#METABOLOMICS WORKBENCH hrani_26_20240617_121348 DATATRACK_ID:4935 STUDY_ID:ST003289 ANALYSIS_ID:AN005387 PROJECT_ID:PR002041 VERSION 1 CREATED_ON 10-16-2024 #PROJECT PR:PROJECT_TITLE Malting Time-course metabolomics study PR:PROJECT_TYPE Untargeted LC-MS and GC-MS analysis PR:PROJECT_SUMMARY Malting, a crucial process for beer production, involves complex biochemical PR:PROJECT_SUMMARY transformations affecting sensory attributes and product quality. Limited PR:PROJECT_SUMMARY knowledge of metabolic alterations during malting hinders the ability to enhance PR:PROJECT_SUMMARY malt quality. This study uses untargeted GC-MS and LC-MS metabolite profiling to PR:PROJECT_SUMMARY characterize metabolic dynamics through the malting process. After data PR:PROJECT_SUMMARY processing, a total of 4980 known metabolites were identified across six stages: PR:PROJECT_SUMMARY dry seed, post-steeping, germination (DOG1, DOG3, DOG5), and kilned, about 82% PR:PROJECT_SUMMARY of these showed significant changes during malting. Statistical analysis PR:PROJECT_SUMMARY revealed stage-dependent shifts in metabolite profiles, highlighting the PR:PROJECT_SUMMARY importance of the first 3 days of germination and kilning in determining the PR:PROJECT_SUMMARY final metabolite content of finished malt. Dynamic changes in chemical classes PR:PROJECT_SUMMARY and metabolic pathways provided insights into processes critical for malt PR:PROJECT_SUMMARY quality and beer production. Additionally, metabolites associated with PR:PROJECT_SUMMARY antimicrobial properties and stress responses were identified, underscoring the PR:PROJECT_SUMMARY interplay between barley and microbial metabolic processes during malting. This PR:PROJECT_SUMMARY comprehensive profiling advances our understanding of malting and suggests PR:PROJECT_SUMMARY potential markers for process monitoring and quality control, ultimately PR:PROJECT_SUMMARY enhancing malt quality and beer production. PR:INSTITUTE USDA Agricultural Research Service PR:DEPARTMENT Cereal Crops Research Unit PR:LABORATORY Whitcomb lab PR:LAST_NAME Rani PR:FIRST_NAME Heena PR:ADDRESS 502 Walnut Street, Madison, WI 53726 PR:EMAIL bansalheena10@gmail.com PR:PHONE 7657759366 PR:FUNDING_SOURCE USDA-ARS PR:PUBLICATIONS https://doi.org/10.1016/j.foodchem.2024.141480 PR:DOI http://dx.doi.org/10.21228/M8SN77 #STUDY ST:STUDY_TITLE Integrative LC-MS and GC-MS Metabolic Profiling Unveils Dynamic Changes during ST:STUDY_TITLE Barley Malting ST:STUDY_TYPE Untargeted LC-MS and GC-MS analysis ST:STUDY_SUMMARY Malting, a crucial process for beer production, involves complex biochemical ST:STUDY_SUMMARY transformations affecting sensory attributes and product quality. Limited ST:STUDY_SUMMARY knowledge of metabolic alterations during malting hinders the ability to enhance ST:STUDY_SUMMARY malt quality. This study uses untargeted GC-MS and LC-MS metabolite profiling to ST:STUDY_SUMMARY characterize metabolic dynamics through the malting process. After data ST:STUDY_SUMMARY processing, a total of 4980 known metabolites were identified across six stages: ST:STUDY_SUMMARY dry seed, post-steeping, germination (DOG1, DOG3, DOG5), and kilned, about 82% ST:STUDY_SUMMARY of these showed significant changes during malting. Statistical analysis ST:STUDY_SUMMARY revealed stage-dependent shifts in metabolite profiles, highlighting the ST:STUDY_SUMMARY importance of the first 3 days of germination and kilning in determining the ST:STUDY_SUMMARY final metabolite content of finished malt. Dynamic changes in chemical classes ST:STUDY_SUMMARY and metabolic pathways provided insights into processes critical for malt ST:STUDY_SUMMARY quality and beer production. Additionally, metabolites associated with ST:STUDY_SUMMARY antimicrobial properties and stress responses were identified, underscoring the ST:STUDY_SUMMARY interplay between barley and microbial metabolic processes during malting. This ST:STUDY_SUMMARY comprehensive profiling advances our understanding of malting and suggests ST:STUDY_SUMMARY potential markers for process monitoring and quality control, ultimately ST:STUDY_SUMMARY enhancing malt quality