#METABOLOMICS WORKBENCH spinelli_lab_20251113_085852 DATATRACK_ID:6688 STUDY_ID:ST004404 ANALYSIS_ID:AN007364 PROJECT_ID:PR002783 VERSION 1 CREATED_ON November 26, 2025, 12:55 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 L-Glutamine-13C5,15N2 stable isotope tracing on cell and 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 test whether the reduced steady-state glutamate levels in FXS neurons were ST:STUDY_SUMMARY due to impaired synthesis, we traced [¹³C₅,¹⁵N₂]-glutamine in 4-week ST:STUDY_SUMMARY induced pluripotent stem cells (iPSC) derived forebrain excitatory neurons ST:STUDY_SUMMARY generated from a Fragile X Syndrome (FXS) and CRISPR edited isogenic control. ST:STUDY_SUMMARY LC/MS polar metabolomics data showed that glutamine was not efficiently ST:STUDY_SUMMARY converted into glutamate and consequently fewer labeled carbons enter the ST:STUDY_SUMMARY oxidative and reductive TCA cycle (reflected by reduced levels of labeled α-KG, ST:STUDY_SUMMARY fumarate, malate, and aspartate). However, the percent labeled glutamate ST:STUDY_SUMMARY secreted to the media by FXS neurons was increased, suggesting altered ST:STUDY_SUMMARY neurotransmission. 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 1 Sample source:Male Homo sapien iPSCs derived neurons cell | Genotype:TD | Sample type:Cultured cells RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S21.raw SUBJECT_SAMPLE_FACTORS - cell iControl 2 Sample source:Male Homo sapien iPSCs derived neurons cell | Genotype:TD | Sample type:Cultured cells RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S22.raw SUBJECT_SAMPLE_FACTORS - cell iControl 3 Sample source:Male Homo sapien iPSCs derived neurons cell | Genotype:TD | Sample type:Cultured cells RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S23.raw SUBJECT_SAMPLE_FACTORS - cell FXS 1 Sample source:Male Homo sapien iPSCs derived neurons cell | Genotype:FXS | Sample type:Cultured cells RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S24.raw SUBJECT_SAMPLE_FACTORS - cell FXS 2 Sample source:Male Homo sapien iPSCs derived neurons cell | Genotype:FXS | Sample type:Cultured cells RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S25.raw SUBJECT_SAMPLE_FACTORS - cell FXS 3 Sample source:Male Homo sapien iPSCs derived neurons cell | Genotype:FXS | Sample type:Cultured cells RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S26.raw SUBJECT_SAMPLE_FACTORS - blank 1 Sample source:blank media | Genotype:NA | Sample type:Blank RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S27.raw SUBJECT_SAMPLE_FACTORS - blank 2 Sample source:blank media | Genotype:NA | Sample type:Blank RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S28.raw SUBJECT_SAMPLE_FACTORS - media iControl 1 Sample source:Male Homo sapien iPSCs derived neurons media | Genotype:TD | Sample type:Culture media RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S07.raw SUBJECT_SAMPLE_FACTORS - media iControl 2 Sample source:Male Homo sapien iPSCs derived neurons media | Genotype:TD | Sample type:Culture media RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S08.raw SUBJECT_SAMPLE_FACTORS - media iControl 3 Sample source:Male Homo sapien iPSCs derived neurons media | Genotype:TD | Sample type:Culture media RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S09.raw SUBJECT_SAMPLE_FACTORS - media FXS 1 Sample source:Male Homo sapien iPSCs derived neurons media | Genotype:FXS | Sample type:Culture media RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S10.raw SUBJECT_SAMPLE_FACTORS - media FXS 2 Sample source:Male Homo sapien iPSCs derived neurons media | Genotype:FXS | Sample type:Culture media RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S11.raw SUBJECT_SAMPLE_FACTORS - media FXS 3 Sample source:Male Homo sapien iPSCs derived neurons media | Genotype:FXS | Sample type:Culture media RAW_FILE_NAME=20250110_MK_Sneha_Neuron_Basal_Metabolomics_and_Glutamine_Tracing_S12.