#METABOLOMICS WORKBENCH michaelsa93_20170413_144440_mwtab.txt DATATRACK_ID:887 STUDY_ID:ST000594 ANALYSIS_ID:AN000910 PROJECT_ID:PR000433 VERSION 1 CREATED_ON April 17, 2017, 3:56 pm #PROJECT PR:PROJECT_TITLE PGD2 and other lipid mediator changes in mouse adipose associated with PR:PROJECT_TITLE administration of an oral inhibitor of H-PGDS (HQL-79) PR:PROJECT_SUMMARY This is an additional experiment being added onto a previous mouse feeding study PR:PROJECT_SUMMARY that aimed to identify changes in metabolites that occur in metabolic tissues in PR:PROJECT_SUMMARY the obese state that are long-lasting and not reversed by weight loss. We PR:PROJECT_SUMMARY observed in the previous mice feeding study that levels of PGD2 increased in HFD PR:PROJECT_SUMMARY fed mice and stayed high after the diet switch. Other members of the PR:PROJECT_SUMMARY Prostaglandin family followed a similar trend (15-deoxy PGJ2, PGJ2) and were PR:PROJECT_SUMMARY specific to adipose tissue. Based on previously published data indicating that PR:PROJECT_SUMMARY central injection of PGD2 stimulates food intake, we attempted to observe this PR:PROJECT_SUMMARY effect using an oral PGD2 inhibitor of H-PGDS (HQL-79). In fact, the oral PR:PROJECT_SUMMARY inhibitor of the H-PGDS (HQL-79) administered peripherally (oral gavage in mice PR:PROJECT_SUMMARY at 30mg/kg dose) reduced daily food intake. Mice were divided into two groups PR:PROJECT_SUMMARY termed Vehicle (Control) and HGL-79 (H-PGDS inhibitor). Each group was analyzed PR:PROJECT_SUMMARY for lipid mediator changes (including PGD2) in adipose tissue by the Newman lab. PR:PROJECT_SUMMARY Analytical results generally met quality control criterion with respect to PR:PROJECT_SUMMARY surrogate recoveries and replicate precision. Surrogate recoveries were good for PR:PROJECT_SUMMARY most oxylipins (58-76%), endocannabinoids (53-75%), and fatty acids (36%). PR:PROJECT_SUMMARY Recovery precision was good for most analytes in these profiles, ranging from PR:PROJECT_SUMMARY 6-28% RSD for most surrogates. The precision for the LTB4 surrogate was higher PR:PROJECT_SUMMARY than most others (38%). Analytical precision was assessed by duplicate analysis PR:PROJECT_SUMMARY of two separate study samples. Analytical precision was 62 - 69% of analytes PR:PROJECT_SUMMARY having <30% RSD for all profiles and correlation analysis for the analytes PR:PROJECT_SUMMARY within these samples ranged from 0.90-0.99 R2. The complete data set is in the PR:PROJECT_SUMMARY associated excel file (Osborn HQL-79 – Deliverable Data Newman Lab.xls). There PR:PROJECT_SUMMARY were few statistically significant differences observed when comparing PR:PROJECT_SUMMARY concentrations (pmol/gr) between the control and HGL-79 treatment groups. PR:PROJECT_SUMMARY However, when we compared ratios we saw numerous differences between