#METABOLOMICS WORKBENCH spinelli_lab_20251113_084720 DATATRACK_ID:6686 STUDY_ID:ST004403 ANALYSIS_ID:AN007361 PROJECT_ID:PR002783 VERSION 1 CREATED_ON November 26, 2025, 2:13 pm #PROJECT PR:PROJECT_TITLE Metabolic Reprogramming during Human Neuron Differentiation Indicates PR:PROJECT_TITLE Glutaminase as a Key Determinant in Fragile X Syndrome PR:PROJECT_SUMMARY Metabolic homeostasis gone awry is a contributor to, if not an underlying cause PR:PROJECT_SUMMARY of, several neurologic disorders. Fragile X syndrome (FXS) is a PR:PROJECT_SUMMARY neurodevelopmental disorder caused by a trinucleotide repeat expansion in FMR1 PR:PROJECT_SUMMARY and consequent loss of the encoded protein FMRP, which results in downstream PR:PROJECT_SUMMARY molecular, neurologic, and mitochondrial deficits that are linked to cognitive PR:PROJECT_SUMMARY impairment. In human postmortem brain, many metabolites and solute carrier PR:PROJECT_SUMMARY proteins are coordinately dysregulated, which also occurs during differentiation PR:PROJECT_SUMMARY of human iPSCs into excitatory neurons. Metabolic tracing in FXS neurons PR:PROJECT_SUMMARY demonstrates a dearth of glutamine deamidation to glutamate, which reduces PR:PROJECT_SUMMARY anaplerosis into the TCA cycle, potentially hindering bioenergetic and PR:PROJECT_SUMMARY biosynthetic functions of mitochondria. Mechanistically, aberrant expression of PR:PROJECT_SUMMARY glutaminase isoforms in FXS is responsible for reduced glutaminolysis, thereby PR:PROJECT_SUMMARY altering glutamate levels which may contribute to FXS. PR:INSTITUTE UMass Chan Medical School PR:LAST_NAME UMass Chan PR:FIRST_NAME Spinelli Lab PR:ADDRESS 55 Lake Avenue North, Worcester, Massachusetts, 01605, USA PR:EMAIL spinellilab@gmail.com PR:PHONE (508) 856-8989 ext. 68148 #STUDY ST:STUDY_TITLE Polar metabolomics on cell pellets and extracellular media of 4-week iPSC ST:STUDY_TITLE derived forebrain excitatory neurons generated from a FXS and CRISPR edited ST:STUDY_TITLE isogenic control. ST:STUDY_SUMMARY To assess whether FMRP influences metabolism during neuron maturation, we ST:STUDY_SUMMARY differentiated isogenic lines into forebrain excitatory neurons and profiled ST:STUDY_SUMMARY intracellular and extracellular metabolites at weeks 4. Polar metabolites during ST:STUDY_SUMMARY neuron differentiation revealed that amino acids, energy/nucleotide metabolism, ST:STUDY_SUMMARY and neurotransmitters were adversely affected in FXS cells and in the culture ST:STUDY_SUMMARY media at this stage. ST:INSTITUTE UMass Chan Medical School ST:LAST_NAME UMass Chan ST:FIRST_NAME Spinelli Lab ST:ADDRESS 55 Lake Avenue North, Worcester, Massachusetts, 01605, USA ST:EMAIL spinellilab@gmail.com ST:PHONE (508) 856-8989 ext. 68148 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:GENDER Male #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 - cell iControl_w4_1 Sample source:Male Homo sapien iPSCs derived neurons | Genotype:TD | Sample type:Cultured cells RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S01.raw SUBJECT_SAMPLE_FACTORS - cell iControl_w4_2 Sample source:Male Homo sapien iPSCs derived neurons | Genotype:TD | Sample type:Cultured cells RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S02.raw SUBJECT_SAMPLE_FACTORS - cell iControl_w4_3 Sample source:Male Homo sapien iPSCs derived neurons | Genotype:TD | Sample type:Cultured cells RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S03.raw SUBJECT_SAMPLE_FACTORS - cell FXS_w4_1 Sample source:Male Homo sapien iPSCs derived neurons | Genotype:FXS | Sample type:Cultured cells RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S04.raw SUBJECT_SAMPLE_FACTORS - cell FXS_w4_2 Sample source:Male Homo sapien iPSCs derived neurons | Genotype:FXS | Sample type:Cultured cells RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S05.raw SUBJECT_SAMPLE_FACTORS - cell FXS_w4_3 Sample source:Male Homo sapien iPSCs derived neurons | Genotype:FXS | Sample type:Cultured cells RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S06.raw SUBJECT_SAMPLE_FACTORS - media iControl_w4_1 Sample source:Male Homo sapien iPSCs derived neurons | Genotype:TD | Sample type:Extracellular media RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S13.raw SUBJECT_SAMPLE_FACTORS - media iControl_w4_2 Sample source:Male Homo sapien iPSCs derived neurons | Genotype:TD | Sample type:Extracellular media RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S14.raw SUBJECT_SAMPLE_FACTORS - media iControl_w4_3 Sample source:Male Homo sapien iPSCs derived neurons | Genotype:TD | Sample type:Extracellular media RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S15.raw SUBJECT_SAMPLE_FACTORS - media FXS_w4_1 Sample source:Male Homo sapien iPSCs derived neurons | Genotype:FXS | Sample type:Extracellular media RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S16.raw SUBJECT_SAMPLE_FACTORS - media FXS_w4_2 Sample source:Male Homo sapien iPSCs derived neurons | Genotype:FXS | Sample type:Extracellular media RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S17.raw SUBJECT_SAMPLE_FACTORS - media FXS_w4_3 Sample source:Male Homo sapien iPSCs derived neurons | Genotype:FXS | Sample type:Extracellular media RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S18.raw SUBJECT_SAMPLE_FACTORS - media blank 1 Sample source:Male Homo sapien iPSCs derived neurons | Genotype:media blank | Sample type:Media Blank RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S19.raw #COLLECTION CO:COLLECTION_SUMMARY iPSC lines were cultured on Matrigel coated plates in mTeSR™1 (Catalog #85850) CO:COLLECTION_SUMMARY with 10uM ROCK inhibitor (Y-27632). Neural induction was performed using the CO:COLLECTION_SUMMARY embryoid body formation protocol from StemCell technologies. Briefly cells were CO:COLLECTION_SUMMARY plated on Matrigel coated AggreWell™800 wells in STEMdiff™ Neural Induction CO:COLLECTION_SUMMARY Medium + SMADi and embryoid bodies were allowed to form for 13 days followed by CO:COLLECTION_SUMMARY Rosette selection as per manufactures instructions. The rosettes were plated on CO:COLLECTION_SUMMARY Poly-L-Ornithine (PLO)/ laminin coated plates for 3 days and then dissociated CO:COLLECTION_SUMMARY into single cells and expanded as NPCs in STEMdiff™ Neural Progenitor Medium. CO:COLLECTION_SUMMARY To generate neuron precursors, NPCs were cultured in STEMdiff™ Forebrain CO:COLLECTION_SUMMARY Neuron Differentiation media (Catalog #08600) for 3 days on PLO/laminin plates CO:COLLECTION_SUMMARY followed by replating at a density of 135,000 cells/cm2 in PLO/laminin plates in CO:COLLECTION_SUMMARY STEMdiff™ Forebrain Neuron Maturation media (Catalog #08605) with Compound E CO:COLLECTION_SUMMARY for 4 weeks. CO:SAMPLE_TYPE Culture Cells and Extracellular Media #TREATMENT TR:TREATMENT_SUMMARY No treatment other than standard culturing conditions and genotypes. