#METABOLOMICS WORKBENCH hienbui_20250306_073923 DATATRACK_ID:5700 STUDY_ID:ST003808 ANALYSIS_ID:AN006261 PROJECT_ID:PR002382 VERSION 1 CREATED_ON March 21, 2025, 11:35 am #PROJECT PR:PROJECT_TITLE Lipidomics of human mammary epithelial stem-like cells (hMECs) overexpressing PR:PROJECT_TITLE Glucose-6-phosphate dehydrogenase (G6PD) PR:PROJECT_SUMMARY Peroxisomes play a crucial role in cellular metabolism. Glucose-6-phosphate PR:PROJECT_SUMMARY dehydrogenase (G6PD), the gatekeeper enzyme of the pentose phosphate pathway, is PR:PROJECT_SUMMARY primarily localized in the cytosol. However, studies have reported its presence PR:PROJECT_SUMMARY in peroxisomes as well. This project aims to determine the function of G6PD on PR:PROJECT_SUMMARY the peroxisomal membrane. To investigate this, we overexpressed G6PD either in PR:PROJECT_SUMMARY the cytosol or on the peroxisomal membrane of human mammary epithelial stem-like PR:PROJECT_SUMMARY cells (hMECs), a cell line that retains characteristics of primary mammary PR:PROJECT_SUMMARY epithelial stem cells, and analyzed their lipidomic profiles for comparison. PR:INSTITUTE University of Helsinki PR:DEPARTMENT Faculty of Biological and Environmental Sciences PR:LABORATORY Katajisto Laboratory PR:LAST_NAME Bui PR:FIRST_NAME Hien PR:ADDRESS Biocenter 1, Viikinkaari 9, Helsinki, Uusimaa, 00790, Finland PR:EMAIL hien.bui@helsinki.fi PR:PHONE +358294159407 #STUDY ST:STUDY_TITLE Glucose-6-phosphate-dehydrogenase on old peroxisomes maintains self-renewal of ST:STUDY_TITLE epithelial stem cells after asymmetric cell division ST:STUDY_SUMMARY Peroxisomes play a crucial role in cellular metabolism. Glucose-6-phosphate ST:STUDY_SUMMARY dehydrogenase (G6PD), the gatekeeper enzyme of the pentose phosphate pathway, is ST:STUDY_SUMMARY primarily localized in the cytosol. However, studies have reported its presence ST:STUDY_SUMMARY in peroxisomes as well. This project aims to determine the function of G6PD on ST:STUDY_SUMMARY the peroxisomal membrane. In this study, we overexpressed G6PD in either the ST:STUDY_SUMMARY cytosol or on the peroxisomal membrane of mammary epithelial stem-like cells ST:STUDY_SUMMARY (hMECs). By comparing their lipidomic profiles, we found that peroxisomal ST:STUDY_SUMMARY membrane-associated G6PD provides NADPH, which feeds into peroxisomal ether ST:STUDY_SUMMARY lipid synthesis. ST:INSTITUTE University of Helsinki ST:DEPARTMENT Faculty of Biological and Environmental Sciences ST:LABORATORY Katajisto Laboratory ST:LAST_NAME Hien ST:FIRST_NAME Bui ST:ADDRESS Biocenter 1, Viikinkaari 9 ST:EMAIL hien.bui@helsinki.fi ST:PHONE +358294159407 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #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 - c-pos-1 Sample source:human mammary epithelial stem-like cell | Sample type:cytosolic G6PD | G6PD expression:Overexpressed Replicate=rep1; RAW_FILE_NAME(Raw_file_name)=c-pos-1_06032024-1.CDF SUBJECT_SAMPLE_FACTORS - c-neg-2 Sample source:human mammary epithelial stem-like cell | Sample type:cytosolic G6PD | G6PD expression:Non-overexpressed Replicate=rep1; RAW_FILE_NAME(Raw_file_name)=c-neg-2_06032024-1.CDF SUBJECT_SAMPLE_FACTORS - p-pos-3 Sample source:human mammary epithelial stem-like cell | Sample type:peroxisomal G6PD | G6PD expression:Overexpressed