#METABOLOMICS WORKBENCH sharmarishi2004_20210220_172201 DATATRACK_ID:2503 STUDY_ID:ST001708 ANALYSIS_ID:AN002782 PROJECT_ID:PR001093
VERSION             	1
CREATED_ON             	February 22, 2021, 3:23 pm
#PROJECT
PR:PROJECT_TITLE                 	Oxylipin biosynthesis reinforces cellular senescence through a RAS/p53 feedback
PR:PROJECT_TITLE                 	loop and allows detection of senolysis
PR:PROJECT_SUMMARY               	Cellular senescence is a stress or damage response that causes a permanent
PR:PROJECT_SUMMARY               	proliferative arrest and secretion of numerous factors with potent biological
PR:PROJECT_SUMMARY               	activities. This senescence-associated secretory phenotype (SASP) has been
PR:PROJECT_SUMMARY               	characterized largely for secreted proteins that participate in embryogenesis,
PR:PROJECT_SUMMARY               	wound healing, inflammation and many age-related pathologies. By contrast, lipid
PR:PROJECT_SUMMARY               	components of the SASP are understudied. We show that senescent cells activate
PR:PROJECT_SUMMARY               	the biosynthesis of several oxylipins that promote segments of the SASP and
PR:PROJECT_SUMMARY               	reinforce the proliferative arrest. Notably, senescent cells synthesize and
PR:PROJECT_SUMMARY               	accumulate an unstudied intracellular prostaglandin,
PR:PROJECT_SUMMARY               	1a,1b-dihomo-15-deoxy-delta-12,14-prostaglandin J2. Released
PR:PROJECT_SUMMARY               	15-deoxy-delta-12,14-prostaglandin J2 is a biomarker of senolysis in culture and
PR:PROJECT_SUMMARY               	in vivo. This and other prostaglandin D2-related lipids promote the senescence
PR:PROJECT_SUMMARY               	arrest and SASP by activating RAS signaling. These data identify an important
PR:PROJECT_SUMMARY               	aspect of cellular senescence and a method to detect senolysis.
PR:INSTITUTE                     	Buck Institute for Research on Aging
PR:LAST_NAME                     	Sharma
PR:FIRST_NAME                    	Rishi
PR:ADDRESS                       	8001 Redwood Blvd, Novato, CA, 94945, USA
PR:EMAIL                         	sharmarishi2004@yahoo.co.in
PR:PHONE                         	5084392367
#STUDY
ST:STUDY_TITLE                   	Oxylipin biosynthesis reinforces cellular senescence through a RAS/p53 feedback
ST:STUDY_TITLE                   	loop and allows detection of senolysis
ST:STUDY_SUMMARY                 	Cellular senescence is a stress or damage response that causes a permanent
ST:STUDY_SUMMARY                 	proliferative arrest and secretion of numerous factors with potent biological
ST:STUDY_SUMMARY                 	activities. This senescence-associated secretory phenotype (SASP) has been
ST:STUDY_SUMMARY                 	characterized largely for secreted proteins that participate in embryogenesis,
ST:STUDY_SUMMARY                 	wound healing, inflammation and many age-related pathologies. By contrast, lipid
ST:STUDY_SUMMARY                 	components of the SASP are understudied. We show that senescent cells activate
ST:STUDY_SUMMARY                 	the biosynthesis of several oxylipins that promote segments of the SASP and
ST:STUDY_SUMMARY                 	reinforce the proliferative arrest. Notably, senescent cells synthesize and
ST:STUDY_SUMMARY                 	accumulate an unstudied intracellular prostaglandin,
ST:STUDY_SUMMARY                 	1a,1b-dihomo-15-deoxy-delta-12,14-prostaglandin J2. Released
ST:STUDY_SUMMARY                 	15-deoxy-delta-12,14-prostaglandin J2 is a biomarker of senolysis in culture and
ST:STUDY_SUMMARY                 	in vivo. This and other prostaglandin D2-related lipids promote the senescence
ST:STUDY_SUMMARY                 	arrest and SASP by activating RAS signaling. These data identify an important
ST:STUDY_SUMMARY                 	aspect of cellular senescence and a method to detect senolysis
ST:INSTITUTE                     	Buck Institute for Research on Aging
ST:LAST_NAME                     	Sharma
ST:FIRST_NAME                    	Rishi
ST:ADDRESS                       	8001 Redwood Blvd, Novato, CA, 94945, USA