PGD2 and PR:PROJECT_SUMMARY its metabolite d15-PGJ2 versus other prostaglandins. Specifically, ratios PR:PROJECT_SUMMARY between PGD2 and other connected pathway metabolites indicate a shift toward PR:PROJECT_SUMMARY PGE2 and PGF2a production instead of PGD2 (Figure 1) with HQL-79 treatment. The PR:PROJECT_SUMMARY PGD2 and PGE2 metabolites ratio of d15-PGJ2/15-keto PGE2 was statistically PR:PROJECT_SUMMARY significant (P<0.01) using a two-tailed t-test. The ratios of PGD2/PGE2 and PR:PROJECT_SUMMARY PGD2/PGF2 had p values of P<0.09 and P=0.07), respectively. Considering that we PR:PROJECT_SUMMARY were predicting changes that indicated less PGD2 production it may be PR:PROJECT_SUMMARY justifiable to use one-tailed tests instead. In order to maintain consistency PR:PROJECT_SUMMARY with the metabolomic data analysis in the previous study, I followed the same PR:PROJECT_SUMMARY statistical protocol that Johannes preformed for the main Pilot study. Using R PR:PROJECT_SUMMARY and Devium log transformed data. Since this was a two group comparision, if the PR:PROJECT_SUMMARY data was normal a 2 tailed t-test was used and if not normal then Mann-Whitney PR:PROJECT_SUMMARY was used. A far as the significance of a shift from PGD2 to PGE2 production, I PR:PROJECT_SUMMARY found a nice review article that discusses in detail the role of prostaglandins PR:PROJECT_SUMMARY in white adipose tissue (Flachs et al. 2013). In the review it cites articles PR:PROJECT_SUMMARY that have shown PGE2 to induce UCP1, modulate lipolysis adipogenesis, and PR:PROJECT_SUMMARY stimulate leptin release. On the other hand, PGD2 was shown to increase PR:PROJECT_SUMMARY adipogenesis and weight gain. Its downstream product d15-PGJ2 has been shown to PR:PROJECT_SUMMARY increase adipogenesis, adipocyte differentiation, and decrease leptin PR:PROJECT_SUMMARY production. This is significant since I also observed that the ratio of d15-PGJ2 PR:PROJECT_SUMMARY to 15-keto PGE2 (the downstream product of PGE2) was also decreased. Another PR:PROJECT_SUMMARY prostaglandin whose ratio versus PGD2 was different in the inhibitor group was PR:PROJECT_SUMMARY PGF2a which has been shown to increase glucose transport in adipose tissue. PR:INSTITUTE University of California, San Diego PR:DEPARTMENT Department of Medicine PR:LAST_NAME Osborn PR:FIRST_NAME Olivia PR:ADDRESS 9500 Gilman Dr., La Jolla, CA 92093 PR:EMAIL oosborn@ucsd.edu PR:PHONE 858-822-6645 PR:FUNDING_SOURCE NIH U24DK097154 #STUDY ST:STUDY_TITLE PGD2 and other lipid mediator changes in mouse adipose associated with ST:STUDY_TITLE administration of an oral inhibitor of H-PGDS (HQL-79) ST:STUDY_SUMMARY This is an additional experiment being added onto a previous mouse feeding study ST:STUDY_SUMMARY that aimed to identify changes in metabolites that occur in metabolic tissues in ST:STUDY_SUMMARY