#METABOLOMICS WORKBENCH patchthr33_20190924_154553_mwtab.txt DATATRACK_ID:1822 STUDY_ID:ST001254 ANALYSIS_ID:AN002083 PROJECT_ID:PR000841 VERSION 1 CREATED_ON September 25, 2019, 5:10 pm #PROJECT PR:PROJECT_TITLE Eicosanoid Expression in Dermal Fibroblasts PR:PROJECT_SUMMARY Quantitative lipidomics studies examining eicosanoid levels in primary dermal PR:PROJECT_SUMMARY fibroblasts PR:INSTITUTE University of South Florida, Tampa PR:LAST_NAME Chalfant PR:FIRST_NAME Charles PR:ADDRESS 4202 E Fowler Ave, Tampa, FL 33620 PR:EMAIL cechalfant@usf.edu PR:PHONE 8139747103 #STUDY ST:STUDY_TITLE Eicosanoid profiles of dermal fibroblasts (part-II) ST:STUDY_SUMMARY The sphingolipid, ceramide-1-phosphate (C1P), directly binds and activates Group ST:STUDY_SUMMARY IVA cytosolic phospholipase A2 (cPLA2) to generate eicosanoids. Due to the role ST:STUDY_SUMMARY of eicosanoids in wound healing, we choose to use our novel genetic mouse model ST:STUDY_SUMMARY expressing cPLA2 with an ablated C1P interaction site (KI) to examine the ST:STUDY_SUMMARY cPLA2/C1P interaction in wound healing. Wound closure rate was not affected, but ST:STUDY_SUMMARY wound maturation was enhanced by loss of the C1P/cPLA2α interaction based on ST:STUDY_SUMMARY the following findings. Wounds in KI mice displayed: i) increased infiltration ST:STUDY_SUMMARY of dermal fibroblasts into the wound environment; ii) increased wound tensile ST:STUDY_SUMMARY strength; and iii) higher Type I/Type III collagen ratios. These findings were ST:STUDY_SUMMARY recapitulated in vitro as primary dermal fibroblasts (pDFs) from KI mice showed ST:STUDY_SUMMARY significantly increased collagen deposition and migration velocity compared to ST:STUDY_SUMMARY WT and KO pDFs. Additionally, the KI showed an altered eicosanoid profile of ST:STUDY_SUMMARY reduced pro-inflammatory prostaglandins (PGE2) and increased levels of specific ST:STUDY_SUMMARY HETE species (5-HETE). Elevated 5-HETE levels promoted increased dermal ST:STUDY_SUMMARY fibroblast migration and collagen deposition. This “gain of function” role ST:STUDY_SUMMARY for the mutant cPLA2 was also linked to differential cellular localization of ST:STUDY_SUMMARY cPLA2α and 5-HETE biosynthetic factors. These studies demonstrate regulation of ST:STUDY_SUMMARY key biological mechanisms by a defined protein:lipid interaction in vivo and ST:STUDY_SUMMARY provide new insights into cPLA2 function. ST:INSTITUTE University of South Florida, Tampa ST:LAST_NAME Chalfant ST:FIRST_NAME Charles ST:ADDRESS 4202 E Fowler Ave, Tampa, FL 33620 ST:EMAIL cechalfant@usf.edu ST:PHONE 8139747103 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - Patrick Media 1 Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 2 Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 3 Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 4 Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 5 Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 6 Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 7 Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 8 Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 9 Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 10 Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 11 Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 12 Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 13 Genotype:cPLA2α KO | Treatment:Wild Type Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 14 Genotype:cPLA2α