ST:EMAIL                         	sharmarishi2004@yahoo.co.in
ST:PHONE                         	5084392367
#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]Raw file names and additional sample data
SUBJECT_SAMPLE_FACTORS           	-	A-121	Factor:Mouse plasma	RAW_FILE_NAME=A-121.d
SUBJECT_SAMPLE_FACTORS           	-	Ctl_61120	Factor:SEN Cells_Ctls	RAW_FILE_NAME=Ctl_61120.d
SUBJECT_SAMPLE_FACTORS           	-	IR_15_60120	Factor:SENCells_IR	RAW_FILE_NAME=IR_15_60120.d
SUBJECT_SAMPLE_FACTORS           	-	IR_DOXOABT_1	Factor:DOXOABT	RAW_FILE_NAME=IR_DOXOABT_1.d
#COLLECTION
CO:COLLECTION_SUMMARY            	Cell culture: Human fetal lung fibroblasts (IMR-90) and HEPG2 cells were
CO:COLLECTION_SUMMARY            	cultured in Dulbecco’s modified eagle medium (DMEM - Gibco) supplemented with
CO:COLLECTION_SUMMARY            	10% FBS and penicillin/streptomycin (Gibco). HUVEC cells were obtained from the
CO:COLLECTION_SUMMARY            	ATCC and cultured using the ATCC protocol and culture media. Quiescence was
CO:COLLECTION_SUMMARY            	induced by replacing culture media with media containing 0.2% FBS for 72 h
CO:COLLECTION_SUMMARY            	before analysis. All cells were cultured at 3% O2, and used prior to 40
CO:COLLECTION_SUMMARY            	population doublings, other than replicative senescent cells, which were
CO:COLLECTION_SUMMARY            	cultured until replicative exhaustion. All cells were mycoplasma free. Mouse
CO:COLLECTION_SUMMARY            	Plasma: Animal experiments were conducted using a protocol approved by the
CO:COLLECTION_SUMMARY            	Institutional Animal Care and Use Committee of the Buck Institute. For DOXO
CO:COLLECTION_SUMMARY            	treatments, 10-16 wk old p16-3MR mice received one intraperitoneal (i.p.)
CO:COLLECTION_SUMMARY            	injection of 10 mg/kg of doxorubicin hydrochloride in PBS, and treated 5 d later
CO:COLLECTION_SUMMARY            	with GCV or vehicle. GCV was administered via daily i.p. injections for 5
CO:COLLECTION_SUMMARY            	consecutive days at 25 mg/kg in PBS. Control mice were injected with an equal
CO:COLLECTION_SUMMARY            	volume of PBS. Mice were euthanized and tissues collected 10 d after DOXO
CO:COLLECTION_SUMMARY            	challenge. For aging studies, C57BL/6 mice were aged for 6 or 24 mo, at which
CO:COLLECTION_SUMMARY            	point mice were euthanized and tissues collected for analysis. For the biomarker
CO:COLLECTION_SUMMARY            	studies, mice with challenged with DOXO as above, and given either vehicle or
CO:COLLECTION_SUMMARY            	ABT-263 6 weeks after DOXO challenge. Whole blood was collected by cardiac
CO:COLLECTION_SUMMARY            	puncture into EDTA-tubes (lavender caps – BD Biosciences) and spun at 2000g
CO:COLLECTION_SUMMARY            	for 5 minutes. For urine collection, mice were placed in plastic containers
CO:COLLECTION_SUMMARY            	prior to euthanasia.
CO:SAMPLE_TYPE                   	Blood (plasma)
#TREATMENT
TR:TREATMENT_SUMMARY             	Cell culture: Human fetal lung fibroblasts (IMR-90) and HEPG2 cells were
TR:TREATMENT_SUMMARY             	cultured in Dulbecco’s modified eagle medium (DMEM - Gibco) supplemented with
TR:TREATMENT_SUMMARY             	10% FBS and penicillin/streptomycin (Gibco). HUVEC cells were obtained from the
TR:TREATMENT_SUMMARY             	ATCC and cultured using the ATCC protocol and culture media. Quiescence was
TR:TREATMENT_SUMMARY             	induced by replacing culture media with media containing 0.2% FBS for 72 h
TR:TREATMENT_SUMMARY             	before analysis. All cells were cultured at 3% O2, and used prior to 40
TR:TREATMENT_SUMMARY             	population doublings, other than replicative senescent cells, which were
TR:TREATMENT_SUMMARY             	cultured until replicative exhaustion. All cells were mycoplasma free. Mouse
TR:TREATMENT_SUMMARY             	Plasma: Animal experiments were conducted using a protocol approved by the
TR:TREATMENT_SUMMARY             	Institutional Animal Care and Use Committee of the Buck Institute. For DOXO
TR:TREATMENT_SUMMARY             	treatments, 10-16 wk old p16-3MR mice received one intraperitoneal (i.p.)