Replicate=rep1; RAW_FILE_NAME(Raw_file_name)=p-pos-3_06032024-1.CDF SUBJECT_SAMPLE_FACTORS - p-neg-4 Sample source:human mammary epithelial stem-like cell | Sample type:peroxisomal G6PD | G6PD expression:Non-overexpressed Replicate=rep1; RAW_FILE_NAME(Raw_file_name)=p-neg-4_06032024-1.CDF SUBJECT_SAMPLE_FACTORS - c-pos-5 Sample source:human mammary epithelial stem-like cell | Sample type:cytosolic G6PD | G6PD expression:Overexpressed Replicate=rep2; RAW_FILE_NAME(Raw_file_name)=c-pos-5_06032024-1.CDF SUBJECT_SAMPLE_FACTORS - c-neg-6 Sample source:human mammary epithelial stem-like cell | Sample type:cytosolic G6PD | G6PD expression:Non-overexpressed Replicate=rep2; RAW_FILE_NAME(Raw_file_name)=c-neg-6_06032024-1.CDF SUBJECT_SAMPLE_FACTORS - p-pos-7 Sample source:human mammary epithelial stem-like cell | Sample type:peroxisomal G6PD | G6PD expression:Overexpressed Replicate=rep2; RAW_FILE_NAME(Raw_file_name)=p-pos-7_06032024-1.CDF SUBJECT_SAMPLE_FACTORS - p-neg-8 Sample source:human mammary epithelial stem-like cell | Sample type:peroxisomal G6PD | G6PD expression:Non-overexpressed Replicate=rep2; RAW_FILE_NAME(Raw_file_name)=p-neg-8_07032024-1.CDF SUBJECT_SAMPLE_FACTORS - c-pos-9 Sample source:human mammary epithelial stem-like cell | Sample type:cytosolic G6PD | G6PD expression:Overexpressed Replicate=rep3; RAW_FILE_NAME(Raw_file_name)=c-pos-9_07032024-1.CDF SUBJECT_SAMPLE_FACTORS - c-neg-10 Sample source:human mammary epithelial stem-like cell | Sample type:cytosolic G6PD | G6PD expression:Non-overexpressed Replicate=rep3; RAW_FILE_NAME(Raw_file_name)=c-neg-10_06032024-1.CDF SUBJECT_SAMPLE_FACTORS - p-pos-11 Sample source:human mammary epithelial stem-like cell | Sample type:peroxisomal G6PD | G6PD expression:Overexpressed Replicate=rep3; RAW_FILE_NAME(Raw_file_name)=p-pos-11_06032024-1.CDF SUBJECT_SAMPLE_FACTORS - p-neg-12 Sample source:human mammary epithelial stem-like cell | Sample type:peroxisomal G6PD | G6PD expression:Non-overexpressed Replicate=rep3; RAW_FILE_NAME(Raw_file_name)=p-neg-12_07032024-1.CDF SUBJECT_SAMPLE_FACTORS - c-pos-13 Sample source:human mammary epithelial stem-like cell | Sample type:cytosolic G6PD | G6PD expression:Overexpressed Replicate=rep4; RAW_FILE_NAME(Raw_file_name)=c-pos-13_07032024-1.CDF SUBJECT_SAMPLE_FACTORS - c-neg-14 Sample source:human mammary epithelial stem-like cell | Sample type:cytosolic G6PD | G6PD expression:Non-overexpressed Replicate=rep4; RAW_FILE_NAME(Raw_file_name)=c-neg-14_06032024-1.CDF SUBJECT_SAMPLE_FACTORS - p-pos-15 Sample source:human mammary epithelial stem-like cell | Sample type:peroxisomal G6PD | G6PD expression:Overexpressed Replicate=rep4; RAW_FILE_NAME(Raw_file_name)=p-pos-15_06032024-1.CDF SUBJECT_SAMPLE_FACTORS - p-neg-16 Sample source:human mammary epithelial stem-like cell | Sample type:peroxisomal G6PD | G6PD expression:Non-overexpressed Replicate=rep4; RAW_FILE_NAME(Raw_file_name)=p-neg-16_07032024-1.CDF #COLLECTION CO:COLLECTION_SUMMARY Human mammary epithelial cells (hMECs) line FL2 (PMID: 21498687) was used for CO:COLLECTION_SUMMARY this study. hMECs were maintained in MEGM mammary epithelial medium (Lonza, CO:COLLECTION_SUMMARY CC-3153). G6PD overexpression plasmids were generated in a pRP backbone by CO:COLLECTION_SUMMARY inserting the G6PD sequence with or without peroxisomal targeting signal CO:COLLECTION_SUMMARY downstream of the hPGK promoter. In addition, the plasmids contain an mCherry CO:COLLECTION_SUMMARY sequence downstream of an IRES sequence. mCherry was used as the selection CO:COLLECTION_SUMMARY marker for transfected cells in FACS sorting. To over-express G6PD in hMECs, CO:COLLECTION_SUMMARY endotoxin-free plasmids were transfected to the cells using jetPRIME (Polyplus, CO:COLLECTION_SUMMARY 101000015). 