#METABOLOMICS WORKBENCH tnuriel_20210602_131536_mwtab.txt DATATRACK_ID:2670 STUDY_ID:ST001820 ANALYSIS_ID:AN002955 PROJECT_ID:000000 VERSION 1 CREATED_ON June 7, 2021, 12:36 pm #PROJECT PR:PROJECT_TYPE Lipidomics analysis PR:PROJECT_SUMMARY Apolipoprotein E ε4 (APOE4) is the primary genetic risk factor for the PR:PROJECT_SUMMARY late-onset form of Alzheimer's disease (AD). Although the reason for this PR:PROJECT_SUMMARY association is not completely understood, researchers have uncovered numerous PR:PROJECT_SUMMARY effects of APOE4 expression on AD-relevant brain processes, including amyloid PR:PROJECT_SUMMARY beta (Aβ) accumulation, lipid metabolism, endosomal-lysosomal processing and PR:PROJECT_SUMMARY bioenergetics. In this study, we aimed to determine the effect of APOE4 allelic PR:PROJECT_SUMMARY dosage on regional brain lipid composition in aged mice, as well as in cultured PR:PROJECT_SUMMARY neurons. We performed a targeted lipidomic analysis on the entorhinal cortex PR:PROJECT_SUMMARY (EC) and primary visual cortex (PVC) from 14–15 month-old APOE3/3, APOE3/4, PR:PROJECT_SUMMARY and APOE4/4 targeted replacement mice, as well as on WT neurons cultured with PR:PROJECT_SUMMARY conditioned media from APOE3/3 or APOE4/4 astrocytes. Our results reveal that PR:PROJECT_SUMMARY the EC possesses increased susceptibility to APOE4-associated lipid alterations PR:PROJECT_SUMMARY compared to the PVC. In the EC, APOE4 expression showed a dominant effect in PR:PROJECT_SUMMARY decreasing diacylglycerol (DAG) levels, and a semi-dominant additive effect in PR:PROJECT_SUMMARY the upregulation of multiple ceramide, glycosylated sphingolipid and PR:PROJECT_SUMMARY bis(monoacylglycerol)phosphate (BMP) species, lipids known to accumulate as a PR:PROJECT_SUMMARY result of endosomal-lysosomal dysfunction and defective lysosomal clearance. PR:PROJECT_SUMMARY Neurons treated with conditioned media from APOE4 vs. APOE3 astrocytes also PR:PROJECT_SUMMARY showed similar alterations of DAG and BMP species as those observed in the mouse PR:PROJECT_SUMMARY EC. Our results suggest that APOE4 expression differentially modulates regional PR:PROJECT_SUMMARY and neuronal lipid signatures, which may underlie the increased susceptibility PR:PROJECT_SUMMARY of EC-localized neurons to AD pathology. PR:INSTITUTE Columbia University PR:FIRST_NAME Tal PR:ADDRESS 630 W 168th St., P&S 12-430 PR:EMAIL tn2283@cumc.columbia.edu PR:PHONE 2123045683 PR:PROJECT_TITLE APOE4-associated differences in lipidomics signatures in mouse brain and PR:PROJECT_TITLE cultured neurons PR:LAST_NAME Nuriel #STUDY ST:STUDY_TITLE WT neurons treated with APOE3/3 and APOE4/4 ACM ST:STUDY_SUMMARY We performed a targeted lipidomic analysis on WT neurons treated with astrocyte ST:STUDY_SUMMARY conditioned media (ACM) from APOE3/3 or APOE4/4 astrocytes. ST:INSTITUTE Columbia University ST:LAST_NAME Nuriel ST:FIRST_NAME Tal ST:ADDRESS 630 W 168th St., P&S 12-430 ST:EMAIL tn2283@cumc.columbia.edu ST:PHONE 2123045683 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENDER Not applicable #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 - 1 Condition:No ACM SUBJECT_SAMPLE_FACTORS - 2 Condition:No ACM SUBJECT_SAMPLE_FACTORS - 3 Condition:No ACM SUBJECT_SAMPLE_FACTORS - 4 Condition:No ACM SUBJECT_SAMPLE_FACTORS - 5 Condition:No ACM SUBJECT_SAMPLE_FACTORS - 6 Condition:No ACM SUBJECT_SAMPLE_FACTORS - 7 Condition:APOE3/3 ACM SUBJECT_SAMPLE_FACTORS - 8 Condition:APOE3/3 ACM SUBJECT_SAMPLE_FACTORS - 9 Condition:APOE3/3 ACM