#METABOLOMICS WORKBENCH Hengqian_Lu_20190225_022736 DATATRACK_ID:1640 STUDY_ID:ST001141 ANALYSIS_ID:AN001874 PROJECT_ID:PR000762 VERSION 1 CREATED_ON February 25, 2019, 7:26 pm #PROJECT PR:PROJECT_TITLE Evaluation of metabolome sample preparation and extraction methodologies for PR:PROJECT_TITLE oleaginous filamentous fungi Mortierella alpina PR:PROJECT_TYPE Method Optimization PR:PROJECT_SUMMARY In this study, based on the method of fast filtration, we evaluated the three PR:PROJECT_SUMMARY metabolomics analysis protocols commonly used for microbial metabolomics PR:PROJECT_SUMMARY analysis in M. alpina and systematically optimised the metabolite extraction PR:PROJECT_SUMMARY solvent. PR:INSTITUTE Jiangnan Unviersity PR:DEPARTMENT School of Food Science and Technology PR:LABORATORY State Key Laboratory of Food Science and Technology PR:LAST_NAME Hengqian PR:FIRST_NAME Lu PR:ADDRESS 1800 Lihu Ave, Wuxi, Jiangsu 214122, P.R. China. PR:EMAIL hengqianlu@163.com PR:PHONE +86 15006176136 PR:FUNDING_SOURCE National Natural Science Foundation of China #STUDY ST:STUDY_TITLE Evaluation of metabolome sample preparation and extraction methodologies for ST:STUDY_TITLE oleaginous filamentous fungi Mortierella alpina ST:STUDY_TYPE method optimization ST:STUDY_SUMMARY In this study, based on the method of fast filtration, we evaluated the three ST:STUDY_SUMMARY metabolomics analysis protocols commonly used for microbial metabolomics ST:STUDY_SUMMARY analysis in M. alpina and systematically optimised the metabolite extraction ST:STUDY_SUMMARY solvent. ST:INSTITUTE Jiangnan Unviersity ST:DEPARTMENT School of Food Science and Technology ST:LABORATORY State Key Laboratory of Food Science and Technology ST:LAST_NAME Hengqian ST:FIRST_NAME Lu ST:ADDRESS 1800 Lihu Ave, Wuxi, Jiangsu 214122, P.R. China. ST:EMAIL hengqianlu@163.com ST:PHONE +86 15006176136 #SUBJECT SU:SUBJECT_TYPE Fungi SU:SUBJECT_SPECIES Mortierella alpina SU:TAXONOMY_ID 64518 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - SM1-1 Treatment:SM1 SUBJECT_SAMPLE_FACTORS - SM1-2 Treatment:SM1 SUBJECT_SAMPLE_FACTORS - SM1-3 Treatment:SM1 SUBJECT_SAMPLE_FACTORS - SM1-4 Treatment:SM1 SUBJECT_SAMPLE_FACTORS - SM1-5 Treatment:SM1 SUBJECT_SAMPLE_FACTORS - SM2-1 Treatment:SM2 SUBJECT_SAMPLE_FACTORS - SM2-2 Treatment:SM2 SUBJECT_SAMPLE_FACTORS - SM2-3 Treatment:SM2 SUBJECT_SAMPLE_FACTORS - SM2-4 Treatment:SM2 SUBJECT_SAMPLE_FACTORS - SM2-5 Treatment:SM2 SUBJECT_SAMPLE_FACTORS - SM3-1 Treatment:SM3 SUBJECT_SAMPLE_FACTORS - SM3-2 Treatment:SM3 SUBJECT_SAMPLE_FACTORS - SM3-3 Treatment:SM3 SUBJECT_SAMPLE_FACTORS - SM3-4 Treatment:SM3 SUBJECT_SAMPLE_FACTORS - SM3-5 Treatment:SM3 SUBJECT_SAMPLE_FACTORS - SM4-1 Treatment:SM4 SUBJECT_SAMPLE_FACTORS - SM4-2 Treatment:SM4 SUBJECT_SAMPLE_FACTORS - SM4-3 Treatment:SM4 SUBJECT_SAMPLE_FACTORS - SM4-4 Treatment:SM4 SUBJECT_SAMPLE_FACTORS - SM4-5 Treatment:SM4 SUBJECT_SAMPLE_FACTORS - SM5-1 Treatment:SM5 SUBJECT_SAMPLE_FACTORS - SM5-2 Treatment:SM5 SUBJECT_SAMPLE_FACTORS - SM5-3 Treatment:SM5 SUBJECT_SAMPLE_FACTORS - SM5-4 Treatment:SM5 SUBJECT_SAMPLE_FACTORS - SM5-5 Treatment:SM5 #COLLECTION CO:COLLECTION_SUMMARY Mycelia were collected by vacuum filtration at 120 h using a paper filter CO:COLLECTION_SUMMARY (Whatman no. 1) and washed with 4°C 0.9% NaCl at room temperature, the sampling CO:COLLECTION_SUMMARY time was less than 45s. After quick-freezing in liquid nitrogen, the frozen cell CO:COLLECTION_SUMMARY pellets were further ground into powder (fresh biomass) and stored at -80°C for CO:COLLECTION_SUMMARY further use. CO:SAMPLE_TYPE fungi #TREATMENT TR:TREATMENT_SUMMARY 1.Sample preparation: Two forms of biomass, including fresh weight and TR:TREATMENT_SUMMARY freeze-dried, have been used for the extraction of metabolites. Three forms of TR:TREATMENT_SUMMARY quantified biomass, including fresh weight, freeze-dried and cell debris, have TR:TREATMENT_SUMMARY been used for data normalisation. 