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 For glutamine flux tracing experiments, cells were cultured for 8hrs in TR:TREATMENT_SUMMARY glutamine free media supplemented with 2mM L-Glutamine-13C5,15N2 TR:TREATMENT_SUMMARY (Catalog#607983, Millipore Sigma). #SAMPLEPREP SP:SAMPLEPREP_SUMMARY For extraction from cell pellets from in-vitro cultures, samples were incubated SP:SAMPLEPREP_SUMMARY 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. For SP:SAMPLEPREP_SUMMARY media, 10ul of sample was added to 90ul of 80% methanol and vortexed for 10mins SP:SAMPLEPREP_SUMMARY before spinning to collect the supernatant containing extracted metabolites and SP:SAMPLEPREP_SUMMARY vacuum dried. The extract was then resuspended 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 NEGATIVE MS:MS_COMMENTS The mass spectrometer had the spray voltage set to 4.0 kV, heated capillary to MS:MS_COMMENTS 350°C, and the HESI probe at 30 °C. The sheath gas flow was set at 10 units, MS:MS_COMMENTS auxiliary gas at 1 units, and sweep gas flow at 1 unit. An additional scan MS:MS_COMMENTS between 220-700 m/z was used to enhance nucleotide detection in the negative MS:MS_COMMENTS mode as well with the maximum injection time set to 80 msec. Data acquired by MS:MS_COMMENTS Thermo Fisher's Xcalibur software and analyzed by their Tracefinder software. MS:MS_COMMENTS The raw files provided contain data from both positive and negative ion mode. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak area MS_METABOLITE_DATA_START Samples cell iControl 1 cell iControl 2 cell iControl 3 cell FXS 1 cell FXS 2 cell FXS 3 media iControl 1 media iControl 2 media iControl 3 media FXS 1 media FXS 2 media FXS 3 blank 1 blank 2 Factors Sample source:Male Homo sapien iPSCs derived neurons cell | Genotype:TD | Sample type:Cultured cells Sample source:Male Homo sapien iPSCs derived neurons cell | Genotype:TD | Sample type:Cultured cells Sample source:Male Homo sapien iPSCs derived neurons cell | Genotype:TD | Sample type:Cultured cells Sample source:Male Homo sapien iPSCs derived neurons cell | Genotype:FXS | Sample type:Cultured cells Sample source:Male Homo sapien iPSCs derived neurons cell | Genotype:FXS | Sample type:Cultured cells Sample source:Male Homo sapien iPSCs derived neurons cell | Genotype:FXS | Sample type:Cultured cells Sample source:Male Homo sapien iPSCs derived neurons media | Genotype:TD | Sample type:Culture media Sample source:Male Homo sapien iPSCs derived neurons media | Genotype:TD | Sample type:Culture media Sample source:Male Homo sapien iPSCs derived neurons media | Genotype:TD | Sample type:Culture media Sample source:Male Homo sapien iPSCs derived neurons media | Genotype:FXS | Sample type:Culture media Sample source:Male Homo sapien iPSCs derived neurons media | Genotype:FXS | Sample type:Culture media Sample source:Male Homo sapien iPSCs derived neurons media | Genotype:FXS | Sample type:Culture media Sample source:blank media | Genotype:NA | Sample type:Blank Sample source:blank media | Genotype:NA | Sample type:Blank 2-Oxoglutaric acid 19310725.3 19630089.02 20208621.35 4207074.683 5245450.266 1641555.119 1664164.338 1080167.031 1145809.266 1324656.7 1365358.423 947446.5108 630115.9682 238549.0084 2-Oxoglutaric acid_13C1 4817381.594 5112402.116 4905949.753 874877.5349 942242.931 233147.93 303276.5263 156216.5899 234222.5653 197853.5988 182804.2804 90948.24957 0 0 2-Oxoglutaric acid_13C2 2961909.429 3194016.036 3149555.037 386897.5145 516981.8367 39298.72149 210610.5237 532617.2305 260250.5301 118605.4554 132133.763 42055.46218 71378.26552 84315.02781 