#METABOLOMICS WORKBENCH prats1988_20210404_005908 DATATRACK_ID:2563 STUDY_ID:ST001779 ANALYSIS_ID:AN002889 PROJECT_ID:PR001132 VERSION 2 CREATED_ON May 8, 2021, 11:06 am #PROJECT PR:PROJECT_TYPE Untargeted quantitative metabolomics analysis PR:PROJECT_SUMMARY A549 lung cancer cells were treated in vitro with control, 0.5 mM ATP and 10 PR:PROJECT_SUMMARY ng/ml TGF-beta for 2, 6 and 12 hours The untargeted metabolomics data was PR:PROJECT_SUMMARY generated from these studies. PR:INSTITUTE Ohio University PR:DEPARTMENT Biological Sciences PR:LABORATORY Dr. Xiaozhuo Chen, Edison biotechnology Institute PR:FIRST_NAME Pratik PR:ADDRESS Room 425, Parks Hall, College of Pharmacy, Ohio State University, Columbus Ohio. PR:ADDRESS 43210 PR:EMAIL ps774614@ohio.edu PR:PHONE 7406033801 PR:FUNDING_SOURCE This study was funded by NIH R15 grant CA242177-01 to Dr. Xiaozhuo Chen. PR:PROJECT_TITLE Untargeted metabolomics analysis of A549 cancer cells treated in control, 0.5 mM PR:PROJECT_TITLE ATP and 10 ng/ml TGF-beta for 2, 6 and 12 hours PR:LAST_NAME Shriwas #STUDY ST:STUDY_TITLE Untargeted Metabolomics analysis of A549 treated with 0.5 mM extracellular ATP ST:STUDY_TITLE and 10 ng/ml TGF-beta ST:STUDY_SUMMARY Control, 0.5 mM extracellular ATP and 10 ng/ml TGF-beta were used to treated 5 ST:STUDY_SUMMARY million A549 lung cancer cells in vitro for 2, 6 and 12 hours. The untargeted ST:STUDY_SUMMARY metabolomics analysis was performed on the cell lysates. The main objective of ST:STUDY_SUMMARY the study was to determine changes in metabolite abundances in lung cancer after ST:STUDY_SUMMARY treatment with extracellular ATP and TGF-beta (a known EMT inducer). ST:INSTITUTE Ohio University ST:DEPARTMENT Biological Sciences ST:LABORATORY Dr. Xiaozhuo Chen, Edison biotechnology Institute ST:LAST_NAME Shriwas ST:FIRST_NAME Pratik ST:ADDRESS Room 425, Parks Hall, College of Pharmacy, Ohio State University, Columbus Ohio. ST:ADDRESS 43210 ST:EMAIL ps774614@ohio.edu ST:NUM_GROUPS 7 ST:STUDY_TYPE Untargeted metabolomics analysis in lung cancer cells ST:PHONE 7406033801 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:CELL_BIOSOURCE_OR_SUPPLIER ATCC SU:CELL_STRAIN_DETAILS Human Lung epithelial A549 cancer cells SU:CELL_PASSAGE_NUMBER 10 SU:CELL_COUNTS 5 million cells #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 - Control_1 Treatment:Control RAW_FILE_NAME=ControlA_1.mzxml SUBJECT_SAMPLE_FACTORS - Control_2 Treatment:Control RAW_FILE_NAME=ControlA_2.mzxml SUBJECT_SAMPLE_FACTORS - Control_3 Treatment:Control RAW_FILE_NAME=ControlB_1.mzxml SUBJECT_SAMPLE_FACTORS - Control_4 Treatment:Control RAW_FILE_NAME=ControlB_1.mzxml SUBJECT_SAMPLE_FACTORS - 2h ATP_1 Treatment:2h ATP RAW_FILE_NAME=2hATP1_1.mzxml SUBJECT_SAMPLE_FACTORS - 2h ATP_2 Treatment:2h ATP RAW_FILE_NAME=2hATP1_2.mzxml SUBJECT_SAMPLE_FACTORS - 2h ATP_3 Treatment:2h ATP RAW_FILE_NAME=2hATP2_1.mzxml SUBJECT_SAMPLE_FACTORS - 2h ATP_4 Treatment:2h ATP RAW_FILE_NAME=2hATP2_2.mzxml