KO | Treatment:Wild Type Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 15 Genotype:cPLA2α KO | Treatment:Wild Type Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 16 Genotype:cPLA2α KO | Treatment:Wild Type Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 17 Genotype:cPLA2α KO | Treatment:Wild Type Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 18 Genotype:cPLA2α KO | Treatment:Wild Type Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 19 Genotype:cPLA2α KO | Treatment:Wild Type Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 20 Genotype:cPLA2α KO | Treatment:Wild Type Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 21 Genotype:cPLA2α KO | Treatment:Wild Type Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 22 Black Genotype:cPLA2α KI | Treatment:Pyrrophenone (50nM) SUBJECT_SAMPLE_FACTORS - Patrick Media 23 Black Genotype:cPLA2α KO | Treatment:Pyrrophenone (50nM) SUBJECT_SAMPLE_FACTORS - Patrick Media 24 Genotype:cPLA2α KO | Treatment:Pyrrophenone (50nM) SUBJECT_SAMPLE_FACTORS - Patrick Media 25 Genotype:cPLA2α KI | Treatment:Pyrrophenone (50nM) SUBJECT_SAMPLE_FACTORS - Patrick Media 26 Genotype:Wild Type | Treatment:Pyrrophenone (50nM) SUBJECT_SAMPLE_FACTORS - Patrick Media 27 Genotype:Wild Type | Treatment:Pyrrophenone (50nM) SUBJECT_SAMPLE_FACTORS - Patrick Media 28 Genotype:cPLA2α KO | Treatment:CMV control Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 29 Genotype:cPLA2α KO | Treatment:CMV control Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 30 Genotype:cPLA2α KO | Treatment:Pyrrophenone (50nM) SUBJECT_SAMPLE_FACTORS - Patrick Media 31 Genotype:Wild Type | Treatment:Pyrrophenone (50nM) SUBJECT_SAMPLE_FACTORS - Patrick Media 32 Genotype:cPLA2α KO | Treatment:CMV control Virus SUBJECT_SAMPLE_FACTORS - Patrick Media 33 Genotype:cPLA2α KI | Treatment:Pyrrophenone (50nM) SUBJECT_SAMPLE_FACTORS - Patrick Media 34 Genotype:Wild Type | Treatment:Untreated SUBJECT_SAMPLE_FACTORS - Patrick Media 37 Genotype:cPLA2α KI | Treatment:Untreated SUBJECT_SAMPLE_FACTORS - Patrick Media 38 Genotype:Wild Type | Treatment:Untreated SUBJECT_SAMPLE_FACTORS - Patrick Media 39 Genotype:Wild Type | Treatment:Untreated SUBJECT_SAMPLE_FACTORS - Patrick Media 40 Genotype:cPLA2α KI | Treatment:Untreated SUBJECT_SAMPLE_FACTORS - Patrick Media 41 Genotype:cPLA2α KI | Treatment:Untreated SUBJECT_SAMPLE_FACTORS - Patrick Media 42 Genotype:cPLA2α KI | Treatment:Untreated SUBJECT_SAMPLE_FACTORS - Patrick Media 43 Genotype:Wild Type | Treatment:Untreated SUBJECT_SAMPLE_FACTORS - Patrick Media KI Genotype:cPLA2α KI | Treatment:Untreated #COLLECTION CO:COLLECTION_SUMMARY pDFs obtained from WT, KI, and KO mice were plated at a density of 2 x 106 on 10 CO:COLLECTION_SUMMARY cm tissue culture plates in high glucose DMEM supplemented with 20% FBS (Gibco) CO:COLLECTION_SUMMARY and 2% penicillin/streptomycin. Following the overnight incubation cells were CO:COLLECTION_SUMMARY rested in 2% FBS (Gibco) 2% penicillin/streptomycin (Bio Whittaker) high glucose CO:COLLECTION_SUMMARY DMEM (Gibco) for 2 hours, then the media was exchanged with 0% FBS 2% CO:COLLECTION_SUMMARY penicillin/ high glucose DMEM, and mechanical trauma was induced on the CO:COLLECTION_SUMMARY monolayer by performing scratched across the diameter of the plate in an CO:COLLECTION_SUMMARY asterisk pattern using 4 x 20 ul pipette tips on a multichannel micro pipettor. CO:COLLECTION_SUMMARY Media was taken for