24hours post-transfection, cells were trypsinized, sorted into CO:COLLECTION_SUMMARY culture media by Fluorescence-Activated Cell Sorting (FACS), centrifuged at 500g CO:COLLECTION_SUMMARY for 5min, washed 1 with cold PBS following by washed 1 with sucrose 0.25M. CO:COLLECTION_SUMMARY Pellets were snap freeze with liquid N2 and store at -70. CO:SAMPLE_TYPE Epithelial cells #TREATMENT TR:TREATMENT_SUMMARY Human mammary epithelial stem like cells (hMECs) were transfected with TR:TREATMENT_SUMMARY endotoxin-free G6PD overexpression plasmids using jetPRIME (Polyplus, TR:TREATMENT_SUMMARY 101000015). Cells were analyzed 24h post-transfection. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY GC-MS: Fatty acyl (FA) and alkenyl chains, and fatty alcohols (FOHs) were SP:SAMPLEPREP_SUMMARY analysed by gas chromatography (GC). Aliquots of cell pellet Folch-extracts were SP:SAMPLEPREP_SUMMARY evaporated near to dryness under nitrogen flow and the extracted lipids were SP:SAMPLEPREP_SUMMARY transmethylated by heating with 1% H2SO4 in methanol under nitrogen atmosphere. SP:SAMPLEPREP_SUMMARY This converts FAs to their methyl esters (FAMEs), phospholipid-derived alkenyl SP:SAMPLEPREP_SUMMARY chains to dimethyl acetals (DMAs) and leaves FOHs (1-ol) intact. After adding SP:SAMPLEPREP_SUMMARY water, the FAMEs, DMAs and FOHs were extracted with hexane, and the sample SP:SAMPLEPREP_SUMMARY solution was dried, and concentrated. LC-MS: Lipids were extracted from cell SP:SAMPLEPREP_SUMMARY pellets (at least from one million cells) according to Folch et al (PMID: SP:SAMPLEPREP_SUMMARY 13428781), and dissolved in chloroform/methanol 1:2 (v/v). Internal standards SP:SAMPLEPREP_SUMMARY [EquiSPLASH® internal standard mixture and Ceramide (Cer) 18:1;O2/17:0, both SP:SAMPLEPREP_SUMMARY from Merck] were added and samples were analyzed with LC-MS/MS. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY The samples were injected into a GC-2010 Plus gas chromatograph (Shimadzu CH:CHROMATOGRAPHY_SUMMARY Scientific Instruments, Kyoto, Japan) with flame-ionization detector (FID) for CH:CHROMATOGRAPHY_SUMMARY quantification of the analytes. Both GC systems were equipped with Zebron ZB-wax CH:CHROMATOGRAPHY_SUMMARY capillary columns (30 m, 0.25 mm ID and film thickness 0.25 μm; Phenomenex, CH:CHROMATOGRAPHY_SUMMARY Torrence CA, USA). GC-FID chromatographic peak areas (in the TXT files; analyte CH:CHROMATOGRAPHY_SUMMARY retention times in Table 1) were converted to mol% using the theoretical CH:CHROMATOGRAPHY_SUMMARY correction factors for FID (6), and the FOHs were calculated as µmol/10^6 CH:CHROMATOGRAPHY_SUMMARY cells. CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Shimadzu GC-2010 CH:COLUMN_NAME Phenomenex Zebron ZB-wax capillary (30m x 0.25mm, 0.25um) CH:SOLVENT_A NA CH:SOLVENT_B NA