#METABOLOMICS WORKBENCH burlab_20190119_200209 DATATRACK_ID:1609 STUDY_ID:ST001140 ANALYSIS_ID:AN001872 PROJECT_ID:PR000761 VERSION 1 CREATED_ON February 21, 2019, 4:41 pm #PROJECT PR:PROJECT_TITLE Changes in the Canine Plasma Lipidome after Short- and Long-Term Excess PR:PROJECT_TITLE Glucocorticoid Exposure PR:PROJECT_SUMMARY Glucocorticoids (GCs) are widely used in veterinary and human medicine. Chromic PR:PROJECT_SUMMARY endogenous or iatrogenic GC overexposure impairs metabolic function and can PR:PROJECT_SUMMARY result in diverse side-effects, including Cushing’s syndrome. This study PR:PROJECT_SUMMARY examines the effects of experimentally induced short-term and long-term GC PR:PROJECT_SUMMARY excess (induced by prednisolone and tetracosactide, respectively) on the plasma PR:PROJECT_SUMMARY lipidome of Beale dogs. Both, long- and short-term GC resulted in significant PR:PROJECT_SUMMARY changes of the plasma lipidome. PR:INSTITUTE National University of Singapore and University of Zurich PR:DEPARTMENT Multiple PR:LABORATORY Singapore Lipidomics Incubator (SLING) PR:LAST_NAME Burla PR:FIRST_NAME Bo PR:ADDRESS 28 Medical Drive, Singapore 117456, Singapore PR:EMAIL bo.burla@nus.edu.sg PR:PHONE +6565166683 #STUDY ST:STUDY_TITLE Changes in the Canine Plasma Lipidome after Short- and Long-Term Excess ST:STUDY_TITLE Glucocorticoid Exposure ST:STUDY_SUMMARY Glucocorticoids (GCs) are widely used in veterinary and human medicine. Chromic ST:STUDY_SUMMARY endogenous or iatrogenic GC overexposure impairs metabolic function and can ST:STUDY_SUMMARY result in diverse side-effects, including Cushing’s syndrome. This study ST:STUDY_SUMMARY examines the effects of experimentally induced short-term and long-term GC ST:STUDY_SUMMARY excess (induced by prednisolone and tetracosactide, respectively) on the plasma ST:STUDY_SUMMARY lipidome of Beale dogs. Both, long- and short-term GC resulted in significant ST:STUDY_SUMMARY changes of the plasma lipidome. ST:INSTITUTE Life Sciences Institute, National University of Singapore ST:LABORATORY Singapore Lipidomics Incubator (SLING) ST:LAST_NAME Burla ST:FIRST_NAME Bo ST:ADDRESS 28 Medical Drive, Singapore 117456, Singapore ST:EMAIL bo.burla@nus.edu.sg ST:PHONE +6565166683 ST:NUM_GROUPS 2 ST:TOTAL_SUBJECTS 14 ST:NUM_MALES 9 ST:NUM_FEMALES 5 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Canis lupus familiaris SU:TAXONOMY_ID 9615 SU:GENOTYPE_STRAIN Beagle SU:AGE_OR_AGE_RANGE 8 to 83 months SU:WEIGHT_OR_WEIGHT_RANGE 12.2 to 18.9 kg SU:GENDER Male and female SU:ANIMAL_ANIMAL_SUPPLIER In-house breeding SU:ANIMAL_FEED Standard pellet/kibble maintenance diet #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS P1 Prednisolone-d0-P1 TreatmentGroup:Prednisolone | TreatmentDuration:0d | SamplingTimePoint:before Sex=F; Age_months=83; Weight_kg=14.7 SUBJECT_SAMPLE_FACTORS P1 Prednisolone-d4-P1 TreatmentGroup:Prednisolone | TreatmentDuration:4d | SamplingTimePoint:after Sex=F; Age_months=89; Weight_kg=12.6 SUBJECT_SAMPLE_FACTORS P2 Prednisolone-d0-P2 TreatmentGroup:Prednisolone | TreatmentDuration:0d | SamplingTimePoint:before Sex=F; Age_months=72; Weight_kg=14 SUBJECT_SAMPLE_FACTORS P2 Prednisolone-d4-P2 TreatmentGroup:Prednisolone | TreatmentDuration:4d | SamplingTimePoint:after Sex=F; Age_months=78; Weight_kg=11.8 SUBJECT_SAMPLE_FACTORS