SUBJECT_SAMPLE_FACTORS - 2h ATP_5 Treatment:2h ATP RAW_FILE_NAME=2hATP3_1.mzxml SUBJECT_SAMPLE_FACTORS - 2h ATP_6 Treatment:2h ATP RAW_FILE_NAME=2hATP3_2.mzxml SUBJECT_SAMPLE_FACTORS - 6h ATP_1 Treatment:6h ATP RAW_FILE_NAME=6hATP1_1.mzxml SUBJECT_SAMPLE_FACTORS - 6h ATP_2 Treatment:6h ATP RAW_FILE_NAME=6hATP1_2.mzxml SUBJECT_SAMPLE_FACTORS - 6h ATP_3 Treatment:6h ATP RAW_FILE_NAME=6hATP2_1.mzxml SUBJECT_SAMPLE_FACTORS - 6h ATP_4 Treatment:6h ATP RAW_FILE_NAME=6hATP2_2.mzxml SUBJECT_SAMPLE_FACTORS - 12h ATP_1 Treatment:12h ATP RAW_FILE_NAME=12hATP1_1.mzxml SUBJECT_SAMPLE_FACTORS - 12h ATP_2 Treatment:12h ATP RAW_FILE_NAME=12hATP1_2.mzxml SUBJECT_SAMPLE_FACTORS - 12h ATP_3 Treatment:12h ATP RAW_FILE_NAME=12hATP2_1.mzxml SUBJECT_SAMPLE_FACTORS - 12h ATP_4 Treatment:12h ATP RAW_FILE_NAME=12hATP2_2.mzxml SUBJECT_SAMPLE_FACTORS - 12h ATP_5 Treatment:12h ATP RAW_FILE_NAME=12hATP3_1.mzxml SUBJECT_SAMPLE_FACTORS - 12h ATP_6 Treatment:12h ATP RAW_FILE_NAME=12hATP3_2.mzxml SUBJECT_SAMPLE_FACTORS - 2h TGF-?_1 Treatment:2h TGF-? RAW_FILE_NAME=2hTGF1_1.mzxml SUBJECT_SAMPLE_FACTORS - 2h TGF-?_2 Treatment:2h TGF-? RAW_FILE_NAME=2hTGF1_2.mzxml SUBJECT_SAMPLE_FACTORS - 2h TGF-?_3 Treatment:2h TGF-? RAW_FILE_NAME=2hTGF2_1.mzxml SUBJECT_SAMPLE_FACTORS - 2h TGF-?_4 Treatment:2h TGF-? RAW_FILE_NAME=2hTGF2_2.mzxml SUBJECT_SAMPLE_FACTORS - 2h TGF-?_5 Treatment:2h TGF-? RAW_FILE_NAME=2hTGF3_1.mzxml SUBJECT_SAMPLE_FACTORS - 2h TGF-?_6 Treatment:2h TGF-? RAW_FILE_NAME=2hTGF3_2.mzxml SUBJECT_SAMPLE_FACTORS - 6h TGF-?_1 Treatment:6h TGF-? RAW_FILE_NAME=6hTGF1_1.mzxml SUBJECT_SAMPLE_FACTORS - 6h TGF-?_2 Treatment:6h TGF-? RAW_FILE_NAME=6hTGF1_2.mzxml SUBJECT_SAMPLE_FACTORS - 6h TGF-?_3 Treatment:6h TGF-? RAW_FILE_NAME=6hTGF2_1.mzxml SUBJECT_SAMPLE_FACTORS - 6h TGF-?_4 Treatment:6h TGF-? RAW_FILE_NAME=6hTGF2_2.mzxml SUBJECT_SAMPLE_FACTORS - 12h TGF-?_1 Treatment:12h TGF-? RAW_FILE_NAME=12hTGF1_1.mzxml SUBJECT_SAMPLE_FACTORS - 12h TGF-?_2 Treatment:12h TGF-? RAW_FILE_NAME=12hTGF1_2.mzxml SUBJECT_SAMPLE_FACTORS - 12h TGF-?_3 Treatment:12h TGF-? RAW_FILE_NAME=12hTGF2_1.mzxml SUBJECT_SAMPLE_FACTORS - 12h TGF-?_4 Treatment:12h TGF-? RAW_FILE_NAME=12hTGF2_2.mzxml SUBJECT_SAMPLE_FACTORS - 12h TGF-?_5 Treatment:12h TGF-? RAW_FILE_NAME=12hTGF3_1.mzxml SUBJECT_SAMPLE_FACTORS - 12h TGF-?_6 Treatment:12h TGF-? RAW_FILE_NAME=12hTGF3_2.mzxml #COLLECTION CO:COLLECTION_SUMMARY A549 lung cancer cells were treated with control, ATP, TGF-beta and then CO:COLLECTION_SUMMARY collected according to sample preparation protocol. CO:COLLECTION_PROTOCOL_FILENAME Collection protocol CO:SAMPLE_TYPE Lung CO:COLLECTION_METHOD Ice cold methanol (80%) using cell scrappers CO:COLLECTION_LOCATION 150mm dish CO:COLLECTION_FREQUENCY Once CO:COLLECTION_DURATION 1-2 sec CO:VOLUMEORAMOUNT_COLLECTED 1 ml CO:STORAGE_CONDITIONS -80? CO:COLLECTION_VIALS Polypropylene 1.5 ml tubes CO:STORAGE_VIALS Polypropylene 1.5 ml CO:COLLECTION_TUBE_TEMP 4C CO:TISSUE_CELL_QUANTITY_TAKEN 5 million #TREATMENT