2-Oxoglutaric acid_13C3 5048286.217 5025743.835 5184467.527 929421.5062 1228124.778 287125.3959 344100.1729 215279.1025 314235.4618 241624.7425 195369.1455 121074.7922 0 0 2-Oxoglutaric acid_13C4 544898.3373 528540.6868 524186.1785 73039.03805 111351.8064 32604.71915 104309.4532 28132.76584 85768.94917 86711.96697 83074.3599 46513.03927 59443.38587 86979.53985 2-Oxoglutaric acid_13C5 9512018.477 9742279.549 10541357.22 1659101.007 1857105.975 188563.5443 1514545.407 1219497.633 1832031.058 781498.882 723774.196 525611.4346 169869.8617 471864.3082 Aspartic acid 89924852.62 73931874.75 75152077.17 54582258.56 60875035.56 45469156.29 1486303.058 1791491.159 1505856.726 1078871.37 1311704.485 2440065.375 1281175.547 1189863.788 Aspartic acid_13C1 20463278.46 16666511.08 17406775.56 11753581 13561496.6 9824187.073 39419.18124 30735.08426 33723.39294 19477.69045 31932.8804 93316.95571 13319.27517 46823.37006 Aspartic acid_13C2 15108471.71 39773445.64 11264310.66 13303271.39 8686552.814 6157303.047 1955037.49 2061212.147 15825266.23 795938.7816 1415252.715 311639.0366 334588.3176 1578430.948 Aspartic acid_13C3 4974127.061 5141899.146 5443714.111 3571309.675 3802427.576 2443160.527 752986.6855 253451.4934 353433.5291 922445.3913 150665.4274 0 476213.7712 198695.4353 Aspartic acid_13C4 26059574.9 22109468.47 24056167.96 19066204.37 18730347.93 18441010.4 4807204.772 3986399.958 4203651.571 4614499.721 4507226.287 4647011.645 4604600.998 3919346.385 Aspartic acid_15N1 21197467.51 16241942.85 17361097.91 13058696.88 17668035.81 11641847.22 15736.23494 72705.38221 87478.34133 10578.76533 9054.171603 0 0 6582.551812 Citric acid 260380530.7 214251540.3 195653967.6 195694060.1 180451182.2 146697774.5 6036208.01 4659400.362 4536968.981 4969056.13 5031381.448 4321409.24 3981188.569 4594753.065 Citric acid_13C1 72306032.87 56918360.97 51925172.41 46688232.2 45771903.52 35383290.21 547389.7476 366848.6457 302318.6968 270710.1673 323337.637 262876.5058 216473.8109 295501.3715 Citric acid_13C2 50312747.75 41910867.5 37468235.33 30481464.59 32971458.19 25695269 155665.6196 96862.46446 60542.81588 46890.50892 26254.24179 13641.68622 9184.058642 19448.11907 Citric acid_13C3 40338741.18 33359458.88 29842891 21481007.83 21543492.34 16909070.61 86270.74487 72074.92199 57035.74985 21453.41433 13067.98069 5804.960908 0 0 Citric acid_13C4 45977060.56 40681607.77 33264909.96 36339376.62 40683646.38 31172762.81 120997.7941 78878.4238 37349.42959 57020.38686 43799.17384 17418.20596 5379.260322 6446.488092 Citric acid_13C5 44104096.86 35106869.35 31064777.09 22781210.08 21011698.12 17834510.84 17105.28458 24748.35033 35002.8361 56235.85088 4760.302582 5296.891409 5766.265217 0 Citric acid_13C6 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Fumaric acid 34583005.51 31087897.26 24200068.96 15644569.53 15775349.97 7898096.901 831116.8509 1158958.18 1028999.639 819854.0067 900169.48 770295.7581 490216.6197 609786.3675 Fumaric acid_13C1 8067657.312 6962054.392 5540735.717 2986491.362 3303829.572 1601428.384 61746.10563 82175.96451 42563.03389 0 19819.31566 28837.06908 6550.726578 9467.438283 Fumaric acid_13C2 7875928.35 6403185.008 4945067.194 2823252.424 2785194.532 1355072.624 40333.85926 72804.6716 100089.792 48462.79284 6248.950099 9974.650657 6465.51008 39086.23066 Fumaric acid_13C3 4103174.258 3769852.594 2712069.532 1261939.701 1046113.491 543542.1771 7327.158885 132715.0547 164324.7806 75958.22748 28926.03331 0 4663.008114 0 Fumaric acid_13C4 8930120.171 8144185.48 6364513.163 4133350.01 4019125.553 2309949.169 99980.87583 73045.53738 101548.602 24815.84337 37638.10773 12731.73768 0 7365.128555 Glutamic acid 238876253.9 192575767.8 165352635.7 119687591 134395639 86728106.64 1690531.298 3715562.131 1177846.959 1690806.133 1349748.111 