#METABOLOMICS WORKBENCH sven_haange_20220819_020051 DATATRACK_ID:3423 STUDY_ID:ST002279 ANALYSIS_ID:AN003723 PROJECT_ID:PR001459 VERSION 1 CREATED_ON August 24, 2022, 9:54 pm #PROJECT PR:PROJECT_TITLE Modulation of blood metabolites by dietary β–glucan in rainbow trout PR:PROJECT_TITLE (Oncorhynchus mykiss) PR:PROJECT_SUMMARY Prebiotics are known to have a positive impact on fish health and growth rate, PR:PROJECT_SUMMARY and β-glucans are among the most used prebiotics on the market. In this study, PR:PROJECT_SUMMARY rainbow trout (Oncorhynchus mykiss) were treated with a β-1,3;1,6-glucan PR:PROJECT_SUMMARY dietary supplement (at a dose of 0 g, 1 g, 10 g, and 50 g β-glucan per kg of PR:PROJECT_SUMMARY feed). After 6 weeks, the effect of the β-glucan was evaluated by determining PR:PROJECT_SUMMARY the changes in the microbiota and the blood serum metabolites in the fish PR:INSTITUTE Helmholtz Centre for Environmental Research - UFZ PR:DEPARTMENT Molecular Systems Biology PR:LABORATORY Functional Metabolomics PR:LAST_NAME Haange PR:FIRST_NAME Sven PR:ADDRESS Permoserstrasse 1, Leipzig, Saxony, 04318, Germany PR:EMAIL sven.haange@ufz.de PR:PHONE 0049 341 2351099 #STUDY ST:STUDY_TITLE Modulation of blood metabolites by dietary β–glucan in rainbow trout ST:STUDY_TITLE (Oncorhynchus mykiss) - Serum amino acids and biogenic amines concentrations ST:STUDY_SUMMARY Measurement of changes in serum amino acids and biogenic amines levels using ST:STUDY_SUMMARY targeted mass spectrometry ST:INSTITUTE Helmholtz Centre for Environmental Research - UFZ ST:LAST_NAME Haange ST:FIRST_NAME Sven ST:ADDRESS Permoserstrasse 1, Leipzig, Saxony, 04318, Germany ST:EMAIL sven.haange@ufz.de ST:PHONE 0049 341 2351099 #SUBJECT SU:SUBJECT_TYPE Fish SU:SUBJECT_SPECIES Oncorhynchus mykiss #FACTORS #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 - 1024025731 Sample time:0h | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_03_0_1_1_00_1024025731.wiff SUBJECT_SAMPLE_FACTORS - 1024025813 Sample time:6_weeks | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_04_0_1_1_00_1024025813.wiff SUBJECT_SAMPLE_FACTORS - 1024025890 Sample time:6_weeks | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_05_0_1_1_00_1024025890.wiff SUBJECT_SAMPLE_FACTORS - 1024028825 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_06_0_1_1_00_1024028825.wiff SUBJECT_SAMPLE_FACTORS - 1024028908 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_07_0_1_1_00_1024028908.wiff SUBJECT_SAMPLE_FACTORS - 1024019677 Sample time:6_weeks | Treatment:5% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_08_0_1_1_00_1024019677.wiff SUBJECT_SAMPLE_FACTORS - 1024019750 Sample time:6_weeks | Treatment:5% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_09_0_1_1_00_1024019750.wiff SUBJECT_SAMPLE_FACTORS - 1024025745 Sample time:0h | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_15_0_1_1_00_1024025745.wiff SUBJECT_SAMPLE_FACTORS - 1024028757 Sample time:6_weeks | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_17_0_1_1_00_1024028757.wiff SUBJECT_SAMPLE_FACTORS - 1024025828 Sample time:6_weeks | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_16_0_1_1_00_1024025828.wiff SUBJECT_SAMPLE_FACTORS - 1024028912 