#METABOLOMICS WORKBENCH purdylab_20250617_065445 DATATRACK_ID:6050 STUDY_ID:ST004000 ANALYSIS_ID:AN006595 PROJECT_ID:PR002505 VERSION 1 CREATED_ON June 24, 2025, 6:50 am #PROJECT PR:PROJECT_TITLE Measuring PC lipids in HCMV infected fibroblasts treated with phosphonoacetic PR:PROJECT_TITLE acid (PAA). PR:PROJECT_SUMMARY Human cytomegalovirus (HCMV) is a common herpesvirus that establishes a lifelong PR:PROJECT_SUMMARY and persistent infection in its human host. HCMV infection in most people does PR:PROJECT_SUMMARY not cause overt disease. However, in immunocompromised individuals, severe PR:PROJECT_SUMMARY CMV-associated disease can lead to permanent disabilities and even death. PR:PROJECT_SUMMARY Additionally, congenital CMV is the leading infectious cause of birth defects. PR:PROJECT_SUMMARY Viruses have evolved to hijack host metabolic pathways to facilitate their PR:PROJECT_SUMMARY replication cycle. We previously reported HCMV infection increases PR:PROJECT_SUMMARY phosphatidylcholine (PC) lipid levels, including PCs with VLCFAs. To expand upon PR:PROJECT_SUMMARY the previously reported PC phenotype in HCMV infection, we determined the PC PR:PROJECT_SUMMARY lipidome of several infected cell types grown under various growth conditions. PR:PROJECT_SUMMARY Additionally, we determined which host pathways HCMV reprograms to induce PC PR:PROJECT_SUMMARY lipid synthesis and describe when during infection PC lipids changes occur. PR:INSTITUTE University of Arizona PR:DEPARTMENT Immunobiology PR:LABORATORY John G. Purdy, PhD PR:LAST_NAME Kline PR:FIRST_NAME Ian PR:ADDRESS 1657 E Helen St, Tucson, AZ 85721 PR:EMAIL ikline@arizona.edu PR:PHONE 5209092596 PR:FUNDING_SOURCE National Institute of Health (NIH) National Institute of Allergy and Infectious PR:FUNDING_SOURCE Disease (NIAID) R01AI162671, R01AI155539, F32AI178919, and National Institute of PR:FUNDING_SOURCE Aging (NIA) T32AG058503 award. #STUDY ST:STUDY_TITLE Measuring PC lipids in HCMV infected fibroblasts treated with phosphonoacetic ST:STUDY_TITLE acid (PAA). ST:STUDY_SUMMARY HCMV viral gene expression is classified into three kinetic classes, immediate ST:STUDY_SUMMARY early (IE), early (E) and late genes (L). HCMV promotes PC lipids by 48 hpi ST:STUDY_SUMMARY relative to uninfected cells, suggesting the involvement of an IE or E viral ST:STUDY_SUMMARY protein in regulating PC lipid levels. Viral DNA synthesis initiates L viral ST:STUDY_SUMMARY gene expression. Phosphonoacetic acid (PAA) inhibits HCMV genome replication and ST:STUDY_SUMMARY consequentially, L gene expression. Therefore, we used PAA to test whether L ST:STUDY_SUMMARY gene expression was required to reprogram PC lipid levels in HCMV infection. We ST:STUDY_SUMMARY find that PAA-treated cells contain similar levels of PC lipids to ST:STUDY_SUMMARY vehicle-treated conditions thus, L HCMV gene expression is not required to ST:STUDY_SUMMARY promote PC levels. ST:INSTITUTE University of Arizona ST:DEPARTMENT Immunobiology ST:LABORATORY John G. Purdy, PhD ST:LAST_NAME Kline ST:FIRST_NAME Ian ST:ADDRESS 1657 E Helen St, Tucson, AZ 85721 ST:EMAIL ikline@arizona.edu ST:PHONE 5209092596 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:GENDER Male SU:CELL_STRAIN_DETAILS HFF