TR:TREATMENT_SUMMARY             	injection of 10 mg/kg of doxorubicin hydrochloride in PBS, and treated 5 d later
TR:TREATMENT_SUMMARY             	with GCV or vehicle. GCV was administered via daily i.p. injections for 5
TR:TREATMENT_SUMMARY             	consecutive days at 25 mg/kg in PBS. Control mice were injected with an equal
TR:TREATMENT_SUMMARY             	volume of PBS. Mice were euthanized and tissues collected 10 d after DOXO
TR:TREATMENT_SUMMARY             	challenge. For aging studies, C57BL/6 mice were aged for 6 or 24 mo, at which
TR:TREATMENT_SUMMARY             	point mice were euthanized and tissues collected for analysis. For the biomarker
TR:TREATMENT_SUMMARY             	studies, mice with challenged with DOXO as above, and given either vehicle or
TR:TREATMENT_SUMMARY             	ABT-263 6 weeks after DOXO challenge. Whole blood was collected by cardiac
TR:TREATMENT_SUMMARY             	puncture into EDTA-tubes (lavender caps – BD Biosciences) and spun at 2000g
TR:TREATMENT_SUMMARY             	for 5 minutes. For urine collection, mice were placed in plastic containers
TR:TREATMENT_SUMMARY             	prior to euthanasia.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Protocol for Lipid Extraction (10.24.2019) Lipid Biomarker for Senolysis Study
SP:SAMPLEPREP_SUMMARY            	1. 80% MeOH was added to each sample 500µL to plasma samples 100µL to urine
SP:SAMPLEPREP_SUMMARY            	sample 2. Transfer the contents to 9 mL glass tube and add 1.0 mL of CHCl3
SP:SAMPLEPREP_SUMMARY            	(containing 200 ng/mL FA 17:0 Saturated). Vortex mix for 5 minutes. 3. Place the
SP:SAMPLEPREP_SUMMARY            	glass tube into a 15mL polypropylene culture tube with Kimi wipes inserted at
SP:SAMPLEPREP_SUMMARY            	bottom as a cushion. Wrap the culture tube with paraffin and place in
SP:SAMPLEPREP_SUMMARY            	centrifuge. This will prevent glass tube being shattered in the centrifuge. 4.
SP:SAMPLEPREP_SUMMARY            	Centrifuge at 4°C for 10 minutes 3000rpm. 5. After spinning there will be two
SP:SAMPLEPREP_SUMMARY            	distinct layers, the top layer is the aqueous layer containing the aqueous
SP:SAMPLEPREP_SUMMARY            	metabolites (80% MeOH) and bottom layer which contains lipids. 6. Transfer the
SP:SAMPLEPREP_SUMMARY            	lower layer into 1.5ml vials. 7. Transfer from the 1.5mL tube the exact amount
SP:SAMPLEPREP_SUMMARY            	of each sample into the MS/HPLC glass vial and dry down using nitrogen gas. 8.
SP:SAMPLEPREP_SUMMARY            	Reconstitute the lipids with 80µL chloroform. 9. Transfer upper layer
SP:SAMPLEPREP_SUMMARY            	(metabolites) into labeled vials and store at -80°C, if possible. Samples
SP:SAMPLEPREP_SUMMARY            	stored in -80°C. Sample volume submitted -Plasma samples=50µL Urine samples=
SP:SAMPLEPREP_SUMMARY            	10µL Sample volume in each vial- Plasma samples µL and Urine samples µL