P3 Prednisolone-d0-P3 TreatmentGroup:Prednisolone | TreatmentDuration:0d | SamplingTimePoint:before Sex=F; Age_months=71; Weight_kg=15.4 SUBJECT_SAMPLE_FACTORS P3 Prednisolone-d4-P3 TreatmentGroup:Prednisolone | TreatmentDuration:4d | SamplingTimePoint:after Sex=F; Age_months=77; Weight_kg=12.2 SUBJECT_SAMPLE_FACTORS P4 Prednisolone-d0-P4 TreatmentGroup:Prednisolone | TreatmentDuration:0d | SamplingTimePoint:before Sex=M; Age_months=23; Weight_kg=16.9 SUBJECT_SAMPLE_FACTORS P4 Prednisolone-d4-P4 TreatmentGroup:Prednisolone | TreatmentDuration:4d | SamplingTimePoint:after Sex=M; Age_months=29; Weight_kg=14.1 SUBJECT_SAMPLE_FACTORS P5 Prednisolone-d0-P5 TreatmentGroup:Prednisolone | TreatmentDuration:0d | SamplingTimePoint:before Sex=M; Age_months=23; Weight_kg=19.8 SUBJECT_SAMPLE_FACTORS P5 Prednisolone-d4-P5 TreatmentGroup:Prednisolone | TreatmentDuration:4d | SamplingTimePoint:after Sex=M; Age_months=29; Weight_kg=16.5 SUBJECT_SAMPLE_FACTORS P6 Prednisolone-d0-P6 TreatmentGroup:Prednisolone | TreatmentDuration:0d | SamplingTimePoint:before Sex=M; Age_months=88; Weight_kg=16.3 SUBJECT_SAMPLE_FACTORS P6 Prednisolone-d4-P6 TreatmentGroup:Prednisolone | TreatmentDuration:4d | SamplingTimePoint:after Sex=M; Age_months=94; Weight_kg=11.9 SUBJECT_SAMPLE_FACTORS P7 Prednisolone-d0-P7 TreatmentGroup:Prednisolone | TreatmentDuration:0d | SamplingTimePoint:before Sex=M; Age_months=58; Weight_kg=12 SUBJECT_SAMPLE_FACTORS P7 Prednisolone-d4-P7 TreatmentGroup:Prednisolone | TreatmentDuration:4d | SamplingTimePoint:after Sex=M; Age_months=58; Weight_kg=13 SUBJECT_SAMPLE_FACTORS P8 Prednisolone-d0-P8 TreatmentGroup:Prednisolone | TreatmentDuration:0d | SamplingTimePoint:before Sex=M; Age_months=43; Weight_kg=13 SUBJECT_SAMPLE_FACTORS P8 Prednisolone-d4-P8 TreatmentGroup:Prednisolone | TreatmentDuration:4d | SamplingTimePoint:after Sex=M; Age_months=43; Weight_kg=14 SUBJECT_SAMPLE_FACTORS T1 Tetracosactide-w00-T1 TreatmentGroup:Tetracosactide | TreatmentDuration:00w | SamplingTimePoint:before Sex=M; Age_months=56; Weight_kg=14 SUBJECT_SAMPLE_FACTORS T1 Tetracosactide-w25-T1 TreatmentGroup:Tetracosactide | TreatmentDuration:25w | SamplingTimePoint:after Sex=M; Age_months=56; Weight_kg=15 SUBJECT_SAMPLE_FACTORS T2 Tetracosactide-w00-T2 TreatmentGroup:Tetracosactide | TreatmentDuration:00w | SamplingTimePoint:before Sex=F; Age_months=43; Weight_kg=15 SUBJECT_SAMPLE_FACTORS T2 Tetracosactide-w25-T2 TreatmentGroup:Tetracosactide | TreatmentDuration:25w | SamplingTimePoint:after Sex=F; Age_months=43; Weight_kg=16 SUBJECT_SAMPLE_FACTORS T3 Tetracosactide-w00-T3 TreatmentGroup:Tetracosactide | TreatmentDuration:00w | SamplingTimePoint:before Sex=F; Age_months=8; Weight_kg=16 SUBJECT_SAMPLE_FACTORS T3 Tetracosactide-w25-T3 TreatmentGroup:Tetracosactide | TreatmentDuration:25w | SamplingTimePoint:after Sex=F; Age_months=8; Weight_kg=17 SUBJECT_SAMPLE_FACTORS T4 Tetracosactide-w00-T4 TreatmentGroup:Tetracosactide | TreatmentDuration:00w | SamplingTimePoint:before Sex=M; Age_months=7; Weight_kg=17 SUBJECT_SAMPLE_FACTORS T4 Tetracosactide-w25-T4 TreatmentGroup:Tetracosactide | TreatmentDuration:25w | SamplingTimePoint:after Sex=M; Age_months=7; Weight_kg=18 SUBJECT_SAMPLE_FACTORS T5 Tetracosactide-w00-T5 TreatmentGroup:Tetracosactide | TreatmentDuration:00w | SamplingTimePoint:before