2. We create a pooled culture and split to TR:TREATMENT_SUMMARY produce identical samples for the different methods tested. Before extraction, TR:TREATMENT_SUMMARY three ways(fresh sample,freeze-dried sample, freeze-dried sample(water added)) TR:TREATMENT_SUMMARY were adopted to prepare the metabolomics analytical sample.To test the effect of TR:TREATMENT_SUMMARY the water contained in the fresh biomass on the extraction of metabolites from TR:TREATMENT_SUMMARY M. alpina, we repeated the freeze-dried biomass with additional water added to TR:TREATMENT_SUMMARY the account for the water content in fresh biomass. 2. Five solvent mixtures TR:TREATMENT_SUMMARY (SM) were used for the extraction of the intracellular metabolites of M. alpina. TR:TREATMENT_SUMMARY The five SM were: SM1, MTBE:methanol:water (20:6:5, vol:vol:vol) at -20°C; SM2, TR:TREATMENT_SUMMARY methanol:acetonitrile:water (2:2:1, vol:vol:vol) at -20°C; SM3, methanol:water TR:TREATMENT_SUMMARY (8:2, vol:vol) at -20°C; SM4, methanol:water (1:1, vol:vol); and SM5, TR:TREATMENT_SUMMARY Ethanol:water (3:1, vol:vol) at -20°C. 3. The mycelia were harvested at 24h and TR:TREATMENT_SUMMARY 120h for metabolomics analysis. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY The obtained supernatants were vacuum dried at 30°C prior to derivatisation. SP:SAMPLEPREP_SUMMARY The extraction solution served as a negative control to subtract from the SP:SAMPLEPREP_SUMMARY background spectrum. The dried extract was derivatised. Briefly, the vacuum SP:SAMPLEPREP_SUMMARY dried samples were resuspended in 100 μL of MeOX (10 mg/mL) in pyridine and SP:SAMPLEPREP_SUMMARY incubated at 37°C for 90 min in a block heater. Then, 40 μL of MSTFA+1%TMCS SP:SAMPLEPREP_SUMMARY was pipetted into each sample and incubated at 37°C for another 30 min. The SP:SAMPLEPREP_SUMMARY resulting solution from each sample was collected and transferred into gas SP:SAMPLEPREP_SUMMARY chromatography (GC) vials for analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Thermo Trace 1310 CH:COLUMN_NAME RTX-5MS column #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo TSQ8000_evo MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE EI MS:ION_MODE POSITIVE MS:MS_COMMENTS The “raw” format files were converted to “abf ” format with the ABF MS:MS_COMMENTS converter. The MSDIAL3.40 equipped with DB_FiehnBinbase-FiehnRI database was MS:MS_COMMENTS used for peaks exaction, retention time adjustment, peak alignment, MS:MS_COMMENTS deconvolution analysis and identification.All metabolite annotations were MS:MS_COMMENTS checked again by using the the TraceFinder 3.3 software (Thermo Fisher MS:MS_COMMENTS Scientific, Waltham, MA, USA), Xcalibur 3.1 and NIST MS search. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak Area MS_METABOLITE_DATA_START Samples SM1-1 SM1-2 SM1-3 SM1-4 SM1-5 SM2-1 SM2-2 SM2-3 SM2-4 SM2-5 SM3-1 SM3-2 SM3-3 SM3-4 SM3-5 SM4-1 SM4-2 SM4-3 SM4-4 SM4-5 SM5-1 SM5-2 SM5-3 SM5-4 SM5-5 Factors Treatment:SM1 Treatment:SM1 Treatment:SM1 Treatment:SM1 Treatment:SM1 Treatment:SM2 Treatment:SM2 Treatment:SM2 Treatment:SM2 Treatment:SM2 Treatment:SM3 Treatment:SM3 Treatment:SM3 Treatment:SM3 Treatment:SM3 Treatment:SM4 Treatment:SM4 Treatment:SM4 Treatment:SM4 Treatment:SM4 Treatment:SM5 Treatment:SM5 Treatment:SM5 Treatment:SM5 Treatment:SM5 Pyruvic acid 1380000.000 806000.000 739000.000 776000.000 1430000.000 1500000.000 1210000.000 1240000.000 1370000.000 626000.000 605000.000 1630000.000 1130000.000 551000.000 1770000.000 1970000.000 1620000.000 1330000.000 1610000.000 1810000.000 1550000.000 1500000.000 1320000.000 1300000.000 619000.000 L-Valine 