TR:TREATMENT_SUMMARY 5 million A549 cells were treated with control (no treatment), 0.5 mM ATP or 10 TR:TREATMENT_SUMMARY ng/ml TGF-beta for 2, 6 and 12 hours. TR:TREATMENT In vitro treatment with small molecules TR:TREATMENT_DOSEVOLUME 10 ml DMEM media containing appropriate compounds TR:CELL_STORAGE 37C; 5% CO2 incubator TR:CELL_GROWTH_CONTAINER 150 mm Dish tissue culture treated TR:CELL_MEDIA DMEM (10% FBS; 1% Pen/Strep) TR:CELL_HARVESTING after 2, 6 and 12 hours TR:CELL_PCT_CONFLUENCE 80% TR:CELL_MEDIA_LASTCHANGED NA TR:TREATMENT_COMPOUND 0.5 mM ATP and 10 ng/ml TGF-beta #SAMPLEPREP SP:SAMPLEPREP_SUMMARY 5 × 106 A549 cells were treated with control (no treatment), 0.5 mM ATP or 10 SP:SAMPLEPREP_SUMMARY ng/ml TGF-beta for 2, 6 and 12 hours. After treatment, cells were washed twice SP:SAMPLEPREP_SUMMARY with deionized water and polar metabolites were then extracted with SP:SAMPLEPREP_SUMMARY cryogenically cold 80% methanol/water mixture. LC-MS grade water, methanol, and SP:SAMPLEPREP_SUMMARY acetonitrile (Fischer Scientific, PA, USA) were used. Methanol-extracted samples SP:SAMPLEPREP_SUMMARY were then sonicated in cycles of sonication phase and rest phase for 10 minutes SP:SAMPLEPREP_SUMMARY (5 second sonication phase and 10 seconds halt). The samples were then SP:SAMPLEPREP_SUMMARY centrifuged at 13,000 rpm for 10 minutes and supernatant was then collected. SP:SAMPLEPREP_SUMMARY Supernatants collected from in vitro and in vivo extraction were then SP:SAMPLEPREP_SUMMARY lyophilized. Briefly, the supernatant was then lyophilized by using a speed SP:SAMPLEPREP_SUMMARY vacuum evaporator. The samples were then dissolved into a mixture of SP:SAMPLEPREP_SUMMARY acetonitrile/water (1:1; v/v). SP:SAMPLEPREP_PROTOCOL_FILENAME Sample preparation protocol SP:PROCESSING_METHOD CEll scrapping Quenching SP:PROCESSING_STORAGE_CONDITIONS -80? SP:EXTRACTION_METHOD Quenching with Ice cold methanol SP:EXTRACT_ENRICHMENT Speed vaccum evaporator SP:EXTRACT_STORAGE -80? SP:SAMPLE_RESUSPENSION Acetonitrile/water (1:1) #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY The entire LC/MS-MS experiment was performed in the Campus Chemical CH:CHROMATOGRAPHY_SUMMARY Instrumentation Center’s Mass Spectrometry and Proteomics facility at The Ohio CH:CHROMATOGRAPHY_SUMMARY State University. Lyophilized samples were dissolved in equal amounts of LC-MS CH:CHROMATOGRAPHY_SUMMARY grade water and acetonitrile and run with LC/MS-MS analysis, using an untargeted CH:CHROMATOGRAPHY_SUMMARY metabolomics approach by utilizing Agilent Q-TOF 6545 mass spectrometer CH:CHROMATOGRAPHY_SUMMARY connected to an Agilent 1290 UHPLC system with a Poroshell 120 SB-C18 (2 x 100 CH:CHROMATOGRAPHY_SUMMARY mm, 2.7 µm particle size) column. The LC gradient consisted of solvent A, H2O CH:CHROMATOGRAPHY_SUMMARY with 0.1 % Formic acid, and solvent B, 100 % acetonitrile at a 200 µL/min flow CH:CHROMATOGRAPHY_SUMMARY rate with an initial 2 % solvent B with a linear ramp to 95 % B at 15 