Sex=M; Age_months=7; Weight_kg=18 SUBJECT_SAMPLE_FACTORS T5 Tetracosactide-w25-T5 TreatmentGroup:Tetracosactide | TreatmentDuration:25w | SamplingTimePoint:after Sex=M; Age_months=7; Weight_kg=19 SUBJECT_SAMPLE_FACTORS T6 Tetracosactide-w00-T6 TreatmentGroup:Tetracosactide | TreatmentDuration:00w | SamplingTimePoint:before Sex=M; Age_months=8; Weight_kg=19 SUBJECT_SAMPLE_FACTORS T6 Tetracosactide-w25-T6 TreatmentGroup:Tetracosactide | TreatmentDuration:25w | SamplingTimePoint:after Sex=M; Age_months=8; Weight_kg=20 #COLLECTION CO:COLLECTION_SUMMARY Blood was collected from the jugular vein after overnight fasting right before CO:COLLECTION_SUMMARY start and at the end of the treatments (12 h after the last prednisolone CO:COLLECTION_SUMMARY administration and 25 weeks after start of tetracosactide infusion, CO:COLLECTION_SUMMARY respectively). Plasma was prepared lithium heparin anticoagulated blood CO:COLLECTION_SUMMARY (Vacuette Greiner Bio-One, Germany) that centrifuged immediately after CO:COLLECTION_SUMMARY collection (1,862 g, 10 min, 4°C). Plasma samples were stored at −80°C until CO:COLLECTION_SUMMARY lipidomic analysis. CO:COLLECTION_PROTOCOL_COMMENTS Blood collected after overnight fasting CO:SAMPLE_TYPE Blood (plasma) CO:COLLECTION_METHOD Venepuncture CO:COLLECTION_LOCATION Jugular vein CO:COLLECTION_FREQUENCY Before and after treatment CO:STORAGE_CONDITIONS -80℃ CO:COLLECTION_VIALS VACUETTE® lithium heparin tubes (Greiner Bio-One, Germany) CO:COLLECTION_TUBE_TEMP Room temperature CO:ADDITIVES Lithium heparin #TREATMENT TR:TREATMENT_SUMMARY In the short-term prednisolone group, dogs were treated with 50 mg prednisolone TR:TREATMENT_SUMMARY (Streuli Pharma AG, Switzerland) orally twice daily for 3 consecutive days. For TR:TREATMENT_SUMMARY the long-term tetracosactide treatment, ALZET osmotic pumps (Durect Corporation, TR:TREATMENT_SUMMARY USA) subcutaneously delivering tetracosactide (synthetic ACTH; Bachem AG, TR:TREATMENT_SUMMARY Switzerland) were implanted into the dorsolateral neck. New pumps were implanted TR:TREATMENT_SUMMARY every 4 weeks. Total infusion time was 25 weeks, with a starting dose of TR:TREATMENT_SUMMARY 1.3–1.95 µg/kg/d tetracosactide, which was gradually increased to a final TR:TREATMENT_SUMMARY dose of 6–10 µg/kg/d. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Plasma lipids were extracted using a single-phase butanol/methanol extraction SP:SAMPLEPREP_SUMMARY method (Alshehry et al, Metabolites 2015 with modifications). After thawing on SP:SAMPLEPREP_SUMMARY ice, a 10 µL aliquot of each plasma sample was transferred into a 2 mL SP:SAMPLEPREP_SUMMARY polypropylene tube (Eppendorf, Germany). Subsequently, 1 µL BHT SP:SAMPLEPREP_SUMMARY (2,6-di-tert-butyl-4-methylphenol, 10 mmol/L in ethanol) and 90 µL of SP:SAMPLEPREP_SUMMARY 1-butanol:methanol (1:1, v/v) containing internal standards was added to each SP:SAMPLEPREP_SUMMARY sample. Following internal standards were added at 50 ng/mL final concentration SP:SAMPLEPREP_SUMMARY in the extraction solvent: ceramide d18:1/17:0 (Avanti 860517P), SP:SAMPLEPREP_SUMMARY Glucosylceramide (Matreya 1533), Lactosylceramide d18:1/16:0 D3 (Matreya 1534), SP:SAMPLEPREP_SUMMARY LPC 20:0 (Avanti 855777P), LPE 14:0 (Avanti 856735P), PI 12:0/13:0 (Avanti SP:SAMPLEPREP_SUMMARY LM-1500), PE 14:0/14:0 (Avanti 