the obese state that are long-lasting and not reversed by weight loss. We ST:STUDY_SUMMARY observed in the previous mice feeding study that levels of PGD2 increased in HFD ST:STUDY_SUMMARY fed mice and stayed high after the diet switch. Other members of the ST:STUDY_SUMMARY Prostaglandin family followed a similar trend (15-deoxy PGJ2, PGJ2) and were ST:STUDY_SUMMARY specific to adipose tissue. Based on previously published data indicating that ST:STUDY_SUMMARY central injection of PGD2 stimulates food intake, we attempted to observe this ST:STUDY_SUMMARY effect using an oral PGD2 inhibitor of H-PGDS (HQL-79). In fact, the oral ST:STUDY_SUMMARY inhibitor of the H-PGDS (HQL-79) administered peripherally (oral gavage in mice ST:STUDY_SUMMARY at 30mg/kg dose) reduced daily food intake. Mice were divided into two groups ST:STUDY_SUMMARY termed Vehicle (Control) and HGL-79 (H-PGDS inhibitor). Each group was analyzed ST:STUDY_SUMMARY for lipid mediator changes (including PGD2) in adipose tissue by the Newman lab. ST:STUDY_SUMMARY Analytical results generally met quality control criterion with respect to ST:STUDY_SUMMARY surrogate recoveries and replicate precision. Surrogate recoveries were good for ST:STUDY_SUMMARY most oxylipins (58-76%), endocannabinoids (53-75%), and fatty acids (36%). ST:STUDY_SUMMARY Recovery precision was good for most analytes in these profiles, ranging from ST:STUDY_SUMMARY 6-28% RSD for most surrogates. The precision for the LTB4 surrogate was higher ST:STUDY_SUMMARY than most others (38%). Analytical precision was assessed by duplicate analysis ST:STUDY_SUMMARY of two separate study samples. Analytical precision was 62 - 69% of analytes ST:STUDY_SUMMARY having <30% RSD for all profiles and correlation analysis for the analytes ST:STUDY_SUMMARY within these samples ranged from 0.90-0.99 R2. The complete data set is in the ST:STUDY_SUMMARY associated excel file (Osborn HQL-79 – Deliverable Data Newman Lab.xls). There ST:STUDY_SUMMARY were few statistically significant differences observed when comparing ST:STUDY_SUMMARY concentrations (pmol/gr) between the control and HGL-79 treatment groups. ST:STUDY_SUMMARY However, when we compared ratios we saw numerous differences between PGD2 and ST:STUDY_SUMMARY its metabolite d15-PGJ2 versus other prostaglandins. Specifically, ratios ST:STUDY_SUMMARY between PGD2 and other connected pathway metabolites indicate a shift toward ST:STUDY_SUMMARY PGE2 and PGF2a production instead of PGD2 (Figure 1) with HQL-79 treatment. The ST:STUDY_SUMMARY PGD2 and PGE2 metabolites ratio of d15-PGJ2/15-keto PGE2 was statistically ST:STUDY_SUMMARY significant (P<0.01) using a two-tailed t-test. The ratios of PGD2/PGE2 and ST:STUDY_SUMMARY PGD2/PGF2 had p values of P<0.09 and P=0.07), respectively. Considering that we ST:STUDY_SUMMARY were predicting changes that