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_19_0_1_1_00_1024028912.wiff SUBJECT_SAMPLE_FACTORS - 1024028830 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_18_0_1_1_00_1024028830.wiff SUBJECT_SAMPLE_FACTORS - 1024019764 Sample time:6_weeks | Treatment:5% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_21_0_1_1_00_1024019764.wiff SUBJECT_SAMPLE_FACTORS - 1024019681 Sample time:6_weeks | Treatment:5% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_20_0_1_1_00_1024019681.wiff SUBJECT_SAMPLE_FACTORS - 1024025750 Sample time:0h | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_27_0_1_1_00_1024025750.wiff SUBJECT_SAMPLE_FACTORS - 1024025832 Sample time:6_weeks | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_28_0_1_1_00_1024025832.wiff SUBJECT_SAMPLE_FACTORS - 1024028927 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_31_0_1_1_00_1024028927.wiff SUBJECT_SAMPLE_FACTORS - 1024028844 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_30_0_1_1_00_1024028844.wiff SUBJECT_SAMPLE_FACTORS - 1024019696 Sample time:6_weeks | Treatment:5% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_32_0_1_1_00_1024019696.wiff SUBJECT_SAMPLE_FACTORS - 1024025764 Sample time:0h | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_39_0_1_1_00_1024025764.wiff SUBJECT_SAMPLE_FACTORS - 1024028859 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_42_0_1_1_00_1024028859.wiff SUBJECT_SAMPLE_FACTORS - 1024028931 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_43_0_1_1_00_1024028931.wiff SUBJECT_SAMPLE_FACTORS - 1024025847 Sample time:6_weeks | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_40_0_1_1_00_1024025847.wiff SUBJECT_SAMPLE_FACTORS - 1024028776 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_41_0_1_1_00_1024028776.wiff SUBJECT_SAMPLE_FACTORS - 1024019707 Sample time:6_weeks | Treatment:5% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_44_0_1_1_00_1024019707.wiff SUBJECT_SAMPLE_FACTORS - 1024025779 Sample time:0h | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_51_0_1_1_00_1024025779.wiff SUBJECT_SAMPLE_FACTORS - 1024019639 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_55_0_1_1_00_1024019639.wiff SUBJECT_SAMPLE_FACTORS - 1024028863 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_54_0_1_1_00_1024028863.wiff SUBJECT_SAMPLE_FACTORS - 1024028781 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_53_0_1_1_00_1024028781.wiff SUBJECT_SAMPLE_FACTORS - 1024025851 Sample time:6_weeks | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_52_0_1_1_00_1024025851.wiff SUBJECT_SAMPLE_FACTORS - 1024019711 Sample time:6_weeks | Treatment:5% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_56_0_1_1_00_1024019711.wiff SUBJECT_SAMPLE_FACTORS - 1024025783 Sample time:0h | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_63_0_1_1_00_1024025783.wiff SUBJECT_SAMPLE_FACTORS - 1024019726 Sample time:6_weeks | Treatment:5% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_68_0_1_1_00_1024019726.wiff SUBJECT_SAMPLE_FACTORS - 1024025866 Sample time:6_weeks | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_64_0_1_1_00_1024025866.wiff SUBJECT_SAMPLE_FACTORS - 1024028795 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_65_0_1_1_00_1024028795.wiff SUBJECT_SAMPLE_FACTORS - 1024028878 