SP:SAMPLEPREP_SUMMARY            	-submitted to mass spec All samples were split in half ½ given to CW for
SP:SAMPLEPREP_SUMMARY            	further testing other 1/2 in -80°C
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Agilent 6520
CH:COLUMN_NAME                   	Phenomenex Kinetex C18 (150 x 2.1mm, 2.6 um)
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Agilent 6520 QTOF
MS:INSTRUMENT_TYPE               	QToF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	Full scan mode was used. Data processed using profinder software
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Peak Area
MS_METABOLITE_DATA_START
Samples	A-121	Ctl_61120	IR_15_60120	IR_DOXOABT_1
Factors	Factor:Mouse plasma	Factor:SEN Cells_Ctls	Factor:SENCells_IR	Factor:DOXOABT
Acetaminophen		611775		
Cer 18:0;3/18:0;1		611775		
DAG 12:0-16:1		94764	191511	
DAG 14:0-16:1			10626	61422
DAG 16:0-18:1			90200	
DAG 18:1-18:1	13960			
Ergosterol		41928		
Gemfibrozil	65073	128741	154892	192382
IPC 18:0;2/18:0;0	8317			18640
IPC 18:0;3/16:0;2			43337	
IPC 18:0;3/22:0;1		46737		
LPA 14:0	38625	57629	67293	91643
LPE 18:0	124148			
LPE 18:1		17297	15216	
LPI 16:0	155580			
LPI 18:0		20462		
LPS 18:1	48179			
Norethynodrel	6425	101459	135195	152762
OOB-PC	81087			
PA 12:0-16:0	15058	9844		
PC 16:0-16:1		113023		
PC 16:0-18:0	140371			
PC(10:0/24:0)	140371			
PC(11:0/23:0)	140371			
PC(12:0/17:2(9Z,12Z))				33376
PC(12:0/19:1(9Z))		172486		
PC(12:0/20:1(11Z))		113023		
PC(12:0/22:0)	140371			
PC(12:0/22:1(11Z))			107070	
PC(13:0/18:1(9Z))		172486		
PC(13:0/18:2(9Z,12Z))		267552	81400	
PC(13:0/19:1(9Z))		113023		
PC(13:0/21:0)	140371			
PC(13:0/22:4(7Z,10Z,13Z,16Z))		374927		
PC(14:0/17:1(9Z))			184966	
PC(14:0/17:2(9Z,12Z))		267552	81400	
PC(14:0/18:1(11Z))		113023		
PC(14:0/18:1(9Z))		113023		
PC(14:0/20:0)	140371			
PC(14:0/20:1(11Z))			107070	
PC(14:0/22:6(4Z,7Z,10Z,13Z,16Z,19Z))			53181	
PC(14:1(9Z)/15:1(9Z))				33376
PC(14:1(9Z)/17:0)		172486		
PC(14:1(9Z)/17:1(9Z))		267552	81400	
PC(14:1(9Z)/18:0)		113023		
PC(14:1(9Z)/20:0)			107070	
PC(14:1(9Z)/22:5(4Z,7Z,10Z,13Z,16Z))			53181	
PC(14:1(9Z)/22:5(7Z,10Z,13Z,16Z,19Z))			53181	
PC(15:0/0:0)		28114	14977	
PC(15:0/16:1(9Z))			184966	
PC(15:0/17:1(9Z))		113023		
PC(15:0/19:0)	140371			
PC(15:0/19:1(9Z))			107070	
PC(15:0/20:4(5Z,8Z,11Z,14Z))			2049489	
PC(15:1(9Z)/0:0)		17297	15216	
PC(15:1(9Z)/14:1(9Z))				33376
PC(15:1(9Z)/16:0)		172486		
PC(15:1(9Z)/16:1(9Z))		267552	81400	
PC(15:1(9Z)/19:0)			107070	
PC(15:1(9Z)/20:3(8Z,11Z,14Z))			2049489	
PC(15:1(9Z)/22:6(4Z,7Z,10Z,13Z,16Z,19Z))				32085
PC(16:0/15:1(14))			184966	
PC(16:0/15:1(9Z))		172486		
PC(16:0/16:1(9Z))		113023		
PC(16:0/18:0)	140371			
PC(16:0/18:1(11E))			107070	
PC(16:0/18:1(11Z))			107070	
PC(16:0/18:1(6E))			107070	
PC(16:0/18:1(6Z))			107070	
PC(16:0/18:1(9E))			107070	
PC(16:0/18:1(9Z))			107070	
PC(16:1(9Z)/15:0)		172486		
PC(16:1(9Z)/15:1(9Z))		267552	81400	
PC(16:1(9Z)/16:0)		113023		