indicated less PGD2 production it may be ST:STUDY_SUMMARY justifiable to use one-tailed tests instead. In order to maintain consistency ST:STUDY_SUMMARY with the metabolomic data analysis in the previous study, I followed the same ST:STUDY_SUMMARY statistical protocol that Johannes preformed for the main Pilot study. Using R ST:STUDY_SUMMARY and Devium log transformed data. Since this was a two group comparision, if the ST:STUDY_SUMMARY data was normal a 2 tailed t-test was used and if not normal then Mann-Whitney ST:STUDY_SUMMARY was used. A far as the significance of a shift from PGD2 to PGE2 production, I ST:STUDY_SUMMARY found a nice review article that discusses in detail the role of prostaglandins ST:STUDY_SUMMARY in white adipose tissue (Flachs et al. 2013). In the review it cites articles ST:STUDY_SUMMARY that have shown PGE2 to induce UCP1, modulate lipolysis adipogenesis, and ST:STUDY_SUMMARY stimulate leptin release. On the other hand, PGD2 was shown to increase ST:STUDY_SUMMARY adipogenesis and weight gain. Its downstream product d15-PGJ2 has been shown to ST:STUDY_SUMMARY increase adipogenesis, adipocyte differentiation, and decrease leptin ST:STUDY_SUMMARY production. This is significant since I also observed that the ratio of d15-PGJ2 ST:STUDY_SUMMARY to 15-keto PGE2 (the downstream product of PGE2) was also decreased. Another ST:STUDY_SUMMARY prostaglandin whose ratio versus PGD2 was different in the inhibitor group was ST:STUDY_SUMMARY PGF2a which has been shown to increase glucose transport in adipose tissue. ST:INSTITUTE U.S.D.A. Western Human Nutrition Research Center, University of California, ST:INSTITUTE Davis ST:DEPARTMENT Nutrition ST:LAST_NAME Newman ST:FIRST_NAME John ST:ADDRESS 430 W. Health Sciences Dr., Davis, CA 95616 ST:EMAIL john.newman@ars.usda.gov ST:PHONE +1-530-752-1009 #SUBJECT SU:SUBJECT_TYPE Animal 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 Osb V1 Veh_01 Treatment:Vehicle Organ=Adipose Tissue; Species=Mus Musculus SUBJECT_SAMPLE_FACTORS Osb V2 Veh_02 Treatment:Vehicle Organ=Adipose Tissue; Species=Mus Musculus SUBJECT_SAMPLE_FACTORS Osb V3 Rep Avg. Veh_03 Treatment:Vehicle Organ=Adipose Tissue; Species=Mus Musculus SUBJECT_SAMPLE_FACTORS Osb V4 Veh_04 Treatment:Vehicle Organ=Adipose Tissue; Species=Mus Musculus SUBJECT_SAMPLE_FACTORS Osb V5 Veh_05 Treatment:Vehicle Organ=Adipose Tissue; Species=Mus Musculus SUBJECT_SAMPLE_FACTORS Osb V6 Veh_06 Treatment:Vehicle Organ=Adipose Tissue; Species=Mus Musculus SUBJECT_SAMPLE_FACTORS Osb V7 Veh_07 Treatment:Vehicle Organ=Adipose Tissue; Species=Mus Musculus SUBJECT_SAMPLE_FACTORS Osb V8 Veh_08 Treatment:Vehicle Organ=Adipose Tissue; Species=Mus Musculus SUBJECT_SAMPLE_FACTORS Osb H1 HQL_01 Treatment:HQL-79 Organ=Adipose Tissue; Species=Mus Musculus SUBJECT_SAMPLE_FACTORS Osb H2 HQL_02 Treatment:HQL-79 