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_66_0_1_1_00_1024028878.wiff SUBJECT_SAMPLE_FACTORS - 1024019643 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_67_0_1_1_00_1024019643.wiff SUBJECT_SAMPLE_FACTORS - 1024025871 Sample time:6_weeks | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_76_0_1_1_00_1024025871.wiff SUBJECT_SAMPLE_FACTORS - 1024028806 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_77_0_1_1_00_1024028806.wiff SUBJECT_SAMPLE_FACTORS - 1024028882 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_78_0_1_1_00_1024028882.wiff SUBJECT_SAMPLE_FACTORS - 1024019658 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_79_0_1_1_00_1024019658.wiff SUBJECT_SAMPLE_FACTORS - 1024025711 Sample time:0h | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_74_0_1_1_00_1024025711.wiff SUBJECT_SAMPLE_FACTORS - 1024025798 Sample time:0h | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_75_0_1_1_00_1024025798.wiff SUBJECT_SAMPLE_FACTORS - 1024025809 Sample time:0h | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_87_0_1_1_00_1024025809.wiff SUBJECT_SAMPLE_FACTORS - 1024025726 Sample time:0h | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_86_0_1_1_00_1024025726.wiff SUBJECT_SAMPLE_FACTORS - 1024019731 Sample time:6_weeks | Treatment:5% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_80_0_1_1_00_1024019731.wiff SUBJECT_SAMPLE_FACTORS - 1024019745 Sample time:6_weeks | Treatment:5% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_92_0_1_1_00_1024019745.wiff SUBJECT_SAMPLE_FACTORS - 1024028811 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_89_0_1_1_00_1024028811.wiff SUBJECT_SAMPLE_FACTORS - 1024025885 Sample time:6_weeks | Treatment:no ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_88_0_1_1_00_1024025885.wiff SUBJECT_SAMPLE_FACTORS - 1024028897 Sample time:6_weeks | Treatment:0,1% ß-glucan RAW_FILE_NAME=KIT2-0-5504_1024019783_90_0_1_1_00_1024028897.wiff #COLLECTION CO:COLLECTION_SUMMARY For the metabolite and microbiota analyses, 10 fish from the control group were CO:COLLECTION_SUMMARY collected on day 0 and 10 fish from each group (control, 0.1, 1.0, 5.0% CO:COLLECTION_SUMMARY β-glucan) at week 6. Fish were euthanized by immersion in an overdose (200 CO:COLLECTION_SUMMARY mg∙L-1) of MS-222 (cat no. A5040, Sigma-Aldrich). The tail was cut, and blood CO:COLLECTION_SUMMARY was sampled from the vena caudalis using Na-heparinised 25 mL and 50 mL CO:COLLECTION_SUMMARY capillary pipettes (Hirschmann Laborgeräte, Germany). Blood samples were CO:COLLECTION_SUMMARY centrifuged at 3,000 g for 10 min at 4 °C, and serum was isolated and stored at CO:COLLECTION_SUMMARY -80 °C. The intestine was aseptically sampled and immediately immersed in CO:COLLECTION_SUMMARY RNAlater® (Sigma-Aldrich) and transferred to -20 °C. CO:SAMPLE_TYPE Blood (serum) CO:STORAGE_CONDITIONS -20℃ #TREATMENT TR:TREATMENT_SUMMARY A commercial pelleted (1.5 mm) trout feed (INICIO 917, BioMar A/S) based on a TR:TREATMENT_SUMMARY mixture of fish and plant protein and containing 47% crude protein, 20% crude TR:TREATMENT_SUMMARY lipid, 18% carbohydrates (NFE), 1.2% fibre, 8.5% ash, and 1.1% phosphorus was TR:TREATMENT_SUMMARY used for preparation of the diets. A β-1,3;1,6-glucan (purity: 