#METABOLOMICS WORKBENCH ryant_20220217_085911 DATATRACK_ID:3095 STUDY_ID:ST002188 ANALYSIS_ID:AN003582 PROJECT_ID:PR001395
VERSION             	1
CREATED_ON             	June 6, 2022, 9:07 pm
#PROJECT
PR:PROJECT_TITLE                 	Metabolomic profiling reveals muscle metabolic changes following iliac
PR:PROJECT_TITLE                 	arteriovenous fistula creation in mice: part 1
PR:PROJECT_TYPE                  	Study of the skeletal muscle metabolome in mice with iliac arteriovenous fistula
PR:PROJECT_TYPE                  	via 1H NMR
PR:PROJECT_SUMMARY               	In the present study, we hypothesize that the creation of an iliac AVF would
PR:PROJECT_SUMMARY               	result in significant alterations to the limb muscle metabolome. Recently, our
PR:PROJECT_SUMMARY               	group developed a new murine model to address the pathophysiology of
PR:PROJECT_SUMMARY               	access-related hand dysfunction (ARHD) in mice, where AVF creation is performed
PR:PROJECT_SUMMARY               	in the iliac artery/vein. Because of the anatomical location of the AVF
PR:PROJECT_SUMMARY               	creation, this model produces clinically relevant changes in the mouse hindlimb
PR:PROJECT_SUMMARY               	including hemodynamic alterations, muscle weakness, and mitochondrial function
PR:PROJECT_SUMMARY               	impairment.
PR:INSTITUTE                     	University of Florida
PR:DEPARTMENT                    	Applied Physiology and Kinesiology
PR:LABORATORY                    	Rm 42 and Rm 43
PR:LAST_NAME                     	Ryan
PR:FIRST_NAME                    	Terence
PR:ADDRESS                       	1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA
PR:EMAIL                         	ryant@ufl.edu
PR:PHONE                         	352-294-1700
PR:FUNDING_SOURCE                	National Institutes of Health (NIH) grant R01HL148597 (STS). Additional salary
PR:FUNDING_SOURCE                	support was provided by NIH grants R01DK119274 (SAB) and R01HL149704 (TER), and
PR:FUNDING_SOURCE                	a postdoctoral fellowship (POST903198) from the American Heart Association (KK).
PR:PROJECT_COMMENTS              	Metabolomic study via NMR
PR:PUBLICATIONS                  	Frontiers
PR:CONTRIBUTORS                  	Ram B. Khattri, Kyoungrae Kim, Erik M. Anderson, Brian Fazzone, Kenneth C
PR:CONTRIBUTORS                  	Harland, Qiongyao Hu, Victoria R. Palzkill, Tomas A. Cort, Kerri A. O’Malle,
PR:CONTRIBUTORS                  	Scott A. Berceli, Terence E. Ryan, Salvatore T. Scali
#STUDY
ST:STUDY_TITLE                   	Metabolomic profiling reveals muscle metabolic changes following iliac
ST:STUDY_TITLE                   	arteriovenous fistula creation in mice
ST:STUDY_TYPE                    	Study of the skeletal muscle metabolome in mice with iliac arteriovenous fistula
ST:STUDY_TYPE                    	via 1H NMR
ST:STUDY_SUMMARY                 	In the present study, we hypothesize that the creation of an iliac AVF would
ST:STUDY_SUMMARY                 	result in significant alterations to the limb muscle metabolome. Recently, our
ST:STUDY_SUMMARY                 	group developed a new murine model to address the pathophysiology of
ST:STUDY_SUMMARY                 	access-related hand dysfunction (ARHD) in mice, where AVF creation is performed
ST:STUDY_SUMMARY                 	in the iliac artery/vein. Because of the anatomical location of the AVF
ST:STUDY_SUMMARY                 	creation, this model produces clinically relevant changes in the mouse hindlimb
ST:STUDY_SUMMARY                 	including hemodynamic alterations, muscle weakness, and mitochondrial function
ST:STUDY_SUMMARY                 	impairment.
ST:INSTITUTE                     	University of Florida
ST:DEPARTMENT                    	Applied Physiology and Kinesiology
ST:LABORATORY                    	Rm 42 and Rm 43
ST:LAST_NAME                     	Ryan
ST:FIRST_NAME                    	Terence
ST:ADDRESS                       	1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA
ST:EMAIL                         	ryant@ufl.edu
ST:PHONE                         	352-294-1700
ST:NUM_GROUPS                    	4
ST:TOTAL_SUBJECTS                	34
ST:NUM_MALES                     	All
ST:STUDY_COMMENTS                	Metabolomic study via NMR
ST:PUBLICATIONS                  	Frontiers
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:GENOTYPE_STRAIN               	C57BL/6J
SU:AGE_OR_AGE_RANGE              	13 weeks
SU:WEIGHT_OR_WEIGHT_RANGE        	(28.4±1.1 g (control SHAM mice) vs 26.4±2.1 g (control AVF mice) vs 23.6±2.1
SU:WEIGHT_OR_WEIGHT_RANGE        	(CKD SHAM mice) vs 24.7±1.3 (CKD AVF mice), N=8-10/group).
