTORS 53 S_53 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=61; RAW_FILE_NAME=S_53.raw SUBJECT_SAMPLE_FACTORS 55 S_55 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=66; RAW_FILE_NAME=S_55.raw SUBJECT_SAMPLE_FACTORS 59 S_59 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=69; RAW_FILE_NAME=S_59.raw SUBJECT_SAMPLE_FACTORS 6 S_6 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=5; RAW_FILE_NAME=S_6.raw SUBJECT_SAMPLE_FACTORS 61 S_61 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=71; RAW_FILE_NAME=S_61.raw SUBJECT_SAMPLE_FACTORS 62 S_62 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=72; RAW_FILE_NAME=S_62.raw SUBJECT_SAMPLE_FACTORS 66 S_66 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=79; RAW_FILE_NAME=S_66.raw SUBJECT_SAMPLE_FACTORS 67 S_67 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=80; RAW_FILE_NAME=S_67.raw SUBJECT_SAMPLE_FACTORS 68 S_68 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=81; RAW_FILE_NAME=S_68.raw SUBJECT_SAMPLE_FACTORS 7 S_7 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=6; RAW_FILE_NAME=S_7.raw SUBJECT_SAMPLE_FACTORS 70 S_70 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=83; RAW_FILE_NAME=S_70.raw SUBJECT_SAMPLE_FACTORS 71 S_71 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=84; RAW_FILE_NAME=S_71.raw SUBJECT_SAMPLE_FACTORS 72 S_72 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=85; RAW_FILE_NAME=S_72.raw SUBJECT_SAMPLE_FACTORS 74 S_74 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=87; RAW_FILE_NAME=S_74.raw SUBJECT_SAMPLE_FACTORS 75 S_75 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=88; RAW_FILE_NAME=S_75.raw SUBJECT_SAMPLE_FACTORS 8 S_8 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=7; RAW_FILE_NAME=S_8.raw SUBJECT_SAMPLE_FACTORS 83 S_83 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=99; RAW_FILE_NAME=S_83.raw SUBJECT_SAMPLE_FACTORS 85 S_85 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=101; RAW_FILE_NAME=S_85.raw SUBJECT_SAMPLE_FACTORS 88 S_88 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=107; RAW_FILE_NAME=S_88.raw SUBJECT_SAMPLE_FACTORS 9 S_9 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=8; RAW_FILE_NAME=S_9.raw SUBJECT_SAMPLE_FACTORS 91 S_91 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=110; RAW_FILE_NAME=S_91.raw SUBJECT_SAMPLE_FACTORS 92 S_92 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=111; RAW_FILE_NAME=S_92.raw SUBJECT_SAMPLE_FACTORS 95 S_95 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=114; RAW_FILE_NAME=S_95.raw SUBJECT_SAMPLE_FACTORS 98 S_98 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=119; RAW_FILE_NAME=S_98.raw SUBJECT_SAMPLE_FACTORS 99 S_99 Sample source:Blood | Phenotype:Life | Sample type:Study sample Analysis Order=120; RAW_FILE_NAME=S_99.raw SUBJECT_SAMPLE_FACTORS NA NIST_1 Sample source:Blood | Phenotype:NA | Sample type:NIST Analysis Order=12; RAW_FILE_NAME=NIST_1.raw SUBJECT_SAMPLE_FACTORS NA NIST_10 Sample source:Blood | Phenotype:NA | Sample type:NIST Analysis Order=129; RAW_FILE_NAME=NIST_10.raw SUBJECT_SAMPLE_FACTORS NA NIST_11 Sample source:Blood | Phenotype:NA | Sample type:NIST Analysis Order=136; RAW_FILE_NAME=NIST_11.raw SUBJECT_SAMPLE_FACTORS NA NIST_2 Sample source:Blood | Phenotype:NA | Sample type:NIST Analysis Order=25; RAW_FILE_NAME=NIST_2.raw SUBJECT_SAMPLE_FACTORS NA NIST_3 Sample source:Blood | Phenotype:NA | Sample type:NIST Analysis Order=38; RAW_FILE_NAME=NIST_3.raw SUBJECT_SAMPLE_FACTORS NA NIST_4 Sample source:Blood | Phenotype:NA | Sample type:NIST Analysis Order=51; RAW_FILE_NAME=NIST_4.raw SUBJECT_SAMPLE_FACTORS NA NIST_5 Sample source:Blood | Phenotype:NA | Sample type:NIST Analysis Order=64; RAW_FILE_NAME=NIST_5.raw SUBJECT_SAMPLE_FACTORS NA NIST_6 Sample source:Blood | Phenotype:NA | Sample type:NIST Analysis Order=77; RAW_FILE_NAME=NIST_6.raw SUBJECT_SAMPLE_FACTORS NA NIST_7 Sample source:Blood | Phenotype:NA | Sample type:NIST Analysis Order=90; RAW_FILE_NAME=NIST_7.raw SUBJECT_SAMPLE_FACTORS NA NIST_8 Sample source:Blood | Phenotype:NA | Sample type:NIST Analysis Order=103; RAW_FILE_NAME=NIST_8.raw SUBJECT_SAMPLE_FACTORS NA NIST_9 Sample source:Blood | Phenotype:NA | Sample type:NIST Analysis Order=116; RAW_FILE_NAME=NIST_9.raw SUBJECT_SAMPLE_FACTORS NA SP_1 Sample source:Blood | Phenotype:NA | Sample type:Study Pool Analysis Order=13; RAW_FILE_NAME=SP_1.raw SUBJECT_SAMPLE_FACTORS NA SP_10 Sample source:Blood | Phenotype:NA | Sample type:Study Pool Analysis Order=130; RAW_FILE_NAME=SP_10.raw SUBJECT_SAMPLE_FACTORS NA SP_11 Sample source:Blood | Phenotype:NA | Sample type:Study Pool Analysis Order=137; RAW_FILE_NAME=SP_11.raw SUBJECT_SAMPLE_FACTORS NA SP_2 Sample source:Blood | Phenotype:NA | Sample type:Study Pool Analysis Order=26; RAW_FILE_NAME=SP_2.raw SUBJECT_SAMPLE_FACTORS NA SP_3 Sample source:Blood | Phenotype:NA | Sample type:Study Pool Analysis Order=39; RAW_FILE_NAME=SP_3.raw SUBJECT_SAMPLE_FACTORS NA SP_4 Sample source:Blood | Phenotype:NA | Sample type:Study Pool Analysis Order=52; RAW_FILE_NAME=SP_4.raw SUBJECT_SAMPLE_FACTORS NA SP_5 Sample source:Blood | Phenotype:NA | Sample type:Study Pool Analysis Order=65; RAW_FILE_NAME=SP_5.raw SUBJECT_SAMPLE_FACTORS NA SP_6 Sample source:Blood | Phenotype:NA | Sample type:Study Pool Analysis Order=78; RAW_FILE_NAME=SP_6.raw SUBJECT_SAMPLE_FACTORS NA SP_7 Sample source:Blood | Phenotype:NA | Sample type:Study Pool Analysis Order=91; RAW_FILE_NAME=SP_7.raw SUBJECT_SAMPLE_FACTORS NA SP_8 Sample source:Blood | Phenotype:NA | Sample type:Study Pool Analysis Order=104; RAW_FILE_NAME=SP_8.raw SUBJECT_SAMPLE_FACTORS NA SP_9 Sample source:Blood | Phenotype:NA | Sample type:Study Pool Analysis Order=117; RAW_FILE_NAME=SP_9.raw SUBJECT_SAMPLE_FACTORS NA B_10 Sample source:Water | Phenotype:NA | Sample type:Blank Analysis Order=128; RAW_FILE_NAME=B_10.raw SUBJECT_SAMPLE_FACTORS NA B_11 Sample source:Water | Phenotype:NA | Sample type:Blank Analysis Order=135; RAW_FILE_NAME=B_11.raw SUBJECT_SAMPLE_FACTORS NA B_2 Sample source:Water | Phenotype:NA | Sample type:Blank Analysis Order=24; RAW_FILE_NAME=B_2.raw SUBJECT_SAMPLE_FACTORS NA B_3 Sample source:Water | Phenotype:NA | Sample type:Blank Analysis Order=37; RAW_FILE_NAME=B_3.raw