lipidomic analysis at 0 h and 2 h. CO:SAMPLE_TYPE Fibroblasts #TREATMENT TR:TREATMENT_SUMMARY Primary Dermal Fibroblasts were plated at a density of 2 x 106 on 10 cm tissue TR:TREATMENT_SUMMARY culture plates in high glucose DMEM supplemented with 20% FBS (Gibco) and 2% TR:TREATMENT_SUMMARY penicillin/streptomycin. Following the overnight incubation cells were rested in TR:TREATMENT_SUMMARY 2% FBS (Gibco) 2% penicillin/streptomycin (Bio Whittaker) high glucose DMEM TR:TREATMENT_SUMMARY (Gibco) for 2 hours, then the media was exchanged with 0% FBS 2% penicillin/ TR:TREATMENT_SUMMARY high glucose DMEM, and mechanical trauma was induced on the monolayer by TR:TREATMENT_SUMMARY performing scratched across the diameter of the plate in an asterisk pattern TR:TREATMENT_SUMMARY using 4 x 20 l pipette tips on a multichannel micro pipettor. Media was taken TR:TREATMENT_SUMMARY for lipidomic analysis at 0 h and 2 h. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Samples were stored at − 80 °C until the time of analysis. Lipids were SP:SAMPLEPREP_SUMMARY extracted for eicosanoids. Eicosanoid Preparation Eicosanoids were extracted SP:SAMPLEPREP_SUMMARY using a modified extraction process from previously described. (1, 2). 4 mL of SP:SAMPLEPREP_SUMMARY media was combined with an IS mixture comprised of comprised of 10% methanol SP:SAMPLEPREP_SUMMARY (400 μl), glacial acetic acid (20 μl), and internal standard (20 μl) SP:SAMPLEPREP_SUMMARY containing the following deuterated eicosanoids (1.5 pmol/μl, 30 pmol total) SP:SAMPLEPREP_SUMMARY (All standards purchased from Cayman Chemicals): (d4) 6keto-Prostaglandin F1α, SP:SAMPLEPREP_SUMMARY (d4) Prostaglandin F2α, (d4) Prostaglandin E2, (d4) Prostaglandin D2, (d8) SP:SAMPLEPREP_SUMMARY 5-Hydroxyeicosa-tetranoic acid (5-HETE), (d8) 12-Hydroxyeicosa-tetranoic acid SP:SAMPLEPREP_SUMMARY (12-HETE) (d8) 15-Hydroxyeicosa-tetranoic acid (15-HETE), (d6) SP:SAMPLEPREP_SUMMARY 20-Hydroxyeicosa-tetranoic acid (20-HETE), (d11) 8,9 Epoxyeicosa-trienoic acid, SP:SAMPLEPREP_SUMMARY (d8) 14,15 Epoxyeicosa-trienoic acid, (d8) Arachidonic acid, (d5) SP:SAMPLEPREP_SUMMARY Eicosapentaenoic acid, (d5) Docosahexaenoic acid, (d4) Prostaglandin A2, (d4) SP:SAMPLEPREP_SUMMARY Leukotriene B4, (d4) Leukotriene C4, (d4) Leukotriene D4, (d4) Leukotriene E4, SP:SAMPLEPREP_SUMMARY (d5) 5(S),6(R)-Lipoxin A4, (d11) 5-iPF2α-VI, (d4) 8-iso Prostaglandin F2α, SP:SAMPLEPREP_SUMMARY (d11) (±)14,15-DHET, (d11) (±)8,9-DHET, (d11) (±)11,12-DHET, (d4) SP:SAMPLEPREP_SUMMARY Prostaglandin E1, (d4) Thromboxane B2, (d6) dihmo gamma linoleic acid, (d5) SP:SAMPLEPREP_SUMMARY Resolvin D2, (d5) Resolvin D1, (d5) Maresin 2, and (d5) Resolvin D3. Samples and SP:SAMPLEPREP_SUMMARY vial rinses (5% MeOH; 2 ml) were applied to Strata-X SPE columns (Phenomenex), SP:SAMPLEPREP_SUMMARY previously washed with methanol (2 ml) and then dH2O (2 ml). Eicosanoids eluted SP:SAMPLEPREP_SUMMARY with isopropanol (2 ml), were dried in vacuuo and reconstituted in EtOH:dH2O SP:SAMPLEPREP_SUMMARY (50:50;100 μl) prior to UPLC ESI-MS/MS analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Shimadzu Nexera X2 CH:COLUMN_NAME Acentis Express C18 column #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME ABI Sciex 5500 QTrap MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE MALDI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Sciex Analyst software was used for analysis #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS pmol of lipid MS_METABOLITE_DATA_START Samples Patrick Media 1 Patrick Media 2 Patrick Media 3 Patrick Media 4 Patrick Media 5 Patrick Media 6 Patrick Media 7 Patrick Media 8 Patrick Media 9 Patrick Media 10 Patrick Media 11 Patrick Media 12 Patrick Media 13 Patrick Media 14 Patrick Media 15 Patrick Media 16 Patrick Media 17 Patrick Media 18 Patrick Media 19 Patrick Media 20 Patrick Media 21 Patrick Media 22 Black Patrick Media 23 Black Patrick Media 24 Patrick Media 25 Patrick Media 26 Patrick Media 27 Patrick Media 28 Patrick Media 29 Patrick Media 30 Patrick Media 31 Patrick Media 32 Patrick Media 33 Patrick Media 34 Patrick Media 37 Patrick Media 38 Patrick Media 39 Patrick Media 40 Patrick Media 41 Patrick Media 42 Patrick Media 43 Patrick Media KI Factors Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus Genotype:cPLA2α KO | Treatment:cPLA2α KI Virus Genotype:cPLA2α KO | Treatment:Wild Type Virus Genotype:cPLA2α KO | Treatment:Wild Type Virus Genotype:cPLA2α KO | Treatment:Wild Type Virus Genotype:cPLA2α KO | Treatment:Wild Type Virus Genotype:cPLA2α KO | Treatment:Wild Type Virus Genotype:cPLA2α KO | Treatment:Wild Type Virus Genotype:cPLA2α KO | Treatment:Wild Type Virus Genotype:cPLA2α KO | Treatment:Wild Type Virus Genotype:cPLA2α KO | Treatment:Wild Type Virus Genotype:cPLA2α KI | Treatment:Pyrrophenone (50nM) Genotype:cPLA2α KO | Treatment:Pyrrophenone (50nM) Genotype:cPLA2α KO | Treatment:Pyrrophenone (50nM) Genotype:cPLA2α KI | Treatment:Pyrrophenone (50nM) Genotype:Wild Type | Treatment:Pyrrophenone (50nM) Genotype:Wild Type | Treatment:Pyrrophenone (50nM) Genotype:cPLA2α KO | Treatment:CMV control Virus Genotype:cPLA2α KO | Treatment:CMV control Virus Genotype:cPLA2α KO | Treatment:Pyrrophenone (50nM) Genotype:Wild Type | Treatment:Pyrrophenone (50nM) Genotype:cPLA2α KO | Treatment:CMV control Virus Genotype:cPLA2α KI | Treatment:Pyrrophenone (50nM) Genotype:Wild Type | Treatment:Untreated Genotype:cPLA2α KI | Treatment:Untreated Genotype:Wild Type | Treatment:Untreated Genotype:Wild Type | Treatment:Untreated Genotype:cPLA2α KI | Treatment:Untreated Genotype:cPLA2α KI | Treatment:Untreated Genotype:cPLA2α KI | Treatment:Untreated Genotype:Wild Type | Treatment:Untreated Genotype:cPLA2α KI | Treatment:Untreated 6-keto PGF1α 0.100724638 0.064583333 0.051931034 0.082774869 0.053736264 0.111882353 0.045522388 0.072162162 0.069695122 0.060193548 0.05961039 0.051910112 16.09411765 17.38636364 16.73684211 17.14285714 17.36842105 17.0877193 16.32124352 17.25806452 17.27272727 0.188684211 0.105691057 0.062 0.183128834 0.195689655 0.194617564 0.237209302 0.226890756 0.058931507 0.207365439 0.307714286 0.217177914 30.68767908 16.41065831 30.08595989 31.48688047 15.67567568 15.55714286 16.05571848 22.47863248 19.89884393 TXB2 2.139130435 2.525 2.553719008 1.911111111 2.22 2.358870968 1.780714286 2.363414634 2.564885496 2.3616 2.295238095 1.996240602 1.911290323 2.032330827 1.971126761 2.001398601 2.049635036 1.988793103 1.909859155 2.032394366 1.984931507 0.225477707 0.217307692 0.120197368 0.255555556 0.269811321 0.261538462 0.267307692 0.244936709 0.114834437 0.253125 0.245962733 0.23877551 0.27375 0.143835616 0.273248408 0.253846154 0.141233766 0.124203822 0.127884615 0.211282051 0.160416667 PGE2 1.469117647 1.185 1.429765886 1.346385542 1.127721335 1.214173228 1.346853147 1.464 1.475482912 1.332786885 1.456168831 1.189189189 11.56398104 12.31671554 11.42857143 11.56968877 