and beer production. ST:INSTITUTE USDA Agricultural Research Service ST:DEPARTMENT Cereal Crops Research Unit ST:LABORATORY Whitcomb lab ST:LAST_NAME Rani ST:FIRST_NAME Heena ST:ADDRESS 502 Walnut Street, Madison, WI 53726 ST:EMAIL bansalheena10@gmail.com ST:PHONE 7657759366 ST:SUBMIT_DATE 2024-06-17 #SUBJECT SU:SUBJECT_TYPE Plant SU:SUBJECT_SPECIES Hordeum vulgare subsp. vulgare SU:TAXONOMY_ID 112509 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - 22-1-blank-blank-Blank Sample source:Seed | Genotype:Conrad | Treatment:BLANK | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_Blank_002_01.mzML,20230210-SW-908_POS_Blank_002_02.mzML SUBJECT_SAMPLE_FACTORS - 39-1-blank-blank-Blank Sample source:Seed | Genotype:Conrad | Treatment:BLANK | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_Blank_001_01.mzML,20230210-SW-908_POS_Blank_001_02.mzML SUBJECT_SAMPLE_FACTORS - 55-1-blank-blank-Blank Sample source:Seed | Genotype:Conrad | Treatment:BLANK | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_Blank_003_01.mzML,20230210-SW-908_POS_Blank_003_02.mzML SUBJECT_SAMPLE_FACTORS - 230327dEGsa01_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa01_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa08_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa08_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa15_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa15_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa17_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa17_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa18_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa18_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa22_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa22_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa37_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa37_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa38_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa38_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa41_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa41_1.cdf SUBJECT_SAMPLE_FACTORS - 13-1-2-dog0-14 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_011_01.mzML,20230210-SW-908_POS_011_02.mzML SUBJECT_SAMPLE_FACTORS - 16-1-2-dog0-10 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_013_01.mzML,20230210-SW-908_POS_013_02.mzML SUBJECT_SAMPLE_FACTORS - 18-1-2-dog0-17 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_015_01.mzML,20230210-SW-908_POS_015_02.mzML SUBJECT_SAMPLE_FACTORS - 27-1-2-dog0-15 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_022_01.mzML,20230210-SW-908_POS_022_02.mzML SUBJECT_SAMPLE_FACTORS - 41-1-2-dog0-18 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_033_01.mzML,20230210-SW-908_POS_033_02.mzML SUBJECT_SAMPLE_FACTORS - 48-1-2-dog0-16 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_039_01.mzML,20230210-SW-908_POS_039_02.mzML SUBJECT_SAMPLE_FACTORS - 56-1-2-dog0-12 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_045_01.mzML,20230210-SW-908_POS_045_02.mzML SUBJECT_SAMPLE_FACTORS - 63-1-2-dog0-11 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_051_01.mzML,20230210-SW-908_POS_051_02.mzML SUBJECT_SAMPLE_FACTORS - 64-1-2-dog0-13 Sample source:Seed | Genotype:Conrad | Treatment:DOG0 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_052_01.mzML,20230210-SW-908_POS_052_02.mzML SUBJECT_SAMPLE_FACTORS - 230327dEGsa04_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa04_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa06_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa06_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa12_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa12_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa23_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa23_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa25_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa25_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa29_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa29_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa47_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa47_1.cdf