Organ=Adipose Tissue; Species=Mus Musculus SUBJECT_SAMPLE_FACTORS Osb H3 HQL_03 Treatment:HQL-79 Organ=Adipose Tissue; Species=Mus Musculus SUBJECT_SAMPLE_FACTORS Osb H4 HQL_04 Treatment:HQL-79 Organ=Adipose Tissue; Species=Mus Musculus SUBJECT_SAMPLE_FACTORS Osb H5 HQL_05 Treatment:HQL-79 Organ=Adipose Tissue; Species=Mus Musculus SUBJECT_SAMPLE_FACTORS Osb H6 HQL_06 Treatment:HQL-79 Organ=Adipose Tissue; Species=Mus Musculus SUBJECT_SAMPLE_FACTORS Osb H7 HQL_07 Treatment:HQL-79 Organ=Adipose Tissue; Species=Mus Musculus SUBJECT_SAMPLE_FACTORS Osb H8 Rep Avg. HQL_08 Treatment:HQL-79 Organ=Adipose Tissue; Species=Mus Musculus #COLLECTION CO:COLLECTION_SUMMARY Mice were sacrificed between 10am and noon (ad libitum fed) and adipose tissue CO:COLLECTION_SUMMARY collected and snap frozen and stored at –80oC. CO:SAMPLE_TYPE Adipose Tissue #TREATMENT TR:TREATMENT_SUMMARY C57BL6 male mice were treated with HQL-79 (H-PDGS inhibitor) or vehicle TR:TREATMENT_SUMMARY (control) by oral gavage at a dose of 30mg/kg for 5 days. TR:TREATMENT_DOSE 30mg/kg TR:TREATMENT_DOSEDURATION 5 days #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Oxylipins, endocannabinoids, and fatty acids were isolated using a Waters Ostro SP:SAMPLEPREP_SUMMARY Sample Preparation Plate (Milford, MA). Adipose samples were pulverized and SP:SAMPLEPREP_SUMMARY aliquoted (~10-15mg) were added to 2mL polypropylene tubes and spiked with a 5 SP:SAMPLEPREP_SUMMARY µL anti-oxidant solution (0.2 mg/ml solution BHT/EDTA in 1:1 MeOH:water) and 10 SP:SAMPLEPREP_SUMMARY μL 1000nM analytical deuterated surrogates. A total of 50 µL of methanol was SP:SAMPLEPREP_SUMMARY added and the tube was placed in a Geno/Grinder for 30 sec. An additional 550µL SP:SAMPLEPREP_SUMMARY isopropanol w/ 10mM ammonium formate & 1% formic acid and 100 uL water was added SP:SAMPLEPREP_SUMMARY and the tube was placed in a Geno/Grinder for 30 sec before being centrifuged at SP:SAMPLEPREP_SUMMARY 10,000g for 5 min at room temp. The supernate was then transferred into the SP:SAMPLEPREP_SUMMARY plate wells and samples were eluted into glass inserts containing 10 μL 20% SP:SAMPLEPREP_SUMMARY glycerol by applying a vacuum at 15 Hg for 10 min. Eluent was dried by speed SP:SAMPLEPREP_SUMMARY vacuum for 35 min at the medium BP setting, before switching to an aqueous SP:SAMPLEPREP_SUMMARY setting for an additional 35 min. Once dry, samples were re-constituted with the SP:SAMPLEPREP_SUMMARY internal standard 1-cyclohexyl ureido, 3-dodecanoic acid (CUDA) and 1-Phenyl SP:SAMPLEPREP_SUMMARY 3-Hexadecanoic Acid Urea (PHAU) at 100 nM (50:50 MeOH:CAN), vortexed 1 min, SP:SAMPLEPREP_SUMMARY transferred to a spin filter (0.1 µm, Millipore, Billerica, MA), centrifuged SP:SAMPLEPREP_SUMMARY for 3 min at 6ºC at <4500g (rcf), before being transferred to 2 mL LC-MS amber SP:SAMPLEPREP_SUMMARY vials. Extracts