#METABOLOMICS WORKBENCH houyuanlong2020_20201209_182755 DATATRACK_ID:2345 STUDY_ID:ST001621 ANALYSIS_ID:AN002656 PROJECT_ID:000000 VERSION 1 CREATED_ON December 11, 2020, 9:55 am #PROJECT PR:PROJECT_TITLE Metabolomics analysis of Intestinal Organoids PR:PROJECT_TYPE Untargeted metabolomics analysis PR:PROJECT_SUMMARY Untargeted metabolomics of intestinal organoids form mouse. We performed PR:PROJECT_SUMMARY untargeted metabolomics of intestinal organoids treated with fructose (10 mM) PR:PROJECT_SUMMARY for 24 hr.At the 5th day of secondary organoid culture, the organoids were PR:PROJECT_SUMMARY treated with fructose (10 mM D-fructose)-containing IntestiCult organoid growth PR:PROJECT_SUMMARY medium for 24 hr and washed with pre-cooled PBS before harvesting. PR:INSTITUTE China Pharmaceutical University PR:DEPARTMENT School of Medicine PR:LABORATORY Metabonomics PR:LAST_NAME Hou PR:FIRST_NAME Yuanlong PR:ADDRESS tongjiaxiang, nanjing, jiangsu, 210000, China PR:EMAIL jian2103@163.com PR:PHONE 18851105337 PR:CONTRIBUTORS Xiao Zheng Haiping Hao #STUDY ST:STUDY_TITLE Untargeted metabolomics of intestinal organoids treated with fructose ST:STUDY_TYPE Metabolite profile and pathway analysis ST:STUDY_SUMMARY We performed untargeted metabolomics of intestinal organoids treated with ST:STUDY_SUMMARY fructose (10 mM) for 24 hr.At the 5th day of secondary organoid culture, the ST:STUDY_SUMMARY organoids were treated with fructose (10 mM D-fructose)-containing IntestiCult ST:STUDY_SUMMARY organoid growth medium for 24 hr and washed with pre-cooled PBS before ST:STUDY_SUMMARY harvesting. ST:INSTITUTE China Pharmaceutical University ST:DEPARTMENT School of Medicine ST:LABORATORY Metabonomics ST:LAST_NAME Hou ST:FIRST_NAME Yuanlong ST:ADDRESS tongjiaxiang, nanjing, jiangsu, 210000, China ST:EMAIL jian2103@163.com ST:PHONE 18851105337 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #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 - C1 Treatment:D-HANKS RAW_FILE_NAME=20200501C1 SUBJECT_SAMPLE_FACTORS - C2 Treatment:D-HANKS RAW_FILE_NAME=20200501C2 SUBJECT_SAMPLE_FACTORS - C3 Treatment:D-HANKS RAW_FILE_NAME=20200501C3 SUBJECT_SAMPLE_FACTORS - F1 Treatment:FRUCTOSE RAW_FILE_NAME=20200501F1 SUBJECT_SAMPLE_FACTORS - F2 Treatment:FRUCTOSE RAW_FILE_NAME=20200501F2 SUBJECT_SAMPLE_FACTORS - F3 Treatment:FRUCTOSE RAW_FILE_NAME=20200501F3 #COLLECTION CO:COLLECTION_SUMMARY At the 5th day of secondary organoid culture, the organoids were treated with CO:COLLECTION_SUMMARY fructose (10 mM D-fructose)-containing IntestiCult organoid growth medium for 24 CO:COLLECTION_SUMMARY hr and washed with pre-cooled PBS before harvesting. CO:SAMPLE_TYPE Intestine #TREATMENT TR:TREATMENT_SUMMARY At the 5th day of secondary organoid culture, the organoids were treated with TR:TREATMENT_SUMMARY fructose (10 mM D-fructose)-containing IntestiCult organoid growth medium for 24 TR:TREATMENT_SUMMARY hr and washed with pre-cooled PBS before harvesting. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY To harvest intracellular metabolites, organoid samples were washed with SP:SAMPLEPREP_SUMMARY pre-cooled PBS for three times and metabolism was quenched with 80% ice-cold SP:SAMPLEPREP_SUMMARY methanol. Pre-cooled methanol (1mL) with para-chlorophenylalanine (1 µM) as the SP:SAMPLEPREP_SUMMARY internal standard was added to each sample and incubated on ice for 10 min. SP:SAMPLEPREP_SUMMARY Sample lysates were then centrifuged at 4 °C for 15 min at 23,000 g. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Waters Acquity H-Class CH:COLUMN_NAME Waters XBridge Amide (100 x 4.6mm, 3.5um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Waters Synapt G2 Si QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Fast data dependent acquisition The Progenesis QI MetaboAnalyst 3.0 #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples C1 C2 C3 F1 F2 F3 Factors Treatment:D-HANKS Treatment:D-HANKS Treatment:D-HANKS Treatment:FRUCTOSE Treatment:FRUCTOSE Treatment:FRUCTOSE Glucose 6-phosphate 12644.10372 14087.75676 11418.07257 20783.99163 28790.92203 30203.18853 Fructose 1-phosphate 762.0955709 788.6198669 743.605603 7068.999219 6373.14956 6371.697486 Fructose 6-phosphate 17288.20485 19048.27364 15925.05821 91985.14749 94056.46654 120045.0314 Fructose 1,6-bisphosphate 2104.069146 2248.145981 1550.202847 3078.57663 2703.838406 3493.326633 Dihydroxyacetone phosphate 303.1253536 264.2506493 247.752083 2224.130089 2283.056874 2951.737738 Glyceraldehyde 3-phosphate 6058.585746 4276.656084 4781.897719 8372.37473 8619.239785 6250.31176 Lactate 516.1052602 234.7216916 536.3705384 7668.107067 8455.227724 8808.426182 Citrate 165945 130783 175618 420126 411037 397250 Pyruvate 8218.903478 7375.238825 8009.08237 13120.39269 14028.13225 12729.35667 Fumarate 261.8074777 245.2576073 302.934605 93.95388134 178.208321 117.2028559 Malate 937.6850971 864.5487737 1063.638912 585.1856076 842.9756427 758.1250044 Glutamine 116139.6936 126593.7299 116429.9857 251950.7706 222506.3608 259687.3082 Glutamate 97532.7393 99028.63588 98228.39923 72683.65286 80750.40915 81294.03385 Histidine 250649.896 270885.8615 250648.3742 305793.401 279519.1384 297345.8882 Valine 12595.41435 13768.96741 13523.76161 16558.05942 14650.073 15459.43082 Cysteine 1126.818776 689.9432173 1229.849509 419.0459888 656.0797165 554.6455393 Proline 23406.93206 19670.47009 26149.49638 26245.0777 29468.76028 28448.4855 Isoleucine 258703.8776 221660.5662 231238.1986 247192.3226 221090.3487 239143.0504 Lysine 175051.474 134089.8535 142708.4022 172520.7501 116206.7447 114502.803 Leucine 36168.27757 30568.23132 28530.96325 29546.41643 33824.85597 35231.18315 Serine 348.2985964 518.3528478 337.6268259 484.0533819 399.6419815 401.0536547 Phenylalanine 324256.2675 343103.3628 326478.576 365596.0566 356844.5044 364129.1819 Tryptophan 67167.41547 79842.97909 69996.73784 98370.62912 89133.69445 96882.98262 Glycine 2798.611908 2791.94616 2948.356601 3416.457321 2908.330064 3300.372599 Thymine 291.3420296 197.6202812 306.2803507 591.0290328 579.0422507 596.4248811 Uridine diphosphate glucose 88126.87942 93354.01236 90959.99078 70528.32666 74538.6483 73776.73149 Oxidized glutathione 102095.9846 92663.30082 85922.34851 36306.57489 38103.71462 48355.57239 Uridine diphosphate-N-acetylgalactosamine 114612.7241 114134.7212 113274.6909 66987.29603 80856.91947 80958.34075 Cyclic AMP 13455.88013 13030.32148 12655.03045 9690.964237 9843.072178 11026.55457 Erucic acid 48164.17261 50026.4396 67974.54884 92901.82676 101775.9258 90563.3891 Uridine diphosphate glucuronic acid 37810.99388 34213.80665 35011.19982 27458.90654 29178.71171 31514.51567 Thymidine 5'-triphosphate 1538.514426 1994.555707 1488.8576 1064.505564 838.4142011 1219.815856 Uridine triphosphate 99815.75462 145733.4883 99758.73919 67873.25521 31119.90505 61278.41795 Glucosamine 6-phosphate 