2550817.571 0 0 Glutamic acid_13C1 69549126.97 56132393.74 51414977.1 33952356.47 36552641.36 22810053.28 320149.462 147851.1205 135751.0226 78261.82874 161758.6078 234269.1179 39408.50313 0 Glutamic acid_13C2 33911190.47 95584225.6 27774700.86 22764739.37 24107590.36 6685458.576 2236295.795 2575941.615 18537639.86 1371273.242 1037944.322 1280283.475 3018305.135 1440173.059 Glutamic acid_13C3 60656901.4 50490255.79 47113373.14 34570329.97 38622805.15 26120818.52 616504.0582 311904.5308 1360762.358 788488.4063 336431.8759 151365.4346 347002.1167 205257.2271 Glutamic acid_13C4 6175480.609 2678935.926 4359495.462 3104783.865 3257470.233 2149563.417 180114.3975 213644.7241 173916.3139 277429.3619 229756.1265 326087.2066 344566.7812 234391.8965 Glutamic acid_13C5 126350817 112124897 96919893.7 75344090.23 70174162.88 57112521.1 6644705.394 5371440.568 6680169.268 6526226.591 6058519.328 5886891.698 6756960.515 6004625.76 Glutamic acid_15N1 84122942.19 60596508.09 62916348.72 46527135.73 45816265.1 31358277.34 436330.9135 0 789772.1111 131198.7834 118213.555 251380.87 62891.36172 86116.14218 Isocitric acid 154804353.2 109825745.1 71400606.2 64645779.89 75002177.56 36074904.1 2900175.892 3675542.461 3586730.06 2438903.026 2310118.214 2084071.442 2936732.302 3339575.225 Isocitric acid_13C1 37274569.17 30027923.86 17688221.62 18336525.48 18679059.45 9259037.191 377474.9361 366848.6457 302318.6968 197412.7855 183134.2415 177164.66 154031.1447 295501.3715 Isocitric acid_13C2 22012801.87 17027212.61 11559453.93 9040444.46 10763349.44 5858126.015 155665.6196 96862.46446 60542.81588 46890.50892 26254.24179 13641.68622 9184.058642 19448.11907 Isocitric acid_13C3 24693833.07 17018214.97 11689725.47 11785170.8 11213235.81 6575148.397 86270.74487 72074.92199 57035.74985 21453.41433 13067.98069 5804.960908 0 0 Isocitric acid_13C4 12119013.52 8886455.03 6177063.959 9612890.173 8823599.868 4423721.272 120997.7941 78878.4238 37349.42959 57020.38686 43799.17384 17418.20596 5379.260322 6446.488092 Isocitric acid_13C5 30214109.27 24790805.79 19374005.84 11431186.48 12438251.6 7136276.23 129489.7087 128236.4062 132597.0384 58785.95903 34477.24105 38157.34404 5795.210262 0 Isocitric acid_13C6 1332450.695 669109.6602 807170.3518 774546.1802 533306.7085 565861.0192 0 0 0 0 0 0 0 0 Malic acid 309119782.7 282545543.9 223481813.1 141268110.1 137869236.3 70576172.9 7087102.104 9557491.679 7312930.881 8814047.661 8042448.478 7577725.329 6092709.131 8008116.389 Malic acid_13C1 73433130.84 64465868.77 51765225.03 28515250.36 27796982.82 14633083.48 828946.402 851728.6544 919289.0385 506841.7273 573140.7722 477169.0948 254992.1111 275553.8828 Malic acid_13C2 69910394.69 60622828.52 49389187.41 28094129.25 26133540.91 14088502.07 417499.0153 385712.0375 658245.1326 603497.3343 195852.4438 166355.7262 149171.7869 64871.54199 Malic acid_13C3 36350400.37 32400366.22 25487050.2 11748025.09 10718724.1 5903390.821 380155.7156 318328.3224 446118.768 165286.5104 72366.83994 36367.07665 8607.645678 6177.629181 Malic acid_13C4 85000837.44 74971710.31 60172289.18 38342119.18 36471593.09 20101032.53 1228264.674 1371143.065 1256272.844 699588.4964 533194.0358 453887.7302 161841.1834 150758.9066 Oxaloacetic acid 105410.4595 81702.96257 82358.13174 132416.6375 164033.6013 100407.0183 0 0 0 8945.840245 0 10011.68717 129954.1845 134796.4497 Oxaloacetic acid_13C1 0 50769.03258 0 0 0 15888.18798 12711.75171 31402.32505 35265.65829 6902.848231 0 0 0 0 Oxaloacetic acid_13C2 0 71940.25205 0 18674.86784 0 19482.58499 8917.414122 63171.47263 59981.47737 29816.62681 0 0 0 0 Oxaloacetic acid_13C3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Oxaloacetic acid_13C4 253044.4352 154681.2688 449450.9239 181161.8466 219626.5618 306578.8271 129051.5565 204604.2807 190359.4905 