SUBJECT_SAMPLE_FACTORS - 10 Condition:APOE3/3 ACM SUBJECT_SAMPLE_FACTORS - 11 Condition:APOE3/3 ACM SUBJECT_SAMPLE_FACTORS - 12 Condition:APOE3/3 ACM SUBJECT_SAMPLE_FACTORS - 13 Condition:APOE4/4 ACM SUBJECT_SAMPLE_FACTORS - 14 Condition:APOE4/4 ACM SUBJECT_SAMPLE_FACTORS - 15 Condition:APOE4/4 ACM SUBJECT_SAMPLE_FACTORS - 16 Condition:APOE4/4 ACM SUBJECT_SAMPLE_FACTORS - 17 Condition:APOE4/4 ACM SUBJECT_SAMPLE_FACTORS - 18 Condition:APOE4/4 ACM #COLLECTION CO:COLLECTION_SUMMARY Astrocyte conditioned media (ACM) was obtained from immortalized astrocyte cell CO:COLLECTION_SUMMARY lines (a gift from Dr. David Holtzman) that were originally generated from CO:COLLECTION_SUMMARY primary astrocytes from P1-2 pups of APOE targeted replacement mice (42). The CO:COLLECTION_SUMMARY immortalized astrocytes were conditioned with Neurobasal media supplemented with CO:COLLECTION_SUMMARY B27, Glutamax-I, Normocin and 1% penicillin/streptomycin for 24 hours. This ACM CO:COLLECTION_SUMMARY was then collected, stored at -80ºC, and thawed prior to use. WT primary CO:COLLECTION_SUMMARY cortical neuronal cultures were obtained from embryonic day 17 (E17) C57Bl/6 CO:COLLECTION_SUMMARY embryos. Briefly, pregnant mice were euthanized by cervical dislocation and the CO:COLLECTION_SUMMARY embryo brains extracted. The meninges were carefully stripped off and the CO:COLLECTION_SUMMARY cortices dissected out. The cortices were then enzymatically dissociated in CO:COLLECTION_SUMMARY 0.25% Trypsin-EDTA and resuspended in Neurobasal media supplemented with B27, CO:COLLECTION_SUMMARY Glutamax-I, Normocin and 1% penicillin/streptomycin. Dissociated cells were CO:COLLECTION_SUMMARY counted and 300K neurons per well were plated directly into ACM in poly-D-lysine CO:COLLECTION_SUMMARY (PDL)-coated 6-well plates. ACM-treated neurons were then incubated at 37⁰C CO:COLLECTION_SUMMARY for 7 days in a humidified chamber with 5% CO2, with 50% media exchange (with CO:COLLECTION_SUMMARY newly thawed ACM) once every 3 days. CO:SAMPLE_TYPE Neurons #TREATMENT TR:TREATMENT_SUMMARY WT neurons were incubated for 7 days in either no ACM, APOE3/3 ACM, or APOE4/4 TR:TREATMENT_SUMMARY ACM. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Lipid and small-molecule metabolite extraction was performed using a methyl SP:SAMPLEPREP_SUMMARY tert-butyl ether (MTBE)/methanol extraction protocol. Briefly, harvested neurons SP:SAMPLEPREP_SUMMARY were homogenized in ice-cold methanol. Following homogenization, samples were SP:SAMPLEPREP_SUMMARY incubated in 1200 ul of MTBE for 1 hr at room temperature to separate SP:SAMPLEPREP_SUMMARY organic-soluble lipids from aqueous-soluble lipids and other small-molecules. SP:SAMPLEPREP_SUMMARY Finally, 360 ul of ultrapure water was added (for a final ratio of 3:1:0.9 SP:SAMPLEPREP_SUMMARY MTBE:methanol:water) to resolve the two liquid phases, and each samples were SP:SAMPLEPREP_SUMMARY centrifuged at 10,000 x g for 10 min. For this experiment, the upper organic SP:SAMPLEPREP_SUMMARY phase was collected from each sample and stored in a separate tube, and the SP:SAMPLEPREP_SUMMARY remaining protein pellets were resuspended in 25 mM ammonium bicarbonate, pH 8, SP:SAMPLEPREP_SUMMARY with 2.5% SDS. A BCA protein assay was performed on each protein fraction, and SP:SAMPLEPREP_SUMMARY the organic phase was normalized to their protein concentration equivalent with SP:SAMPLEPREP_SUMMARY 100% methanol. All samples were then stored at -80°C prior to analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Reverse