were stored at -20ºC until analysis by UPLC-MS/MS. The internal SP:SAMPLEPREP_SUMMARY standard was used to quantify the recovery of surrogate standards. SP:SAMPLEPREP_PROTOCOL_FILENAME HQL-79_Lipid_Mediator_Data_Report.docx #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Waters Acquity BEH C18 (150 x 2.1mm, 1.7um) CH:FLOW_GRADIENT See protocol/methods file CH:FLOW_RATE 0.25 CH:COLUMN_TEMPERATURE 60 °C CH:SOLVENT_A 0.1% acetic acid CH:SOLVENT_B 90% ACN / 10% IPA CH:INTERNAL_STANDARD See protocol/methods file CH:RETENTION_TIME See protocol/methods file CH:SAMPLE_INJECTION 5 uL CH:ANALYTICAL_TIME 16 min CH:WEAK_WASH_SOLVENT_NAME 20% methanol, 10% isopropanol CH:WEAK_WASH_VOLUME 600 µL CH:STRONG_WASH_SOLVENT_NAME 50:50 Acetonitrile:Methanol CH:STRONG_WASH_VOLUME 600 µL CH:SAMPLE_LOOP_SIZE 17 uL #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME ABI Sciex API 4000 QTrap MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE NEGATIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Concentration pmol/g MS_METABOLITE_DATA_START Samples Veh_01 Veh_02 Veh_03 Veh_04 Veh_05 Veh_06 Veh_07 Veh_08 HQL_01 HQL_02 HQL_03 HQL_04 HQL_05 HQL_06 HQL_07 HQL_08 Factors Treatment:Vehicle Treatment:Vehicle Treatment:Vehicle Treatment:Vehicle Treatment:Vehicle Treatment:Vehicle Treatment:Vehicle Treatment:Vehicle Treatment:HQL-79 Treatment:HQL-79 Treatment:HQL-79 Treatment:HQL-79 Treatment:HQL-79 Treatment:HQL-79 Treatment:HQL-79 Treatment:HQL-79 6-keto PGF1a 0.213 2.6 1.891 0.89 1.15 4.95 9.59 3.83 1.14 9.97 1.66 2.01 1.35 3.69 1.57 1.86 TXB2 0.768 1.68 0.8525 0.627 1.13 3.03 4.1 3.61 1.92 5.34 1.81 1.16 1.36 1.93 1.91 3.825 PGE1 2.11 1.02 1.1615 3.76 0.841 1.97 4.4 1.17 0.763 8.92 1.01 1.92 1.27 6.18 0.959 1.46 PGE2 25 17.2 13.7 32.2 10 50.1 52.6 25.8 22.5 78.4 23 23.8 34 67.4 18 26 15-Keto PGE2 0.433 0.216 0.1074 0.17 0.081 0.352 0.838 0.665 0.707 2.1 0.6 0.489 1.53 2.51 0.105 0.1821 PGD2 72.7 75.7 84.3 153 102 253 163 324 92.6 351 101 55.2 126 268 65 95.45 15-deoxy PGJ2 3.46 2.35 2.105 3.68 3.07 5.41 7.48 10.5 3.98 11.8 3.52 1.63 3.51 10.3 2.12 2.81 PGF2a 6.14 7.32 4.275 5.25 4.51 9.57 8.56 11.1 3.75 22 6.1 5.88 8.6 18.6 7.64 7.68 11-HETE 51.8 54.8 56.7 47.6 58.4 117 189 181 64.9 154 70.2 18.7 68.2 81 37.9 75.9 Lipoxin A4 31.8 10 6.67 34.4 7.55 20.6 27.7 23.2 17.4 38.8 16.9 15.9 27 46.1 4.51 8.195 6-trans-LTB4 2.3 1.39 2.08 1.91 1.82 2.66 10.3 6.36 3.37 4.77 2.74 0.917 3.78 2.99 1.72 3.57 5_15-DiHETE 0.706 0.676 1.3545 1.9 1.76 2.27 4.32 0.698 1.56 1.46 0.587 0.606 2.82 1.01 1.05 2.47 8_15-DiHETE 6.61 3.75 4.29 4.96 4.82 5.63 21.4 12.3 4.9 7.06 5.12 2.9 6.9 3.8 2.38 3.965 9_12_13-TriHOME 974 500 192.5 1350 165 695 620 775 573 1570 743 1380 1040 3570 112 207 9_10-e-DiHO 1480 631 255 1520 151 979 832 1070 613 2260 799 1370 943 3900 128 173.5 12_13-DiHOME 24.2 12.5 6.29 28.6 4.88 17.2 13.9 19 20.1 35.4 15.4 