#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 Å)
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