SUBJECT_SAMPLE_FACTORS - 230328dEGsa01_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230328dEGsa01_1.cdf SUBJECT_SAMPLE_FACTORS - 230328dEGsa03_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230328dEGsa03_1.cdf SUBJECT_SAMPLE_FACTORS - 14-1-3-dog1-22 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_012_01.mzML,20230210-SW-908_POS_012_02.mzML SUBJECT_SAMPLE_FACTORS - 20-1-3-dog1-26 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_017_01.mzML,20230210-SW-908_POS_017_02.mzML SUBJECT_SAMPLE_FACTORS - 21-1-3-dog1-23 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_018_01.mzML,20230210-SW-908_POS_018_02.mzML SUBJECT_SAMPLE_FACTORS - 28-1-3-dog1-21 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_023_01.mzML,20230210-SW-908_POS_023_02.mzML SUBJECT_SAMPLE_FACTORS - 29-1-3-dog1-24 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_024_01.mzML,20230210-SW-908_POS_024_02.mzML SUBJECT_SAMPLE_FACTORS - 4-1-3-dog1-20 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_003_01.mzML,20230210-SW-908_POS_003_02.mzML SUBJECT_SAMPLE_FACTORS - 51-1-3-dog1-27 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_042_01.mzML,20230210-SW-908_POS_042_02.mzML SUBJECT_SAMPLE_FACTORS - 53-1-3-dog1-25 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_043_01.mzML,20230210-SW-908_POS_043_02.mzML SUBJECT_SAMPLE_FACTORS - 61-1-3-dog1-19 Sample source:Seed | Genotype:Conrad | Treatment:DOG1 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_049_01.mzML,20230210-SW-908_POS_049_02.mzML SUBJECT_SAMPLE_FACTORS - 230327dEGsa02_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa02_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa07_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa07_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa09_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa09_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa19_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa19_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa24_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa24_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa32_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa32_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa34_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa34_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa35_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa35_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa43_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa43_1.cdf SUBJECT_SAMPLE_FACTORS - 10-1-4-dog3-36 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_008_01.mzML,20230210-SW-908_POS_008_02.mzML SUBJECT_SAMPLE_FACTORS - 17-1-4-dog3-30 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_014_01.mzML,20230210-SW-908_POS_014_02.mzML SUBJECT_SAMPLE_FACTORS - 2-1-4-dog3-29 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_001_01.mzML,20230210-SW-908_POS_001_02.mzML SUBJECT_SAMPLE_FACTORS - 25-1-4-dog3-31 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_020_01.mzML,20230210-SW-908_POS_020_02.mzML SUBJECT_SAMPLE_FACTORS - 36-1-4-dog3-33 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_030_01.mzML,20230210-SW-908_POS_030_02.mzML SUBJECT_SAMPLE_FACTORS - 42-1-4-dog3-28 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_034_01.mzML,20230210-SW-908_POS_034_02.mzML SUBJECT_SAMPLE_FACTORS - 5-1-4-dog3-32 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_004_01.mzML,20230210-SW-908_POS_004_02.mzML SUBJECT_SAMPLE_FACTORS - 57-1-4-dog3-34 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_046_01.mzML,20230210-SW-908_POS_046_02.mzML SUBJECT_SAMPLE_FACTORS - 65-1-4-dog3-35 Sample source:Seed | Genotype:Conrad | Treatment:DOG3 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_053_01.mzML,20230210-SW-908_POS_053_02.mzML SUBJECT_SAMPLE_FACTORS - 