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY For media, 10ul of sample was added to 90ul of 80% methanol and vortexed for SP:SAMPLEPREP_SUMMARY 10mins before spinning to collect the supernatant containing extracted SP:SAMPLEPREP_SUMMARY metabolites and vacuum dried. The extract was then resuspended in LC/MS grade SP:SAMPLEPREP_SUMMARY water. For extraction from cell pellets from in-vitro cultures, samples were SP:SAMPLEPREP_SUMMARY incubated on dry ice with cold 80% methanol. The extracts were spun down and the SP:SAMPLEPREP_SUMMARY supernatant was vacuum dried before resuspending in LC/MS grade water. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Merck SeQuant ZIC-HILIC (150 x 2.1mm,5um) CH:SOLVENT_A 100% water; 0.1% ammonium hydroxide; 20mM ammonium carbonate CH:SOLVENT_B 100% acetonitrile CH:FLOW_GRADIENT 20 min, 80% - 20% B; 0.5 min, 20% - 80% B; 7.5min, 80% B CH:FLOW_RATE 0.15ml/min CH:COLUMN_TEMPERATURE 25 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Plus Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS The mass spectrometer was set to full scan (70-1000 m/z), with the spray voltage MS:MS_COMMENTS set to 4.0 kV, heated capillary to 350°C, and the HESI probe at 30 °C. The MS:MS_COMMENTS sheath gas flow was set at 10 units, auxiliary gas at 1 units, and sweep gas MS:MS_COMMENTS flow at 1 unit. The resolution of scan was set to 70,000, AGC target to 1x106, MS:MS_COMMENTS and maximum injection time at 20 msec. Data acquired by Thermo Fisher's Xcalibur MS:MS_COMMENTS software and analyzed by their Tracefinder software. The raw files provided MS:MS_COMMENTS contain data from both positive and negative ion mode. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS log2FC(peak area) MS_METABOLITE_DATA_START Samples cell iControl_w4_1 cell iControl_w4_2 cell iControl_w4_3 cell FXS_w4_1 cell FXS_w4_2 cell FXS_w4_3 media blank 1 media iControl_w4_1 media iControl_w4_2 media iControl_w4_3 media FXS_w4_1 media FXS_w4_2 media FXS_w4_3 Factors Sample source:Male Homo sapien iPSCs derived neurons | Genotype:TD | Sample type:Cultured cells Sample source:Male Homo sapien iPSCs derived neurons | Genotype:TD | Sample type:Cultured cells Sample source:Male Homo sapien iPSCs derived neurons | Genotype:TD | Sample type:Cultured cells Sample source:Male Homo sapien iPSCs derived neurons | Genotype:FXS | Sample type:Cultured cells Sample source:Male Homo sapien iPSCs derived neurons | Genotype:FXS | Sample type:Cultured cells Sample source:Male Homo sapien iPSCs derived neurons | Genotype:FXS | Sample type:Cultured cells Sample source:Male Homo sapien iPSCs derived neurons | Genotype:media blank | Sample type:Media Blank Sample source:Male Homo sapien iPSCs derived neurons | Genotype:TD | Sample type:Extracellular media Sample source:Male Homo sapien iPSCs derived neurons | Genotype:TD | Sample type:Extracellular media Sample source:Male Homo sapien iPSCs derived neurons | Genotype:TD | Sample type:Extracellular media Sample source:Male Homo sapien iPSCs derived neurons | Genotype:FXS | Sample type:Extracellular media Sample source:Male Homo sapien iPSCs derived neurons | Genotype:FXS | Sample type:Extracellular media Sample source:Male Homo sapien iPSCs derived neurons | Genotype:FXS | Sample type:Extracellular media 1-Aminocyclopropanecarboxylic acid 330433645.1 452601418.4 438177362.5 603801051.1 494664730.2 573771204 380978440 57265259.44 51794393.81 -42440982.14 -11227170.06 -169871824.7 -10286238.4 1-Methyl nicotinamide 268392045.7 526496120.6 362825632.5 214735777 162336241.5 196536342.4 111250.2105 77628237.53 73235292.82 41964863.22 11400123.69 6753800.379 14339478.68 2,6-Diaminopimelic acid 6133042.335 13144494.88 10913953.32 14915403.14 14879213.1 14496392.11 32704125.29 -14037088.97 -13203985.29 -12842132.92 -7436827.253 -20160265.07 -2767107.296 2-Amino-2-deoxy-D-mannose 453946.1097 550142.1488 536912.8914 1199260.32 816952.3939 960621.7963 0 0 0 0 0 0 0 3-Aminoisobutanoic acid 856004986 1525926414 1101453319 1096296633 1066605866 886208507 321115.2837 3956886.706 6604710.964 2174357.172 3947232.433 2743286.387 3366833.466 3-Dehydroxycarnitine 125819970 46399969.27 43770990.6 527037115.7 410101728.3 428919791.1 623600.0831 12407156.14 11661896.46 7701863.566 4977763.622 2399002.558 2650702.625 4a-Carbinolamine tetrahydrobiopterin 608273734 1240201866 937490621.9 1596025038 1640287947 1507297854 2321057819 309028908.7 259237259.8 -98306772.3 -18400868.01 -741246486.3 96479731.21 4-Guanidinobutanoic acid 76492213.81 121889866.4 102107949.9 60979422.13 56265550.56 54194677.84 4282201.498 6280605.943 11029854.8 5017841.686 2343844.235 -1500551.415 1407407.313 5-Amino-levulinic acid 214550263.4 298578315.7 272800635.2 422980563.6 287671471.1 434626720.5 584920273.8 132862622.8 134147155.1 4483578.297 24764761.44 -230787083.3 17186761.59 5'-Methylthioadenosine 7394619.398 12399137.55 9725065.501 10426189.29 15593496.72 10901104.23 6-Hydroxydopamine 998298.6735 854060.8013 1135453.644 1434454.011 1529554.087 1652789.891 0 0 1546734.706 1292309.868 2266311.149 839500.4272 1519741.849 Acetylcholine 9105005.807 3014216.696 3305584.163 40410845.18 31472068.95 31572547.02 0 675237.8862 695855.3035 462960.2953 126810.9274 0 0 Acetyl-CoA 144357.5245 554755.6301 134038.8711 428424.9259 452883.1402 162138.0785 0 0 0 0 0 0 0 Adenine 39549902.2 51257488.71 34620211.46 28326165.29 29861580.42 23269285.81 0 0 0 0 0 0 0 Alanine 465359187.1 721722430.9 600405052.3 738610265 696638126.4 661148121.6 4899783.031 65698916.39 95090375.81 104023487.3 103410923.6 51048452.92 109956489.3 Allothreonine 1627445951 2317621025 2203468034 2949417176 2453534296 2781425826 1869762701 220181742.6 262601784.6 -205083681.8 -24788953.8 -893873974.7 -50273771.21 alpha-Aminoadipic acid 198152706.9 327382414.1 357455774 316182069.8 244667323.5 266857280.5 790362.7697 4399396.94 4596125.359 6639732.043 4798110.522 1656946.586 3752700.352 Aminoisobutyric acid 856004986 1525926414 1101453319 1096296633 1066605866 886208507 0 0 0 0 0 0 0 Arginine 843078827 1607865193 1324424679 1876403616 1759988031 1655444298 2892031302 -1237648703 -925319777.6 -919561777.9 -860498249.6 -1749946824 -185214284.1 Argininosuccinic acid 3254661.459 7884120.51 6384435.374 5539630.916 5327494.601 5503479.914 0 99263.16027 284947.5184 41265.87331 0 0 0 Asparagine 71900110.83 101547399.6 59094314.46 52370361.39 51275828.98 49634716.77 36858008.4 9562841.088 8580434.951 -11036737.86 -10307234.98 -23359551.89 -7623682.363 beta-Alanine 465359187.1 721722430.9 600405052.3 738610265 696638126.4 661148121.6 3509681.449 67089017.97 96480477.39 105413588.9 104801025.2 52438554.51 111346590.8 Betaine 245156537.7 240500190.1 390446975.5 179366662.7 152622171.6 201466809.4 21811995.7 319959688.3 286352274.1 286561066.5 128219286 38859130.82 61408645.53 Biotin 19747705.4 44594655.16 27598402.32 41378474.84 39397145.5 32069697.54 16841238.44 3666312.612 2090712.696 -1571532.585 -778955.5959 -7956748.868 1671715.266 CAR 2:0 573908247.2 1090499384 1951565979 1735269836 1633641890 1972582604 1948437.152 154016658.4 176895131.1 141081791.6 153317611 91357201.95 123234101.1 CAR DC5:0 7410952.184 11711038.91 10698448.83 10827097.45 7533461.086 8715059.128 36902.77711 348594.3119 273897.2448 111043.7881 340103.0301 94796.33741 25495.93514 Carnitine 