850745P), PS 14:0/14:0 (Avanti 840033P), SM SP:SAMPLEPREP_SUMMARY d18:1/12:0 (Avanti 860583P), and at 100 ng/mL: DG 12:0_12:0 (Avanti 800812P), PC SP:SAMPLEPREP_SUMMARY 14:0/14:0 (Avanti 850345P), TAG 16:0_16:0_16:0 d5 (CDN Isotopes D5815). Samples SP:SAMPLEPREP_SUMMARY were then vortexed (30 sec) and sonicated in an ultrasound water bath for 30 min SP:SAMPLEPREP_SUMMARY (20°C). After centrifugation (14,000 g, 10 min, 4°C), 90 µL of supernatant SP:SAMPLEPREP_SUMMARY were transferred into 1.5 mL polypropylene tubes (Sarstedt, Germany) and dried SP:SAMPLEPREP_SUMMARY under a nitrogen stream at 37°C. Dried extracts were stored at −80°C until SP:SAMPLEPREP_SUMMARY LC-MS analysis. Dried samples were reconstituted with 90 µL 1-butanol:methanol SP:SAMPLEPREP_SUMMARY (1:1, v/v) and sonication in an ultrasound water for 10 min. After SP:SAMPLEPREP_SUMMARY centrifugation for 10 min at 20,800 g (4°C), supernatants (80 µL) were SP:SAMPLEPREP_SUMMARY transferred into autosampler vials with glass inserts (Agilent Technologies, SP:SAMPLEPREP_SUMMARY USA). For the analysis of sphingosine-1-phosphate (S1P), lipid extracts were SP:SAMPLEPREP_SUMMARY derivatized according to the method described by Narayanaswamy et al. (Anal. SP:SAMPLEPREP_SUMMARY Chem., 2014). To 50 µL lipid extract (see above), 50 µL 13C2D2–S1P d18:1 SP:SAMPLEPREP_SUMMARY internal standard solution (Toronto Research Chemicals, Canada; 20 ng/mL in SP:SAMPLEPREP_SUMMARY 1-butanol:methanol 1:1 [v/v]) was added. Derivatization was performed by adding SP:SAMPLEPREP_SUMMARY 20 µL TMS-diazomethane (2 mol/L in hexanes; Acros Organics, Thermo Fisher SP:SAMPLEPREP_SUMMARY Scientific, USA) and subsequent incubation for 20 min at 25°C and 700 rpm SP:SAMPLEPREP_SUMMARY (Thermomixer, Eppendorf, Germany). To stop the reaction and inactivate TMS, 1 SP:SAMPLEPREP_SUMMARY µL acetic acid 100% (glacial) was added. After centrifugation for 10 min at SP:SAMPLEPREP_SUMMARY 20,800 g (7°C), supernatants were transferred into autosampler vials for LC-MS SP:SAMPLEPREP_SUMMARY analysis. Process Quality Control (PQC) samples were generated by pooling equal SP:SAMPLEPREP_SUMMARY volumes of plasma samples within an experimental group. For Blank samples, SP:SAMPLEPREP_SUMMARY plasma was omitted. PQC and Blank were processed together with the study plasma SP:SAMPLEPREP_SUMMARY samples with the same procedures. SP:EXTRACTION_METHOD Liquid–liquid extraction using butanol:methanol SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Agilent 1290 Infinity CH:COLUMN_NAME Waters Acquity BEH HILIC (100 x 2.1mm,1.7 µm, 130 Ã…) CH:FLOW_GRADIENT 99.9% B: 0 - 5 min; 40% B: 5 - 5.5 min; 10% B: 5.5 - 6.6 min and 99.9% B: 6.6 - CH:FLOW_GRADIENT 9.6 min (total run time 9.6 min) CH:FLOW_RATE 0.4 mL/min CH:COLUMN_TEMPERATURE 60 CH:SOLVENT_A Acetonitrile:ammonium formate 25 mmol/L in water pH 4.6 1:1 (v:v) CH:SOLVENT_B Acetonitrile:ammonium formate 25 mmol/L in water pH 4.6 95:5 (v:v) CH:SAMPLE_INJECTION 2 µL #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME Singapore Lipidomics Incubator AN:SOFTWARE_VERSION . MassHunter Data Acquisition B.06 AN:DATA_FORMAT .d #MS MS:INSTRUMENT_NAME Agilent 6490 QQQ MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Derivatized S1P species were measured with an Agilent 6490 triple quadrupole MS:MS_COMMENTS mass spectrometer in