phase CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Agilent 1260 CH:COLUMN_NAME Agilent Eclipse XDB-C18 (100 x 3.0mm) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6490 QQQ MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Lipid profiling was performed using an Agilent 1260 HPLC coupled to an Agilent MS:MS_COMMENTS 6490 triple quadrupole (QQQ) mass spectrometer (41). Each sample was run through MS:MS_COMMENTS three separate chromatographic conditions (reverse-phase negative mode, MS:MS_COMMENTS reverse-phase positive mode and normal-phase positive mode) for the effective MS:MS_COMMENTS quantification of over 337 distinct lipids from over 28 lipid subclasses, as MS:MS_COMMENTS previously described (41). Individual lipid species were measured by multiple MS:MS_COMMENTS reaction monitoring transitions. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS area under the curve MS_METABOLITE_DATA_START Samples 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Factors Condition:No ACM Condition:No ACM Condition:No ACM Condition:No ACM Condition:No ACM Condition:No ACM Condition:APOE3/3 ACM Condition:APOE3/3 ACM Condition:APOE3/3 ACM Condition:APOE3/3 ACM Condition:APOE3/3 ACM Condition:APOE3/3 ACM Condition:APOE4/4 ACM Condition:APOE4/4 ACM Condition:APOE4/4 ACM Condition:APOE4/4 ACM Condition:APOE4/4 ACM Condition:APOE4/4 ACM Cer d18:1/16:0 0.73017 0.57609 0.34963 0.44577 0.28386 0.54156 0.64483 0.34346 0.36564 0.38758 0.46827 0.34631 0.78287 0.57645 0.61650 0.83883 0.98733 0.73438 Cer d18:1/16:1 0.10681 0.09229 0.08725 0.08412 0.07677 0.08197 0.10998 0.09236 0.09426 0.07315 0.07817 0.08482 0.10395 0.09373 0.08893 0.08982 0.08686 0.08399 Cer d18:1/18:0 2.39533 1.54941 0.92958 1.39400 0.85257 1.53768 0.85218 0.50465 0.51728 0.55274 0.65264 0.46479 1.02039 0.65747 0.73230 0.99205 1.03761 0.89678 Cer d18:1/18:1 0.01435 0.01070 0.00623 0.00905 0.00603 0.00978 0.01258 0.00729 0.01005 0.00995 0.01056 0.00837 0.01248 0.00704 0.01019 0.01197 0.01543 0.01385 Cer d18:1/20:0 0.04830 0.03552 0.02205 0.03281 0.02196 0.03319 0.02976 0.01826 0.01965 0.01987 0.02390 0.01593 0.04195 0.02527 0.03028 0.03415 0.03516 0.03308 Cer d18:1/20:1 0.00210 0.00173 0.00128 0.00115 0.00077 0.00143 0.00221 0.00166 0.00189 0.00109 0.00224 0.00151 0.00339 0.00189 0.00210 0.00348 0.00302 0.00189 Cer d18:1/22:0 0.03529 0.02723 0.01612 0.02013 0.01518 0.02470 0.04451 0.02376 0.02706 0.02530 0.02593 0.01944 0.04956 0.02881 0.03098 0.03303 0.03689 0.04134 Cer d18:1/22:1 0.00741 0.00549 0.00307 0.00500 0.00267 0.00465 0.01409 0.00896 0.01066 0.00860 0.00912 0.00677 0.01244 0.00727 0.00876 0.01073 0.00995 0.01107 Cer d18:1/24:0 0.02983 0.01825 0.01362 0.01585 0.01059 0.01630 0.03897 0.02404 0.02798 0.02293 0.02722 0.01935 0.04035 0.02656 0.03355 0.03863 0.03729 0.03958 Cer d18:1/24:1 0.07870 0.05398 0.03755 0.04159 0.02985 0.05053 0.09316 0.05134 0.06739 0.05507 0.06208 0.05627 0.09829 0.06879 0.07232 0.08856 0.07801 0.09821 SM d18:1/16:0 1.05615 1.20233 1.03028 1.10057 0.68982 1.08090 3.27254 1.56607 1.78255 1.99436 2.14729 0.93978 2.18198 1.28287 2.28218 2.20512 2.63729 2.21768 SM d18:1/16:1 0.01133 0.01305 0.01267 0.01419 0.01126 0.01453 0.08025 0.03845 0.04556 0.05755 0.04620 0.02304 0.09933 0.06605 0.10460 0.10503 0.10462 0.08129 SM d18:1/18:0 2.29590 2.11651 1.45786 1.72017 1.01627 1.60009 2.64115 1.36354 1.60577 1.59330 1.77727 0.85666 1.48472 0.94337 1.66839 2.01295 2.41688 1.81281 SM d18:1/18:1 0.06422 0.06689 0.06544 0.06982 0.04440 0.06605 