3548.452087 3716.197787 3411.590605 4023.321974 3874.556806 4589.59318 3'-AMP 124145.8808 79831.68214 124894.6813 147512.0861 147361.6562 147434.2019 Nicotinuric acid 337.9742028 413.3790557 458.0755707 570.0496544 503.3642239 466.3753628 Guanosine monophosphate 13249.95064 11264.25504 13446.56828 15880.51262 13625.05249 15127.24209 Cytidine diphosphate 211.5097361 312.7273875 269.7533794 233.8142048 107.9846077 170.0832167 Tyrosine 302333.7718 332942.1006 311428.335 360934.5262 447140.6723 337718.8929 Pyridoxine (Vitamin B6) 617587.7838 620455.5197 628734.1435 1197741.357 698551.5825 759262.4309 Alanine 13804.91373 15347.22321 14096.55356 16789.09071 15189.87988 14971.96293 LysoPC(15:0) 57030.35302 67707.95201 63265.63109 84011.22583 60869.76883 77742.08494 Uridine 5'-diphosphate 18906.34621 26122.83152 22075.12098 39725.14164 20956.72018 30994.53595 Adenosine triphosphate 254649.1473 385421.5319 215247.4585 260730.3643 94588.9627 209664.1704 Glutathione 713910.6156 780007.5547 622952.1258 606071.4043 612027.3893 702930.2326 Pantothenic Acid 280892.1484 307915.791 281270.7198 301399.8264 297863.177 298175.3489 Hypoxanthine 69.75555464 60.47433245 51.90481288 290.0091876 59.31845882 47.75747598 Folate 15305.50116 17650.59376 16151.60512 16674.12688 16921.57631 17139.36111 Adenosine monophosphate 1499.004498 1361.498902 1479.709047 1420.008414 1418.236339 1664.770582 Uridine 31148.51935 34063.07543 29306.16067 29542.60669 34359.36126 31175.27749 Cytidine 281265.005 271611.6965 279309.747 230017.8076 286943.4738 277656.3298 FAD 1936.671236 1785.197828 1855.415508 1429.123982 1851.309498 2006.00683 ADP 371940.9685 388802.0233 337694.7364 402068.5637 202658.4566 312194.4235 Xanthine 12791.19497 11999.24874 16468.28361 12583.32183 13605.54787 11031.46124 Guanosine diphosphate 11998.70855 13847.20203 11112.94969 21238.7989 7626.604889 12265.04158 Riboflavin (Vitamin B2) 6069.945124 6357.712874 6528.482871 6935.663438 5686.386554 6302.587684 ADP-glucose 412.6623609 331.6560322 408.237823 328.3056059 402.7740689 364.4257595 dTDP 379.3967035 415.6184023 403.9449953 429.9916545 409.6051343 254.8654791 Adenine 1142.460705 1839.658915 794.574443 1662.440403 597.9712293 874.7677519 NAD 17120.84452 16650.8865 15678.381 16399.6468 15745.42549 16103.82667 NADH 1702.087769 1244.500509 1684.163913 1189.174553 1778.172059 1663.069613 Nicotinamide adenine dinucleotide (NAD) 105.3622831 140.0420891 174.5223942 171.5164209 118.0205058 105.6547674 Guanosine triphosphate 7936.500061 12894.51639 7168.199075 15139.99739 3341.157148 7001.370564 LysoPE(0:0/20:0) 15560.95124 16274.28668 18831.8843 23468.89522 20920.15099 23942.99719 PE(18:1/20:1) 57045.36498 60522.02824 52633.49418 38789.28229 47111.29721 41988.78499 PE(18:2/18:0) 95828.74861 129637.0718 79731.1466 43287.5589 54316.9092 46724.14277 LysoPE(0:0/18:1) 33723.25362 39160.22859 35261.68179 51742.7613 41164.21171 48928.07558 LysoPE(0:0/22:1) 2386.985105 2829.894195 3054.963962 3471.355038 3200.995988 3544.065416 LysoPE(0:0/20:1) 37738.47069 43179.85854 38834.45817 52273.72906 43098.95213 52428.01795 PE(16:1/18:0) 34835.86231 46014.12597 33427.1747 27555.32667 27508.18932 28227.15807 LysoPE(0:0/16:0) 5774.2203 6811.334527 6460.100011 10479.76809 6545.27835 8677.595737 PG(16:0/0:0) 4615.279846 5706.174138 5004.308419 11422.61087 8587.220785 9431.103223 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z HMDB ID 3'-AMP 8.408283333 