81.6%; mean TR:TREATMENT_SUMMARY particle size: 37.7 µm; supplementary table S2) purified from yeast TR:TREATMENT_SUMMARY (Saccharomyces cerevisiae) (Biorigin, Brazil) was supplemented to this diet at TR:TREATMENT_SUMMARY varying doses: 0 g, 1 g, 10 g, and 50 g β-glucan were added to 1 kg of INICIO TR:TREATMENT_SUMMARY 917, respectively, during continuous mixing and sealed to the pellets by TR:TREATMENT_SUMMARY spraying with 30 mL organic, cold-pressed rapeseed oil (Gyldenmark), resulting TR:TREATMENT_SUMMARY in a control diet without β-glucan and three experimental diets of 0.1%, 1.0%, TR:TREATMENT_SUMMARY and 5.0% Wt/Wt β-glucan. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Sample preparation for MetIDQ p180 Kit measurement Solvents: Acetonitril (Merck SP:SAMPLEPREP_SUMMARY KGaA, Darmstadt, Germany hypergrade for LC-MS) Water MiliQ, Extracting agent - SP:SAMPLEPREP_SUMMARY ACN / H2O (1:1) Equipment 4 steel balls size M Eppendorf Tubes 2mL Tissue slicer SP:SAMPLEPREP_SUMMARY (Rettberg, Germany) Centrifuge (Sigma) Work steps Approximately 100 mg of sample SP:SAMPLEPREP_SUMMARY and 4 steel balls of size M into each tube. Per mg of sample 5 µL of extracting SP:SAMPLEPREP_SUMMARY agent was added. Shake the samples for 10 minutes at 30 Hz in the tissue slicer SP:SAMPLEPREP_SUMMARY and centrifuge for 2 minutes at 14000 rpm. 10µL of supernatant was used for the SP:SAMPLEPREP_SUMMARY targeted analytics. For blood sample analysis 10 µL plasma was taken. Kit SP:SAMPLEPREP_SUMMARY reparation The analysis was performed using the validated MetIDQ p180 Kit and SP:SAMPLEPREP_SUMMARY described in Siskos et al. [1]. Data processing was carried out with the SP:SAMPLEPREP_SUMMARY provided quantitation method Kit (Biocrates Life Sciences AG, Innsbruck, SP:SAMPLEPREP_SUMMARY Austria). 1 Siskos AP, Jain P, Römisch-Margl W, Bennett M, Achaintre D, Asad Y, SP:SAMPLEPREP_SUMMARY et al. Interlaboratory Reproducibility of a Targeted Metabolomics Platform for SP:SAMPLEPREP_SUMMARY Analysis of Human Serum and Plasma. Anal Chem 2017;89:656-65. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY LC - Instrument Parameters AbsoluteIDQ® p180 Kit (Biocrates Life Science AG, CH:CHROMATOGRAPHY_SUMMARY Innsbruck, Austria) System: UPLC (Waters Acquity, Waters Corporation, Milford, CH:CHROMATOGRAPHY_SUMMARY USA) Column: Agilent, Zorbax Eclipse XDB C18, 3.0 x 100 mm, 3.5 µM, Agilent CH:CHROMATOGRAPHY_SUMMARY Waldbronn, Germany Precolumn: Security Guard, Phenomenex, C18, 4 x 3 mm; CH:CHROMATOGRAPHY_SUMMARY Phenomenex, Aschaffenburg, Germany Materials: Water MiliQ, Methanol (Merck KGaA, CH:CHROMATOGRAPHY_SUMMARY Darmstadt, Germany, hypergrade for LC-MS) Acetonitril (Merck KGaA, Darmstadt, CH:CHROMATOGRAPHY_SUMMARY Germany hypergrade for LC-MS) Ammonium Acetate (Honeywell - Fluka, Seelze, CH:CHROMATOGRAPHY_SUMMARY Germany) Formic Acid (Honeywell, Fluka, Seelze, Germany) Running Solvent: 5mM CH:CHROMATOGRAPHY_SUMMARY ammonium acetat in methanol FIA: B: Mix running solvent and MiliQ water 1:1 CH:CHROMATOGRAPHY_SUMMARY Gradient Table FIA Time (min) Flow Rate (mL/min) %A %B Curve Initial 0.030 0.0 CH:CHROMATOGRAPHY_SUMMARY 100.0 Initial 1.60 0.030 0.0 100.0 6 2.40 0.200 0.0 100.0 6 2.80 0.200 0.0 100.0 CH:CHROMATOGRAPHY_SUMMARY 6 3.00 0.030 0.0 100.0 