0.14202 0.06734 0.07516 0.07922 0.08535 0.04152 0.10752 0.06313 0.11160 0.10343 0.12959 0.10968 SM d18:1/20:0 0.17117 0.15922 0.10129 0.10515 0.06507 0.09943 0.36209 0.16305 0.17239 0.17623 0.19379 0.08840 0.23150 0.12009 0.19892 0.21858 0.28090 0.19763 SM d18:1/20:1 0.00705 0.00645 0.00510 0.00510 0.00425 0.00536 0.02775 0.01324 0.01172 0.01294 0.01483 0.00684 0.01626 0.01089 0.01549 0.01667 0.01993 0.01449 SM d18:1/22:0 0.17414 0.17863 0.09543 0.11161 1.72077 2.21982 0.49381 0.23171 0.24671 0.28036 0.27384 0.12648 0.29471 0.15943 0.25669 0.29235 0.37694 0.27223 SM d18:1/22:1 0.02142 0.01897 0.01246 0.01476 0.00941 0.01316 0.11373 0.05396 0.05068 0.06047 0.06598 0.03323 0.05470 0.03240 0.05230 0.05749 0.07182 0.04931 SM d18:1/24:0 0.15916 0.14337 0.08306 0.09661 0.05738 0.09118 0.40520 0.20019 0.19962 0.23396 0.22448 0.10816 0.32818 0.16184 0.26211 0.31157 0.41425 0.31159 SM d18:1/24:1 0.25595 0.23856 0.14510 0.14748 0.09482 0.15831 0.84097 0.42030 0.45428 0.48395 0.49529 0.21906 0.62196 0.30303 0.51687 0.62712 0.72444 0.62531 SM d18:1/26:0 0.00117 0.00158 0.00070 0.00093 0.00040 0.00061 0.00355 0.00137 0.00148 0.00159 0.00161 0.00076 0.00342 0.00149 0.00237 0.00323 0.00381 0.00269 SM d18:1/26:1 0.00186 0.00193 0.00102 0.00137 0.00067 0.00138 0.00737 0.00317 0.00315 0.00392 0.00416 0.00158 0.00584 0.00308 0.00507 0.00654 0.00732 0.00585 dhSM d18:0/16:0 0.04385 0.04916 0.03123 0.03206 0.01981 0.00168 0.10140 0.04795 0.05624 0.05786 0.07377 0.03053 0.05089 0.03101 0.05387 0.06215 0.07920 0.05051 dhSM d18:0/18:0 0.08889 0.09752 0.05697 0.05900 0.03375 0.05345 0.12178 0.05572 0.07061 0.06787 0.07498 0.03644 0.06294 0.04058 0.06165 0.07810 0.11202 0.07341 dhSM d18:0/18:1 0.00656 0.00807 0.00484 0.00660 0.00379 0.00455 0.04722 0.02038 0.02191 0.02528 0.02846 0.01044 0.02523 0.01260 0.01927 0.01987 0.02144 0.02039 dhSM d18:0/22:0 0.00260 0.00247 0.00162 0.00194 0.00111 0.00153 0.00696 0.00384 0.00410 0.00466 0.00506 0.00174 0.00472 0.00342 0.00513 0.00431 0.00598 0.00519 dhSM d18:0/24:0 0.00180 0.00210 0.00087 0.00118 0.00067 0.00093 0.00543 0.00244 0.00208 0.00291 0.00310 0.00152 0.00319 0.00176 0.00277 0.00380 0.00483 0.00315 dhSM d18:0/24:1 0.01348 0.01033 0.00533 0.00653 0.00440 0.00701 0.05304 0.02472 0.02333 0.02361 0.02767 0.01296 0.02616 0.01428 0.02445 0.03232 0.04249 0.03190 GalCer d18:0/16:0 0.00181 0.00175 0.00125 0.00144 0.00111 0.00218 0.00376 0.00250 0.00274 0.00289 0.00309 0.00257 0.00284 0.00229 0.00281 0.00367 0.00366 0.00281 GalCer d18:0/16:1 0.00040 0.00032 0.00034 0.00038 0.00025 0.00042 0.00077 0.00050 0.00061 0.00056 0.00072 0.00048 0.00065 0.00049 0.00063 0.00074 0.00079 0.00071 GalCer d18:0/18:0 0.01182 0.01098 0.00706 0.00975 0.00724 0.01256 0.01280 0.00867 0.00960 0.01008 0.01261 0.00921 0.00962 0.00689 0.00863 0.01149 0.01221 0.01027 GalCer d18:0/18:1 0.00316 0.00260 0.00170 0.00236 0.00176 0.00275 0.00294 0.00189 0.00238 0.00211 0.00268 0.00216 0.00221 0.00157 0.00178 0.00222 0.00317 0.00253 GalCer d18:0/20:0 0.00063 0.00070 0.00054 0.00057 0.00056 0.00066 0.00133 0.00090 0.00115 0.00103 0.00123 0.00083 0.00102 0.00053 0.00101 0.00107 0.00112 0.00114 GalCer d18:0/22:0 0.00099 0.00080 0.00066 0.00079 0.00078 0.00091 0.00294 0.00200 0.00228 0.00221 0.00274 0.00198 0.00211 0.00165 0.00176 0.00274 0.00258 0.00251 GalCer d18:0/24:0 0.00068 0.00057 0.00047 0.00045 0.00039 0.00064 0.00155 0.00094 0.00099 0.00103 0.00140 0.00111 0.00172 0.00116 0.00119 0.00185 0.00217 0.00177 GalCer d18:0/24:1 0.00065 0.00056 0.00039 0.00050 0.00043 