SU:CELL_PASSAGE_NUMBER <30 #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 rep_1_PAA_studies_pos_HCMV_H2O_72_a 20241023_pos_HCMV_h2O_72_R1_a Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:H20 Time (hpi)=72; Infection status=HCMV; RAW_FILE_NAME(.mzxML file name)=20241023_pos_HCMV_h2O_72_R1_a SUBJECT_SAMPLE_FACTORS rep_1_PAA_studies_pos_HCMV_H2O_72_b 20241023_pos_HCMV_h2O_72_R1_b Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:H20 Time (hpi)=72; Infection status=HCMV; RAW_FILE_NAME(.mzxML file name)=20241023_pos_HCMV_h2O_72_R1_b SUBJECT_SAMPLE_FACTORS rep_1_PAA_studies_pos_HCMV_PAA_100ug_72_a 20241023_pos_HCMV_PAA_72_R1_a Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:PAA 100ug/mL Time (hpi)=72; Infection status=HCMV; RAW_FILE_NAME(.mzxML file name)=20241023_pos_HCMV_PAA_72_R1_a SUBJECT_SAMPLE_FACTORS rep_1_PAA_studies_pos_HCMV_PAA_100ug_72_b 20241023_pos_HCMV_PAA_72_R1_b Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:PAA 100ug/mL Time (hpi)=72; Infection status=HCMV; RAW_FILE_NAME(.mzxML file name)=20241023_pos_HCMV_PAA_72_R1_b SUBJECT_SAMPLE_FACTORS rep_2_PAA_studies_pos_HCMV_H2O_72_a 20241023_pos_HCMV_h2O_72_R2_a Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:H20 Time (hpi)=72; Infection status=HCMV; RAW_FILE_NAME(.mzxML file name)=20241023_pos_HCMV_h2O_72_R2_a SUBJECT_SAMPLE_FACTORS rep_2_PAA_studies_pos_HCMV_H2O_72_b 20241023_pos_HCMV_h2O_72_R2_b Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:H20 Time (hpi)=72; Infection status=HCMV; RAW_FILE_NAME(.mzxML file name)=20241023_pos_HCMV_h2O_72_R2_b SUBJECT_SAMPLE_FACTORS rep_2_PAA_studies_pos_HCMV_PAA_100ug_72_a 20241023_pos_HCMV_PAA_72_R2_a Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:PAA 100ug/mL Time (hpi)=72; Infection status=HCMV; RAW_FILE_NAME(.mzxML file name)=20241023_pos_HCMV_PAA_72_R2_a SUBJECT_SAMPLE_FACTORS rep_2_PAA_studies_pos_HCMV_PAA_100ug_72_b 20241023_pos_HCMV_PAA_72_R2_b Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:PAA 100ug/mL Time (hpi)=72; Infection status=HCMV; RAW_FILE_NAME(.mzxML file name)=20241023_pos_HCMV_PAA_72_R2_b SUBJECT_SAMPLE_FACTORS rep_3_PAA_studies_pos_HCMV_H2O_72_a 20241023_pos_HCMV_h2O_72_R3_a Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:H20 Time (hpi)=72; Infection status=HCMV; RAW_FILE_NAME(.mzxML file name)=20241023_pos_HCMV_h2O_72_R3_a SUBJECT_SAMPLE_FACTORS rep_3_PAA_studies_pos_HCMV_H2O_72_b 20241023_pos_HCMV_h2O_72_R3_b Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:H20 Time (hpi)=72; Infection status=HCMV; RAW_FILE_NAME(.mzxML file name)=20241023_pos_HCMV_h2O_72_R3_b SUBJECT_SAMPLE_FACTORS rep_3_PAA_studies_pos_HCMV_PAA_100ug_72_a 20241023_pos_HCMV_PAA_72_R3_a Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:PAA 100ug/mL Time (hpi)=72; Infection status=HCMV; RAW_FILE_NAME(.mzxML file name)=20241023_pos_HCMV_PAA_72_R3_a SUBJECT_SAMPLE_FACTORS rep_3_PAA_studies_pos_HCMV_PAA_100ug_72_b 20241023_pos_HCMV_PAA_72_R3_b Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:PAA 100ug/mL Time (hpi)=72; Infection status=HCMV; RAW_FILE_NAME(.mzxML file name)=20241023_pos_HCMV_PAA_72_R3_b #COLLECTION CO:COLLECTION_SUMMARY General Notes: -Work over ice when possible (during scraping and between vortex CO:COLLECTION_SUMMARY steps etc.). -Chloroform