28.1 21.3 74.6 3.26 5.28 9_10-DiHOME 31.5 15.8 8.25 37 6.67 24.7 20.3 23.7 21.7 46 20.6 34.5 24 86.6 5.17 7.32 15_16-DiHODE 0.59 1.37 2.21 1.51 4.28 0.84 4.95 0.74 2.02 0.6 1.38 1.67 0.47 1.16 9_10-DiHODE 0.49 0.29 0.3 0.32 0.17 0.64 0.33 0.63 0.47 0.37 0.45 0.41 0.3 0.73 0.12 0.2 14_15-DiHETrE 0.923 0.595 0.5355 0.887 0.443 1.17 1.59 2.17 0.613 1.11 0.569 0.418 0.737 1.42 0.361 0.426 11_12-DiHETrE 1 0.426 0.342 0.709 0.394 0.979 1.07 1.13 0.473 1.02 0.457 0.251 0.547 0.987 0.209 0.3575 8_9-DiHETrE 0.846 0.716 0.382 1.21 0.273 1.04 1.18 0.84 0.356 0.913 0.304 0.375 0.602 0.911 0.739 0.4045 5_6-DiHETrE 0.362 0.233 0.1405 0.309 0.222 0.191 0.252 0.806 0.174 0.227 0.0808 0.12 0.357 0.233 0.0468 0.0904 17_18-DiHETE 31.1 30.7 23.2 39.4 30.1 34.3 39 39.9 29.9 47.6 30 21.7 54.6 22.5 29.4 19.3 19_20-DiHDoPA 1.59 1.46 1.995 1.25 1.42 2.87 4.26 7.77 0.929 2.26 0.994 0.732 0.605 1.63 0.88 1.915 13-HODE 1580 701 835.5 1430 934 1480 3790 2750 1110 2310 1460 847 1520 2370 406 1067 9-HODE 1370 560 583.5 1170 584 1130 1980 1950 997 1620 1130 727 1020 2070 328 700.5 13-HOTE 40.3 17.9 39.35 26.7 31.8 42.1 151 77.6 36.8 77.6 38.1 16.2 52.3 34 15.4 74.05 9-HOTE 2.3 1.25 1.81 1.53 0.931 1.66 1.51 5 3.63 3.02 1.15 0.714 1.46 3.57 1.19 2.41 20-HETE 0.712 1.415 1.68 1 2.82 4.68 7.46 0.434 0.1 1.23 1.36 15-HETE 112 81.2 89.8 104 139 200 583 310 84.8 279 125 37.8 145 131 60.7 148 12-HETE 179 297 215.5 262 382 349 2020 928 163 960 273 172 413 270 172 348.5 9-HETE 52.5 46.7 57.5 37.1 50.9 84.4 137 153 71.6 88.6 70.5 9.91 64.9 53.3 29.6 65.3 8-HETE 42 32.8 33.8 33.1 40.7 57.4 128 108 45.9 80.8 49.5 9.29 50.9 44 22.9 47.1 5-HETE 71.8 43.2 37.75 51.4 49.9 82 135 151 54.4 117 61.7 16.7 67.1 82.3 22.3 50.25 15-HEPE 8.02 3.36 8.635 6.75 10.3 12.6 47.2 20 7.48 21.5 8.61 2.84 14.1 6.54 4.32 15.45 12-HEPE 7.24 15.9 11.14 10.3 13.2 19.1 40.7 41.9 7.52 43.8 11.7 8.27 14.2 15.9 7.02 18.25 9-HEPE 4.74 5.96 6.26 3.66 5.52 6.48 6.45 9.27 8.7 6.28 6.01 1.02 5.79 4.04 2.14 8.89 5-HEPE 6.67 4.05 3.63 4.24 3.25 7.18 7.65 10.1 3.13 7.1 5.21 1.84 7.09 4.75 1.73 4.95 17-HDoHE 29.3 17.9 37.4 33.2 46.7 45.4 280 115 49.1 116 55.2 22.3 66.8 45 36.1 87.25 14-HDoHE 98.7 38.6 116.05 90.9 110 125 1210 365 74.7 313 115 38 191 64.8 52.2 237.45 4-HDoHE 8.35 2.26 3.46 3.87 3.95 6.57 11 15.2 4.58 10.6 6.92 2.47 5.39 8.97 3.01 3.98 9_10-EpO 1570 345 135 721 63.3 417 580 982 451 1020 705 732 523 3420 72.7 113 12(13)-EpOME 49.2 17.8 30.3 27.9 12.7 29 32.9 63.2 59.7 50.9 36.6 30.1 36.1 101 19.2 32.5 9(10)-EpOME 61.3 19.6 34.35 34.5 14.2 30.5 37.1 67.8 65.3 66.3 41.1 38.7 41.7 127 20.3 32.8 12(13)-Ep-9-KODE 122 57 27.35 86.1 16.5 67.3 77.2 167 51.8 161 77.9 104 67.9 285 10.3 23.8 15(16)-EpODE 6.82 2.87 4.475 2.69 1.71 6.26 6.07 11.1 7.33 11.1 5.86 3.13 4.47 12.2 1.94 4.455 12(13)-EpODE 1.27 0.394 0.671 0.769 0.555 0.756 0.564 1.52 2.11 0.881 0.797 0.74 0.729 1.19 0.591 