0.00069 0.00222 0.00138 0.00162 0.00161 0.00229 0.00145 0.00210 0.00129 0.00170 0.00252 0.00231 0.00205 GalCer d18:1/16:0 0.06500 0.05728 0.03990 0.05232 0.03656 0.07189 0.13312 0.08768 0.09690 0.09377 0.12405 0.08675 0.11042 0.08031 0.10850 0.12955 0.15029 0.12120 GalCer d18:1/16:1 0.00017 0.00020 0.00024 0.00021 0.00021 0.00034 0.00067 0.00040 0.00062 0.00060 0.00071 0.00055 0.00075 0.00043 0.00050 0.00082 0.00090 0.00081 GalCer d18:1/18:0 0.44230 0.40830 0.26443 0.34631 0.25711 0.42538 0.44885 0.29099 0.33013 0.30767 0.40720 0.28649 0.36276 0.23208 0.29212 0.35351 0.44202 0.34090 GalCer d18:1/18:1 0.00491 0.00616 0.00607 0.00643 0.00375 0.00412 0.00854 0.00758 0.00863 0.00842 0.00658 0.00508 0.00647 0.00653 0.00770 0.00885 0.00590 0.00523 GalCer d18:1/20:0 0.02028 0.01819 0.01319 0.01621 0.01269 0.01998 0.03939 0.02465 0.02733 0.02539 0.03162 0.02416 0.03090 0.01993 0.02507 0.03217 0.03109 0.02851 GalCer d18:1/20:1 0.00042 0.00039 0.00049 0.00043 0.00022 0.00024 0.00117 0.00084 0.00098 0.00089 0.00101 0.00084 0.00078 0.00077 0.00084 0.00110 0.00093 0.00095 GalCer d18:1/22:0 0.03576 0.03235 0.02533 0.02953 0.02472 0.03500 0.10977 0.06825 0.07608 0.07530 0.09241 0.07194 0.07939 0.05424 0.06690 0.08830 0.08734 0.08026 GalCer d18:1/22:1 0.00191 0.00177 0.00118 0.00131 0.00114 0.00176 0.01209 0.00748 0.00824 0.00813 0.01068 0.00764 0.00700 0.00471 0.00628 0.00848 0.00777 0.00694 GalCer d18:1/24:0 0.02790 0.02255 0.01611 0.02074 0.01621 0.02464 0.06965 0.04045 0.04564 0.04380 0.05521 0.04038 0.07044 0.04260 0.05410 0.07241 0.07309 0.06948 GalCer d18:1/24:1 0.02623 0.02066 0.01320 0.01751 0.01274 0.02326 0.09674 0.05264 0.05939 0.05611 0.07355 0.05400 0.08044 0.04864 0.06290 0.07888 0.08452 0.07335 GalCer d18:1/26:0 0.00191 0.00208 0.00186 0.00203 0.00123 0.00170 0.00317 0.00230 0.00284 0.00260 0.00243 0.00179 0.00258 0.00242 0.00289 0.00307 0.00261 0.00264 GalCer d18:1/26:1 0.00050 0.00041 0.00028 0.00041 0.00025 0.00034 0.00130 0.00081 0.00097 0.00095 0.00114 0.00093 0.00138 0.00093 0.00116 0.00131 0.00146 0.00114 LacCer d18:1/16:0 0.06248 0.04486 0.02970 0.03544 0.02235 0.06132 0.07608 0.03830 0.04443 0.04289 0.05454 0.04348 0.05882 0.02801 0.04653 0.05670 0.08218 0.05676 LacCer d18:1/18:0 0.13474 0.10214 0.05324 0.07650 0.04427 0.11359 0.10434 0.04779 0.04950 0.05467 0.07229 0.05136 0.07388 0.03456 0.04972 0.00222 0.10476 0.06377 LacCer d18:1/20:0 0.00921 0.00510 0.00238 0.00436 0.00268 0.00637 0.01243 0.00676 0.00763 0.00688 0.00656 0.00855 0.00795 0.00353 0.00530 0.00574 0.01012 0.00653 LacCer d18:1/22:0 0.01286 0.00753 0.00442 0.00604 0.00256 0.00833 0.02611 0.01376 0.01233 0.01364 0.01667 0.01435 0.01501 0.00652 0.01169 0.01087 0.02094 0.01332 LacCer d18:1/22:1 0.00220 0.00124 0.00101 0.00126 0.00112 0.00236 0.00558 0.00303 0.00260 0.00253 0.00542 0.00697 0.00325 0.00131 0.00200 0.00301 0.00498 0.00242 LacCer d18:1/24:0 0.00850 0.00409 0.00392 0.00513 0.00266 0.00851 0.02665 0.01152 0.01470 0.01762 0.01697 0.01529 0.02172 0.00919 0.01353 0.01337 0.02818 0.01921 LacCer d18:1/24:1 0.01612 0.00959 0.00536 0.00767 0.00477 0.01268 0.03957 0.01882 0.01877 0.02182 0.03078 0.02319 0.02473 0.01352 0.01599 0.02315 0.03524 0.02661 PC 30:0 19.65318 18.42581 18.01096 19.83852 16.00068 22.53957 16.91211 12.52783 12.16847 16.68752 16.62289 12.30422 15.91423 11.84651 15.08348 16.39606 18.02509 17.20752 PC 32:0 26.70829 24.36462 21.46745 22.56379 19.52500 27.02059 15.76329 14.46393 15.10026 15.54107 18.36383 14.85217 16.09385 15.36206 16.75229 16.02953 17.91921 15.24026 