CH:FLOW_GRADIENT NA CH:FLOW_RATE 1.8 ml min–1 for FID and 1.0 ml min–1 for MSD CH:COLUMN_TEMPERATURE 280 °C for FID and 210 °C for MSD. #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME Helsinki University Lipidomics Unit (HiLIPID) #MS MS:INSTRUMENT_NAME Shimadzu QP2010 Plus MS:INSTRUMENT_TYPE GC-FID & GC-MSD MS:MS_TYPE EI MS:ION_MODE POSITIVE MS:MS_COMMENTS The samples were injected into a GC-2010 Plus gas chromatograph (Shimadzu MS:MS_COMMENTS Scientific Instruments, Kyoto, Japan) with flame-ionization detector (FID) for MS:MS_COMMENTS quantification of the analytes. Both GC systems were equipped with Zebron ZB-wax MS:MS_COMMENTS capillary columns (30 m, 0.25 mm ID and film thickness 0.25 μm; Phenomenex, MS:MS_COMMENTS Torrence CA, USA). GC-FID chromatographic peak areas were converted to mol% MS:MS_COMMENTS using the theoretical correction factors for FID, and the FOHs were calculated MS:MS_COMMENTS as µmol/1 million cells (ebook ISBN:9780429127557). The identifications were MS:MS_COMMENTS performed using a GCMS-QP2010 Ultra (Shimadzu Scientific Instruments, Kyoto, MS:MS_COMMENTS Japan) with mass selective detector (MSD). The spectra of the analytes were MS:MS_COMMENTS compared with the spectra of several authentic standard mixtures of FAMEs MS:MS_COMMENTS (Supelco CRM47885, 47033, 47085-U, 47015-U and 47080-U) and published reference MS:MS_COMMENTS mass spectra (https://lipidmaps.org/resources/lipid_web?page=ms/masspec.html). #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples c-pos-1 c-neg-2 p-pos-3 p-neg-4 c-pos-5 c-neg-6 p-pos-7 p-neg-8 c-pos-9 c-neg-10 p-pos-11 p-neg-12 c-pos-13 c-neg-14 p-pos-15 p-neg-16 Factors Sample source:human mammary epithelial stem-like cell | Sample type:cytosolic G6PD | G6PD expression:Overexpressed Sample source:human mammary epithelial stem-like cell | Sample type:cytosolic G6PD | G6PD expression:Non-overexpressed Sample source:human mammary epithelial stem-like cell | Sample type:peroxisomal G6PD | G6PD expression:Overexpressed Sample source:human mammary epithelial stem-like cell | Sample type:peroxisomal G6PD | G6PD expression:Non-overexpressed Sample source:human mammary epithelial stem-like cell | Sample type:cytosolic G6PD | G6PD expression:Overexpressed Sample source:human mammary epithelial stem-like cell | Sample type:cytosolic G6PD | G6PD expression:Non-overexpressed Sample source:human mammary epithelial stem-like cell | Sample type:peroxisomal G6PD | G6PD expression:Overexpressed Sample source:human mammary epithelial stem-like cell | Sample type:peroxisomal G6PD | G6PD expression:Non-overexpressed Sample source:human mammary epithelial stem-like cell | Sample type:cytosolic G6PD | G6PD expression:Overexpressed Sample source:human mammary epithelial stem-like cell | Sample type:cytosolic G6PD | G6PD expression:Non-overexpressed Sample source:human mammary epithelial stem-like cell | Sample type:peroxisomal G6PD | G6PD expression:Overexpressed Sample source:human mammary epithelial stem-like cell | Sample type:peroxisomal G6PD | G6PD expression:Non-overexpressed Sample source:human mammary epithelial stem-like cell | Sample type:cytosolic G6PD | G6PD expression:Overexpressed Sample source:human mammary epithelial stem-like cell | Sample type:cytosolic G6PD | G6PD expression:Non-overexpressed