1.18 9(10)-EpODE 3.29 1.67 2.64 2.5 1.38 2.18 2.95 6.49 4.01 5.34 3.09 2.01 2.61 6.44 1.79 3.195 14(15)-EpETrE 6.76 4.62 6.765 5.26 3.52 5.38 5.64 11.6 14.9 8.79 4.07 3.63 5.75 10.7 5.8 6.965 11(12)-EpETrE 4.78 2.74 3.395 2.69 2.19 2.58 3.86 7.19 9.56 4.64 3.74 1.81 3.64 6.7 3.21 3.41 8(9)-EpETrE 4.78 2.3 3.555 2.95 2.19 3.01 3.48 9.32 10.1 5.53 4.1 2.44 3.77 6.6 2.84 3.01 19(20)-EpDPE 5.73 2.62 3.17 3.62 1.54 2.69 4.2 9.93 5.11 4.4 2.67 2.36 1.37 6.68 1.69 2.915 13-KODE 42.5 28.7 17.25 44.1 18 37.5 48.3 60.9 49.9 73.9 66.4 40.6 50.2 93 16.4 40.2 9-KODE 125 68.3 42.35 126 41.7 91.1 107 109 142 163 186 108 130 251 31.1 84.05 15-KETE 2.92 1.78 2.13 2.76 1.26 1.91 7.12 6.24 2.4 4.89 2.99 0.583 3.12 3.26 0.977 2.645 5-KETE 2.37 2.44 0.715 1.51 0.975 1.87 2.88 5.57 1.69 3.02 2.26 0.736 2.71 3.57 0.519 1.485 10-Nitrooleate 3.19 1.54 3.375 3.75 2.22 1.05 5.71 13.1 0.928 7.94 3.09 0.976 7.58 7.23 2.71 2.834 PGE3 0.167 0.665 0.45375 0.163 0.00609 1.09 1.09 0.332 0.258 1.78 0.43 0.288 0.451 0.781 0.431 LTE4 LTB4 LTB5 Resolvin D1 12_13-DiHODE 14_15-DiHETE 17(18)-EpETE 14(15)-EpETE 11(12)-EpETE 16(17)-EpDPE 15-HpETE screen 12-HpETE screen 5-HpETE screen 9-Nitrooleate 10-Nitrolinoleate PGF3a MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z PubChem ID KEGG ID CID 10-Nitrooleate 9-Nitrooleate 10-Nitrolinoleate 6-keto PGF1a C05961 5280888 TXB2 5283137 PGE1 C04741 5280723 PGE2 C00584 5280360 15-Keto PGE2 PGD2 C00696 448457 15-deoxy PGJ2 C14717 5311211 PGF2a 53481911 11-HETE 5312981 Lipoxin A4 C06314 5280914 6-trans-LTB4 5283128 5_15-DiHETE 5283158 8_15-DiHETE 53480358 9_12_13-TriHOME C14833 9858729 9_10-e-DiHO 441460 12_13-DiHOME C14829 10236635 9_10-DiHOME C14828 9966640 15_16-DiHODE 16061068 9_10-DiHODE 16061066 14_15-DiHETrE C14775 5283147 11_12-DiHETrE C14774 5283146 8_9-DiHETrE C14773 5283144 5_6-DiHETrE C14772 5283142 17_18-DiHETE 16061120 19_20-DiHDoPA 16061148 13-HODE C14762 6443013 9-HODE 5282945 13-HOTE 10469728 9-HOTE 53480359 20-HETE 35027640 15-HETE C04742 5280724 12-HETE 5312983 9-HETE 5312978 8-HETE C14776 5283154 5-HETE C04805 5280733 15-HEPE 53480357 12-HEPE 10041593 9-HEPE 5283187 5-HEPE 6439678 17-HDoHE 6439179 14-HDoHE 11566378 4-HDoHE 11566378 9_10-EpO 15868 12(13)-EpOME 5356421 9(10)-EpOME C14828 9966640 12(13)-Ep-9-KODE 15(16)-EpODE 16061062 12(13)-EpODE 16061061 9(10)-EpODE 16061060 14(15)-EpETrE C14813 11954058 11(12)-EpETrE 8(9)-EpETrE C14769 5283203 19(20)-EpDPE 11631565 13-KODE C14765 6446027 9-KODE C14766 9839084 15-KETE C04577 5280701 5-KETE C14732 5283159 PGE3 C06439 5280937 LTE4 5280749 LTB4 C05961 5280888 LTB5 5283125 Resolvin D1 16061135 12_13-DiHODE 16061067 14_15-DiHETE 16061119 17(18)-EpETE 16061089 14(15)-EpETE 16061088 11(12)-EpETE 16061087 16(17)-EpDPE 14392758 15-HpETE screen C05966 5280893 12-HpETE screen C05965 5280892 5-HpETE screen 5283171 PGF3a METABOLITES_END #END