6 CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME UPLC (Waters Acquity, Waters Corporation, Milford, USA) CH:COLUMN_NAME Agilent, Zorbax Eclipse XDB C18, 3.0 x 100 mm, 3.5 µM, Agilent Waldbronn, CH:COLUMN_NAME Germany #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME ABI Sciex 5500 QTrap MS:INSTRUMENT_TYPE QTRAP MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS MS - Instrument Parameter AbsoluteIDQ® p180 Kit (Biocrates Life Science AG, MS:MS_COMMENTS Innsbruck, Austria) MS -Q-Trap 5500 Sciex Ion Source: Turbo Spray Curtian Gas: MS:MS_COMMENTS 20 CAD Gas: Medium Ion Spray Voltage: 5500 V Temperature: 500°C Ion Source Gas MS:MS_COMMENTS 1: 40psi Ion Source Gas 2: 50 psi #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS µM MS_METABOLITE_DATA_START Samples 1024025731 1024025813 1024025890 1024028825 1024028908 1024019677 1024019750 1024025745 1024028757 1024025828 1024028912 1024028830 1024019764 1024019681 1024025750 1024025832 1024028927 1024028844 1024019696 1024025764 1024028859 1024028931 1024025847 1024028776 1024019707 1024025779 1024019639 1024028863 1024028781 1024025851 1024019711 1024025783 1024019726 1024025866 1024028795 1024028878 1024019643 1024025871 1024028806 1024028882 1024019658 1024025711 1024025798 1024025809 1024025726 1024019731 1024019745 1024028811 1024025885 1024028897 Factors Sample time:0h | Treatment:no ß-glucan Sample time:6_weeks | Treatment:no ß-glucan Sample time:6_weeks | Treatment:no ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:5% ß-glucan Sample time:6_weeks | Treatment:5% ß-glucan Sample time:0h | Treatment:no ß-glucan Sample time:6_weeks | Treatment:no ß-glucan Sample time:6_weeks | Treatment:no ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:5% ß-glucan Sample time:6_weeks | Treatment:5% ß-glucan Sample time:0h | Treatment:no ß-glucan Sample time:6_weeks | Treatment:no ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:5% ß-glucan Sample time:0h | Treatment:no ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:no ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:5% ß-glucan Sample time:0h | Treatment:no ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:no ß-glucan Sample time:6_weeks | Treatment:5% ß-glucan Sample time:0h | Treatment:no ß-glucan Sample time:6_weeks | Treatment:5% ß-glucan Sample time:6_weeks | Treatment:no ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:no ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:0h | Treatment:no ß-glucan Sample time:0h | Treatment:no ß-glucan Sample time:0h | Treatment:no ß-glucan Sample time:0h | Treatment:no ß-glucan Sample time:6_weeks | Treatment:5% ß-glucan Sample time:6_weeks | Treatment:5% ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Sample time:6_weeks | Treatment:no ß-glucan Sample time:6_weeks | Treatment:0,1% ß-glucan Ala 558 630 608 774 771 655 596 483 551 628 704 703 734 622 629 509 440 674 702 613 735 603 821 1030 839 516 824 707 743 578 923 528 673 565 624 633 433 547 490 688 695 500 569 375 548 679 588 497 534 673 Arg 136 196 158 145 135 167 90.3 136 150 148 187 163 97.9 144 124 160 85.8 151 156 138 168 141 197 181 157 120 119 117 117 181 128 144 158 156 112 195 83.8 111 67.2 119 83.5 113 127 112 175 143 110 185 161 108 Asn 14.7 21.4 12 