Sample source:human mammary epithelial stem-like cell | Sample type:peroxisomal G6PD | G6PD expression:Overexpressed Sample source:human mammary epithelial stem-like cell | Sample type:peroxisomal G6PD | G6PD expression:Non-overexpressed FA 12:0 738 4289 13601 6041 18427 6804 7384 10366 11646 515 7993 7077 11182 9245 9742 10964 FA 13:0 2377 141 322 3607 5517 4438 2741 3131 3039 2721 4456 2966 3017 3378 347 3501 FA 14:0 iso 3034 2377 6748 2052 2632 3135 388 2819 2365 208 2488 2684 2388 2452 2795 3212 FA 14:0 58745 40553 201451 42744 102106 46681 13387 64558 75148 52241 70466 90063 82853 75327 80649 97421 FA 14:1n-5 3256 496 19279 3662 5644 4032 13757 6367 3329 3218 4637 6199 2636 4805 4274 6608 FA 15:0 iso 12146 4945 31862 9674 12751 7129 20213 24914 3483 1332 4015 3295 2792 3965 3195 450 FA 15:0 anteiso 4454 233 16243 3671 7468 2265 7977 863 3185 1934 2726 4258 2786 4343 3924 5859 FA 15:0 9578 9225 43043 10086 20961 8651 25208 12534 9945 9257 9967 10572 7223 6083 7923 10589 FA 16:0 iso 3945 3244 25182 398 559 3541 10542 4908 3439 2718 3252 6674 2525 3933 3671 5558 DMA 16:0 84444 54241 85679 54369 96374 68139 8087 78957 124457 69399 113386 10917 145327 121255 150477 126182 DMA 16:1n-7 1178 879 1929 38 1664 54 8897 1034 985 672 1275 31588 1253 801 1048 1031 FA 16:0 683939 459091 1924283 483034 1017129 513837 1135187 69372 848724 58824 786394 857364 965793 869902 990763 957149 FA 16:1n-9 590 4974 39434 5378 13057 7535 25849 11024 15008 6863 897 29556 18027 12372 15523 16928 FA 16:1n-7 309938 281913 103308 280646 320236 284395 701515 378483 408034 367355 393565 506712 448008 483842 479657 522887 FA 16:1n-5 4365 3507 24456 443 4162 5453 11851 6283 630 4042 5768 4664 6755 7104 7773 6468 FA 17:0 iso 10038 5566 44075 7495 10635 681 2758 8372 2601 6041 9055 26654 3328 7458 728 7881 FA 17:0 anteiso 4951 2016 22054 2552 6454 2155 11914 5746 3972 2098 3999 1914 3986 3612 5348 4914 FA 17:0 4492 2552 31853 3011 10734 260 14907 582 4916 3207 504 5303 5457 4674 4995 5845 FA 17:1n-8 4279 3316 46096 3038 6598 3046 25555 7966 5011 4522 534 3972 5476 5401 694 3759 DMA 18:0 52638 31326 56558 30887 6296 35336 50709 46621 75758 37201 67982 58232 87547 67668 93848 72092 DMA 18:1n-9 18384 15526 42966 14899 23916 16124 22032 2332 24589 15698 22841 56858 28148 26746 30775 38111 DMA 18:1n-7 2668 1979 33739 20075 32789 2133 28746 28986 31723 19131 3189 33494 40544 35245 43474 38517 FA 18:0 497598 287118 840659 311724 656542 329353 518829 424989 604286 361595 532256 495523 697744 580967 682358 637232 FA 18:1n-11 8998 5914 11868 6107 10929 6325 13283 7659 10143 5542 9443 3679 12992 10439 13454 6548 FA 18:1n-9 1219006 857028 344594 88365 1407047 963638 235757 1335317 1613146 1130209 1560506 159728 1888278 1685981 2011249 1792481 FA 18:1n-7 413109 314231 745118 332269 471511 367742 584957 469276 529137 360851 539113 527913 688775 616748 735699 655824 FA 18:1n-5 17293 1534 32664 18606 21459 18705 32436 26371 23828 19616 26234 22085 3411 30885 3380 25453 FA 18:2n-6 49102 25422 80231 25233 55936 32812 66463 46762 48704 24959 50004 48168 70201 59612 76872 70833 FA 19:0 66 228 3592 217 1937 201 2515 829 367 868 799 418 44 464 249 205 18:2NMI1 9833 7177 39871 6911 14519 6927 15544 10478 10756 6074 8481 28778 10938 8974 11784 17342 18:2NMI2 2362 3508 4637 2892 3298 