230327dEGsa03_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa03_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa05_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa05_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa10_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa10_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa28_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa28_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa31_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa31_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa33_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa33_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa36_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa36_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa39_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa39_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa46_1 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa46_1.cdf SUBJECT_SAMPLE_FACTORS - 11-1-5-dog5-42 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_009_01.mzML,20230210-SW-908_POS_009_02.mzML SUBJECT_SAMPLE_FACTORS - 19-1-5-dog5-45 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_016_01.mzML,20230210-SW-908_POS_016_02.mzML SUBJECT_SAMPLE_FACTORS - 24-1-5-dog5-37 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_019_01.mzML,20230210-SW-908_POS_019_02.mzML SUBJECT_SAMPLE_FACTORS - 26-1-5-dog5-38 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_021_01.mzML,20230210-SW-908_POS_021_02.mzML SUBJECT_SAMPLE_FACTORS - 38-1-5-dog5-40 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_031_01.mzML,20230210-SW-908_POS_031_02.mzML SUBJECT_SAMPLE_FACTORS - 40-1-5-dog5-44 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_032_01.mzML,20230210-SW-908_POS_032_02.mzML SUBJECT_SAMPLE_FACTORS - 47-1-5-dog5-43 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_038_01.mzML,20230210-SW-908_POS_038_02.mzML SUBJECT_SAMPLE_FACTORS - 62-1-5-dog5-41 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_050_01.mzML,20230210-SW-908_POS_050_02.mzML SUBJECT_SAMPLE_FACTORS - 66-1-5-dog5-39 Sample source:Seed | Genotype:Conrad | Treatment:DOG5 | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_054_01.mzML,20230210-SW-908_POS_054_02.mzML SUBJECT_SAMPLE_FACTORS - 230327dEGsa13_1 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa13_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa14_1 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa14_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa16_2 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa16_2.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa21_1 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa21_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa26_1 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa26_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa27_1 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa27_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa30_1 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa30_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa40_1 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa40_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa42_1 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa42_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa48_1 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa48_1.cdf SUBJECT_SAMPLE_FACTORS - 35-1-1-dry-4 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_029_01.mzML,20230210-SW-908_POS_029_02.mzML SUBJECT_SAMPLE_FACTORS - 43-1-1-dry-2 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_035_01.mzML,20230210-SW-908_POS_035_02.mzML SUBJECT_SAMPLE_FACTORS - 44-1-1-dry-8 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_036_01.mzML,20230210-SW-908_POS_036_02.mzML SUBJECT_SAMPLE_FACTORS - 46-1-1-dry-9 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_037_01.mzML,20230210-SW-908_POS_037_02.mzML SUBJECT_SAMPLE_FACTORS - 49-1-1-dry-1 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_040_01.mzML,20230210-SW-908_POS_040_02.mzML SUBJECT_SAMPLE_FACTORS - 50-1-1-dry-7 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_041_01.mzML,20230210-SW-908_POS_041_02.mzML SUBJECT_SAMPLE_FACTORS - 54-1-1-dry-3 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_044_01.mzML,20230210-SW-908_POS_044_02.mzML SUBJECT_SAMPLE_FACTORS - 58-1-1-dry-6 