11.74698795 11.17241379 11.1637931 11.64305949 11.6781293 0.996240602 1.041871921 0.566489362 1.5046875 1.44895288 1.479619565 1.408244681 1.384303112 0.513833992 1.458278146 1.474375821 1.462707182 18.36340206 10.03551136 18.56767411 19.30013459 9.658634538 9.409090909 8.732876712 13.72214941 11.08600583 PGF2α 0.14440678 0.191538462 0.173800738 0.185172414 0.198634812 0.168265683 0.139464883 0.255555556 0.249295775 0.15498155 0.211636364 0.180836237 0.77601476 0.832525952 0.835494881 0.822711864 0.831010453 0.849790795 0.833333333 0.842805755 0.790635452 0.073773585 0.049880597 0.036509434 0.17032967 0.16 0.162264151 0.177358491 0.137142857 0.040263158 0.131775701 0.145820433 0.163087248 1.186708861 0.602040816 1.147335423 1.213375796 0.611464968 0.597178683 0.612149533 0.715457413 0.813993174 PGE1 0.126119403 0.145689655 0.155898876 0.132989691 0.120153061 0.129395604 0.1657289 0.17027027 0.180672269 0.167751479 0.193965517 0.153824363 1.366071429 1.424581006 1.398963731 1.434108527 1.471698113 1.380191693 1.3671875 1.433070866 1.464379947 0.112142857 0.105924171 0.04886747 0.176521739 0.17027027 0.162561576 0.188613861 0.15403423 0.049411765 0.155072464 0.175362319 0.142931937 1.101449275 0.586253369 1.085574572 1.129584352 0.555147059 0.54679803 0.548693587 0.793269231 0.807377049 Resolvin D1 0.216342857 0.226707132 0.225853659 0.224133504 0.204769001 0.205053191 0.219809069 0.205740181 0.251226994 0.2273021 0.241521395 0.242633229 0.23392283 0.240620384 0.231238447 0.230390921 0.233015267 0.233870968 0.214519056 0.238830585 0.214059754 0.240606061 0.218723849 0.235514019 0.235542857 0.238918919 0.225714286 0.246055046 0.231818182 0.237286432 0.232767857 0.224482759 0.234769539 0.230086957 0.113632075 0.233275862 0.238648649 0.118016529 0.12 0.115769231 0.245948276 0.118446602 PGA2 0.511312217 0.462441315 0.483412322 0.488269795 0.46359584 0.45 0.482608696 0.47275405 0.468481375 0.462222222 0.469985359 0.431004367 0.834621329 0.790598291 0.882857143 0.794238683 0.936416185 0.882661996 0.855337079 0.873103448 0.901709402 0.442307692 0.460947503 0.415446072 0.405417957 0.429697766 0.447058824 0.432539683 0.425806452 0.419915254 0.414852753 0.453621347 0.394092827 0.74522293 0.403521127 0.725490196 0.734693878 0.354085603 0.386658031 0.374519846 0.859569649 0.524373259 (±)14,15-DHET 0.186764706 0.226943005 0.225 0.170792079 0.207575758 0.215217391 0.154411765 0.1921875 0.2203125 0.216923077 0.220812183 0.19119171 0.203743316 0.215625 0.196039604 0.209405941 0.225 0.185294118 0.199009901 0.208823529 0.210606061 0.064736842 0.057906977 0.047368421 0.046540541 0.0571875 0.048947368 0.057567568 0.054776786 0.027681159 0.051615721 0.049380531 0.057548077 0.066753247 0.034316038 0.063053097 0.069147982 0.0338 0.03277533 0.040394737 0.051527778 0.039073171 (±)11,12-DHET 0.1475 0.17261194 0.174586466 0.118510638 0.144125874 0.145846154 0.110896552 0.146793893 0.151521739 0.152888889 0.139051095 0.123724138 0.141724138 0.15 0.149224138 0.12953271 0.129226069 0.129073857 0.127864078 0.138857143 0.139322034 0.045971223 0.035869565 0.032627737 0.043636364 0.04943662 0.038482759 0.039657534 0.040416667 0.016125 0.039657534 0.042567568 0.041307692 0.051636364 0.0212 0.038148148 0.031132075 0.019082278 0.02015625 0.021634615 0.035912409 0.021590909 (±)8,9-DHET 0.055701754 0.090350877 0.079459459 0.05340176 0.074302326 0.093301887 0.066017192 0.061791045 0.093134328 0.063153153 0.1 0.087617647 0.068888889 0.071462687 