PC 32:1 12.05411 11.35981 12.92399 11.75117 12.22501 14.35371 18.56892 16.59494 15.30673 16.65190 18.48896 19.47778 15.61215 16.71923 17.61408 14.75333 16.85297 17.10418 PC 34:0 7.37749 6.43733 4.86225 5.88305 5.70816 6.32896 3.16618 2.89611 2.72900 2.85831 3.11670 3.18965 3.53889 3.27934 3.74628 4.17802 4.09242 3.54692 PC 34:1 18.39201 19.44021 15.54132 16.99821 14.92485 19.51176 18.92329 17.55026 18.07867 18.84128 18.45054 17.45543 19.67609 17.70267 18.54464 19.61154 21.01967 17.68418 PC 34:2 4.87851 5.06862 4.53061 4.72635 4.88067 4.90707 6.90070 5.60410 6.44735 6.62550 6.22043 6.51308 5.62970 5.50117 6.35795 5.65255 5.54042 6.47311 PC 36:0 0.85794 0.70097 0.51265 0.65381 0.63305 0.73923 0.02659 0.46711 0.45195 0.46029 0.52452 0.47822 0.59825 0.57206 0.64243 0.59755 0.64362 0.57986 PC 36:1 7.45454 6.29749 5.01665 5.91575 5.39408 6.75010 5.13331 4.49333 4.34109 5.00016 5.32479 4.87522 5.99700 6.05381 5.96264 5.88877 7.11639 6.20388 PC 36:2 5.02996 4.52519 3.56905 4.78039 3.92845 5.13480 7.67702 7.24447 6.15567 7.79207 8.00636 7.21895 7.69506 7.55969 7.75408 7.91845 9.01805 8.11928 PC 36:3 3.73583 3.33693 3.19998 3.54804 3.26632 3.85433 4.06323 3.23911 3.03916 3.31570 4.06048 3.07646 3.42305 3.17745 3.35721 3.46357 3.52616 3.74691 PC 36:4 9.72948 9.71531 8.66753 9.18610 8.84039 10.51945 2.93129 2.39515 1.93219 2.06034 2.80132 1.83302 2.88865 2.51414 3.11323 2.74357 3.43175 2.87573 PC 38:1 0.09867 0.07790 0.05252 0.06861 0.06571 0.08285 0.28846 0.22252 0.18268 0.19203 0.28086 0.19063 0.30575 0.24760 0.26024 0.31867 0.33433 0.29509 PC 38:2 0.39223 0.30651 0.23616 0.26118 0.26065 0.31651 1.17001 0.92374 0.80794 0.83209 1.04561 0.86544 1.18388 1.02975 1.11447 1.23924 1.40897 1.21886 PC 38:3 1.63126 1.45071 1.19439 1.37343 1.34258 1.75068 2.73532 1.91084 1.62702 1.80454 2.42112 1.61043 2.65357 2.08503 2.63759 2.71194 2.93734 2.86340 PC 38:4 8.67344 7.50546 5.94828 6.41234 6.08284 8.69393 3.37462 2.56733 2.35352 2.46236 2.98878 2.12700 4.07282 3.35820 3.45587 4.01099 4.97434 3.76008 PC 38:5 5.15909 4.21053 3.81759 4.75476 3.85009 4.71435 1.90630 1.24903 0.96941 1.30866 1.55700 1.04393 2.07584 1.57043 1.83562 1.72250 2.28124 1.80860 PC 38:6 2.32146 2.33422 2.33622 2.29512 2.03624 2.55322 1.43013 0.96987 0.84357 0.88245 1.30277 0.91348 1.29802 1.13380 1.32180 1.33627 1.61788 1.47454 PC 40:4 1.31259 0.93662 0.65485 0.78411 0.73860 0.94256 0.51964 0.31287 0.23211 0.26952 0.38988 0.23200 0.65906 0.37316 0.42149 0.53382 0.64057 0.53017 PC 40:5 2.63733 2.03941 1.53073 1.66457 1.37980 1.99320 0.70382 0.37188 0.34236 0.38325 0.51379 0.33020 0.88826 0.48818 0.64115 0.74747 0.92481 0.75370 PC 40:6 3.30520 2.97379 2.09652 2.73793 1.75922 3.58178 1.03128 0.60350 0.43646 0.52243 0.78073 0.41229 1.24079 0.70993 0.94750 1.18239 1.41779 1.08892 PC 40:7 1.24743 1.08287 0.82522 1.01759 0.65337 1.06102 0.54654 0.24971 0.21525 0.25442 0.36480 0.19629 0.70186 0.36477 0.52611 0.61828 0.86728 0.55119 PC 42:5 0.53441 0.44617 0.22496 0.39214 0.22748 0.44056 0.10207 0.05966 0.04745 0.05196 0.07264 0.04345 0.12793 0.07243 0.08828 0.12205 0.16740 0.10371 PC 42:6 0.44472 0.38071 0.20316 0.31980 0.17766 0.39704 0.07976 0.04606 0.03133 0.03888 0.05093 0.02947 0.11257 0.05259 0.06632 0.09071 0.13417 0.07527 PC 42:7 0.28116 0.25571 0.13197 0.22159 0.11599 0.29255 0.06482 0.03561 0.02962 0.02859 0.04640 0.02261 0.08522 0.04429 0.05501 0.07096 0.09542 0.07345 PCe 30:0 0.33905 0.31892 0.28412 0.34767 0.28778 0.39184 1.15349 0.57403 0.72649 0.87744 0.95468 0.46559 0.64999 0.44127 0.57035 0.71624 