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_047_01.mzML,20230210-SW-908_POS_047_02.mzML SUBJECT_SAMPLE_FACTORS - 7-1-1-dry-5 Sample source:Seed | Genotype:Conrad | Treatment:DRY | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_006_01.mzML,20230210-SW-908_POS_006_02.mzML SUBJECT_SAMPLE_FACTORS - 230327dEGsa11_1 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa11_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa20_1 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa20_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa44_1 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa44_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa45_1 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa45_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa49_1 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa49_1.cdf SUBJECT_SAMPLE_FACTORS - 230327dEGsa50_1 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230327dEGsa50_1.cdf SUBJECT_SAMPLE_FACTORS - 230328dEGsa02_1 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230328dEGsa02_1.cdf SUBJECT_SAMPLE_FACTORS - 230328dEGsa04_1 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230328dEGsa04_1.cdf SUBJECT_SAMPLE_FACTORS - 12-1-6-kilned-54 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_010_01.mzML,20230210-SW-908_POS_010_02.mzML SUBJECT_SAMPLE_FACTORS - 31-1-6-kilned-53 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_025_01.mzML,20230210-SW-908_POS_025_02.mzML SUBJECT_SAMPLE_FACTORS - 3-1-6-kilned-46 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_002_01.mzML,20230210-SW-908_POS_002_02.mzML SUBJECT_SAMPLE_FACTORS - 32-1-6-kilned-50 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_026_01.mzML,20230210-SW-908_POS_026_02.mzML SUBJECT_SAMPLE_FACTORS - 33-1-6-kilned-47 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_027_01.mzML,20230210-SW-908_POS_027_02.mzML SUBJECT_SAMPLE_FACTORS - 34-1-6-kilned-48 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_028_01.mzML,20230210-SW-908_POS_028_02.mzML SUBJECT_SAMPLE_FACTORS - 59-1-6-kilned-49 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_048_01.mzML,20230210-SW-908_POS_048_02.mzML SUBJECT_SAMPLE_FACTORS - 6-1-6-kilned-51 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_005_01.mzML,20230210-SW-908_POS_005_02.mzML SUBJECT_SAMPLE_FACTORS - 9-1-6-kilned-52 Sample source:Seed | Genotype:Conrad | Treatment:KILNED | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_007_01.mzML,20230210-SW-908_POS_007_02.mzML SUBJECT_SAMPLE_FACTORS - 230328dEGsa05_1 Sample source:Seed | Genotype:Conrad | Treatment:QC_GC | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230328dEGsa05_1.cdf SUBJECT_SAMPLE_FACTORS - 230328dEGsa06_1 Sample source:Seed | Genotype:Conrad | Treatment:QC_GC | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230328dEGsa06_1.cdf SUBJECT_SAMPLE_FACTORS - 230328dEGsa07_1 Sample source:Seed | Genotype:Conrad | Treatment:QC_GC | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230328dEGsa07_1.cdf SUBJECT_SAMPLE_FACTORS - 230328dEGsa08_1 Sample source:Seed | Genotype:Conrad | Treatment:QC_GC | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230328dEGsa08_1.cdf SUBJECT_SAMPLE_FACTORS - 230328dEGsa09_1 Sample source:Seed | Genotype:Conrad | Treatment:QC_GC | Platform:GC-MS RAW_FILE_NAME(Raw file name)=230328dEGsa09_1.cdf SUBJECT_SAMPLE_FACTORS - 1-1-qc-qc-QC1 Sample source:Seed | Genotype:Conrad | Treatment:QC_LC | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_QC_001_01.mzML, 20230210-SW-908_POS_QC_001_02.mzML SUBJECT_SAMPLE_FACTORS - 15-1-qc-qc-QC3 Sample source:Seed | Genotype:Conrad | Treatment:QC_LC | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_QC_003_01.mzML,20230210-SW-908_POS_QC_003_02.mzML SUBJECT_SAMPLE_FACTORS - 23-1-qc-qc-QC4 Sample source:Seed | Genotype:Conrad | Treatment:QC_LC | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_QC_004_01.mzML,20230210-SW-908_POS_QC_004_02.mzML SUBJECT_SAMPLE_FACTORS - 30-1-qc-qc-QC5 Sample source:Seed | Genotype:Conrad | Treatment:QC_LC | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_QC_005_01.mzML,20230210-SW-908_POS_QC_005_02.mzML SUBJECT_SAMPLE_FACTORS - 