0.060685714 0.062406877 0.074955752 0.063402062 0.086440678 0.070916905 0.063088235 0.01572973 0.01059322 0.00661157 0.010315457 0.015281501 0.014755435 0.015616438 0.012752044 0 0 0.017734807 0.012209302 0.01697479 0.009009009 0.024830508 0.016324786 0.009375 0.007352941 0.006829609 0.016512968 0.01379822 20-HETE 17.56097561 15.79710145 16.86567164 16.53658537 17.1 16.57894737 17.63033175 17.75510204 18.05970149 19.47115385 18.41584158 19.55223881 21.3368984 20.625 22.61306533 22.92307692 21.12244898 22.54545455 22.82608696 23.33333333 27.52747253 39.94897959 38.08290155 33.37078652 34.04907975 29.53367876 30.78125 31.72774869 31.10526316 32.61627907 31.0880829 30.47619048 33.22033898 35.15151515 18.37912088 37.0212766 36.2755102 18.07894737 19.17525773 18.40206186 38.02139037 21.06557377 15 HETE 0.991836735 1.155737705 1.168421053 0.95625 1.079497908 1.115973742 0.923076923 1.135483871 1.158663883 1.071868583 1.090336134 0.989878543 0.55921659 0.603870968 0.632026144 0.638443936 0.650985222 0.570304569 0.552078775 0.587846482 0.662291169 0.270449679 0.268677494 0.140425532 0.400554017 0.373362445 0.380405405 0.396226415 0.365584416 0.132167832 0.357543103 0.405681818 0.347630332 0.389938398 0.222695035 0.382258065 0.412095032 0.194516129 0.200210526 0.213525499 0.320712695 0.258352941 12 HETE 11.96629213 14.15929204 14.47647951 10.71428571 12.99703264 12.91021672 8.958333333 12.39329268 12.80752533 13.41853035 11.88841202 10.01432665 5.882352941 5.925925926 5.664335664 5.502645503 6.352201258 5.513347023 5.55353902 5.671378092 6.222222222 0.659913169 0.664086687 0.371602787 0.892605634 0.812130178 0.818941504 0.896103896 0.8125 0.270899471 0.887573964 0.918083462 0.834405145 0.222188906 0.117391304 0.210046368 0.223111111 0.109495549 0.110480349 0.110949464 0.192215569 0.147196262 (±)14(15)-EET 0.084080717 0.094736842 0.099315068 0.078472222 0.094090909 0.092137592 0.078801843 0.096774194 0.131060606 0.10125 0.096821516 0.085085575 0.048065395 0.049453125 0.061526718 0.051688654 0.053395225 0.047926829 0.048903061 0.048467337 0.074634146 0.023606557 0.025690141 0.017492711 0.0305625 0.0353125 0.038692308 0.03689295 0.011220159 0.010120482 0.039481865 0.036832461 0.016147309 0.011560976 0.020303867 0.04047486 0.025623342 0.012532637 0.017772152 0.011701571 0.015163043 0.023323944 5 HETE 0.527455919 0.594414894 0.620215633 0.49379845 0.58230563 0.570422535 0.51171875 0.728455285 0.714441417 0.6824 0.6416 0.532283465 0.282075472 0.276854599 0.253353659 0.211764706 0.221779141 0.226027397 0.238575668 0.215652174 0.279936306 0.026553525 0.039806094 0.051311475 0.126959248 0.079572193 0.096946565 0.100519481 0.076675325 0.036333333 0.08125 0.092670157 0.140782123 0.033178295 0.036306818 0.042152975 0.048179348 2.907754011 2.888297872 2.964659686 0.032591623 0.044606414 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Retention Time m/z PubChem ID KEGG ID 6-keto PGF1α 4.33 369.2 5280888 C05961 TXB2 5.09 369.2 5283137 C05963 PGE2 4.56 351.2 5280360 C00584 PGF2α 5.5 353.2 5280636 C00639 PGE1 5.63 353.2 5280723 C04741 Resolvin D1 6.09 375.201 44251266 C18178 PGA2 6.4 333.2 5280880 C05953 (±)14,15-DHET 7.52 337.2 5283147 C14775 (±)11,12-DHET 7.61 337.2 129590 C14774 (±)8,9-DHET 7.73 337.2 5283144 C14773 20-HETE 7.75 319.2 5283157 C14748 15 HETE 7.99 319.2 9966861 C04742 12 HETE 8.15 319.2 13786989 C14777 (±)14(15)-EET 8.33 319.2 5283205 C14771 5 HETE 8.41 319.2 5280733 C04805 METABOLITES_END #END