39.9 36.3 16.5 27.9 20.2 13.6 26.6 18 20 11.9 29 11.5 17.9 23.4 20.1 22.4 13.3 18.1 18.5 15.9 25.3 40.5 15.9 17.9 25 31.8 16.7 16.3 15 39.3 33.7 19.3 22 16.8 18.6 16.2 19 15.4 10.3 17.9 10.3 13.6 26.5 13.6 25.9 16.7 26 Asp 153 179 134 110 180 125 113 147 216 135 206 164 199 95.9 182 121 85.1 151 179 133 168 114 236 183 199 122 157 169 166 166 128 164 137 117 201 204 97.7 232 65.6 175 168 118 197 121 128 116 116 215 149 169 Cit 16.6 15.3 11.5 28.9 8.29 9.64 9.49 19.4 12.4 15.3 18.8 16.7 10.9 10.2 20.4 15.8 7.77 13.9 8.8 16.3 20.7 12.1 25.2 11.3 20.1 22.2 13 14.5 13.4 18.5 13.9 14.3 62.4 9.43 14.3 12.9 7.67 18.6 9.75 8.79 8.5 22.1 21.6 15.6 25.9 14.3 12.7 27.4 19.7 9.58 Gln 317 441 413 528 560 504 485 426 415 525 289 447 396 497 322 290 302 558 433 310 489 431 616 720 603 336 413 364 414 405 467 207 683 409 511 542 329 401 321 495 427 282 308 177 356 475 464 434 441 406 Glu 184 220 136 122 147 129 115 146 169 180 220 150 187 125 184 126 83.4 155 153 232 155 117 223 141 165 164 190 177 181 142 180 172 127 184 189 163 106 176 114 155 147 150 188 113 156 118 158 162 167 129 Gly 2920 2920 1870 1520 2360 1680 1670 2140 1990 1880 2240 2540 1670 1220 2250 2650 1680 1690 2390 2220 2130 1260 1920 2320 1920 2640 2260 1940 2390 1740 2750 1990 2080 1870 1970 2060 1300 1870 1050 1720 1740 2370 1810 1670 1790 1590 1560 2250 1850 1510 His 147 171 119 122 182 121 175 120 125 120 105 166 121 133 125 138 103 170 142 131 124 114 178 171 146 142 118 165 123 118 156 100 144 102 131 180 145 99.1 96 178 144 134 126 86.2 126 164 126 157 101 124 Ile 115 102 136 212 181 161 146 98.5 152 129 127 166 120 151 119 90.2 106 200 199 146 174 115 189 291 173 124 111 154 131 121 150 114 198 134 145 104 109 116 98.8 137 139 130 115 60.3 135 152 134 108 149 164 Leu 211 208 276 367 333 268 223 190 299 272 255 293 243 283 246 171 170 344 353 271 319 237 417 485 317 237 196 292 240 265 275 223 359 250 262 218 204 262 172 241 249 248 215 121 249 264 246 238 302 303 Lys 261 265 298 250 274 220 260 325 290 229 285 272 234 282 222 254 224 271 255 289 376 276 562 372 344 187 241 231 273 349 290 204 335 271 310 509 250 248 128 256 204 197 246 165 274 253 247 315 263 270 Met 112 104 124 115 104 98.5 125 126 119 89.2 106 153 94.1 95.4 150 115 89.8 126 102 95.4 137 88.2 150 133 121 126 101 95.7 86.8 118 109 91.6 152 110 99.5 90.5 103 79.4 70.8 112 98.1 124 105 91.3 154 107 114 186 109 116 Orn 31.7 32.1 37 34 29.4 26.6 41.1 31.1 25.7 25.4 26.8 23.3 42.5 42.6 13.1 29.3 67.2 35.5 30 15.2 29.8 20.6 81.1 40.3 58.2 12.4 48.2 22.9 36.9 23.2 37.7 13.6 65 25.6 43.4 46.2 36.1 35.8 19.3 57.3 38.1 14.5 13 8.43 17.5 38.8 27.1 29.1 24.7 47.2 Phe 188 156 171 180 172 159 154 180 128 159 162 198 111 131 157 137 121 210 149 175 253 154 204 180 158 141 148 145 182 215 179 149 180 196 132 126 118 127 142 141 130 172 159 144 188 123 229 202 152 201 Pro 132 96.1 113 143 143 99.4 51.8 130 126 92.7 106 128 133 88.7 162 111 82.8 103 108 121 138 101 172 236 168 137 98 86.7 110 102 141 87.5 159 91.6 149 135 42.4 126 40.9 107 99.1 111 112 87.1 94.1 88.2 91.1 90.9 119 95 Ser 218 272 202 198 276 205 201 242 205 222 229 164 195 213 227 233 196 275 164 196 257 182 286 255 253 175 160 263 154 260 285 130 203 248 258 142 130 140 112 140 149 202 206 115 230 191 188 471 172 175 Thr 159 249 309 