min, CH:CHROMATOGRAPHY_SUMMARY holding at 95% B for 1 minutes, and back to 2 % B from 16 min and equilibration CH:CHROMATOGRAPHY_SUMMARY of 2 % B until min 32. A 5 µL volume sample was injected for each run and the CH:CHROMATOGRAPHY_SUMMARY top 5 ions were selected for data-dependent analysis with a 15 second exclusion CH:CHROMATOGRAPHY_SUMMARY window. CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Agilent 1290 CH:COLUMN_NAME Poroshell 120 SB-C18 (2.1 x 100 mm x 2.7 µm) CH:FLOW_GRADIENT an initial 2 % solvent B with a linear ramp to 95 % B at 15 min, holding at 95% CH:FLOW_GRADIENT B for 1 minutes, and back to 2 % B from 16 min and equilibration of 2 % B until CH:FLOW_GRADIENT min 32. CH:FLOW_RATE 200 µL/min CH:COLUMN_TEMPERATURE 40 CH:SOLVENT_A H2O with 0.1 % Formic acid CH:SOLVENT_B 100 % acetonitrile #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME Campus Center for Chemical Instrumentation (CCIC) #MS MS:INSTRUMENT_NAME Agilent 6545 QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Masshunter software was used to collect the raw data. Progenesis was used for MS:MS_COMMENTS peak intergration. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS normalized MS_METABOLITE_DATA_START Samples Control_1 Control_2 Control_3 Control_4 2h ATP_1 2h ATP_2 2h ATP_3 2h ATP_4 2h TGF-?_1 2h TGF-?_2 2h TGF-?_3 2h TGF-?_4 Factors Treatment:Control Treatment:Control Treatment:Control Treatment:Control Treatment:2h ATP Treatment:2h ATP Treatment:2h ATP Treatment:2h ATP Treatment:2h TGF-? Treatment:2h TGF-? Treatment:2h TGF-? Treatment:2h TGF-? D-glucose 180871.632 155945.6778 108028.4886 104402.9947 78839.74905 85242.05611 60614.97795 59749.29751 62000.47622 69113.92096 61821.18276 63305.78447 fructose-6-phosphate 108652.7831 113442.9838 101617.833 107619.6798 157699.2985 138380.9583 127504.1583 139665.6394 86733.57814 80194.96147 83990.47581 84488.61864 Fructose 1,6-bisphosphate 168828.2037 149177.5662 665758.3249 630921.2057 698882.5107 691736.3535 631672.2715 652749.9638 200882.3149 186185.3748 308911.3652 190311.5511 Dihydroxyacetone phosphate 403254.5628 444441.2613 545070.6032 649191.3628 634123.3158 634123.3158 286943.5153 294878.0796 280738.9848 279249.4544 2-Phosphoglycerate 47073.16003 51566.05788 57316.24788 58308.27312 47550.38252 47969.96964 50910.56659 41519.16996 43143.44111 33455.92417 33608.83334 Pyruvic acid 76072.8415 84177.61543 60142.82689 65608.99229 61622.93244 64340.00074 69347.5298 50249.44972 58455.82322 53227.70587 58297.79217 L-Lactic acid 90771.13666 98022.23754 122993.4062 134833.0798 41344.11362 58963.75902 58963.75902 28261.79525 55811.73775 56920.39771 31009.62537 69319.53517 L-Glutamic acid 710496.604 676775.9899 1116337.183 1075631.557 659082.8565 774086.3579 694834.8322 640777.2211 439033.9826 513353.3698 405548.8024 547766.1365 Oxoglutaric acid 23653.57037 22139.81353 28315.34195 11732.14612 13215.39986 11806.56515 9978.930114 13090.35466 9834.016906 9087.907292 6916.593616 L-Aspartic