leeches plastics. Avoid contact with plastic caps, CO:COLLECTION_SUMMARY tubes and gloves around glass vial tops. -Work in manageable batch numbers. A CO:COLLECTION_SUMMARY batch of 8-16 samples at a time is common. -Clean syringes using chloroform CO:COLLECTION_SUMMARY before and after collection. Use a unique syringe for each sample (the same CO:COLLECTION_SUMMARY syringe can be used if A/B technical replicates are used. Recommend a quick CO:COLLECTION_SUMMARY flush of the syringe using 500uL chloroform before moving from A to B). Begin by CO:COLLECTION_SUMMARY counting cell numbers for each sample using a dedicated well meant for MS CO:COLLECTION_SUMMARY normalization based on cell count. In a 6-well plate, wash cells 2x with cold CO:COLLECTION_SUMMARY PBS. Add 1mL of cold 50% methanol to each well and scrape cells into glass CO:COLLECTION_SUMMARY vials. Add 500uL of chloroform. Vortex on low setting (careful to avoid CO:COLLECTION_SUMMARY splashing chloroform onto plastic caps and liners). Centrifuge @1000g for 5 CO:COLLECTION_SUMMARY mins. There should be a clear phase separation of methanol and cell debris on CO:COLLECTION_SUMMARY top, while the lower phase contains chloroform and lipids. Use a syringe to CO:COLLECTION_SUMMARY carefully extract the lower phase without transferring cell debris from the top CO:COLLECTION_SUMMARY layer. Transfer to clean vial and place on ice. Once all lipids have been CO:COLLECTION_SUMMARY extracted, add 500uL of chloroform and repeat the process again once more. In CO:COLLECTION_SUMMARY total each sample should contain ~1mL of chloroform and lipids from two CO:COLLECTION_SUMMARY extractions. Carefully, dry lipids under nitrogen gas. Avoid direct high CO:COLLECTION_SUMMARY pressure air flow onto chloroform:lipid solution as it can splash high up in CO:COLLECTION_SUMMARY vial walls. Store at -80C for up to 30 days. Samples may become unstable and CO:COLLECTION_SUMMARY degrade over time. CO:SAMPLE_TYPE Cultured cells CO:COLLECTION_LOCATION Fume hood required when working with chloroform. CO:COLLECTION_FREQUENCY Perform extraction twice on each sample. CO:COLLECTION_DURATION 1-2 hours dependent upon batch size CO:VOLUMEORAMOUNT_COLLECTED ~1mL chloroform:lipid soluition CO:STORAGE_CONDITIONS -80℃ CO:COLLECTION_VIALS Glass vials, no plastic! (plastic cap is okay) CO:STORAGE_VIALS Glass vials, no plastic! (plastic cap is okay) CO:COLLECTION_TUBE_TEMP 25 CO:ADDITIVES UHPLC 100% chloroform #TREATMENT TR:TREATMENT_SUMMARY Cells were grown to full confluence in Dulbecco's modified eagle medium (DMEM) TR:TREATMENT_SUMMARY with 10% fetal bovine serum (FBS). Cells were growth arrested at full confluence TR:TREATMENT_SUMMARY for 3 days and then switched to serum-free DMEM for 24 hours. Cells were TR:TREATMENT_SUMMARY HCMV-infected for 1 hour in serum-free DMEM. At 1 hpi, cells were washed of TR:TREATMENT_SUMMARY unattached virus and maintained in serum-free DMEM with either 100ug/mL PAA or TR:TREATMENT_SUMMARY vehicle H2O. PAA or H2O was replaced every 24 h and medium was refreshed at 48 TR:TREATMENT_SUMMARY hpi. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Before starting, calculate the volume of 1:1:1 choloroform:methanol:isopropanol SP:SAMPLEPREP_SUMMARY resuspension buffer needed for each sample. Volume of 1:1:1 is dependent on SP:SAMPLEPREP_SUMMARY normalize cell count from experiment. Use 200uL of 1:1:1 for every 2E5 cells. SP:SAMPLEPREP_SUMMARY When ready to start resuspension, remove dried lipids from -80C storage and SP:SAMPLEPREP_SUMMARY resuspend in volume of 1:1:1 solution calculated for each sample. No cell SP:SAMPLEPREP_SUMMARY conditions use 200uL. Use gentle vortex to allow for dried lipids from vial wall SP:SAMPLEPREP_SUMMARY to get into solution. Prepare "blank" vials of 1:1:1 for buffer background SP:SAMPLEPREP_SUMMARY analysis. Store lipids in autosampler between 4-7C. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Phenomenex Kinetex C18 (100 x 2.1mm,2.6um) CH:SOLVENT_A 40:60 water-methanol, plus 10mM ammonium formate and 0.1% formic acid CH:SOLVENT_B 10:90 methanol-isopropanol, plus 10mM ammonium formate and 0.1% formic acid CH:FLOW_GRADIENT 75% solvent A–25% solvent B for 2 min, 35% solvent A–65% solvent B for 2min CH:FLOW_GRADIENT at a curve value of 4, a hold at 35% solvent A–65% solvent B for 1min, 0% CH:FLOW_GRADIENT solvent A–100% solvent B for 11min at a curve value of 4, and a hold at 0% CH:FLOW_GRADIENT solvent A–100% solvent B for 4 min. CH:FLOW_RATE 0.25-mL/min CH:COLUMN_TEMPERATURE 60 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Orbitrap Exploris 240 MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS PCs were identified in positive mode using parent lipid m/z, RT and choline head MS:MS_COMMENTS fragment of 184.073 m/z. Confirmation of PCs was performed by FA tail analysis MS:MS_COMMENTS in negative mode. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak Area MS_METABOLITE_DATA_START Samples 20241023_pos_HCMV_h2O_72_R1_a 20241023_pos_HCMV_h2O_72_R1_b 20241023_pos_HCMV_PAA_72_R1_a 20241023_pos_HCMV_PAA_72_R1_b 20241023_pos_HCMV_h2O_72_R2_a 20241023_pos_HCMV_h2O_72_R2_b 20241023_pos_HCMV_PAA_72_R2_a 20241023_pos_HCMV_PAA_72_R2_b 20241023_pos_HCMV_h2O_72_R3_a 20241023_pos_HCMV_h2O_72_R3_b 20241023_pos_HCMV_PAA_72_R3_a 20241023_pos_HCMV_PAA_72_R3_b Factors Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:H20 Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:H20 Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:PAA 100ug/mL Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:PAA 100ug/mL Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:H20 Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:H20 Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:PAA 100ug/mL Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:PAA 100ug/mL Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:H20 Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:H20 Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:PAA 100ug/mL Sample source:Primary human fibroblast cells (HFF) | DNA synthesis inhibtor:PAA 100ug/mL LPC 16:0 5185726 6367798 7133064 8403642 5430408 7249794 6897566 7919348 4979436 6535584 5966332 9353482 LPC 16:1 331704.5 461382.1 415175.6 610813.4 325537.8 547836.5 385221.4 595142.6 288546.5 496601 393396.2 687385.1 LPC 18:0 4422182 4433122 6567258 7291986 4202418 5114269 5838232 6047132 4480598 4667238 4671272 7078982 LPC 18:1 4066760 6102904 4720424 6901478 3735432 6856838 5360356 7526454 3872364 6144606 