SU:GENDER                        	Male
SU:ANIMAL_ANIMAL_SUPPLIER        	Jackson Labs (Stock # 000664)
SU:ANIMAL_HOUSING                	Housed in a temperature of 22 oC
SU:ANIMAL_LIGHT_CYCLE            	12-hour light/12-hour dark
SU:ANIMAL_FEED                   	Ad libitum (Casein control diet vs. adenine-supplemented diet to induce CKD)
SU:ANIMAL_WATER                  	free access to food and water (3-5 animals per cage).
SU:ANIMAL_INCLUSION_CRITERIA     	(3-5 animals per cage).
#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           	-	Sample name	Factor1***:Group	RAW_FILE_NAME=raw data file names
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS18_Quad_Org	Factor1***:CKD_AVF	RAW_FILE_NAME=Terence_MS18_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS26_Quad_Org	Factor1***:CKD_AVF	RAW_FILE_NAME=Terence_MS26_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS27_Quad_Org	Factor1***:CKD_AVF	RAW_FILE_NAME=Terence_MS27_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS28_Quad_Org	Factor1***:CKD_AVF	RAW_FILE_NAME=Terence_MS28_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS29_Quad_Org	Factor1***:CKD_AVF	RAW_FILE_NAME=Terence_MS29_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS30_Quad_Org	Factor1***:CKD_AVF	RAW_FILE_NAME=Terence_MS30_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS36_Quad_Org	Factor1***:CKD_AVF	RAW_FILE_NAME=Terence_MS36_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS37_Quad_Org	Factor1***:CKD_AVF	RAW_FILE_NAME=Terence_MS37_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS39_Quad_Org	Factor1***:CKD_AVF	RAW_FILE_NAME=Terence_MS39_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS40_Quad_Org	Factor1***:CKD_AVF	RAW_FILE_NAME=Terence_MS40_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS6_Quad_Org	Factor1***:CKD_SHAM	RAW_FILE_NAME=Terence_MS6_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS7_Quad_Org	Factor1***:CKD_SHAM	RAW_FILE_NAME=Terence_MS7_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS8_Quad_Org	Factor1***:CKD_SHAM	RAW_FILE_NAME=Terence_MS8_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS9_Quad_Org	Factor1***:CKD_SHAM	RAW_FILE_NAME=Terence_MS9_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS10_Quad_Org	Factor1***:CKD_SHAM	RAW_FILE_NAME=Terence_MS10_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS16_Quad_Org	Factor1***:CKD_SHAM	RAW_FILE_NAME=Terence_MS16_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS17_Quad_Org	Factor1***:CKD_SHAM	RAW_FILE_NAME=Terence_MS17_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS20_Quad_Org	Factor1***:CKD_SHAM	RAW_FILE_NAME=Terence_MS20_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS14_Quad_Org	Factor1***:CON_AVF	RAW_FILE_NAME=Terence_MS14_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS22_Quad_Org	Factor1***:CON_AVF	RAW_FILE_NAME=Terence_MS22_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS23_Quad_Org	Factor1***:CON_AVF	RAW_FILE_NAME=Terence_MS23_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS24_Quad_Org	Factor1***:CON_AVF	RAW_FILE_NAME=Terence_MS24_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS25_Quad_Org	Factor1***:CON_AVF	RAW_FILE_NAME=Terence_MS25_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS32_Quad_Org	Factor1***:CON_AVF	RAW_FILE_NAME=Terence_MS32_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS33_Quad_Org	Factor1***:CON_AVF	RAW_FILE_NAME=Terence_MS33_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS35_Quad_Org	Factor1***:CON_AVF	RAW_FILE_NAME=Terence_MS35_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS1_Quad_Org	Factor1***:CON_SHAM	RAW_FILE_NAME=Terence_MS1_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS2_Quad_Org	Factor1***:CON_SHAM	RAW_FILE_NAME=Terence_MS2_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS3_Quad_Org	Factor1***:CON_SHAM	RAW_FILE_NAME=Terence_MS3_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS4_Quad_Org	Factor1***:CON_SHAM	RAW_FILE_NAME=Terence_MS4_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS5_Quad_Org	Factor1***:CON_SHAM	RAW_FILE_NAME=Terence_MS5_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS11_Quad_Org	Factor1***:CON_SHAM	RAW_FILE_NAME=Terence_MS11_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS12_Quad_Org	Factor1***:CON_SHAM	RAW_FILE_NAME=Terence_MS12_Quad_Org.raw
SUBJECT_SAMPLE_FACTORS           	-	Terence_MS15_Quad_Org	Factor1***:CON_SHAM	RAW_FILE_NAME=Terence_MS15_Quad_Org.raw
#COLLECTION
CO:COLLECTION_SUMMARY            	Two weeks after surgery, mice were anesthetized using isoflurane and the left
CO:COLLECTION_SUMMARY            	quadriceps muscle was rapidly dissected and immediately frozen in liquid
CO:COLLECTION_SUMMARY            	nitrogen and stored at -80oC for metabolite extraction. Euthanasia was carried
CO:COLLECTION_SUMMARY            	out by thoracotomy followed by cervical dislocation.