110368.5191 144050.9562 0 85129.74049 12865.32765 Succinic acid 8277203.338 8252137.252 4507804.827 6435362.483 7661055.737 3845859.732 396313.7834 584338.4287 3264795.207 523459.8475 276090.224 372374.4043 0 987108.903 Succinic acid_13C1 2631902.092 1279932.14 1264374.054 1828517.494 2080615.442 981515.6815 28210.62951 113573.945 166499.6092 66748.20755 24390.01974 28332.93849 39580.91409 18891.71526 Succinic acid_13C2 2501308.931 12763452.75 678527.168 2924117.617 1264006.448 474992.5309 576527.3493 1799622.366 5379801.093 0 491756.4434 82217.67583 218059.9253 542849.682 Succinic acid_13C3 199556.8296 445353.051 41484.06044 397501.3949 129203.2984 180480.8033 174822.9291 658432.4679 1051963.288 137030.2858 29660.9511 4666.868934 62789.48601 29299.47671 Succinic acid_13C4 4167308.267 2943940.235 2035551.831 2640382.786 3658962.552 1254949.026 0 0 0 0 0 0 0 0 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Retention index Quantified m/z PubChem ID KEGG ID Polarity 2-Oxoglutaric acid 9.3 145.01425 51 C00026 Negative 2-Oxoglutaric acid_13C1 9.3 146.0176 51 C00026 Negative 2-Oxoglutaric acid_13C2 9.3 147.02096 51 C00026 Negative 2-Oxoglutaric acid_13C3 9.3 148.02431 51 C00026 Negative 2-Oxoglutaric acid_13C4 9.3 149.02767 51 C00026 Negative 2-Oxoglutaric acid_13C5 9.3 150.03102 51 C00026 Negative Aspartic acid 8.6 132.03023 5960 C00049 Negative Aspartic acid_13C1 8.6 133.03359 5960 C00049 Negative Aspartic acid_13C2 8.6 134.03694 5960 C00049 Negative Aspartic acid_13C3 8.6 135.0403 5960 C00049 Negative Aspartic acid_13C4 8.6 136.04365 5960 C00049 Negative Aspartic acid_15N1 8.6 136.04365 5960 C00049 Negative Citric acid 10.9 191.01973 311 C00158 Negative Citric acid_13C1 10.9 192.02308 311 C00158 Negative Citric acid_13C2 10.9 193.02644 311 C00158 Negative Citric acid_13C3 10.9 194.02979 311 C00158 Negative Citric acid_13C4 10.9 195.03315 311 C00158 Negative Citric acid_13C5 10.9 196.0365 311 C00158 Negative Citric acid_13C6 10.9 197.03985 311 C00158 Negative Fumaric acid 9.5 115.00368 444972 C00122 Negative Fumaric acid_13C1 9.5 116.00704 444972 C00122 Negative Fumaric acid_13C2 9.5 117.01039 444972 C00122 Negative Fumaric acid_13C3 9.5 118.01375 444972 C00122 Negative Fumaric acid_13C4 9.5 119.0171 444972 C00122 Negative Glutamic acid 8.5 146.0459 33032 C00025 Negative Glutamic acid_13C1 8.5 147.04292 33032 C00025 Negative Glutamic acid_13C2 8.5 147.04292 33032 C00025 Negative Glutamic acid_13C3 8.5 147.04292 33032 C00025 Negative Glutamic acid_13C4 8.5 147.04292 33032 C00025 Negative Glutamic acid_13C5 8.5 147.04292 33032 C00025 Negative Glutamic acid_15N1 8.5 147.04292 33032 C00025 Negative Isocitric acid 10.8 191.01973 1198 C00311 Negative Isocitric acid_13C1 10.8 192.02308 1198 C00311 Negative Isocitric acid_13C2 10.8 193.02644 1198 C00311 Negative Isocitric acid_13C3 10.8 194.02979 1198 C00311 Negative Isocitric acid_13C4 10.8 195.03315 1198 C00311 Negative Isocitric acid_13C5 10.8 196.0365 1198 C00311 Negative Isocitric acid_13C6 10.8 197.03985 1198 C00311 Negative Malic acid 9.5 133.01425 222656 C00149 Negative Malic acid_13C1 9.5 134.0176 222656 C00149 Negative Malic acid_13C2 9.5 135.02096 222656 C00149 Negative Malic acid_13C3 9.5 136.02431 222656 C00149 Negative Malic acid_13C4 9.5 137.02767 222656 C00149 Negative Oxaloacetic acid 9 130.9986 Negative Oxaloacetic acid_13C1 9 132.00195 Negative Oxaloacetic acid_13C2 9 133.00531 Negative Oxaloacetic acid_13C3 9 134.00866 Negative Oxaloacetic acid_13C4 9 135.01202 Negative Succinic acid 8.8 117.01933 1110 C00042 Negative Succinic acid_13C1 8.8 118.02269 1110 C00042 Negative Succinic acid_13C2 8.8 119.02604 1110 C00042 Negative Succinic acid_13C3 8.8 120.0294 1110 C00042 Negative Succinic acid_13C4 8.8 121.03275 1110 C00042 Negative METABOLITES_END #END