4353664 7757708 LPC 20:1 392895.2 427901 523547.8 595389.3 382833.5 411826 526534.4 505143.4 375869.2 374248.1 384317.7 582976.3 LPC 20:3 205100 268713.4 209208.6 339241.2 199881.5 443297.4 265685 424805 305414.9 521860.5 293353.7 654619.8 LPC 20:4 173017.5 231291.5 157038.1 256986.3 166962.4 348033.3 248683.1 356868.4 297166 508647.2 273208.1 599457.7 LPC 22:3 64683.11 130443.3 51372.51 84008.09 55863.2 123018.5 45055.88 155168.2 140108.3 222773.8 72250.11 210719.9 PC 28:0 2.35E+07 2.23E+07 2.84E+07 3.59E+07 1.80E+07 2.25E+07 2.20E+07 2.03E+07 1.74E+07 1.78E+07 1.60E+07 2.46E+07 PC 28:1 974233.9 948296.8 1511576 2027957 696132 755717.2 1179098 1116359 729708 695759.7 1176078 1502707 PC 30:0 6.41E+07 6.59E+07 8.08E+07 1.08E+08 5.82E+07 7.44E+07 7.21E+07 8.00E+07 5.80E+07 6.12E+07 5.28E+07 8.43E+07 PC 30:1 4.10E+07 3.72E+07 5.41E+07 6.05E+07 3.29E+07 3.77E+07 4.05E+07 4.01E+07 2.88E+07 2.78E+07 2.93E+07 4.01E+07 PC 32:1 7.81E+08 8.54E+08 8.97E+08 1.22E+09 8.22E+08 9.48E+08 9.26E+08 9.72E+08 6.85E+08 7.05E+08 5.96E+08 9.03E+08 PC 32:2 4.70E+07 4.60E+07 5.83E+07 7.49E+07 4.13E+07 4.92E+07 4.92E+07 5.37E+07 3.30E+07 3.22E+07 2.84E+07 4.52E+07 PC 34:0 8045582 7513726 9409505 1.16E+07 7153784 9208187 8595884 8821065 7044670 6551180 4903770 8778914 PC 34:1 1.04E+09 1.06E+09 1.17E+09 1.40E+09 1.06E+09 1.13E+09 1.13E+09 1.17E+09 9.03E+08 9.27E+08 7.26E+08 9.95E+08 PC 34:2 2.52E+08 2.63E+08 2.89E+08 3.96E+08 2.24E+08 3.03E+08 2.68E+08 2.94E+08 1.84E+08 1.93E+08 1.55E+08 2.38E+08 PC 34:3 1.62E+07 2.01E+07 1.84E+07 2.65E+07 1.70E+07 2.63E+07 2.18E+07 2.89E+07 1.86E+07 2.29E+07 1.88E+07 2.47E+07 PC 36:1 1.99E+08 2.00E+08 2.13E+08 3.01E+08 1.97E+08 2.53E+08 2.23E+08 2.60E+08 1.68E+08 1.69E+08 1.33E+08 2.15E+08 PC 36:2 4.60E+08 4.48E+08 5.28E+08 6.08E+08 4.35E+08 5.82E+08 4.90E+08 5.28E+08 3.66E+08 3.62E+08 3.00E+08 4.63E+08 PC 36:3 6.06E+07 6.53E+07 7.49E+07 9.37E+07 6.13E+07 8.00E+07 8.13E+07 8.57E+07 5.56E+07 5.82E+07 4.96E+07 7.35E+07 PC 36:4 4.83E+07 5.23E+07 5.68E+07 7.92E+07 5.04E+07 6.96E+07 6.40E+07 6.95E+07 6.05E+07 6.44E+07 5.40E+07 8.62E+07 PC 36:5 4834340 5374158 6690084 7768420 5069330 6162544 6436760 6810132 5462582 6126342 5177888 8474441 PC 38:1 2380515 2608645 2738412 3475706 2649214 3110495 2996423 3297149 2421128 2429887 1854217 3121906 PC 38:2 2.47E+07 2.78E+07 2.72E+07 3.81E+07 2.57E+07 3.61E+07 2.89E+07 3.12E+07 2.15E+07 2.30E+07 1.72E+07 2.66E+07 PC 38:3 2.69E+07 3.09E+07 3.14E+07 4.51E+07 3.08E+07 4.06E+07 4.13E+07 4.28E+07 2.95E+07 2.90E+07 2.43E+07 3.84E+07 PC 38:4 3.02E+07 3.19E+07 3.74E+07 4.71E+07 3.38E+07 4.13E+07 4.31E+07 4.53E+07 3.98E+07 3.95E+07 3.56E+07 5.45E+07 PC 38:5 3.17E+07 3.31E+07 4.03E+07 4.78E+07 3.30E+07 4.68E+07 4.44E+07 5.11E+07 3.65E+07 4.16E+07 3.43E+07 5.90E+07 PC 38:6 1.29E+07 1.34E+07 1.60E+07 1.73E+07 1.50E+07 1.86E+07 1.70E+07 1.72E+07 1.60E+07 1.49E+07 1.51E+07 1.98E+07 PC 40:1 674446.6 629174.9 778697.3 956202.8 561756.9 681460.7 740141 690922.2 554969.2 574824.2 534080.3 788089.8 PC 40:2 2299116 2320936 2545514 3375664 2271001 3051538 2388918 2911266 1954437 2140099 1743274 2662982 PC 40:3 2749615 2966366 3854713 4757136 3085240 4427247 4060214 4710238 3154731 3591568 2780278 4812044 PC 40:4 6221206 6378956 6899826 1.01E+07 6999438 9028721 8572437 