37-1-qc-qc-QC6 Sample source:Seed | Genotype:Conrad | Treatment:QC_LC | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_QC_006_01.mzML,20230210-SW-908_POS_QC_006_02.mzML SUBJECT_SAMPLE_FACTORS - 45-1-qc-qc-QC7 Sample source:Seed | Genotype:Conrad | Treatment:QC_LC | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_QC_007_01.mzML,20230210-SW-908_POS_QC_007_02.mzML SUBJECT_SAMPLE_FACTORS - 52-1-qc-qc-QC8 Sample source:Seed | Genotype:Conrad | Treatment:QC_LC | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_QC_008_01.mzML,20230210-SW-908_POS_QC_008_02.mzML SUBJECT_SAMPLE_FACTORS - 60-1-qc-qc-QC9 Sample source:Seed | Genotype:Conrad | Treatment:QC_LC | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_QC_009_01.mzML,20230210-SW-908_POS_QC_009_02.mzML SUBJECT_SAMPLE_FACTORS - 67-1-qc-qc-QC10 Sample source:Seed | Genotype:Conrad | Treatment:QC_LC | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_QC_010_01.mzML,20230210-SW-908_POS_QC_010_02.mzML SUBJECT_SAMPLE_FACTORS - 8-1-qc-qc-QC2 Sample source:Seed | Genotype:Conrad | Treatment:QC_LC | Platform:LC-MS RAW_FILE_NAME(Raw file name)=20230210-SW-908_POS_QC_002_01.mzML,20230210-SW-908_POS_QC_002_02.mzML #COLLECTION CO:COLLECTION_SUMMARY 9 samples were collected (three per micro-malting replicate) at 6 different CO:COLLECTION_SUMMARY stages: unmalted mature grain (DRY), out of steep/post-step (DOG0), on CO:COLLECTION_SUMMARY alternating days of germination (DOG1, DOG3, and DOG5), and at the end of CO:COLLECTION_SUMMARY kilning (KILNED). Samples were collected in Eppendorf tubes, immediately flash CO:COLLECTION_SUMMARY frozen in liquid nitrogen and subsequently stored at −80°C until grinding. CO:SAMPLE_TYPE Barley seeds #TREATMENT TR:TREATMENT_SUMMARY 9 samples were collected (three per micro-malting replicate) at 6 different TR:TREATMENT_SUMMARY stages: unmalted mature grain (DRY), out of steep/post-step (DOG0), on TR:TREATMENT_SUMMARY alternating days of germination (DOG1, DOG3, and DOG5), and at the end of TR:TREATMENT_SUMMARY kilning (KILNED) #SAMPLEPREP SP:SAMPLEPREP_SUMMARY For grinding, the seeds were lyophilized, placed in 5 ml polycarbonate vials SP:SAMPLEPREP_SUMMARY along with three stainless steel balls (6.3 mm), and cooled in liquid nitrogen. SP:SAMPLEPREP_SUMMARY The frozen tissue was ground to a fine powder in a 2010 Geno/Grinder (Spex SP:SAMPLEPREP_SUMMARY SamplePrep) with intermittent rests, rapidly re-cooled in liquid nitrogen, and SP:SAMPLEPREP_SUMMARY transferred to their final storage vials. The ground samples were stored at SP:SAMPLEPREP_SUMMARY −80°C until further analysis Preparation of samples for GC-MS analysis Finely SP:SAMPLEPREP_SUMMARY ground samples (4 mg) were extracted using 1 ml of 3:3:2 isopropanol SP:SAMPLEPREP_SUMMARY (IPA)/acetonitrile (ACN)/water (v/v/v) for 6 min at 4°C. After centrifugation, SP:SAMPLEPREP_SUMMARY the supernatant was aliquoted into two equal portions and dried. One aliquot was SP:SAMPLEPREP_SUMMARY derivatized with methoxamine, a mixture of fatty acid methyl esters (FAMEs) SP:SAMPLEPREP_SUMMARY ranging from C08 to C30 was added to each sample, and finally derivatized with SP:SAMPLEPREP_SUMMARY N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA). The prepared samples were SP:SAMPLEPREP_SUMMARY then transferred to vials, sealed, and loaded into the GC-MS instrument. Quality SP:SAMPLEPREP_SUMMARY control (QC) samples were prepared by equally pooling ground powder from all SP:SAMPLEPREP_SUMMARY biological samples and were processed as described above. Method blanks were SP:SAMPLEPREP_SUMMARY prepared using the same 3:3:2 IPA/ACN/water mixture without the addition of SP:SAMPLEPREP_SUMMARY biological sample and processed using the same extraction and processing SP:SAMPLEPREP_SUMMARY procedures as the other samples. Preparation of samples for LC-MS analysis SP:SAMPLEPREP_SUMMARY Finely ground samples (100-130mg) were extracted with 800 μl of 80% IPA in SP:SAMPLEPREP_SUMMARY water for 10 minutes at room temperature. After centrifugation, 700 μl of the SP:SAMPLEPREP_SUMMARY supernatant was transferred to a fresh vial. This process was repeated with SP:SAMPLEPREP_SUMMARY