2907336659 3760717739 3415109531 3295950383 2585257180 2708559653 1988521378 -262613293 -151343113.5 -353978355.5 -239189058.8 -1106280836 -223007738.9 CDP-choline 35789605.51 61597799.07 33731302.62 37761948.13 38674008.49 22518218.52 0 0 0 0 0 0 0 Choline 24827470.16 48406108.96 47506148.53 64337715.96 46446603.74 70067025.03 298392481.1 -280923156.3 -281286248.4 -247552503.5 -209544365.6 -270493463.3 -169271265.9 Choline phosphate 10959227491 21462253950 21362703026 31078656124 27560494839 24576412937 2594572.708 261930106.9 270016610 202317078.3 214265666.5 108896679.6 181643110.5 cis-4-Hydroxy-D-proline 206755456.8 295450990.8 273338194.8 436067452.9 285476190.5 457995166.1 582649630.1 134309966.4 134907206.3 6946037.223 23307037.93 -228747515.9 16929060.06 Citrulline 3423231.386 4516335.674 1420402.482 9204431.059 7147198.925 7096493.383 4807659.526 -117051.4831 4188523.466 1039945.581 1022346.505 -2247831.627 -89169.79928 Creatine 28315752527 42021445974 31924396980 39371453183 38617673679 33258236115 66404025.03 1106938456 1031490354 491380010.3 591477899 243043680.6 432002014.5 Creatinine 95510252.54 153992526.6 112748444.9 123555385.5 112841403.8 104120659.2 17686367.78 137394627.9 123137281.4 82480457.07 50013608.57 17699094.57 37500119.62 Cys-Gly 1554783.597 4034895.678 1366620.449 8516861.979 2598747.617 4458116.113 0 0 0 0 0 0 0 Cystathionine 61983635.06 133091647.8 72519627.81 45918713.91 62291762.46 32875856.8 0 2040301.065 2897719.006 1206842.809 1253198.321 715738.0925 664145.841 Cystine 480127.9557 105406.1249 188637.273 0 111337.6065 0 19081587.83 -13988058.33 -12340637.91 -6710061.547 -6505002.948 -11302081.28 -5142633.653 Cytosine 39700599.39 59330650.21 72164673.36 70354177.81 62471501.15 73642909.47 4277336.005 84483359.88 103674463.1 101648714.1 32331215.29 12468374.78 32956822.21 D-Alanine 465359187.1 721722430.9 600405052.3 738610265 696638126.4 661148121.6 3314679.958 67284019.46 96675478.88 105608590.4 104996026.7 52633556 111541592.3 Dethiobiotin 10424128.79 9913996.88 11149433 9839008.21 7973254.034 7733401.712 491763.0464 3193189.397 2443365.074 2299573.413 1664479.018 735251.438 2943554.602 Diethanolamine 65949001.69 75265210.54 89454196.64 92445468.93 92749873.03 109263569.2 62722213.04 -4029466.348 675925.453 -17062277.38 -6916199.226 -33062378.37 -8824917.97 Dihydrouracil 11309134.11 15315501.11 13802258.86 15543639.24 15905505.71 15375313.41 0 0 0 0 0 0 0 Dimethylarginine 54441526.44 90461792.08 61915318.73 32276011.94 27434654.25 24835920.01 438843.1276 112904674.8 90664130.74 68653380.62 28511291.3 15146008 24330954.23 D-Ornithine 61884096.84 95989208.13 51669862.04 54154512.86 53344909.09 45470650.51 4569824.735 111018589.2 109936118.2 57401994.97 57673390.09 28094460.43 55171497.36 Galactosamine 453946.1097 550142.1488 536912.8914 1199260.32 816952.3939 960621.7963 0 0 0 0 0 0 0 Glucosamine 439103.6141 550142.1488 536912.8914 1199260.32 816952.3939 960621.7963 862844.4026 -565414.5445 -250715.0711 32219.87594 -424560.9579 -399303.1678 -304810.1686 Glutamine 1760127499 2620425391 2217464259 5131037516 4765066631 5039948167 6364953.003 1405531995 1694938471 1261811341 2064155774 1104515635 2158687238 Glutathione 95846802.21 286884928.4 111352969.7 519832042.5 223824246.2 256719631.1 0 0 0 0 0 0 0 Glycine 38470736.22 53541634.46 43879650.72 63084851.27 53803017.77 56358779.71 4736447.365 4799903.278 1989728.97 1859178.472 446985.9109 -2130269.953 1326427.936 Guanidoacetic acid 1511405422 1750733164 1473130434 1387879676 1221861741 1332126765 