PC(16:1(9Z)/18:0)			107070	
PC(16:1(9Z)/20:5(5Z,8Z,11Z,14Z,17Z))			53181	
PC(17:0/0:0)	89286			
PC(17:0/14:1(9Z))		172486		
PC(17:0/15:1(9Z))		113023		
PC(17:0/17:0)	140371			
PC(17:0/17:1(9Z))			107070	
PC(17:0/18:4(6Z,9Z,12Z,15Z))			2049489	
PC(17:1(9Z)/14:0)			184966	
PC(17:1(9Z)/14:1(9Z))		267552	81400	
PC(17:1(9Z)/15:0)		113023		
PC(17:1(9Z)/17:0)			107070	
PC(17:1(9Z)/18:3(6Z,9Z,12Z))		374927		
PC(17:1(9Z)/18:3(9Z,12Z,15Z))		374927		
PC(17:2(9Z,12Z)/0:0)	100924			
PC(17:2(9Z,12Z)/12:0)				33376
PC(17:2(9Z,12Z)/14:0)		267552	81400	
PC(17:2(9Z,12Z)/18:2(9Z,12Z))			2049489	
PC(17:2(9Z,12Z)/20:5(5Z,8Z,11Z,14Z,17Z))				32085
PC(18:0/14:1(9Z))		113023		
PC(18:0/16:0)	140371			
PC(18:0/16:1(9Z))			107070	
PC(18:1(11Z)/14:0)		113023		
PC(18:1(11Z)/16:0)			107070	
PC(18:1(9Z)/13:0)		172486		
PC(18:1(9Z)/14:0)		113023		
PC(18:1(9Z)/16:0)			107070	
PC(18:2(9Z,12E)/17:2(9Z,11E))			2049489	
PC(18:2(9Z,12Z)/13:0)		267552	81400	
PC(18:2(9Z,12Z)/17:2(9Z,12Z))			2049489	
PC(18:2(9Z,12Z)/18:4(6Z,9Z,12Z,15Z))			53181	
PC(18:3(6Z,9Z,12Z)/17:1(9Z))			2049489	
PC(18:3(6Z,9Z,12Z)/18:3(6Z,9Z,12Z))			53181	
PC(18:3(6Z,9Z,12Z)/18:3(9Z,12Z,15Z))			53181	
PC(18:3(9E,11E,13E)/18:3(9E,11E,13E))			53181	
PC(18:3(9Z,12Z,15Z)/17:1(9Z))			2049489	
PC(18:3(9Z,12Z,15Z)/18:3(6Z,9Z,12Z))			53181	
PC(18:3(9Z,12Z,15Z)/18:3(9Z,12Z,15Z))			53181	
PC(18:4(6Z,9Z,12Z,15Z)/17:0)			2049489	
PC(18:4(6Z,9Z,12Z,15Z)/18:2(9Z,12Z))			53181	
PC(18:4(9E,11E,13E,15E)/18:2(9Z,12Z))			53181	
PC(19:0/15:0)	140371			
PC(19:0/15:1(9Z))			107070	
PC(19:1(9Z)/12:0)		172486		
PC(19:1(9Z)/13:0)		113023		
PC(19:1(9Z)/15:0)			107070	
PC(20:0/14:0)	140371			
PC(20:0/14:1(9Z))			107070	
PC(20:1(11Z)/12:0)		113023		
PC(20:1(11Z)/14:0)			107070	
PC(20:3(8Z,11Z,14Z)/15:1(9Z))			2049489	
PC(20:4(5Z,8Z,11Z,14Z)/15:0)		374927		
PC(20:5(5Z,8Z,11Z,14Z,17Z)/16:1(9Z))			53181	
PC(20:5(5Z,8Z,11Z,14Z,17Z)/17:2(9Z,12Z))				32085
PC(21:0/13:0)	140371			
PC(22:0/12:0)	140371			
PC(22:1(11Z)/12:0)			107070	
PC(22:4(7Z,10Z,13Z,16Z)/13:0)			2049489	
PC(22:5(4Z,7Z,10Z,13Z,16Z)/14:1(9Z))			53181	
PC(22:5(7Z,10Z,13Z,16Z,19Z)/14:1(9Z))			53181	
PC(22:6(4Z,7Z,10Z,13Z,16Z,19Z)/14:0)			53181	
PC(22:6(4Z,7Z,10Z,13Z,16Z,19Z)/15:1(9Z))				32085
PC(24:1(15Z)/0:0)				15667
PC(36:0/16:0)				6911
PC(O-1:0/16:0)	89286			
PC(O-14:0/18:1(9Z))				10957
PC(O-14:0/22:0)	100655			70051
PC(O-15:0/2:0)	89286			
PC(O-16:0/1:0)	89286			
PC(O-16:0/16:1(9Z))				10957
PC(O-16:0/17:2(9Z,12Z))			23115	
PC(O-16:0/20:0)	100655			70051
PC(O-18:0/14:1(9Z))				10957
PC(O-20:0/16:0)	100655			70051
PC(O-20:0/22:6(4Z,7Z,10Z,13Z,16Z,19Z))		1350914		
PC(O-22:0/22:3(10Z,13Z,16Z))	14337			
PC(O-22:1(13Z)/22:2(13Z,16Z))	14337			
PC(P-15:0/0:0)		57898	63112	
PC(P-16:0/16:0)				10957
PC(P-16:0/17:1(9Z))			23115	
PC(P-16:0/20:5(5Z,8Z,11Z,14Z,17Z))		509910		
PC(P-17:0/0:0)		37673		
PC(P-18:0/14:0)				10957
PC(P-18:0/15:1(9Z))			23115	
PC(P-20:0/0:0)				310050
PC(P-20:0/12:0)				10957
PC(P-20:0/19:1(9Z))	3749		26277	67393