95600.000 56100.000 64400.000 46100.000 83100.000 69800.000 83500.000 73200.000 96700.000 124000.000 78500.000 131000.000 105000.000 79700.000 95500.000 147000.000 152000.000 119000.000 171000.000 133000.000 75400.000 136000.000 90100.000 91500.000 61700.000 L-Alanine 5880000.000 9620000.000 7960000.000 8450000.000 7680000.000 5250000.000 4220000.000 9530000.000 4040000.000 1920000.000 4660000.000 3890000.000 5200000.000 8110000.000 9570000.000 9170000.000 7300000.000 6660000.000 2550000.000 6950000.000 6940000.000 1510000.000 8570000.000 7550000.000 10300000.000 Hydroxylamine 50300.000 49200.000 39100.000 55400.000 55700.000 63700.000 48700.000 46100.000 52000.000 36200.000 42800.000 58500.000 32300.000 40400.000 77300.000 59500.000 65500.000 61300.000 44300.000 65500.000 57500.000 19400.000 51000.000 68300.000 53600.000 Glycine 209000.000 297000.000 310000.000 253000.000 265000.000 182000.000 172000.000 407000.000 171000.000 85700.000 224000.000 152000.000 278000.000 336000.000 427000.000 527000.000 335000.000 242000.000 157000.000 286000.000 213000.000 94700.000 265000.000 256000.000 267000.000 L-Isoleucine 485000.000 337000.000 430000.000 285000.000 500000.000 268000.000 336000.000 358000.000 442000.000 485000.000 336000.000 573000.000 456000.000 418000.000 481000.000 799000.000 756000.000 679000.000 841000.000 602000.000 273000.000 582000.000 304000.000 378000.000 359000.000 Methanolphosphate 789000.000 460000.000 463000.000 425000.000 860000.000 103000.000 110000.000 137000.000 117000.000 174000.000 193000.000 199000.000 216000.000 233000.000 189000.000 1940000.000 1920000.000 379000.000 1990000.000 2160000.000 8180.000 9080.000 5720.000 9050.000 7110.000 L-Valine 2 986000.000 1100000.000 1280000.000 1200000.000 1340000.000 682000.000 647000.000 1360000.000 601000.000 197000.000 1040000.000 345000.000 852000.000 1230000.000 1240000.000 1370000.000 752000.000 1290000.000 329000.000 1320000.000 956000.000 205000.000 906000.000 1190000.000 1690000.000 Urea 24900.000 32900.000 24800.000 27600.000 29400.000 39600.000 1270.000 48500.000 33900.000 673.000 182.000 213.000 602.000 503.000 60200.000 584.000 561.000 469.000 473.000 523.000 50200.000 190.000 49500.000 76800.000 37900.000 L-Leucine 662000.000 813000.000 925000.000 940000.000 851000.000 297000.000 312000.000 515000.000 255000.000 128000.000 429000.000 214000.000 430000.000 565000.000 636000.000 901000.000 423000.000 880000.000 258000.000 888000.000 476000.000 114000.000 675000.000 561000.000 1070000.000 Ethanolamine 5100000.000 4740000.000 4960000.000 4800000.000 4990000.000 4890000.000 5230000.000 4810000.000 5060000.000 5070000.000 4900000.000 5070000.000 4950000.000 5790000.000 5270000.000 5110000.000 4850000.000 5480000.000 5130000.000 5360000.000 4700000.000 5140000.000 4850000.000 4720000.000 5310000.000 Phosphate 17000000.000 17800000.000 17200000.000 16700000.000 17600000.000 17500000.000 16200000.000 17100000.000 17900000.000 17500000.000 17300000.000 17700000.000 17300000.000 17100000.000 17700000.000 17500000.000 17600000.000 18200000.000 18000000.000 17700000.000 16600000.000 17200000.000 16800000.000 16500000.000 17900000.000 Succinic acid 685000.000 1220000.000 910000.000 729000.000 631000.000 714000.000 901000.000 1320000.000 925000.000 2360000.000 2050000.000 951000.000 1420000.000 2080000.000 885000.000 831000.000 1530000.000 1490000.000 1410000.000 1070000.000 697000.000 979000.000 989000.000 1120000.000 1640000.000 Glyceric acid 16500.000 8550.000 10400.000 7950.000 16500.000 7880.000 7460.000 9080.000 8540.000 8030.000 7500.000 11800.000 9900.000 7640.000 10300.000 13700.000 11800.000 10500.000 12000.000 12100.000 9840.000 10800.000 9320.000 10300.000 9300.000 Uracil 347000.000 285000.000 287000.000 235000.000 389000.000 289000.000 296000.000 