SUBJECT_SAMPLE_FACTORS NA B_4 Sample source:Water | Phenotype:NA | Sample type:Blank Analysis Order=50; RAW_FILE_NAME=B_4.raw SUBJECT_SAMPLE_FACTORS NA B_5 Sample source:Water | Phenotype:NA | Sample type:Blank Analysis Order=63; RAW_FILE_NAME=B_5.raw SUBJECT_SAMPLE_FACTORS NA B_6 Sample source:Water | Phenotype:NA | Sample type:Blank Analysis Order=76; RAW_FILE_NAME=B_6.raw SUBJECT_SAMPLE_FACTORS NA B_7 Sample source:Water | Phenotype:NA | Sample type:Blank Analysis Order=89; RAW_FILE_NAME=B_7.raw SUBJECT_SAMPLE_FACTORS NA B_8 Sample source:Water | Phenotype:NA | Sample type:Blank Analysis Order=102; RAW_FILE_NAME=B_8.raw SUBJECT_SAMPLE_FACTORS NA B_9 Sample source:Water | Phenotype:NA | Sample type:Blank Analysis Order=115; RAW_FILE_NAME=B_9.raw #COLLECTION CO:COLLECTION_SUMMARY N/A CO:SAMPLE_TYPE Blood (plasma) CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY N/A #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Study samples were shipped from Human Performance Laboratory (Nieman Laboratory) SP:SAMPLEPREP_SUMMARY at Appalachan State University to the UNC NRI Metabolomics and Exposome SP:SAMPLEPREP_SUMMARY Laboratory (MEL) on dry ice. The MEL thawed and transferred 50 µL of the study SP:SAMPLEPREP_SUMMARY samples to a new set of tubes and used them for the analysis. An additional 10 SP:SAMPLEPREP_SUMMARY µL was taken from the original study sample and transferred to another tube to SP:SAMPLEPREP_SUMMARY make the total study pool for this project, and then distributed with 50 µL per SP:SAMPLEPREP_SUMMARY aliquot, used as quality control study pools (QCSP) throughout the whole SP:SAMPLEPREP_SUMMARY analysis. All sample aliquots (50 µL each) and QC study pools (50 µL each) SP:SAMPLEPREP_SUMMARY were stored at -80° C until the day of sample preparation. NIST plasma (SRM SP:SAMPLEPREP_SUMMARY 1950) reference material (50 µL each) were provided by the MEL. LC-MS grade SP:SAMPLEPREP_SUMMARY water (50 µL) was used as blanks. All samples were thawed at 4°C overnight SP:SAMPLEPREP_SUMMARY before the preparation. Samples, including study samples, QCSP samples, NIST SP:SAMPLEPREP_SUMMARY reference plasma, 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 4℃ 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_SUMMARY Reversed phase CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) 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:INTERNAL_STANDARD Tryptophan-d5 CH:SOLVENT_A 100% water; 0.1% formic acid CH:SOLVENT_B 100% methanol; 0.1% formic acid 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 CH:CHROMATOGRAPHY_TYPE Reversed phase #ANALYSIS AN:ANALYSIS_TYPE MS AN:ACQUISITION_DATE 11/09/2211/11/2022 AN:SOFTWARE_VERSION Xcalibur 4.1.31.9 AN:DETECTOR_TYPE Orbitrap AN:DATA_FORMAT Profile #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:FRAGMENTATION_METHOD CID MS:ION_SPRAY_VOLTAGE 3.5kV MS:IONIZATION ES+ MS:MASS_ACCURACY 5 ppm MS:DATAFORMAT Profile MS:DESOLVATION_GAS_FLOW 55 MS:DESOLVATION_TEMPERATURE 400 ℃ MS:MS_RESULTS_FILE ST003110_AN005093_Results.txt UNITS:Normalized Intensity Has m/z:Yes Has RT:Yes RT units:Minutes #END