MRM mode. The ESI source settings were: polarity: positive, MS:MS_COMMENTS dry gas temperature 200°C, dry gas flow 12 L/min, nebulizer pressure: 25 psi, MS:MS_COMMENTS sheath gas temperature: 400°C, sheath gas flow: 12 L/min, capillary voltage: MS:MS_COMMENTS 3500 V, noozle: 500, iFunnel high pressure RF: 200, iFunnel high pressure RF: MS:MS_COMMENTS 110. MRM transitions with collision energies are detailed in the attached MS:MS_COMMENTS protocol. Data were processed with Agilent MassHunter QQQ Quantitative Analysis MS:MS_COMMENTS (version B.08). The m/z 60 fragments were used as quantifiers, the m/z 103 MS:MS_COMMENTS fragments as qualifiers. Normalised peak areas were calculated by dividing the MS:MS_COMMENTS peak areas of the S1P species with the internal standard. Relative abundance was MS:MS_COMMENTS obtained by multiplying the normalised peak areas with the molar concentration MS:MS_COMMENTS of the corresponding internal standard. S1P species with a median peak area in MS:MS_COMMENTS the PQC samples below 250 or less than 5 times of the highest signal in Blank MS:MS_COMMENTS samples were excluded. Additionally, the coefficient of variation (CV) of the MS:MS_COMMENTS normalised peak area was calculated for each lipid species in the PQC samples of MS:MS_COMMENTS each experimental group. Species with a CV higher than 25% in any of the two MS:MS_COMMENTS groups were excluded from subsequent evaluation. MS:CAPILLARY_VOLTAGE 3500 V MS:COLLISION_GAS Nitrogen MS:DRY_GAS_FLOW 12 L/min MS:DRY_GAS_TEMP 200°C MS:FRAGMENT_VOLTAGE 135 V MS:NEBULIZER 25 psi #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS µmol/L MS_METABOLITE_DATA_START Samples Tetracosactide-w00-T1 Tetracosactide-w25-T1 Tetracosactide-w00-T2 Tetracosactide-w25-T2 Tetracosactide-w00-T3 Tetracosactide-w25-T3 Tetracosactide-w00-T4 Tetracosactide-w25-T4 Tetracosactide-w00-T5 Tetracosactide-w25-T5 Tetracosactide-w00-T6 Tetracosactide-w25-T6 Prednisolone-d0-P1 Prednisolone-d4-P1 Prednisolone-d0-P2 Prednisolone-d4-P2 Prednisolone-d0-P3 Prednisolone-d4-P3 Prednisolone-d0-P4 Prednisolone-d4-P4 Prednisolone-d0-P5 Prednisolone-d4-P5 Prednisolone-d0-P6 Prednisolone-d4-P6 Prednisolone-d0-P7 Prednisolone-d4-P7 Prednisolone-d0-P8 Prednisolone-d4-P8 Factors TreatmentGroup:Tetracosactide | TreatmentDuration:00w | SamplingTimePoint:before TreatmentGroup:Tetracosactide | TreatmentDuration:25w | SamplingTimePoint:after TreatmentGroup:Tetracosactide | TreatmentDuration:00w | SamplingTimePoint:before TreatmentGroup:Tetracosactide | TreatmentDuration:25w | SamplingTimePoint:after TreatmentGroup:Tetracosactide | TreatmentDuration:00w | SamplingTimePoint:before TreatmentGroup:Tetracosactide | TreatmentDuration:25w | SamplingTimePoint:after TreatmentGroup:Tetracosactide | TreatmentDuration:00w | SamplingTimePoint:before TreatmentGroup:Tetracosactide | TreatmentDuration:25w | SamplingTimePoint:after TreatmentGroup:Tetracosactide | TreatmentDuration:00w | SamplingTimePoint:before TreatmentGroup:Tetracosactide | TreatmentDuration:25w | SamplingTimePoint:after TreatmentGroup:Tetracosactide | TreatmentDuration:00w | SamplingTimePoint:before TreatmentGroup:Tetracosactide | TreatmentDuration:25w | SamplingTimePoint:after TreatmentGroup:Prednisolone | TreatmentDuration:0d | SamplingTimePoint:before TreatmentGroup:Prednisolone | TreatmentDuration:4d | SamplingTimePoint:after TreatmentGroup:Prednisolone | TreatmentDuration:0d | SamplingTimePoint:before TreatmentGroup:Prednisolone | TreatmentDuration:4d | SamplingTimePoint:after TreatmentGroup:Prednisolone | TreatmentDuration:0d | SamplingTimePoint:before TreatmentGroup:Prednisolone | TreatmentDuration:4d | SamplingTimePoint:after TreatmentGroup:Prednisolone | TreatmentDuration:0d | SamplingTimePoint:before TreatmentGroup:Prednisolone | TreatmentDuration:4d | SamplingTimePoint:after TreatmentGroup:Prednisolone | TreatmentDuration:0d | SamplingTimePoint:before TreatmentGroup:Prednisolone | TreatmentDuration:4d | SamplingTimePoint:after TreatmentGroup:Prednisolone | TreatmentDuration:0d | SamplingTimePoint:before TreatmentGroup:Prednisolone | TreatmentDuration:4d | SamplingTimePoint:after TreatmentGroup:Prednisolone | TreatmentDuration:0d | SamplingTimePoint:before TreatmentGroup:Prednisolone | TreatmentDuration:4d | SamplingTimePoint:after TreatmentGroup:Prednisolone | TreatmentDuration:0d | SamplingTimePoint:before TreatmentGroup:Prednisolone | TreatmentDuration:4d | SamplingTimePoint:after S1P(d16:1) 0.03053038 0.021999781 0.031793158 0.011081207 0.017417576 0.013812035 0.030268205 0.008473094 0.031182421 0.011611322 0.026833769 0.016772678 0.026328643 0.026511247 0.014278956 0.023005666 0.02429985 0.027249574 0.021473677 0.037118307 0.023880851 0.035752253 0.022927944 0.025359393 0.027288279 0.040338016 0.025877229 0.029729852 S1P(d17:1) 0.032917529 0.043864593 0.044004943 0.020463986 0.033636408 0.019901624 0.040730924 0.012608884 0.04927733 0.02358778 0.034830213 0.025937343 0.029466973 0.032528449 0.017492635 0.020271122 0.032327213 0.032375429 0.018735485 0.033145711 0.027407969 0.042930161 0.030299921 0.026241459 0.022787217 0.045512132 0.026453725 0.034755545 S1P(d18:0) 0.060280001 0.076184233 0.12808202 0.050735359 0.06070656 0.07422768 0.154258797 0.040292774 0.105052895 0.056825581 0.058037796 0.24469347 0.09157628 0.122281139 0.037176413 0.085335534 0.06312022 0.107938479 0.060442384 0.104968209 0.080066647 0.157541334 0.083728507 0.083722749 0.087042469 0.263447933 0.057379942 0.106388175 S1P(d18:1) 0.68042723 0.887040705 1.200982994 0.538867716 0.682747521 0.680007382 1.257255366 0.431695959 1.286934951 0.823976137 0.725764433 1.242879121 0.837128659 1.045439635 0.356389605 0.632389623 0.669893896 1.097279421 0.548024422 1.04427325 0.777864253 1.794947476 0.949059934 0.787309094 0.743530431 2.074911176 0.58139615 1.275602045 S1P(d18:2) 0.228968878 0.290066951 0.345980891 0.184849035 0.169944375 0.171353754 0.274958298 0.097666077 0.325366945 0.134651693 0.195314798 0.204004006 0.149066039 0.263268943 0.098619749 0.156369158 0.159597056 0.249954211 0.140934217 0.236705604 0.201390863 0.376330221 0.21230929 0.167478166 0.211531147 0.448761681 0.172855797 0.27186832 S1P(d19:1) 0.01122678 0.02103424 0.02344332 0.010625531 0.013302484 0.013744799 0.022212402 0.010296296 0.023819273 0.015502628 0.011556698 0.022586931 0.019940892 0.01910906 0.008938042 0.01561675 0.015540026 0.01979008 0.014508745 0.018324872 0.01669718 0.026583752 0.018457314 0.012422427 0.012704461 0.02939702 0.014486785 0.020310213 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name S1P(d16:1) S1P(d17:1) S1P(d18:0) S1P(d18:1) S1P(d18:2) S1P(d19:1) METABOLITES_END #END