319000.000 317000.000 213000.000 264000.000 472000.000 371000.000 258000.000 470000.000 550000.000 397000.000 479000.000 394000.000 491000.000 459000.000 339000.000 461000.000 458000.000 400000.000 Fumaric acid 639000.000 579000.000 475000.000 403000.000 700000.000 403000.000 429000.000 552000.000 475000.000 518000.000 512000.000 661000.000 695000.000 524000.000 598000.000 1140000.000 1560000.000 905000.000 1550000.000 1230000.000 447000.000 577000.000 499000.000 537000.000 395000.000 L-Alanine 2 75500.000 52800.000 87400.000 56300.000 102000.000 762000.000 507000.000 295000.000 1080000.000 1070000.000 1280000.000 1370000.000 862000.000 439000.000 368000.000 412000.000 620000.000 250000.000 1330000.000 1010000.000 539000.000 2490000.000 355000.000 506000.000 158000.000 L-Serine 483000.000 797000.000 670000.000 713000.000 572000.000 294000.000 284000.000 463000.000 250000.000 105000.000 616000.000 394000.000 447000.000 662000.000 647000.000 853000.000 503000.000 754000.000 119000.000 857000.000 541000.000 55700.000 698000.000 599000.000 972000.000 L-Threonine 193000.000 253000.000 261000.000 263000.000 236000.000 134000.000 127000.000 197000.000 120000.000 62700.000 242000.000 220000.000 188000.000 256000.000 257000.000 315000.000 277000.000 286000.000 60700.000 304000.000 223000.000 29800.000 316000.000 252000.000 352000.000 Hydrocinnamic acid 139000.000 82000.000 117000.000 68200.000 161000.000 58600.000 74900.000 88700.000 84800.000 115000.000 112000.000 72900.000 138000.000 101000.000 133000.000 300000.000 134000.000 201000.000 328000.000 236000.000 86800.000 208000.000 38500.000 120000.000 89900.000 Glutaric acid 9540.000 11900.000 9810.000 65900.000 7590.000 11900.000 29600.000 12100.000 11900.000 20800.000 22400.000 9530.000 13000.000 19300.000 42400.000 9250.000 7700.000 9210.000 18800.000 23000.000 13300.000 18100.000 12000.000 34700.000 15700.000 Aspartate 139000.000 87800.000 151000.000 90100.000 144000.000 86600.000 110000.000 119000.000 124000.000 312000.000 334000.000 59800.000 126000.000 226000.000 115000.000 261000.000 156000.000 180000.000 294000.000 252000.000 81800.000 166000.000 44100.000 85000.000 59400.000 Beta alanine 229000.000 260000.000 208000.000 195000.000 270000.000 242000.000 174000.000 272000.000 208000.000 299000.000 320000.000 390000.000 250000.000 309000.000 294000.000 321000.000 404000.000 296000.000 346000.000 369000.000 321000.000 244000.000 340000.000 335000.000 336000.000 Malate 7240.000 5950.000 3520.000 2760.000 4470.000 6430.000 9130.000 7160.000 11300.000 24600.000 11300.000 15000.000 20500.000 6890.000 7660.000 10200.000 13700.000 5960.000 23200.000 14500.000 7430.000 18600.000 7380.000 6150.000 7260.000 L-Methionine 199000.000 265000.000 225000.000 197000.000 236000.000 77800.000 69100.000 122000.000 62200.000 159000.000 138000.000 339000.000 113000.000 169000.000 168000.000 332000.000 520000.000 276000.000 260000.000 316000.000 145000.000 65200.000 290000.000 172000.000 257000.000 L-5-Oxoproline 2700000.000 2010000.000 1930000.000 910000.000 1610000.000 1890000.000 1850000.000 2080000.000 2380000.000 3950000.000 3740000.000 3950000.000 3550000.000 2320000.000 2420000.000 2850000.000 3600000.000 3060000.000 4960000.000 3360000.000 1950000.000 5540000.000 1910000.000 1910000.000 2130000.000 Aspartic acid 116000.000 208000.000 188000.000 162000.000 157000.000 61900.000 45500.000 95200.000 40700.000 306000.000 290000.000 176000.000 63400.000 269000.000 100000.000 208000.000 446000.000 159000.000 201000.000 264000.000 94800.000 30700.000 196000.000 84400.000 122000.000 4-Aminobutyric acid 1770000.000 2620000.000 2060000.000 1820000.000 2530000.000 0.000 2760000.000 1220000.000 3250000.000 1720000.000 1430000.000 2780000.000 2590000.000 1920000.000 1330000.000 1920000.000 