228 293 286 213 158 187 233 240 388 241 307 169 225 200 254 321 176 229 218 350 521 319 182 257 209 256 202 398 133 254 201 340 346 203 160 167 319 225 186 164 103 191 304 196 274 242 293 Trp 35.5 30.1 29.3 33.9 34.2 33 29.7 33 24.1 31.2 24.1 31.3 27.8 30.9 24.2 22.8 26.5 27.2 36.6 29.3 30.9 28.9 34.4 35.5 34.2 15.8 27.9 31.6 25.3 33.5 31.8 19.3 39.2 31.4 29.3 35.3 26.9 23.8 22.6 29.7 26.8 26.1 22.9 17.3 30.6 26.2 30.1 36.7 26.1 28.9 Tyr 110 74.1 88.4 74.3 79.9 82.1 84.3 117 79.3 79.9 85.5 89.7 54.3 85.1 95.9 61.5 57.9 79.7 65.8 65.9 132 57 112 81 90.1 69.9 77.5 64.7 75.3 97.8 61 52.9 100 63 49 51.1 83 47.5 61.8 69.1 57.9 95.7 84.3 85.1 76.1 61.5 134 112 56.4 102 Val 284 257 349 454 418 332 287 281 381 329 296 401 272 361 299 252 233 413 390 306 393 258 504 574 353 313 254 354 336 314 412 278 400 344 339 273 283 336 232 341 316 306 293 184 329 320 312 330 380 371 Ac-Orn 0.059 0.139 0.045 0.098 0.017 0.104 0.04 0.051 0.058 0.051 0.013 0.029 0 0.123 0.107 0.122 0.104 0.095 0.176 0.09 0.164 0.046 0.123 0 0.23 0.031 0.019 0.118 0.104 0.028 0.098 0.072 0.099 0 0.016 0.037 0.02 0.002 0.046 0.06 0.168 0.002 0 0 0.071 0.026 0.009 0.036 0.077 0 ADMA 2.64 1.55 1.48 0 0.787 0.168 0.024 0 0.648 0.345 1.33 1.92 2.49 1.96 1.39 0.317 0.013 0.68 2.11 0.224 1.05 0.867 0.985 1.69 0.952 2.23 2.27 0.334 1.03 0.127 1.77 1.74 0.873 0.718 0.881 0.277 1.05 0.694 0 0.545 0.218 0.166 1.03 0.164 1.84 2.41 1.45 1.75 0 1.28 alpha-AAA 0.958 0.311 0.608 0.665 0.368 0.743 0.07 0.709 1.29 0.497 0.523 0.362 0.632 1.33 0.795 0.854 1.03 0.549 0.531 1.54 1.45 0.551 1.32 0.464 0.508 0.692 0.877 0.705 0.601 0.763 0.874 0.957 0.596 0.388 0.378 0.66 0.83 1.03 0.129 0.748 0.837 0.453 0.667 0.719 0.699 0.735 1.12 0.562 0.641 0.818 c4-OH-Pro 0.005 0 0.002 0.003 0.005 0.005 0 0 0 0.009 0.013 0 0.007 0 0.016 0.019 0 0 0.007 NA 0 0.006 NA 0.007 0.021 0.018 0 0.007 0 0.008 0 0.005 0 0 0.008 0 NA 0 0 0 0.003 0 0.006 0 0 0.007 0.006 0.034 0.004 0.006 Carnosine 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Creatinine 1.77 10.9 17.3 46.3 2.48 11 9.89 5.32 21.7 9.44 14.1 8.13 10.2 11.6 18.4 15.8 7.98 15.4 21.6 4.5 7.4 15.7 9.12 4.42 8.55 5.8 13.6 18.3 9.24 17.5 9.21 4.09 59.5 10.7 10.8 2.01 6.99 8.47 4.48 2.2 8.88 7.04 11.4 6.03 7.44 13.7 8.94 13.7 37.7 11.7 DOPA 0.07 0 0 0.095 0.208 0.013 0.176 0.007 0.1 0.238 0.186 0.209 0.444 0.186 0 0.035 0.19 0.133 0.184 0 0.206 0.032 0.081 0.287 0.324 0.111 0 0.167 0.015 0.037 0.295 0 0.05 0.174 0.281 0.209 0.065 0.028 0.075 0.116 0.058 0.121 0.053 0.153 0 0.274 0.075 0 0.106 0.099 Dopamine 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Histamine 0.304 0.15 0.164 0.197 0.183 0.152 0.164 0.252 0.157 0.161 0.198 0.162 0.152 0.2 0.192 0.165 0.157 0.224 0.155 0.199 0.187 0.222 0.17 0.203 0.149 0.228 0.149 0.144 0.158 0.158 0.181 0.186 0.183 0.174 0.158 0.202 0.158 0.165 0.158 0.186 0.17 0.49 0.173 0.164 0.288 0.232 0.187 0.202 0.166 0.162 Kynurenine 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Met-SO 7.88 7.17 11.4 16.7 11.5 10.9 11.2 8.98 11 6.72 15 18.9 12.5 17.1 12.3 12.6 4.84 19.1 13.1 10.2 11.8 25.2 10.9 15 11.3 23.4 9.08 13.8 9.49 19.2 10.5 9.3 14.7 9.85 17.1 6.66 5.44 6.87 6.62 8.74 13.7 10.7 12.8 10.6 21.6 13.7 15.9 35.9 9.77 11.5 Nitro-Tyr 0 0 0.003 0.013 0.006 0 0.015 0 0 0 0.007 0 0.001 0 0 0 