869148.0651 615580604.1 520188926.9 384773535.2 285664918.1 122043078.7 291366172.5 Guanine 14595258.25 22449548.49 22606197.09 26789374.1 23868745.56 24097552.09 33119489.22 -5652390.491 -4623876.537 -6540170.222 -2403448.662 -16245076.63 -1174827.799 Homoserine 1668784006 2327315572 2219110132 2970006164 2474529200 2830467715 1903311662 226076496.2 271929096.7 -202688478.9 -17817709.23 -907079770.4 -51920429.31 Hypotaurine 75245347.17 166535020.6 68703730.46 464350235 314657412.2 303573961.4 0 132012.0378 779692.3039 121148.347 2156421.627 927977.0885 2405905.957 Hypoxanthine 229757320.7 233038465.2 141995490.2 91374762.24 87389555.66 78969764.68 1321322.553 1414397629 1072713875 703286504.9 483074616.5 267970438.5 433997983.4 Isoleucine 2746400677 5140066489 4159736019 5357097453 4700911982 4923839226 8288381211 -637768522.5 -585952657.9 -1102612847 -562719893.1 -4124492210 -324952324.8 Kynurenine 13734737.06 15980539.71 27561242.89 3692234.083 4349309.6 2877310.818 595455.145 68749282.12 50286769.83 69275596.94 8095525.506 4298931.537 4849677.91 Leucine 259240722.3 384902612.5 402819279.1 387891611.1 511652757.8 406443508.3 1446861780 -285705694.4 -246703741.4 -386434840.5 -69564925.16 -816592072.6 -141781006.3 L-Homocysteine thiolactone 67738.03748 42154.02381 152733.6547 244055.7623 525558.0045 409931.2386 0 209267.7354 2220479.262 75061.4375 60031.7833 71054.88564 56857.50534 Lysine 173183742.7 391852197.4 309018352.9 395646781.2 391758585 362063876 925234804.9 -425290762.9 -362464380.8 -372722533.3 -252632356.4 -602070627.1 -19172821.72 Methionine 300514200 489946697.4 455035677.8 548118210.7 473850004.1 500906967.5 730304316.4 -61509207.52 -69014094.54 -112697970.3 -28808779.48 -349609063.2 15247210.87 N(2)-Acetyllysine 1502345.036 2060271.412 2030545.707 2738902.237 2380701.088 2047780.602 0 0 0 0 0 0 0 N-6-Trimethyllysine 24210933.48 50063693.77 30208856.23 28336470.42 23157739.08 22631217.25 244455.5208 12541366.83 11891411.55 6702431.226 2823315.524 1547123.326 2354793.396 N-Acetylasparagine 45999.52056 64364.4747 69844.41175 165468.396 222167.7032 127029.4844 0 0 0 0 0 0 0 N-Acetyl-D-galactosamine 1199671.476 1552990.18 1619270.611 549438.798 883940.8906 1127028.625 0 0 0 0 0 0 0 N-Acetyl-D-glucosamine 0 0 0 0 0 174374.3117 137226.5961 8424541.171 12931730.95 12690970.33 867835.3864 -21778.63513 3358621.224 N-Acetylneuraminic acid 3369028.598 10546497.92 4271812.832 14499602.31 18276404.47 9487922.379 1045927.957 7982289.526 7676118.931 2953237.06 2810255.584 1110713.304 2754900.002 N-Acetylserotonin 4724067.679 5857210.761 3973621.184 460358.3154 815402.0531 285720.2349 0 0 0 0 0 0 0 NAD+ 59349284.84 112363490.5 85658325 116655636.4 117979592.1 101904869.3 0 0 0 0 0 0 0 NADP+ 603718.1665 1634320.001 732437.2575 925888.8685 1391376.722 732665.3355 0 24802.36928 0 0 0 0 0 Niacinamide 30744421.34 48878964.69 33419876.22 121005349 70948992.79 86586440.76 730109988.5 -320704924.1 -240813904.6 -265219465 -48644322.74 -417805871.3 144927803.7 Ophthalmic acid 855048.7448 1471837.333 622418.4374 468296.1219 507023.7506 510524.1524 0 0 0 0 0 0 0 Ornithine 14888.40648 9234.166717 13626.89828 343039.0634 62842.24967 0 107489.7623 -57552.34567 -71950.97645 -107489.7623 -51940.24081 99761.95021 -15915.92834 Oxidized glutathione 101057678.4 185855411 133095914.2 225541907.5 255547563.1 242331474.5 162340392.8 21069458.37 10166439.2 -14891405.3 -5950977.314 -76427483.1 6313436.118 Pantothenic acid 728800244.8 1586483510 960217478.2 1259419900 1323531556 1087415885 11258124686 1827594432 