2250000.000 2920000.000 2200000.000 1860000.000 1350000.000 2300000.000 1690000.000 1910000.000 3880000.000 Glutamate 1460000.000 2620000.000 2060000.000 1820000.000 2540000.000 1740000.000 2760000.000 1870000.000 3240000.000 1720000.000 1980000.000 2780000.000 2590000.000 1660000.000 2090000.000 1970000.000 2250000.000 14400.000 1860000.000 1280000.000 1350000.000 2300000.000 1690000.000 1690000.000 3880000.000 L-Cysteine 111000.000 246000.000 139000.000 217000.000 197000.000 127000.000 70800.000 207000.000 46200.000 388000.000 160000.000 668000.000 105000.000 206000.000 207000.000 227000.000 752000.000 202000.000 459000.000 292000.000 193000.000 92800.000 709000.000 184000.000 318000.000 Alpha-ketoglutarate 429000.000 201000.000 195000.000 175000.000 446000.000 523000.000 497000.000 512000.000 577000.000 665000.000 344000.000 641000.000 445000.000 186000.000 534000.000 644000.000 582000.000 341000.000 697000.000 594000.000 518000.000 703000.000 484000.000 466000.000 236000.000 L-Ornithine 10800.000 700000.000 194000.000 768000.000 65500.000 8650.000 9270.000 37400.000 4450.000 269000.000 332000.000 1060000.000 60200.000 500000.000 292000.000 435000.000 1200000.000 265000.000 480000.000 994000.000 244000.000 34600.000 971000.000 348000.000 753000.000 Phenylalanine 196000.000 243000.000 224000.000 217000.000 268000.000 58900.000 53100.000 83600.000 38000.000 142000.000 106000.000 189000.000 85600.000 137000.000 115000.000 220000.000 301000.000 229000.000 229000.000 237000.000 101000.000 79600.000 169000.000 121000.000 227000.000 L-Glutamic acid 4140000.000 4780000.000 3990000.000 3840000.000 4560000.000 4060000.000 3010000.000 5280000.000 2560000.000 6030000.000 5470000.000 6220000.000 4590000.000 5760000.000 5440000.000 6080000.000 6850000.000 4840000.000 6400000.000 6550000.000 5020000.000 4150000.000 6090000.000 5120000.000 4750000.000 Tartaric acid 1030.000 4510.000 4490.000 1850.000 3610.000 2890.000 2390.000 619.000 1470.000 3680.000 2720.000 4570.000 2760.000 3430.000 6020.000 4320.000 2410.000 4920.000 3450.000 1620.000 335.000 802.000 3380.000 2480.000 82.900 Asparagine 120000.000 241000.000 194000.000 235000.000 200000.000 101000.000 50100.000 263000.000 5520.000 250000.000 352000.000 451000.000 213000.000 399000.000 457000.000 626000.000 603000.000 284000.000 395000.000 610000.000 353000.000 136000.000 450000.000 437000.000 362000.000 Ribose 128000.000 55200.000 88100.000 62500.000 136000.000 47100.000 47500.000 53500.000 53200.000 45800.000 43400.000 61000.000 49500.000 39100.000 57800.000 76800.000 76800.000 44900.000 83700.000 75100.000 58600.000 71400.000 57000.000 59500.000 46100.000 Alpha-aminoadipic acid 115000.000 163000.000 130000.000 117000.000 135000.000 130000.000 79700.000 214000.000 51600.000 270000.000 284000.000 310000.000 216000.000 289000.000 274000.000 296000.000 296000.000 186000.000 260000.000 299000.000 224000.000 175000.000 255000.000 264000.000 205000.000 Arabitol 131000.000 124000.000 111000.000 94900.000 142000.000 154000.000 152000.000 198000.000 180000.000 220000.000 222000.000 218000.000 209000.000 184000.000 195000.000 211000.000 218000.000 165000.000 220000.000 211000.000 186000.000 234000.000 188000.000 209000.000 175000.000 Putrescine 256000.000 413000.000 305000.000 297000.000 369000.000 81800.000 48500.000 99600.000 84200.000 152000.000 280000.000 230000.000 157000.000 259000.000 176000.000 226000.000 197000.000 277000.000 293000.000 261000.000 167000.000 175000.000 160000.000 225000.000 215000.000 Glycerol-alpha-phosphate 2130000.000 2390000.000 2090000.000 2000000.000 2180000.000 2600000.000 1680000.000 2810000.000 1750000.000 2500000.000 3360000.000 3540000.000 2020000.000 2760000.000 2450000.000 2200000.000 2640000.000 2580000.000 2130000.000 57700.000 3260000.000 1930000.000 