0.75900 0.67852 PCe 32:0 1.62574 1.35633 1.22088 1.48083 1.21222 1.61558 4.32233 3.14248 3.20683 3.52968 3.82706 3.02988 2.52896 1.99600 2.61698 2.67837 2.96751 2.59219 PCe 32:1 0.29245 0.27120 0.25466 0.28387 0.27054 0.32313 1.26259 1.00518 1.06170 1.26103 1.22351 1.28709 0.82486 0.81296 0.98113 0.78207 0.92947 0.89713 PCe 34:0 0.58357 0.46511 0.41723 0.43682 0.46958 0.52047 0.69362 0.61538 0.62637 0.64324 0.74880 0.67532 0.58941 0.54616 0.60389 0.65317 0.69155 0.59900 PCe 34:1 1.42630 1.32204 1.11715 1.30006 1.11101 1.44045 2.35057 2.16675 2.03119 2.35224 2.36654 2.29634 1.85131 1.77311 1.95918 2.00924 2.06400 1.90942 PCe 34:2 0.22910 0.19039 0.18301 0.21781 0.17264 0.22300 0.62336 0.50867 0.49538 0.50405 0.59071 0.54954 0.41544 0.39304 0.42315 0.44570 0.44991 0.42023 PCe 36:0 0.07500 0.06147 0.05772 0.06352 0.05979 0.07759 0.10455 0.08810 0.08648 0.09561 0.10940 0.09849 0.09265 0.08701 0.09880 0.10015 0.10482 0.09614 PCe 36:1 0.22812 0.17540 0.15697 0.17094 0.16011 0.22189 0.74208 0.60050 0.63323 0.65484 0.71342 0.68614 0.56246 0.52403 0.58963 0.59664 0.65025 0.61939 PCe 36:2 0.20310 0.12570 0.11040 0.14492 0.12239 0.14094 0.77030 0.65296 0.61863 0.69893 0.76951 0.64077 0.60468 0.49736 0.56021 0.53949 0.59078 0.57304 PCe 36:3 0.12378 0.09015 0.08018 0.09285 0.07673 0.09158 0.33862 0.26091 0.27623 0.29796 0.31583 0.26616 0.21221 0.17897 0.21817 0.21455 0.22920 0.21426 PCe 36:4 0.17896 0.16233 0.14886 0.16767 0.14113 0.19380 0.16473 0.11974 0.09432 0.12968 0.13592 0.09958 0.11737 0.09637 0.11497 0.11085 0.13520 0.10072 PCe 38:0 0.28299 0.29770 0.22728 0.27585 0.22089 0.33771 0.15827 0.10656 0.08683 0.10565 0.13763 0.09159 0.17206 0.12517 0.13615 0.14863 0.19258 0.15241 PCe 38:1 0.05291 0.04932 0.03759 0.04347 0.04225 0.05964 0.08286 0.07188 0.07211 0.07528 0.08042 0.06664 0.08429 0.07558 0.07178 0.08624 0.08960 0.07520 PCe 38:2 0.04482 0.03087 0.02465 0.02831 0.02716 0.03093 0.21092 0.15178 0.14323 0.14470 0.17987 0.14293 0.15880 0.13829 0.15465 0.17566 0.17231 0.16599 PCe 38:3 0.10067 0.06474 0.05818 0.06875 0.05559 0.07660 0.38638 0.29654 0.24308 0.31641 0.37089 0.24865 0.30471 0.23460 0.25578 0.30079 0.31688 0.25759 PCe 38:4 0.32430 0.25223 0.21070 0.25522 0.20381 0.28346 0.35140 0.22675 0.19676 0.23197 0.26845 0.19785 0.26800 0.19598 0.21165 0.27015 0.29612 0.24779 PCe 38:5 0.27673 0.21268 0.16715 0.24827 0.16849 0.29168 0.16438 0.09801 0.08538 0.09083 0.12339 0.08673 0.15864 0.09768 0.11651 0.13634 0.16895 0.12783 PCe 38:6 0.12937 0.09942 0.08634 0.11140 0.07384 0.11550 0.08167 0.04777 0.04467 0.04779 0.06114 0.04115 0.07577 0.04471 0.05991 0.06569 0.08681 0.06416 PCe 40:4 0.15240 0.10694 0.07346 0.08955 0.07030 0.11777 0.10561 0.06321 0.05566 0.05761 0.08022 0.05010 0.09113 0.06030 0.07211 0.08924 0.10490 0.08868 PCe 40:5 0.25773 0.17989 0.12573 0.17325 0.10059 0.19246 0.07951 0.04535 0.03826 0.04111 0.05747 0.03704 0.07980 0.04665 0.05770 0.07679 0.09173 0.06693 PCe 40:6 0.21522 0.14639 0.10942 0.14534 0.09275 0.16223 0.09097 0.05298 0.04128 0.04508 0.06759 0.04071 0.08426 0.04956 0.06861 0.07409 0.09322 0.07594 PCe 40:7 0.16433 0.11432 0.07735 0.12270 0.06480 0.13541 0.07996 0.04459 0.03362 0.03960 0.05246 0.03317 0.08489 0.05078 0.05696 0.07350 0.10329 0.07321 PCe 42:5 0.04070 0.02944 0.01707 0.02332 0.01645 0.03342 0.01748 0.00772 0.00712 0.00796 0.01148 0.00754 0.01611 0.00994 0.01104 0.01697 0.01604 0.01445 PCe 42:6 0.04370 0.03354 0.01895 0.02935 0.01793 0.03635 0.01903 0.00936 0.00854 0.00880 0.01334 0.00703 0.01897 0.01181 0.01383 