PE 16:0-18:1		172486		
PE 16:1-16:1				33376
PE 16:1-18:1		267552	81400	
PE 18:0-16:1			184966	
PECPC	25693			
PG 16:0-18:1	6305			
PG 18:1-18:1			53213	
PGJ2_Class_1	71597			
PGJ2_Class		42637	40417	41091
PHHdiA-PC	30840			
PHODiA-PC	81602			
PI 12:0-18:0	37722			
PI 14:0-16:0	37722			
PI 14:0-18:1		49225		
PI 16:0-16:1		49225		
P-IsoPGA2-PC			479985	
POB-PC	1517760			
Procainamide	118550	394882	456817	709023
Progesterone	348339	802691	979669	1258979
PS 18:0-18:1			278533	
Quinidine	24157	36410	24277	31701
Quinine	24157	36410	24277	31701
TAG 14:0-16:0-16:0		7329		
TAG 14:0-16:1-22:0	138085			
TAG 14:0-18:1-20:0	138085			
TAG 16:0-16:0-18:0			13687	
TAG 16:0-16:1-26:0	9990			
TAG 16:0-18:0-18:1	138085			
TAG 16:0-18:1-24:0	9990			
TAG 16:1-18:0-18:0	138085			
TAG 18:0-18:1-22:0	9990			
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	[ret] A-121	[ret] Ctl_61120	[ret] IR_15_60120	[ret] IR_DOXOABT_1	Mass (avg)	RSD (Mass, ppm)
Acetaminophen		28.9			599.5445
Cer 18:0;3/18:0;1		28.9			599.5445
DAG 12:0-16:1		26.274	26.211		510.4251	1.51
DAG 14:0-16:1			30.807	30.765	538.4513	10.02
DAG 16:0-18:1			31.313		594.5274
DAG 18:1-18:1	29.661				620.5387
Ergosterol		27.133			396.3371
Gemfibrozil	15.267	15.314	15.252	15.31	250.1568	1.26
IPC 18:0;2/18:0;0	22.994			22.987	809.582	6.71
IPC 18:0;3/16:0;2			31.464		829.5313
IPC 18:0;3/22:0;1		27.739			897.6362
LPA 14:0	18.499	18.597	18.585	18.442	382.2151	2.04
LPE 18:0	16.126				481.3153
LPE 18:1		16.173	16.161		479.2968	3.35
LPI 16:0	16.126				572.2957
LPI 18:0		32.284			600.3231
LPS 18:1	29.712				523.2907
Norethynodrel	18.398	18.496	18.332	18.341	298.194	0.93
OOB-PC	16.176				605.3643
PA 12:0-16:0	22.742	22.89			592.4121	4.07
PC 16:0-16:1		30.921			731.5454
PC 16:0-18:0	28.096				761.5908
PC(10:0/24:0)	28.096				761.5908
PC(11:0/23:0)	28.096				761.5908
PC(12:0/17:2(9Z,12Z))				30.31	687.4837
PC(12:0/19:1(9Z))		13.243			717.532
PC(12:0/20:1(11Z))		30.921			731.5454
PC(12:0/22:0)	28.096				761.5908
PC(12:0/22:1(11Z))			30.959		759.5805
PC(13:0/18:1(9Z))		13.243			717.532
PC(13:0/18:2(9Z,12Z))		32.032	32.121		715.5123	0.42
PC(13:0/19:1(9Z))		30.921			731.5454
PC(13:0/21:0)	28.096				761.5908
PC(13:0/22:4(7Z,10Z,13Z,16Z))		12.94			767.5468
PC(14:0/17:1(9Z))			32.777		717.5322
PC(14:0/17:2(9Z,12Z))		32.032	32.121		715.5123	0.42
PC(14:0/18:1(11Z))		30.921			731.5454
PC(14:0/18:1(9Z))		30.921			731.5454
PC(14:0/20:0)	28.096				761.5908
PC(14:0/20:1(11Z))			30.959		759.5805
PC(14:0/22:6(4Z,7Z,10Z,13Z,16Z,19Z))			24.141		777.5325
PC(14:1(9Z)/15:1(9Z))				30.31	687.4837
PC(14:1(9Z)/17:0)		13.243			717.532
PC(14:1(9Z)/17:1(9Z))		32.032	32.121		715.5123	0.42
PC(14:1(9Z)/18:0)		30.921			731.5454
PC(14:1(9Z)/20:0)			30.959		759.5805
PC(14:1(9Z)/22:5(4Z,7Z,10Z,13Z,16Z))			24.141		777.5325
PC(14:1(9Z)/22:5(7Z,10Z,13Z,16Z,19Z))			24.141		777.5325
PC(15:0/0:0)		17.486	17.474		481.3148	0.54
PC(15:0/16:1(9Z))			32.777		717.5322
PC(15:0/17:1(9Z))		30.921			731.5454