2910000.000 3430000.000 3600000.000 Glucose-1-phosphate 71900.000 80600.000 113000.000 78200.000 133000.000 409000.000 344000.000 567000.000 509000.000 573000.000 664000.000 608000.000 483000.000 608000.000 673000.000 194000.000 180000.000 275000.000 205000.000 196000.000 360000.000 527000.000 388000.000 318000.000 556000.000 Phosphoethanolamine 362000.000 437000.000 363000.000 370000.000 379000.000 118000.000 102000.000 148000.000 91800.000 174000.000 262000.000 274000.000 183000.000 209000.000 220000.000 452000.000 438000.000 195000.000 429000.000 487000.000 381000.000 314000.000 350000.000 410000.000 291000.000 L-Ornithine 2 453000.000 919000.000 551000.000 684000.000 674000.000 223000.000 116000.000 320000.000 66700.000 221000.000 663000.000 531000.000 363000.000 619000.000 512000.000 783000.000 782000.000 662000.000 629000.000 1160000.000 518000.000 184000.000 445000.000 621000.000 662000.000 3-Phosphoglycerate 117000.000 69700.000 80700.000 46300.000 117000.000 25700.000 21700.000 28300.000 21400.000 22800.000 29500.000 49900.000 42400.000 24300.000 45900.000 61200.000 12400.000 26300.000 19900.000 32500.000 28000.000 32000.000 25800.000 25000.000 17000.000 L-Citrulline 170000.000 513000.000 406000.000 497000.000 212000.000 115000.000 91200.000 350000.000 16400.000 276000.000 641000.000 698000.000 313000.000 759000.000 828000.000 1030000.000 904000.000 472000.000 457000.000 892000.000 481000.000 105000.000 657000.000 689000.000 520000.000 Citric acid 917000.000 1120000.000 870000.000 834000.000 1050000.000 393000.000 388000.000 504000.000 448000.000 729000.000 1180000.000 797000.000 804000.000 948000.000 713000.000 1580000.000 1910000.000 1630000.000 1920000.000 1900000.000 459000.000 599000.000 506000.000 470000.000 486000.000 Adenine 498000.000 740000.000 561000.000 452000.000 530000.000 100000.000 83800.000 184000.000 12800.000 244000.000 358000.000 342000.000 208000.000 319000.000 318000.000 799000.000 796000.000 568000.000 676000.000 920000.000 322000.000 200000.000 328000.000 442000.000 280000.000 Glucose 4450000.000 4470000.000 4350000.000 4530000.000 4570000.000 4400000.000 4340000.000 4310000.000 4510000.000 4360000.000 4350000.000 4370000.000 4310000.000 4260000.000 4380000.000 4280000.000 4340000.000 4610000.000 4520000.000 4360000.000 4140000.000 4460000.000 4320000.000 4200000.000 4610000.000 L-Lysine 5190000.000 6610000.000 5460000.000 5170000.000 5200000.000 3350000.000 2730000.000 3990000.000 1880000.000 4600000.000 6640000.000 6400000.000 5540000.000 6240000.000 6060000.000 7100000.000 7010000.000 6540000.000 7210000.000 7290000.000 4560000.000 3960000.000 4780000.000 5340000.000 5130000.000 Tyrosine 410000.000 495000.000 540000.000 432000.000 405000.000 187000.000 140000.000 314000.000 68400.000 448000.000 517000.000 493000.000 331000.000 554000.000 427000.000 604000.000 609000.000 460000.000 534000.000 630000.000 375000.000 250000.000 363000.000 416000.000 489000.000 Pantothenic acid 50000.000 47400.000 43700.000 35300.000 57600.000 66300.000 60800.000 78100.000 73700.000 89100.000 92100.000 94400.000 83500.000 78200.000 82500.000 83700.000 88200.000 73200.000 92600.000 88100.000 72300.000 99300.000 72900.000 84600.000 67400.000 Gluconic acid 1690.000 1160.000 1260.000 1190.000 1960.000 1960.000 1760.000 2280.000 2110.000 2710.000 2220.000 2240.000 2430.000 1750.000 1950.000 2430.000 2440.000 1860.000 3300.000 2540.000 2000.000 2530.000 1620.000 1940.000 1680.000 Palmitic acid 1270000.000 918000.000 1000000.000 934000.000 1090000.000 596000.000 483000.000 673000.000 719000.000 635000.000 655000.000 644000.000 589000.000 459000.000 515000.000 390000.000 397000.000 631000.000 407000.000 407000.000 564000.000 717000.000 523000.000 649000.000 720000.000 N-acetylgalactosamine 25700.000 20900.000 