acid 43981.80493 41190.39788 68638.19089 71916.92879 76292.68749 76818.31479 69401.10106 52174.61891 51486.70749 61880.86784 64954.8084 Oxalacetic acid 101658.5072 92233.32717 84399.79539 98249.65837 66683.74156 69797.4527 73238.44135 73405.35754 95316.12473 92734.22456 112109.3542 127271.5843 Glutathione 7272884.285 7744367.879 10062766.55 10803037.98 11356232.01 12674648.53 10861963.7 12120642.76 12933273.8 12025747.79 Oxidized Glutathione 2673139.343 3034161.983 6773703.111 7030776.01 7607149.483 5659046.245 6548066.536 10217288.06 9067432.331 10132139.43 Glycine 27101.20389 45294.35487 34886.29831 31376.84759 30309.63276 36557.42001 19168.23792 21590.36787 20516.32801 26790.12944 Acetyl-CoA 4850.471688 6219.514105 11252.96551 15272.18273 13267.02367 15524.25492 12122.90255 20454.71726 19825.77331 19254.21466 22086.7727 Citric acid 4374210.795 4879821.671 3230198.559 3139679.891 4030733.622 4270204.053 4100164.069 4204844.563 4810954.617 4392899.705 4112454.902 4106534.708 cis-Aconitic acid 292954.0616 264251.4851 285104.851 227440.616 331380.5934 341854.6757 334824.6748 330093.3035 512163.9311 505435.555 521079.7336 472515.5865 Succinyl-CoA 3223.100333 3552.717209 2037.363439 1191.078626 2806.222126 3704.565211 1732.297693 3823.77642 5184.842766 5258.99672 3906.539424 Succinic acid 800496.8317 875186.4758 1160137.074 1112538.082 953844.2706 968193.6116 869383.2121 949124.6734 525286.7966 582278.7404 624104.5299 641490.8725 fumaric acid 265758.3716 323465.8319 260131.2553 221480.2787 134217.9263 112933.9642 126089.5124 183695.081 195164.108 181610.488 114675.3273 205552.4569 Malic acid 1303503.138 1577512.156 1679568.861 1908545.329 2049257.638 1717979.85 1880342.162 1412869.815 1371362.035 1188249.46 1406914.553 Coenzyme A 6380.182928 7527.09858 9593.579425 11454.71803 16146.2739 11525.29601 18622.78236 16800.47879 13613.77461 11634.7926 Guanosine diphosphate 167361.5949 235830.6054 342440.9087 350654.8394 116708.6638 115731.8189 171885.3336 186910.7322 167951.078 184681.6131 184430.5036 276832.077 NAD 277420.5361 315257.3727 253265.2725 288909.4183 395627.274 342643.8471 348114.0684 451238.5385 180779.8092 254751.3093 236546.8188 223521.776 FAD 440240.8346 426299.8268 916454.6465 1080478.074 634732.413 674630.689 672205.6323 638229.664 849485.6005 888096.6359 753230.1234 815346.4388 Phosphorylcholine 42639.38468 30362.53348 36300.32893 10925.85392 16726.28758 19164.74603 7138.525219 9828.463308 8837.477535 LysoPC(16:0) 12971.20699 21149.74965 15265.30071 4956.081214 5321.371999 6827.593009 2846.79835 3477.906443 2835.742259 Phosphatidylinositol-3,4,5-trisphosphate 16213.49502 19233.0199 16570.40632 20457.97856 22589.74084 20885.26642 19284.32375 20474.10801 23773.70479 Arachidonic acid 162714.8731 139235.8577 159440.3653 271258.2418 279712.4534 122324.4237 160067.8528 116332.6167 178420.0286 Thromboxane a2 52850.78953 31920.78269 99193.19941 35414.27619 49127.56733 74768.51556 119487.0977 100616.1232 103719.9255 