PC(15:0/19:0)	28.096				761.5908
PC(15:0/19:1(9Z))			30.959		759.5805
PC(15:0/20:4(5Z,8Z,11Z,14Z))			28.282		767.5453
PC(15:1(9Z)/0:0)		16.173	16.161		479.2968	3.35
PC(15:1(9Z)/14:1(9Z))				30.31	687.4837
PC(15:1(9Z)/16:0)		13.243			717.532
PC(15:1(9Z)/16:1(9Z))		32.032	32.121		715.5123	0.42
PC(15:1(9Z)/19:0)			30.959		759.5805
PC(15:1(9Z)/20:3(8Z,11Z,14Z))			28.282		767.5453
PC(15:1(9Z)/22:6(4Z,7Z,10Z,13Z,16Z,19Z))				25.866	789.5309
PC(16:0/15:1(14))			32.777		717.5322
PC(16:0/15:1(9Z))		13.243			717.532
PC(16:0/16:1(9Z))		30.921			731.5454
PC(16:0/18:0)	28.096				761.5908
PC(16:0/18:1(11E))			30.959		759.5805
PC(16:0/18:1(11Z))			30.959		759.5805
PC(16:0/18:1(6E))			30.959		759.5805
PC(16:0/18:1(6Z))			30.959		759.5805
PC(16:0/18:1(9E))			30.959		759.5805
PC(16:0/18:1(9Z))			30.959		759.5805
PC(16:1(9Z)/15:0)		13.243			717.532
PC(16:1(9Z)/15:1(9Z))		32.032	32.121		715.5123	0.42
PC(16:1(9Z)/16:0)		30.921			731.5454
PC(16:1(9Z)/18:0)			30.959		759.5805
PC(16:1(9Z)/20:5(5Z,8Z,11Z,14Z,17Z))			24.141		777.5325
PC(17:0/0:0)	17.641				509.3473
PC(17:0/14:1(9Z))		13.243			717.532
PC(17:0/15:1(9Z))		30.921			731.5454
PC(17:0/17:0)	28.096				761.5908
PC(17:0/17:1(9Z))			30.959		759.5805
PC(17:0/18:4(6Z,9Z,12Z,15Z))			28.282		767.5453
PC(17:1(9Z)/14:0)			32.777		717.5322
PC(17:1(9Z)/14:1(9Z))		32.032	32.121		715.5123	0.42
PC(17:1(9Z)/15:0)		30.921			731.5454
PC(17:1(9Z)/17:0)			30.959		759.5805
PC(17:1(9Z)/18:3(6Z,9Z,12Z))		12.94			767.5468
PC(17:1(9Z)/18:3(9Z,12Z,15Z))		12.94			767.5468
PC(17:2(9Z,12Z)/0:0)	15.671				505.3158
PC(17:2(9Z,12Z)/12:0)				30.31	687.4837
PC(17:2(9Z,12Z)/14:0)		32.032	32.121		715.5123	0.42
PC(17:2(9Z,12Z)/18:2(9Z,12Z))			28.282		767.5453
PC(17:2(9Z,12Z)/20:5(5Z,8Z,11Z,14Z,17Z))				25.866	789.5309
PC(18:0/14:1(9Z))		30.921			731.5454
PC(18:0/16:0)	28.096				761.5908
PC(18:0/16:1(9Z))			30.959		759.5805
PC(18:1(11Z)/14:0)		30.921			731.5454
PC(18:1(11Z)/16:0)			30.959		759.5805
PC(18:1(9Z)/13:0)		13.243			717.532
PC(18:1(9Z)/14:0)		30.921			731.5454
PC(18:1(9Z)/16:0)			30.959		759.5805
PC(18:2(9Z,12E)/17:2(9Z,11E))			28.282		767.5453
PC(18:2(9Z,12Z)/13:0)		32.032	32.121		715.5123	0.42
PC(18:2(9Z,12Z)/17:2(9Z,12Z))			28.282		767.5453
PC(18:2(9Z,12Z)/18:4(6Z,9Z,12Z,15Z))			24.141		777.5325
PC(18:3(6Z,9Z,12Z)/17:1(9Z))			28.282		767.5453
PC(18:3(6Z,9Z,12Z)/18:3(6Z,9Z,12Z))			24.141		777.5325
PC(18:3(6Z,9Z,12Z)/18:3(9Z,12Z,15Z))			24.141		777.5325
PC(18:3(9E,11E,13E)/18:3(9E,11E,13E))			24.141		777.5325
PC(18:3(9Z,12Z,15Z)/17:1(9Z))			28.282		767.5453
PC(18:3(9Z,12Z,15Z)/18:3(6Z,9Z,12Z))			24.141		777.5325
PC(18:3(9Z,12Z,15Z)/18:3(9Z,12Z,15Z))			24.141		777.5325
PC(18:4(6Z,9Z,12Z,15Z)/17:0)			28.282		767.5453
PC(18:4(6Z,9Z,12Z,15Z)/18:2(9Z,12Z))			24.141		777.5325
PC(18:4(9E,11E,13E,15E)/18:2(9Z,12Z))			24.141		777.5325
PC(19:0/15:0)	28.096				761.5908
PC(19:0/15:1(9Z))			30.959		759.5805
PC(19:1(9Z)/12:0)		13.243			717.532
PC(19:1(9Z)/13:0)		30.921			731.5454