1.02E+07 6392088 7129700 5700360 9488771 PC 40:5 6233968 7271900 8180752 1.10E+07 7964758 1.07E+07 9755973 1.17E+07 8303590 8276782 7875574 1.30E+07 PC 40:7 6162426 6600646 6710300 9009809 6383536 7928772 8083088 8096036 6323334 6692818 5492510 9322595 PC 42:1 763512.2 816800.3 1095830 1345358 728639.2 882178.2 914613.7 949276.7 824679.9 862846.8 747486.5 1097333 PC 42:2 1553045 1587344 1969994 2528864 1528560 1637553 1895568 1937522 1314708 1484040 1262375 1805559 PC 42:3 693892.6 738086 851650.3 1229194 664715.4 1002529 988498.3 1008279 742412 789756.3 700960 1019442 PC 42:4 705037.6 719981.2 991698.1 1104190 825499.3 1149284 1103510 1132108 965411.2 997785.7 889898.3 1334676 PC 42:5 641836.1 788551.2 849540.7 1213834 620995.5 997451.7 619336.5 1015834 944139.7 971518.4 690565.2 1132947 PC 44:1 734292.3 681852.9 954162.2 1192523 627203.8 764865 778027.3 740275.1 663382.4 708699.5 645153.8 991577 PC 44:2 1397082 1383841 1933797 2171701 1031853 1413856 1534981 1475307 1198828 1250692 1191074 1717678 PC 44:3 436572.8 427452.3 560366.1 631822.4 411591.2 495612.8 574245.3 596524.9 375225.9 419304 457777.8 578454.2 PC 44:4 290392.4 329020.4 418684.8 423918 329888.5 352552.8 446978.5 471855.9 316610.8 371473.9 367838.8 516376.2 PC 44:5 347439 330642.3 486770.5 654840.8 341893.5 442599.4 514040.5 528751.7 395939.5 534967.6 456464.7 731945.2 PC 46:2 540095.3 578321.2 705105.3 928762 518866.7 539768.4 554814.8 545495.5 448116.6 455670.5 472690.8 675801.9 PC 46:5 309734 412472.8 525859.8 642687.7 291452.4 423536.7 514652.4 451940 421645.8 506096.2 480081 742436.5 PC 48:2 198288.1 171005.2 273391.1 335927.6 147443.5 161294.4 180800.2 189887.9 149852 164280.8 158359.5 216030.7 PC 48:5 125278.6 126161.5 183142.2 204923.7 136092.2 143304.3 158929.8 160658.7 128767.5 152632.7 186323.3 312595.2 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name m/z RT LPC 16:0 496.339478 4.605876 LPC 16:1 494.324219 4.059206 LPC 18:0 524.370911 5.164821 LPC 18:1 522.354675 4.68758 LPC 20:1 550.386597 5.254297 LPC 20:3 546.355225 4.587824 LPC 20:4 544.339905 4.270367 LPC 22:3 574.386414 5.065664 PC 28:0 678.506592 6.483876 PC 28:1 676.49115 6.248266 PC 30:0 706.537781 6.955493 PC 30:1 704.52124 6.626201 PC 32:1 732.552979 7.100755 PC 32:2 730.53772 6.767225 PC 34:0 762.599609 7.939215 PC 34:1 760.584473 7.541438 PC 34:2 758.569275 7.207694 PC 34:3 756.552734 7.045916 PC 36:1 788.615601 8.05116 PC 36:2 786.600098 7.647182 PC 36:3 784.584717 7.421206 PC 36:4 782.56897 7.203283 PC 36:5 780.552368 6.87941 PC 38:1 816.647217 8.559466 PC 38:2 814.631958 8.167977 PC 38:3 812.615723 7.917977 PC 38:4 810.600098 7.687399 PC 38:5 808.584412 7.318213 PC 38:6 806.568115 7.119165 PC 40:1 844.67926 9.077656 PC 40:2 842.663635 8.665079 PC 40:3 840.647522 8.348866 PC 40:4 838.631104 8.038808 PC 40:5 836.61554 7.785725 PC 40:7 832.583801 7.2558 PC 42:1 872.7099 9.618421 PC 42:2 870.694458 9.164523 PC 42:3 868.678833 8.81037 PC 42:4 866.662231 8.475049 PC 42:5 864.646729 8.278852 PC 44:1 900.741211 10.15312 PC 44:2 898.725891 9.670039 PC 44:3 896.709412 9.28294 PC 44:4 894.693604 8.989198 PC 44:5 892.678772 8.763958 PC 46:2 926.756531 10.20806 PC 46:5 920.710266 9.282605 PC 48:2 954.787354 10.72064 PC 48:5 948.741699 9.7409 METABOLITES_END #END