346.0598624 HMDB03540 Adenine 8.6904 134.0477183 HMDB00034 Adenosine monophosphate 8.271666667 346.0566973 HMDB00045 Adenosine triphosphate 8.6904 505.9923787 HMDB00538 ADP 8.578266667 426.0262025 HMDB01341 ADP-glucose 8.271666667 588.0759788 HMDB06557 Alanine 7.9774 88.0399149 HMDB00161 Citrate HMDB00094 Cyclic AMP 8.653983333 328.0487637 HMDB00058 Cysteine 9.15145 120.0120357 HMDB00574 Cytidine 9.4093 242.0803235 HMDB00089 Cytidine diphosphate 8.930033333 402.0095626 HMDB01546 Dihydroxyacetone phosphate 8.511483333 168.9946699 HMDB0001473 dTDP 8.174533333 401.0170609 HMDB01274 Erucic acid 2.32205 337.3110975 HMDB02068 FAD 8.308083333 784.1536139 HMDB01248 Folate 8.314166667 440.1370476 HMDB00121 Fructose 1,6-bisphosphate 8.593266667 338.9913085 HMDB0003971 Fructose 1-phosphate 8.371666667 259.0210061 HMDB01076 Fructose 6-phosphate 8.511483333 259.0250744 HMDB00124 Fumarate 8.3263 115.0044482 HMDB00134 Glucosamine 6-phosphate 8.462916667 258.0420738 HMDB01254 Glucose 6-phosphate 8.617566667 259.0251329 Glutamate 8.432566667 146.0472089 HMDB0060475 Glutamine 9.27895 145.0619953 HMDB0000641 Glutathione 8.420416667 306.0795223 HMDB00125 Glyceraldehyde 3-phosphate 8.623633333 168.9991288 HMDB0001473 Glycine 3.408216667 74.02473116 HMDB00123 Guanosine diphosphate 8.830033333 442.0173981 HMDB01201 Guanosine monophosphate 8.578266667 362.0536763 HMDB01397 Guanosine triphosphate 8.930033333 521.9846723 HMDB01273 Histidine 9.403233333 154.062489 HMDB0000177 Hypoxanthine 7.61635 135.0313536 HMDB00157 Isoleucine 8.025966667 130.0870803 HMDB00172 Lactate 6.087333333 89.02417117 HMDB00190 Leucine 8.192733333 130.0872199 HMDB00687 Lysine 8.0956 145.0980739 HMDB00182 LysoPC(15:0) 3.426433333 480.3114577 HMDB10381 LysoPE(0:0/16:0) 3.4325 452.2784638 HMDB11473 LysoPE(0:0/18:1) 3.426433333 478.2951447 HMDB11475 LysoPE(0:0/20:0) 3.426433333 508.3417462 HMDB11481 LysoPE(0:0/20:1) 3.426433333 506.3243971 HMDB11482 LysoPE(0:0/22:1) 3.426433333 534.3461711 HMDB11491 Malate 8.320233333 133.015283 HMDB00156 NAD 8.653983333 662.1067795 HMDB0000902 NADH 8.314166667 664.1219258 HMDB01487 Nicotinamide adenine dinucleotide (NAD) 8.475066667 662.099886 HMDB0000902 Nicotinuric acid 2.713616667 161.0369993 HMDB03269 Oxidized glutathione 8.578266667 611.1503584 HMDB03337 Pantothenic Acid 7.9774 218.1035031 HMDB0000210 PE(16:1/18:0) 3.25965 716.5264828 HMDB08958 PE(18:1/20:1) 3.180733333 770.573796 HMDB09032 PE(18:2/18:0) 3.21715 742.543029 HMDB09090 PG(16:0/0:0) 2.6529 483.2759989 HMDB0011503 Phenylalanine 7.971316667 164.0712961 HMDB00159 Proline 8.74505 114.0563499 HMDB00162 Pyridoxine (Vitamin B6) 4.6673 150.0558494 HMDB0000239 Pyruvate 8.61755 87.0088086 HMDB00243 Riboflavin (Vitamin B2) 7.234083333 375.1306632 HMDB0000244 Serine 9.385016667 104.0352952 HMDB00187 Thymidine 5'-triphosphate 8.593266667 480.9859097 HMDB01342 Thymine 9.27895 125.0360073 HMDB00262 Tryptophan 8.001683333 203.0823546 HMDB00929 Tyrosine 8.523633333 180.0681141 HMDB0000158 Uridine 8.062383333 243.0637919 HMDB00296 Uridine 5'-diphosphate 8.74505 402.9983568 HMDB00295 Uridine diphosphate glucose 8.3263 565.0575166 HMDB00286 Uridine diphosphate glucuronic acid 8.3838 579.033172 HMDB00935 Uridine diphosphate-N-acetylgalactosamine 8.314166667 606.0805491 HMDB00304 Uridine triphosphate 8.74505 482.9645559 HMDB00285 Valine 8.553983333 116.0723148 HMDB00883 Xanthine 7.215833333 151.0253379 HMDB00292 METABOLITES_END #END