PC(19:1(9Z)/15:0)			30.959		759.5805
PC(20:0/14:0)	28.096				761.5908
PC(20:0/14:1(9Z))			30.959		759.5805
PC(20:1(11Z)/12:0)		30.921			731.5454
PC(20:1(11Z)/14:0)			30.959		759.5805
PC(20:3(8Z,11Z,14Z)/15:1(9Z))			28.282		767.5453
PC(20:4(5Z,8Z,11Z,14Z)/15:0)		12.94			767.5468
PC(20:5(5Z,8Z,11Z,14Z,17Z)/16:1(9Z))			24.141		777.5325
PC(20:5(5Z,8Z,11Z,14Z,17Z)/17:2(9Z,12Z))				25.866	789.5309
PC(21:0/13:0)	28.096				761.5908
PC(22:0/12:0)	28.096				761.5908
PC(22:1(11Z)/12:0)			30.959		759.5805
PC(22:4(7Z,10Z,13Z,16Z)/13:0)			28.282		767.5453
PC(22:5(4Z,7Z,10Z,13Z,16Z)/14:1(9Z))			24.141		777.5325
PC(22:5(7Z,10Z,13Z,16Z,19Z)/14:1(9Z))			24.141		777.5325
PC(22:6(4Z,7Z,10Z,13Z,16Z,19Z)/14:0)			24.141		777.5325
PC(22:6(4Z,7Z,10Z,13Z,16Z,19Z)/15:1(9Z))				25.866	789.5309
PC(24:1(15Z)/0:0)				28.341	605.4407
PC(36:0/16:0)				28.745	1013.8766
PC(O-1:0/16:0)	17.641				509.3473
PC(O-14:0/18:1(9Z))				27.28	717.5661
PC(O-14:0/22:0)	12.843			12.835	775.6427	1.52
PC(O-15:0/2:0)	17.641				509.3473
PC(O-16:0/1:0)	17.641				509.3473
PC(O-16:0/16:1(9Z))				27.28	717.5661
PC(O-16:0/17:2(9Z,12Z))			29.949		729.5628
PC(O-16:0/20:0)	12.843			12.835	775.6427	1.52
PC(O-18:0/14:1(9Z))				27.28	717.5661
PC(O-20:0/16:0)	12.843			12.835	775.6427	1.52
PC(O-20:0/22:6(4Z,7Z,10Z,13Z,16Z,19Z))		28.446			847.6458
PC(O-22:0/22:3(10Z,13Z,16Z))	29.661				881.7239
PC(O-22:1(13Z)/22:2(13Z,16Z))	29.661				881.7239
PC(P-15:0/0:0)		17.991	18.029		465.3202	0.63
PC(P-16:0/16:0)				27.28	717.5661
PC(P-16:0/17:1(9Z))			29.949		729.5628
PC(P-16:0/20:5(5Z,8Z,11Z,14Z,17Z))		31.931			763.5523
PC(P-17:0/0:0)		20.466			493.3523
PC(P-18:0/14:0)				27.28	717.5661
PC(P-18:0/15:1(9Z))			29.949		729.5628
PC(P-20:0/0:0)				29.957	535.396
PC(P-20:0/12:0)				27.28	717.5661
PC(P-20:0/19:1(9Z))	22.843		22.928	22.836	813.6711	12.13
PE 16:0-18:1		13.243			717.532
PE 16:1-16:1				30.31	687.4837
PE 16:1-18:1		32.032	32.121		715.5123	0.42
PE 18:0-16:1			32.777		717.5322
PECPC	16.126				809.5141
PG 16:0-18:1	23.045				748.5243
PG 18:1-18:1			32.121		774.5407
PGJ2_Class_1	17.54				344.2327
PGJ2_Class		18.092	17.979	17.987	344.2314	3.57
PHHdiA-PC	8.55				651.3765
PHODiA-PC	17.691				665.3904
PI 12:0-18:0	23.449				782.496
PI 14:0-16:0	23.449				782.496
PI 14:0-18:1		22.991			808.5122
PI 16:0-16:1		22.991			808.5122
P-IsoPGA2-PC			31.868		811.5366
POB-PC	15.671				579.3534
Procainamide	17.085	17.082	17.171	17.027	235.1651	0.4
Progesterone	18.853	18.951	19.039	18.947	314.2247	0.93
PS 18:0-18:1			12.979		789.5514
Quinidine	15.267	15.365	15.403	15.31	324.1844	3.57
Quinine	15.267	15.365	15.403	15.31	324.1844	3.57
TAG 14:0-16:0-16:0		32.89			778.7046
TAG 14:0-16:1-22:0	31.328				860.7816
TAG 14:0-18:1-20:0	31.328				860.7816
TAG 16:0-16:0-18:0			31.414		834.7664
TAG 16:0-16:1-26:0	29.207				944.8771
TAG 16:0-18:0-18:1	31.328				860.7816
TAG 16:0-18:1-24:0	29.207				944.8771
TAG 16:1-18:0-18:0	31.328				860.7816
TAG 18:0-18:1-22:0	29.207				944.8771
METABOLITES_END
#END