another 700 µl of 80% IPA, and the resulting 1.4 ml of combined supernatant was SP:SAMPLEPREP_SUMMARY discarded. The pellet was re-extracted twice more using 700 µl of 80% methanol. SP:SAMPLEPREP_SUMMARY The 1.4 ml of combined methanol extract was dried under nitrogen and then SP:SAMPLEPREP_SUMMARY reconstituted in 100 µl of methanol. The reconstituted extracts were then SP:SAMPLEPREP_SUMMARY transferred to autosampler vials for UPLC-MS analysis. QCs were prepared by SP:SAMPLEPREP_SUMMARY equally pooling ground powder from all samples and were processed as described SP:SAMPLEPREP_SUMMARY above. Method blanks were prepared and processed in the same manner without the SP:SAMPLEPREP_SUMMARY addition of biological sample. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Injection temperature: 50°C ramped to 250°C by 12°C s-1. Oven temperature CH:CHROMATOGRAPHY_SUMMARY program: 50°C for 1 min, then ramped at 20°C min-1 to 330°C, held constant CH:CHROMATOGRAPHY_SUMMARY for 5 min. CH:INSTRUMENT_NAME Agilent 7890A CH:COLUMN_NAME Agilent DB5-MS (30m x 0.25mm, 0.25um) CH:COLUMN_TEMPERATURE 50-330°C CH:FLOW_GRADIENT N/A CH:FLOW_RATE 1 ml/min CH:SOLVENT_A Mobile phase: Helium CH:SOLVENT_B N/A CH:CHROMATOGRAPHY_TYPE GC #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Leco Pegasus IV TOF MS:INSTRUMENT_TYPE GC-TOF MS:MS_TYPE EI MS:MS_COMMENTS R package XCMS v3.20.0 was used to process raw data. XCMS steps included: peak MS:MS_COMMENTS detection (CentWave), peak grouping (PeakDensity), retention time correction MS:MS_COMMENTS (PeakGroups), peak regrouping (PeakDensity), and missing peak filling MS:MS_COMMENTS (FillChromPeaks). R package RAMClustR v1.2.4 (Broeckling et al., 2014) was used MS:MS_COMMENTS to normalize, filter, cluster features into spectra, and infer molecular MS:MS_COMMENTS weights. Missing values were replaced with small values simulating noise: for MS:MS_COMMENTS each feature, the replacement value was equal to the absolute value of 0.5 times MS:MS_COMMENTS the minimum detected value of that feature. Features were normalized by linearly MS:MS_COMMENTS regressing run order versus QC feature intensities to account for instrument MS:MS_COMMENTS signal intensity drift. Only features showing significant regression (p < MS:MS_COMMENTS 0.05, r-squared > 0.1) were corrected. Features failing to demonstrate at MS:MS_COMMENTS least a 2-fold greater signal intensity in QC samples compared to blanks were MS:MS_COMMENTS removed. Subsequent filtering based on QC sample coefficient of variation (CV) MS:MS_COMMENTS values retained only features with CV values ≤ 0.5 in MS or MSMS data sets. MS:MS_COMMENTS Features were clustered using the ramclustR algorithm (Broeckling et al., 2014). MS:MS_COMMENTS MSFinder v 3.52 (Tsugawa et al., 2016) was used for spectral matching, formula MS:MS_COMMENTS inference, and tentative structure assignment. MSFinder results were imported MS:MS_COMMENTS into the RAMClustR object where a total score was computed based on the product MS:MS_COMMENTS scores from the findmain function and the MSFinder formula and structure scores. MS:MS_COMMENTS Spectra matches took precedence over computational inference-based annotations. MS:MS_COMMENTS CHEBI and COCONUT were set as priority databases, and matches to these databases MS:MS_COMMENTS were given a priority factor value of 1. Matches to databases other than the MS:MS_COMMENTS priority databases were assigned a priority factor of 0.9. A custom database of MS:MS_COMMENTS compounds found in barley was generated based on associations from FooDB and MS:MS_COMMENTS PubChem. The list of 8858 InChIKey structures in this custom barley database was MS:MS_COMMENTS also used for annotation prioritization. Annotations with InChIKey(s) that MS:MS_COMMENTS didn’t match those in the custom barley database were assigned an InChIKey MS:MS_COMMENTS priority factor of 0.9 to decrease scores for these annotations). The annotation MS:MS_COMMENTS with the highest total score was selected for each compound. MS:ION_MODE POSITIVE MS:MS_RESULTS_FILE ST003289_AN005387_Results.txt UNITS:Peak Height Has m/z:Yes Has RT:Yes RT units:Retention index #END