0.01689 0.01992 0.01488 LPC 16:0 0.49682 0.44950 0.25068 0.27061 0.27336 0.34082 0.47837 0.20584 0.24336 0.27744 0.30428 0.14076 0.54551 0.24457 0.34781 0.34294 0.46549 0.31160 LPC 16:1 0.03011 0.03247 0.01454 0.01634 0.01753 0.02419 0.06668 0.03428 0.03837 0.04177 0.05365 0.02480 0.04465 0.02676 0.04049 0.03875 0.06655 0.04035 LPC 18:0 0.18942 0.19819 0.14814 0.13081 0.11904 0.13795 0.13329 0.05864 0.07260 0.07920 0.07833 0.03699 0.21109 0.09331 0.14585 0.12281 0.21747 0.13802 LPC 18:1 0.09713 0.10346 0.05893 0.06141 0.05607 0.08311 0.17343 0.08584 0.10100 0.10837 0.11582 0.06324 0.19299 0.09876 0.14929 0.14543 0.24084 0.14993 LPC 20:0 0.00137 0.00191 0.00139 0.00121 0.00130 0.00149 0.00278 0.00110 0.00150 0.00198 0.00185 0.00100 0.00292 0.00165 0.00345 0.00231 0.00553 0.00339 LPC 20:1 0.00326 0.00418 0.00245 0.00239 0.00220 0.00309 0.00806 0.00394 0.00485 0.00613 0.00612 0.00309 0.01077 0.00576 0.00936 0.00853 0.01493 0.00880 LPC 20:2 0.00241 0.00293 0.00177 0.00176 0.00181 0.00225 0.00431 0.00236 0.00262 0.00290 0.00319 0.00166 0.00425 0.00262 0.00394 0.00388 0.00672 0.00458 LPC 20:3 0.00596 0.00679 0.00332 0.00371 0.00376 0.00565 0.00797 0.00406 0.00500 0.00547 0.00587 0.00295 0.00725 0.00502 0.00681 0.00670 0.01120 0.00734 LPC 20:4 0.02461 0.02923 0.01281 0.01433 0.01539 0.02476 0.00637 0.00368 0.00451 0.00451 0.00543 0.00278 0.00664 0.00413 0.00633 0.00604 0.01011 0.00607 LPCe 16:0 0.00278 0.00243 0.00134 0.00146 0.00159 0.00205 0.00986 0.00486 0.00493 0.00650 0.00699 0.00370 0.00558 0.00327 0.00518 0.00476 0.00715 0.00488 LPCe 18:0 0.00692 0.00667 0.00410 0.00402 0.00362 0.00501 0.02031 0.00865 0.01011 0.01316 0.01348 0.00592 0.01597 0.00864 0.01280 0.01293 0.01808 0.01233 LPCe 18:1 0.00079 0.00083 0.00041 0.00043 0.00060 0.00089 0.00533 0.00256 0.00308 0.00316 0.00372 0.00175 0.00309 0.00192 0.00277 0.00254 0.00419 0.00294 LPCe 20:0 0.00061 0.00069 0.00063 0.00059 0.00056 0.00055 0.00148 0.00074 0.00079 0.00093 0.00096 0.00066 0.00117 0.00075 0.00123 0.00109 0.00144 0.00117 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Cer d18:1/16:0 Cer d18:1/16:1 Cer d18:1/18:0 Cer d18:1/18:1 Cer d18:1/20:0 Cer d18:1/20:1 Cer d18:1/22:0 Cer d18:1/22:1 Cer d18:1/24:0 Cer d18:1/24:1 SM d18:1/16:0 SM d18:1/16:1 SM d18:1/18:0 SM d18:1/18:1 SM d18:1/20:0 SM d18:1/20:1 SM d18:1/22:0 SM d18:1/22:1 SM d18:1/24:0 SM d18:1/24:1 SM d18:1/26:0 SM d18:1/26:1 dhSM d18:0/16:0 dhSM d18:0/18:0 dhSM d18:0/18:1 dhSM d18:0/22:0 dhSM d18:0/24:0 dhSM d18:0/24:1 GalCer d18:0/16:0 GalCer d18:0/16:1 GalCer d18:0/18:0 GalCer d18:0/18:1 GalCer d18:0/20:0 GalCer d18:0/22:0 GalCer d18:0/24:0 GalCer d18:0/24:1 GalCer d18:1/16:0 GalCer d18:1/16:1 GalCer d18:1/18:0 GalCer d18:1/18:1 GalCer d18:1/20:0 GalCer d18:1/20:1 GalCer d18:1/22:0 GalCer d18:1/22:1 GalCer d18:1/24:0 GalCer d18:1/24:1 GalCer d18:1/26:0 GalCer d18:1/26:1 LacCer d18:1/16:0 LacCer d18:1/18:0 LacCer d18:1/20:0 LacCer d18:1/22:0 LacCer d18:1/22:1 LacCer d18:1/24:0 LacCer d18:1/24:1 PC 30:0 PC 32:0 PC 32:1 PC 34:0 PC 34:1 PC 34:2 PC 36:0 PC 36:1 PC 36:2 PC 36:3 PC 36:4 PC 38:1 PC 38:2 PC 38:3 PC 38:4 PC 38:5 PC 38:6 PC 40:4 PC 40:5 PC 40:6 PC 40:7 PC 42:5 PC 42:6 PC 42:7 PCe 30:0 PCe 32:0 PCe 32:1 PCe 34:0 PCe 34:1 PCe 34:2 PCe 36:0 PCe 36:1 PCe 36:2 PCe 36:3 PCe 36:4 PCe 38:0 PCe 38:1 PCe 38:2 PCe 38:3 PCe 38:4 PCe 38:5 PCe 38:6 PCe 40:4 PCe 40:5 PCe 40:6 PCe 40:7 PCe 42:5 PCe 42:6 LPC 16:0 LPC 16:1 LPC 18:0 LPC 18:1 LPC 20:0 LPC 20:1 LPC 20:2 LPC 20:3 LPC 20:4 LPCe 16:0 LPCe 18:0 LPCe 18:1 LPCe 20:0 METABOLITES_END #END