Prostaglandin D2 3152.854781 3145.715439 3340.243838 1564.785459 3406.361938 3809.758901 2372.503629 630.2070965 862.7063276 Prostaglandin E2 360889.8949 337384.3162 484712.8096 379676.8793 334910.0197 487043.3875 303542.7643 221641.0789 275253.326 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START Metabolite_name KEGG ID HMDB ID PUBCHEM ID Mode retention time_m/z m/z D-glucose C00221 HMDB0000122 5793 Negative 1.40_215.0338m/z 215.0337908 fructose-6-phosphate C00085 HMDB0000124 69507 Negative 1.43_260.0313n 260.0312509 Fructose 1,6-bisphosphate C05378 HMDB0001058 10267 Negative 2.02_338.9909m/z 338.9908581 Dihydroxyacetone phosphate C00111 HMDB0001473 668 Negative 1.60_169.9992n 338.9908202 2-Phosphoglycerate C00631 HMDB0000362 59 Negative 14.75_370.9787m/z 370.9786937 Pyruvic acid C00022 HMDB0000243 1060 Negative 1.93_87.0093m/z 87.00933104 L-Lactic acid C00186 HMDB0000190 107689 Negative 1.31_179.0572m/z 179.0572277 L-Glutamic acid C00025 HMDB0000148 33032 Negative 1.34_146.0470m/z 146.0469686 Oxoglutaric acid C00026 HMDB0000208 51 Negative 1.83_145.0148m/z 145.0148367 L-Aspartic acid C00049 HMDB0000191 5960 Negative 1.34_132.0311m/z 132.0310582 Oxalacetic acid C00036 HMDB0000223 970 Negative 13.87_262.9934m/z 262.9934231 Glutathione C00051 HMDB0062697 745 Negative 2.33_307.0861n 306.0788438 Oxidized Glutathione C00127 HMDB0003337 65359 Negative 4.92_612.1551n 611.1478583 Glycine C00037 HMDB0000123 750 Negative 1.29_74.0254m/z 74.02537056 Acetyl-CoA C00024 HMDB0001206 444493 Negative 8.38_808.0797m/z 808.0796604 Citric acid C00158 HMDB0000094 311 Negative 14.28_226.9929m/z 226.9928819 cis-Aconitic acid C00417 HMDB0000072 643757 Negative 3.37_173.0100m/z 173.0100124 Succinyl-CoA C00091 HMDB0001022 439161 Negative 12.08_906.0920m/z 906.0920187 Succinic acid C00042 HMDB0000254 1110 Negative 3.16_117.0201m/z 117.0201424 fumaric acid C00122 HMDB0000134 444972 Negative 2.93_115.0043m/z 115.0043117 Malic acid C03668 HMDB0000744 525 Negative 1.72_133.0152m/z 133.0152414 Coenzyme A C00010 HMDB0001423 87642 Negative 9.71_766.1110m/z 766.1110235 Guanosine diphosphate C00035 HMDB0001201 8977 Negative 3.35_444.0296m/z 444.0296018 NAD C00003 HMDB0000902 5893 Negative 3.87_663.1158m/z 663.1157954 FAD C00016 HMDB0001248 643975 Negative 9.09_784.1532m/z 784.1532207 Phosphorylcholine C00588 HMDB0001565 1014 Negative 7.92_219.0456m/z 219.0455678 Choline C00114 HMDB0000097 305 Positive 10.21_143.0694m/z 143.0693893 LysoPC(16:0) C04230 HMDB0010382 460602 Negative 15.74_530.2924m/z 530.2924336 Phosphatidylinositol-3,4,5-trisphosphate C05981 HMDB0004249 53477782 Negative 15.12_642.9822m/z 642.9821693 Arachidonic acid C00219 HMDB0001043 444899 Negative 16.50_303.2350m/z 303.2350214 Thromboxane a2 C02198 HMDB0001452 5280497 Negative 13.83_352.2272n 351.2199179 Prostaglandin D2 C00696 HMDB0001403 448457 Negative 13.86_387.1986m/z 387.1986467 Prostaglandin E2 C00584 HMDB0001220 5283116 Negative 14.10_351.2200m/z 351.2199929 METABOLITES_END #END