23500.000 26400.000 45800.000 30300.000 30000.000 39300.000 38300.000 32500.000 33400.000 50900.000 37700.000 29200.000 43700.000 37200.000 35400.000 30900.000 34900.000 39400.000 42900.000 46400.000 35500.000 45200.000 43100.000 Myo-inositol 222000.000 268000.000 158000.000 183000.000 277000.000 189000.000 167000.000 211000.000 226000.000 244000.000 262000.000 278000.000 241000.000 212000.000 225000.000 529000.000 550000.000 261000.000 593000.000 557000.000 219000.000 284000.000 187000.000 227000.000 178000.000 Ribose-5-phosphate 13400.000 11200.000 9550.000 10100.000 17000.000 16400.000 17500.000 27100.000 16300.000 35800.000 34500.000 35700.000 32000.000 29000.000 28200.000 36100.000 40100.000 15900.000 56200.000 30700.000 18900.000 17400.000 18600.000 18900.000 10200.000 Ribulose-5-phosphate 14100.000 9940.000 7000.000 9830.000 18000.000 22300.000 22700.000 32200.000 21600.000 33800.000 30800.000 45700.000 42800.000 28100.000 35300.000 45300.000 54600.000 30300.000 70900.000 39100.000 23900.000 31300.000 20600.000 19200.000 8410.000 Glutathione 29200.000 17500.000 25200.000 11400.000 33200.000 52800.000 41800.000 33500.000 32700.000 187000.000 93100.000 80500.000 83000.000 16900.000 43100.000 37700.000 26700.000 85300.000 128000.000 43900.000 26400.000 236000.000 27200.000 13900.000 41100.000 Linoleic acid 11300.000 12100.000 11600.000 12200.000 20100.000 12300.000 14100.000 15400.000 12700.000 14200.000 7600.000 10900.000 11300.000 8900.000 9940.000 1380.000 3200.000 6130.000 3960.000 4210.000 10800.000 18600.000 13400.000 14700.000 9460.000 Spermidine 4810.000 4020.000 4680.000 3730.000 9950.000 1310.000 1340.000 1960.000 311.000 2850.000 5740.000 6220.000 4190.000 5000.000 5720.000 2260.000 1890.000 2660.000 2390.000 2030.000 1020.000 3040.000 908.000 1240.000 966.000 Oleic acid 607000.000 537000.000 521000.000 543000.000 824000.000 501000.000 468000.000 482000.000 514000.000 529000.000 313000.000 378000.000 386000.000 308000.000 309000.000 66200.000 87000.000 307000.000 117000.000 138000.000 417000.000 647000.000 488000.000 512000.000 453000.000 Tryptophan 41700.000 102000.000 183000.000 153000.000 7900.000 12000.000 11200.000 272000.000 1180.000 255000.000 1680000.000 2110000.000 229000.000 1370000.000 1670000.000 238000.000 283000.000 90600.000 146000.000 192000.000 751000.000 25200.000 1090000.000 1090000.000 347000.000 Stearic acid 14700.000 31900.000 42400.000 29400.000 28300.000 20900.000 17400.000 12400.000 27300.000 27800.000 32600.000 23300.000 18300.000 24600.000 26000.000 14900.000 14100.000 21500.000 18100.000 15200.000 29800.000 29800.000 31100.000 35600.000 42900.000 Fructose-6-phosphate 356000.000 334000.000 195000.000 317000.000 656000.000 82700.000 81300.000 113000.000 68400.000 116000.000 154000.000 242000.000 216000.000 159000.000 179000.000 1270000.000 1380000.000 474000.000 1620000.000 1160000.000 118000.000 162000.000 108000.000 123000.000 75700.000 Glucose-6-phosphate 841000.000 928000.000 865000.000 1050000.000 1290000.000 197000.000 201000.000 282000.000 170000.000 271000.000 281000.000 501000.000 421000.000 328000.000 384000.000 1840000.000 2050000.000 1430000.000 2380000.000 1800000.000 256000.000 338000.000 241000.000 256000.000 165000.000 Inositol-4-monophosphate 1150000.000 1110000.000 991000.000 880000.000 1090000.000 458000.000 407000.000 540000.000 374000.000 707000.000 804000.000 832000.000 579000.000 555000.000 594000.000 353000.000 359000.000 189000.000 405000.000 418000.000 1480000.000 1750000.000 1430000.000 1520000.000 1010000.000 Uridine 1030000.000 1160000.000 28600.000 642000.000 1330000.000 647000.000 720000.000 807000.000 916000.000 608000.000 416000.000 718000.000 750000.000 353000.000 673000.000 969000.000 688000.000 818000.000 989000.000 669000.000 557000.000 925000.000 