0 0.007 0 0 0 0 0 0 0 0 0 0 0.003 0 0 0 0 0 0 0 0 0 0.005 0 0 0 0 0 0 0 0 0 0 0 PEA 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Putrescine 39.1 66.3 39.5 23.1 34.9 24.8 36.6 30.1 50.2 37.6 74.2 39.7 70 27.9 26.7 49.4 38.4 33.5 52.2 49.4 50.1 33.7 60.8 62.7 33.9 26.4 55.4 51.4 64.9 46.6 37.9 55.6 43.8 56.6 49.1 60.8 49.9 38 46.8 45.4 43 29.6 33.6 18.9 43.5 29.9 52.4 34.4 52.2 45.3 SDMA 1.35 2.59 2.94 0.863 2.46 2.19 3.74 0.397 1.15 2.64 2.73 1.18 2.89 3.19 3.24 0.952 3.36 1.81 2.17 2.11 1.65 1.22 3.06 2.71 3.44 1.99 3.11 2 1.55 2.55 2.96 1.24 2.25 3.08 1.33 3.09 2.25 1.85 1.84 1.43 3.96 1.04 1.89 0.93 2.7 1.44 2.14 1.47 2.16 3.24 Serotonin 0.068 0.057 0.029 0.036 0.036 0.023 0.049 0.017 0.028 0.022 0.033 0.037 0.039 0.037 0.033 0.026 0.03 0.028 0.016 0.07 0.024 0.035 0.019 0.05 0.017 0.037 0.024 0.028 0.031 0.043 0.037 0.036 0.043 0.033 0.029 0.042 0.034 0.063 0.029 0.029 0.045 0.03 0.059 0.036 0.031 0.014 0.027 0.023 0.053 0.013 Spermidine 14.7 73.6 10.3 11.2 23.1 18 24.4 24.8 33.7 47.4 33 27 51.5 24.5 20.5 38.1 39.2 33.5 47.8 39.8 37.9 24.7 37.2 41.3 24.4 9.75 54.4 35 66.9 26.3 30.6 19.7 36 49 45 54.6 48.2 42.1 20.3 35.1 31.6 18.7 20.6 12.9 25.5 29.9 30.8 31.1 50.4 31.9 Spermine 1.13 3.37 0.313 0 0 0 0 1.3 0.672 0 0.702 0 0 0 0 5.05 4.02 0 3.32 3.05 0.132 0.354 1.56 0 0 3.13 0.386 1.62 0 0.463 0.467 0.562 0.475 0.437 0 0 NA 0 0.861 0.853 0.638 0.626 4.16 0.657 2.36 0.499 0.604 0 3.24 0 t4-OH-Pro 273 270 92.8 96.6 103 118 62.5 199 164 157 75.5 105 236 79.7 207 144 46.9 73.2 158 191 104 118 148 146 302 243 116 125 108 127 109 135 115 112 125 227 47.9 99 29.5 65.4 92.6 217 167 149 143 102 68.8 190 79.6 46.6 Taurine 728 759 799 773 746 673 682 682 717 794 656 654 746 773 695 708 760 716 747 746 801 694 729 724 716 678 737 734 679 789 797 662 693 716 713 745 684 721 735 736 693 734 719 691 649 697 645 743 712 682 Total DMA 1.51 3.9 2.32 2.15 2.65 3.01 3.57 0.891 1.76 2.84 3.58 2.32 2.09 3.18 2.43 1.7 1.95 1.61 2.08 2.4 2.47 1.41 2.85 3.23 2.29 1.84 3.58 2.11 2.15 2.88 3.26 1.79 2.55 2.65 2.33 3.89 3.16 1.37 1.58 1.74 2.28 1.56 1.62 1.56 1.94 2.88 2.76 1.9 2.08 3.11 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Biochemical Name KEGG Ala Alanine C00041 Arg Arginine C00062 Asn Asparagine C00152 Asp Aspartate C00049 Cit Citrulline C00327 Gln Glutamine C00064 Glu Glutamate C00025 Gly Glycine C00037 His Histidine C00135 Ile Isoleucine C00407 Leu Leucine C00123 Lys Lysine C00047 Met Methionine C00073 Orn Ornithine C00077 Phe Phenylalanine C00079 Pro Proline C00148 Ser Serine C00065 Thr Threonine C00188 Trp Tryptophan C00078 Tyr Tyrosine C00082 Val Valine C00183 Ac-Orn Acetylornithine C00437 ADMA Asymmetric dimethylarginine C03626 alpha-AAA Alpha- Aminoadipic acid C00956 c4-OH-Pro cis-Hydroxyproline C01015 Carnosine Carnosine C00386 Creatinine Creatinine C00791 DOPA DOPA C00355 Dopamine Dopamine C03758 Histamine Histamine C00388 Kynurenine Kynurenine C00328 Met-SO Methioninesulfoxide C02989 Nitro-Tyr Nitrotyrosine Not available PEA Phenylethylamine C02455 Putrescine Putrescine C00134 SDMA Sarcosine C21189 Serotonin Serotonine C00780 Spermidine Spermidine C00315 Spermine Spermine C00750 t4-OH-Pro trans-Hydroxyproline C01157 Taurine Taurine C00245 Total DMA Total dimethylarginine C03626 METABOLITES_END #END