2682 3917 346 2855 2189 2134 2171 215 333 3704 2251 FA 20:0 12185 7378 24887 6656 45018 8188 14846 11351 13753 7995 12981 8364 15024 1320 15228 10963 FA 20:1n-11 4855 275 18381 3269 23044 380 12811 5975 6877 5019 6557 894 10343 7337 10445 63302 FA 20:1n-9 37941 21831 128977 23363 56954 27749 79927 42951 45459 26504 4721 82203 6046 43841 63984 63588 FA 20:1n-7 27088 1844 52655 19446 36022 23777 38609 30131 33445 21393 34474 37915 46666 35471 49543 38419 FA 20:2n-6 3593 1918 7955 1646 767 3032 7239 4127 3042 122 4209 16328 5938 5009 5838 14265 20:3NMI 6698 4639 7064 4802 11284 6613 9123 8044 886 7794 8336 2898 11813 10355 12498 7539 FA 20:3n-6 6436 3463 8769 3921 5455 4308 5789 5766 5787 3531 6312 12056 6862 586 9396 11121 FA 21:0 521 253 845 177 1713 79 264 333 426 31 187 149 35 414 53 388 FA 20:4n-6 35343 19436 42239 18375 3876 22801 29171 34316 35436 18155 35345 42915 49927 42993 61538 53379 FA 20:5n-3 7458 4085 14325 3359 7284 4767 6349 667 6605 3975 754 14697 9617 9346 12527 19558 FA 22:0 26164 19658 28536 16874 52558 21551 24997 26266 30872 19718 27733 31534 32154 31528 34663 31638 FA 22:1n-11 5073 3174 12421 1898 10456 3132 10591 5035 416 3166 4812 4154 5876 322 4768 4001 FA 22:1n-9 66151 45362 10837 38348 54242 43093 53553 83054 85924 37787 44847 77467 80471 43765 46719 79791 FA 22:1n-7 829 6345 7597 5961 7733 8014 9213 8357 5357 4273 7249 4959 6986 656 6992 611 FA 22:2n-9 3976 187 4477 2422 7289 3497 6206 333 3785 1797 4612 2718 5494 2943 5693 4327 FA 23:0 1747 2457 11379 241 11898 3109 9045 4719 1542 2275 2256 13488 3494 2123 3213 7543 FA 22:4n-6 5371 3288 4164 1861 4223 305 3331 3433 3436 1698 4341 3721 6603 5599 6944 5915 FA 22:5n-3 21017 10417 25583 11096 20848 13174 15477 21736 19355 8334 1884 20531 28837 25878 34432 29654 FA 24:0 43794 20425 39405 18985 89132 28964 61234 31494 54547 28422 44295 35537 5739 43438 56442 37289 FA 22:6n-3 6043 2606 7494 1908 45168 3705 36822 7871 4516 2385 4148 17876 7632 6824 9162 24095 FA 24:1n-9 48312 32228 72754 27238 103651 36734 80685 50722 50395 2758 44265 37825 63896 52518 65809 51731 FA 24:1n-7 1027 8506 16702 6224 7455 10122 14527 1750 11147 5375 12214 6009 11448 12897 17244 9808 FA 26:0 701 285 1765 116 191 1198 562 16 1061 6417 4633 355 13254 8901 861 2731 FA 26:1n-9 412 905 19194 316 692 343 761 825 5301 261 3373 1647 4755 5393 7295 3977 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Retention time FA 12:0 2.82 FA 13:0 3.29 FA 14:0 iso 3.62 FA 14:0 3.99 FA 14:1n-5 4.35 FA 15:0 iso 4.47 FA 15:0 anteiso 4.63 FA 15:0 4.98 FA 16:0 iso 5.66 DMA 16:0 5.87 DMA 16:1n-7 6.24 FA 16:0 6.41 FA 16:1n-9 6.77 FA 16:1n-7 6.89 FA 16:1n-5 7.13 FA 17:0 iso 7.37 FA 17:0 anteiso 7.7 FA 17:0 8.43 FA 17:1n-8 8.96 DMA 18:0 9.99 DMA 18:1n-9 10.39 DMA 18:1n-7 10.6 FA 18:0 10.86 FA 18:1n-11 11.17 FA 18:1n-9 11.37 FA 18:1n-7 11.55 FA 18:1n-5 11.9 FA 18:2n-6 12.54 FA 19:0 13.48 18:2NMI1 14.78 18:2NMI2 15.7 FA 20:0 16.13 FA 20:1n-11 16.51 FA 20:1n-9 16.62 FA 20:1n-7 16.85 FA 20:2n-6 17.85 20:3NMI 17.99 FA 20:3n-6 18.57 FA 21:0 18.8 FA 20:4n-6 19.2 FA 20:5n-3 21.25 FA 22:0 22.01 FA 22:1n-11 22.43 FA 22:1n-9 22.7 FA 22:1n-7 23.05 FA 22:2n-9 23.32 FA 23:0 24.8 FA 22:4n-6 26.87 FA 22:5n-3 30.3 FA 24:0 31.44 FA 22:6n-3 32.03 FA 24:1n-9 32.6 FA 24:1n-7 33.27 FA 26:0 48.56 FA 26:1n-9 51.11 METABOLITES_END #END