1877930542 194490774.1 544604794.1 -5037238203 1077890151 Phenylalanine 3750418772 6883044446 5045998670 6964420470 6612623116 6465785411 0 7841185.532 7081671.94 6265654.933 6725458.524 3319750.273 7145996.832 Phosphoethanolamine 141699655.9 228342520.8 156363052.2 141509297.9 137479148.6 170013107.1 0 0 0 0 0 0 0 Proline 25095931002 35551819689 33457454564 40035130126 37934355840 37910290749 27698153495 1583223499 1128753392 298209443.2 1248440164 -10294217461 2331428330 Pyridoxal 30341747.23 40859875.77 29353695.51 29951178.54 37425227.32 33878433.15 3926656.541 137120034.4 123656093.1 70299094.3 60440100.71 25606974.5 49303772.84 Riboflavin 0 0 0 0 149597.4759 213006.2952 3396879.16 -462610.5305 -464547.8888 -1410793.071 -952203.9002 -2316472.623 -1084207.432 S-Adenosylhomocysteine 2956988.299 5115289.131 3959989.138 3222908.811 3177582.918 2588430.402 0 0 0 0 0 0 0 S-Adenosylmethionine 6487603.462 7039728.481 6540975.596 3356825.998 5211091.884 4757728.03 0 0 0 0 0 0 0 Sarcosine 465359187.1 721722430.9 600405052.3 738610265 696638126.4 661148121.6 3349003.792 67249695.63 96641155.05 105574266.6 104961702.9 52599232.16 111507268.5 Serine 103867186.3 181681671.7 168155706.8 310572601.2 236936228.8 251538680.1 124379383.6 -28664337.58 -3211038.495 -29975757.04 -12119520.92 -67892291.68 -2001029.536 Taurine 24644769.73 34043741.76 25730065.08 24592904.63 25961745.73 29690972.16 1580960.311 776684.9364 2507074.138 -1156718.627 2113935.511 -981840.6891 -1168726.781 Thiamine 15005508.34 25368689.51 16251060.99 19693653.62 18595549.74 17511894.25 144216066.7 -1033592.936 -1977673.625 -18796124.48 -14525441 -77514007.97 -16890609.9 Threonine 1668784006 2327315572 2219110132 2970006164 2474529200 2830467715 1903311662 226076496.2 271929096.7 -202688478.9 -17817709.23 -907079770.4 -51920429.31 Trigonelline 7473157.564 9487789.892 7399124.044 13867884.89 4961690.302 7131037.91 12075568.77 8102243.921 30637658.78 6229291.704 40619094.65 -5036027.952 5430043.204 Tryptophan 381843873.3 701710139.5 485591079.8 688519867.3 630027878.9 651498755.8 1270676522 35658873.26 13563375.12 -163575396.8 -107064321.5 -646813189.3 50611740.41 Tyrosine 101176917 176141010.7 126425953.2 199932728.4 181461953.7 178653134.8 283680876.5 44478298.27 49732768.14 -2061804.064 31372495.71 -113977350.7 51224946.35 Valine 1599694144 2311607259 1844180226 2403237890 2337253745 2221065748 5399371073 214724715.1 214784049.5 -191732190.7 -208611838.6 -2354622580 413302132.6 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Retention index Quantified m/z PubChem ID KEGG ID Polarity 1-Aminocyclopropanecarboxylic acid 7.6 102.05496 535 C01234 Positive 1-Methyl nicotinamide 12.5 138.07876 457 C02918 Positive 2,6-Diaminopimelic acid 11.8 191.10263 99290 C00666 Positive 2-Amino-2-deoxy-D-mannose 7.81 180.08665 440049 C03570 Positive 3-Aminoisobutanoic acid 8.5 104.07061 64956 C05145 Positive 3-Dehydroxycarnitine 7.16 146.11756 134 C01181 Positive 4a-Carbinolamine tetrahydrobiopterin 6.5 240.10912 119055 C02953 Positive 4-Guanidinobutanoic acid 8.1 146.0924 500 C01035 Positive 5-Amino-levulinic acid 7.66 132.06552 137 C00430 Positive 5'-Methylthioadenosine 3.3 298.09684 439176 C00170 Positive 6-Hydroxydopamine 7.42 170.08117 4624 0 Positive Acetylcholine 7 147.12538 187 C01996 Positive Acetyl-CoA 8.1 810.13305 444493 C00024 Positive Adenine 4.6 136.06177 190 C00147 Positive Alanine 7.9 90.05496 5950 C00041 Positive Allothreonine 7.5 120.06552 99289 C05519 Positive alpha-Aminoadipic acid 8.7 162.07608 469 C00956 Positive Aminoisobutyric