537000.000 473000.000 354000.000 1-Monopalmitin 56600.000 62600.000 67200.000 65700.000 115000.000 83400.000 87300.000 89100.000 105000.000 87900.000 52500.000 52900.000 61300.000 47100.000 50900.000 29500.000 35300.000 49200.000 42000.000 36200.000 60900.000 97900.000 60700.000 60000.000 57700.000 Adenosine 2380000.000 2420000.000 2320000.000 2210000.000 2100000.000 2310000.000 2380000.000 3000000.000 976000.000 3020000.000 3100000.000 3660000.000 3050000.000 2940000.000 3390000.000 3420000.000 3480000.000 3360000.000 3490000.000 3440000.000 3200000.000 3250000.000 3440000.000 3430000.000 3360000.000 Trehalose 58300.000 59000.000 50400.000 46200.000 62300.000 66900.000 64100.000 76000.000 73500.000 87600.000 82700.000 92200.000 77900.000 76000.000 79100.000 78000.000 83600.000 75300.000 88200.000 87200.000 77700.000 87500.000 78100.000 77500.000 69400.000 1-Monoolein 152000.000 146000.000 148000.000 140000.000 1130000.000 642000.000 658000.000 695000.000 1230000.000 195000.000 57000.000 60500.000 183000.000 72900.000 78600.000 4000.000 20200.000 125000.000 43100.000 40600.000 125000.000 198000.000 132000.000 119000.000 100000.000 Lanosterol 442000.000 437000.000 350000.000 350000.000 385000.000 91100.000 102000.000 116000.000 104000.000 137000.000 79000.000 81900.000 66400.000 69700.000 69800.000 7260.000 6470.000 7240.000 7070.000 8410.000 269000.000 334000.000 290000.000 296000.000 223000.000 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name HMDB ID KEGG ID Pyruvic acid HMDB0000243 C00022 L-Valine HMDB0000883 C00183 L-Alanine HMDB0000161 C00041 Hydroxylamine HMDB0003338 C00192 Glycine HMDB0000123 C00037 L-Isoleucine HMDB0000172 C00407 Methanolphosphate NA NA L-Valine 2 NA NA Urea HMDB0000294 C00086 L-Leucine HMDB0000687 C00123 Ethanolamine HMDB0000149 C00189 Phosphate HMDB0001429 C00009 Succinic acid HMDB0000254 C00042 Glyceric acid HMDB0000139 C00258 Uracil HMDB0000300 C00106 Fumaric acid HMDB0000134 C00122 L-Alanine 2 NA NA L-Serine HMDB0000187 C00065 L-Threonine HMDB0000167 C00188 Hydrocinnamic acid HMDB0000764 C05629 Glutaric acid HMDB0000661 C00489 Aspartate HMDB0000191 C00049 Beta alanine HMDB0000056 C00099 Malate HMDB0000156 C00149 L-Methionine HMDB0000696 C00073 L-5-Oxoproline HMDB0000267 C01879 Aspartic acid HMDB0000191 C00049 4-Aminobutyric acid HMDB0000112 C00334 Glutamate HMDB0000148 C00025 L-Cysteine HMDB0000574 C00097 Alpha-ketoglutarate HMDB0000208 C00026 L-Ornithine HMDB0000214 C00077 Phenylalanine HMDB0000159 C00079 L-Glutamic acid HMDB0000148 C00025 Tartaric acid NA NA Asparagine HMDB0000168 C00152 Ribose HMDB0000283 C00121 Alpha-aminoadipic acid HMDB0000510 C00956 Arabitol HMDB0000568 C01904 Putrescine HMDB0001414 C00134 Glycerol-alpha-phosphate NA NA Glucose-1-phosphate HMDB0001586 C00103 Phosphoethanolamine HMDB0000224 C00346 L-Ornithine 2 HMDB0000214 C00077 3-Phosphoglycerate HMDB0000807 C00597 L-Citrulline HMDB0000904 C00327 Citric acid HMDB0000094 C00158 Adenine HMDB0000034 C00147 Glucose HMDB0000122 C00031 L-Lysine HMDB0000182 C00047 Tyrosine HMDB0000158 C00082 Pantothenic acid HMDB0000210 C00864 Gluconic acid HMDB0000625 C00257 Palmitic acid HMDB0000220 C00249 N-acetylgalactosamine HMDB0000212 C01074 Myo-inositol HMDB0000211 C00137 Ribose-5-phosphate HMDB0001548 C00117 Ribulose-5-phosphate HMDB0001548 C00117 Glutathione HMDB0000125 C00051 Linoleic acid HMDB0000673 C01595 Spermidine HMDB0001257 C00315 Oleic acid HMDB0000207 C00712 Tryptophan HMDB0000929 C00078 Stearic acid HMDB0000827 C01530 Fructose-6-phosphate HMDB0000124 C00085 Glucose-6-phosphate HMDB0001401 C00668 Inositol-4-monophosphate NA NA Uridine HMDB0000296 C00299 1-Monopalmitin HMDB0011564 Adenosine HMDB0000050 C00212 Trehalose HMDB0000975 C01083 1-Monoolein HMDB0011567 Lanosterol HMDB0001251 C01724 METABOLITES_END #END