acid 7.45 104.07061 6119 C03665 Positive Arginine 12.3 175.11895 6322 C00062 Positive Argininosuccinic acid 10.22 291.12991 16950 C03406 Positive Asparagine 8.26 133.06077 6267 C00152 Positive beta-Alanine 7.8 90.05496 239 C00099 Positive Betaine 5.6 118.08626 247 C00719 Positive Biotin 4.5 245.09544 171548 C00120 Positive CAR 2:0 5.5 204.12303 7045767 C02571 Positive CAR DC5:0 7.17 276.14416 71317118 0 Positive Carnitine 7 162.11247 10917 C00318 Positive CDP-choline 8.8 490.12243 13804 C00307 Positive Choline 9.75 105.11482 305 C00114 Positive Choline phosphate 8.5 184.07332 1014 C00588 Positive cis-4-Hydroxy-D-proline 7.6 132.06552 440014 C03440 Positive Citrulline 8.6 176.10297 9750 C00327 Positive Creatine 8 132.07675 586 C00300 Positive Creatinine 4.7 114.06619 588 C00791 Positive Cys-Gly 9.15 179.04849 439498 C01419 Positive Cystathionine 9.5 223.0747 439258 C02291 Positive Cystine 9 241.03112 67678 C00491 Positive Cytosine 5.45 112.05054 597 C00380 Positive D-Alanine 7.9 90.05496 71080 C00133 Positive Dethiobiotin 4.6 215.13902 445027 C01909 Positive Diethanolamine 7.5 106.08626 8113 C06772 Positive Dihydrouracil 7.8 115.0502 649 C00429 Positive Dimethylarginine 10.1 203.15025 123831 C03626 Positive D-Ornithine 11 133.09715 71082 C00515 Positive Galactosamine 8.21 180.08665 3034366 0 Positive Glucosamine 7.88 180.08665 439213 C00329 Positive Glutamine 8.1 147.07642 5961 C00064 Positive Glutathione 8.5 308.09108 124886 C00051 Positive Glycine 8.5 76.0393 750 C00037 Positive Guanidoacetic acid 8.5 118.0611 763 C00581 Positive Guanine 6.5 152.05669 135398634 C00242 Positive Homoserine 7.7 120.06552 12647 C00263 Positive Hypotaurine 8.15 110.02703 107812 C00519 Positive Hypoxanthine 5.21 137.04579 135398638 C00262 Positive Isoleucine 5.5 132.10191 6306 C00407 Positive Kynurenine 5.4 209.09207 161166 C00328 Positive Leucine 5.3 132.10191 6106 C00123 Positive L-Homocysteine thiolactone 3.05 118.03211 134505 0 Positive Lysine 11.9 147.1128 5962 C00047 Positive Methionine 5.7 150.05833 6137 C00073 Positive N(2)-Acetyllysine 8 189.12337 192590 C12989 Positive N-6-Trimethyllysine 10.8 189.15975 440120 C03793 Positive N-Acetylasparagine 6.2 175.07133 99715 0 Positive N-Acetyl-D-galactosamine 6.02 222.09721 35717 C01074 Positive N-Acetyl-D-glucosamine 6.34 222.09721 439174 C00140 Positive N-Acetylneuraminic acid 7.5 310.11326 3568 C00270 Positive N-Acetylserotonin 2.86 219.1128 903 C00978 Positive NAD+ 8 664.1164 5893 C00003 Positive NADP+ 10 744.08273 5886 C00006 Positive Niacinamide 3.33 123.05529 936 C00153 Positive Ophthalmic acid 7.5 290.13466 7018721 C21016 Positive Ornithine 11 133.09715 6262 C00077 Positive Oxidized glutathione 10.5 613.15924 65359 C00127 Positive Pantothenic acid 4.5 220.11795 6613 C00864 Positive Phenylalanine 4.87 166.08626 6140 C00079 Positive Phosphoethanolamine 9.3 142.02637 1015 C00346 Positive Proline 6.76 116.07061 145742 C00148 Positive Pyridoxal 4 168.06552 1050 C00250 Positive Riboflavin 4 377.14556 493570 C00255 Positive S-Adenosylhomocysteine 7 385.12886 439155 C00021 Positive S-Adenosylmethionine 8.4 400.15234 34756 C00019 Positive Sarcosine 7.97 90.05496 1088 C00213 Positive Serine 8.5 106.04987 5951 C00716 Positive Taurine 8.1 126.02194 1123 C00245 Positive Thiamine 10 266.11958 1130 C00378 Positive Threonine 7.8 120.06552 6288 C00188 Positive Trigonelline 5.94 138.05496 5570 C01004 Positive Tryptophan 5.9 205.09715 6305 C00078 Positive Tyrosine 6.65 182.08117 6057 C00082 Positive Valine 5.7 118.08626 6287 C00183 Positive METABOLITES_END #END