CO:SAMPLE_TYPE                   	Muscle
CO:COLLECTION_METHOD             	While under isoflurane anesthesia, tissues were rapidly dissected and snap
CO:COLLECTION_METHOD             	frozen in liquid nitrogen and stored at -80°C until metabolite extraction
CO:COLLECTION_LOCATION           	University of Florida, Applied Physiology and Kinesiology, 1864 stadium RD,
CO:COLLECTION_LOCATION           	Gainesville, FL 32611
CO:STORAGE_CONDITIONS            	-80℃
CO:STORAGE_VIALS                 	cryovials
#TREATMENT
TR:TREATMENT_SUMMARY             	Briefly, mice were acclimated to a standard casein-based chow (Control Diet) for
TR:TREATMENT_SUMMARY             	7 days. Thereafter, the animals were randomly assign to either remain on Control
TR:TREATMENT_SUMMARY             	Diet (non-CKD groups) or receive adenine-supplemented diet to induce CKD.
TR:TREATMENT_SUMMARY             	Following three weeks of the casein/adenine diet, animals were randomly assigned
TR:TREATMENT_SUMMARY             	to receive either an arteriovenous fistula surgery (AVF) or sham surgery (SHAM)
TR:ANIMAL_VET_TREATMENTS         	none
TR:ANIMAL_ANESTHESIA             	isoflurane
TR:ANIMAL_FASTING                	non-fasted
TR:ANIMAL_ENDP_EUTHANASIA        	Euthanasia was carried out by thoracotomy followed by cervical dislocation.
TR:ANIMAL_ENDP_TISSUE_COLL_LIST  	Skeletal muscle (quadriceps)
TR:ANIMAL_ENDP_TISSUE_PROC_METHOD	Two weeks after surgery, mice were anesthetized using isoflurane and the left
TR:ANIMAL_ENDP_TISSUE_PROC_METHOD	quadriceps muscle was rapidly dissected and immediately frozen in liquid
TR:ANIMAL_ENDP_TISSUE_PROC_METHOD	nitrogen and stored at -80oC for metabolite extraction.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Weights of frozen quadriceps specimens were weighed using a microbalance
SP:SAMPLEPREP_SUMMARY            	(Mettler-Toledo, Columbus, OH, USA). Next, a slightly modified FOLCH extraction
SP:SAMPLEPREP_SUMMARY            	was performed to extract aqueous and lipid phase metabolites. The aqueous phase
SP:SAMPLEPREP_SUMMARY            	was lyophilized overnight (Labconco Corporation, Kansas, MO, USA) and the lipid
SP:SAMPLEPREP_SUMMARY            	phase was dried by passing inert nitrogen gas. The resulting aqueous and lipid
SP:SAMPLEPREP_SUMMARY            	phase dry powders were stored at -80oC until analysis using nuclear magnetic
SP:SAMPLEPREP_SUMMARY            	resonance (NMR). The dry powder of aqueous phase samples was dissolved in 50 µL
SP:SAMPLEPREP_SUMMARY            	of phosphate buffer system (50 mM, pH 7.2) consisting of 0.5 mM D6-DSS, 2 mM
SP:SAMPLEPREP_SUMMARY            	EDTA and 0.2% NaN2. Lipid phase dry powders were dissolved in 70 µL of CDCl3
SP:SAMPLEPREP_SUMMARY            	supplemented with 10 mM of pyrazine (as internal NMR standard). All samples were
SP:SAMPLEPREP_SUMMARY            	loaded into 1.5 mm optical density (O.D.) NMR tubes.
SP:PROCESSING_METHOD             	Lyophilization and Homogenization
SP:PROCESSING_STORAGE_CONDITIONS 	-80℃
SP:EXTRACTION_METHOD             	Modified FOLCH extraction
SP:EXTRACT_STORAGE               	-80℃
SP:SAMPLE_RESUSPENSION           	Deuterated chloroform (80 microliter) with 10 mM pyrazine was used to re-suspend
SP:SAMPLE_RESUSPENSION           	organic phase samples.
SP:SAMPLE_SPIKING                	10 mM of pyrazine for organic phase samples.
#ANALYSIS
AN:DATA_FORMAT                   	fid, 1r
#NMR
NM:INSTRUMENT_NAME               	Bruker Avance III Cryo800 MHz 54 mm
NM:INSTRUMENT_TYPE               	FT-NMR
NM:NMR_EXPERIMENT_TYPE           	1D-1H
NM:FIELD_FREQUENCY_LOCK          	Deuterated chloroform
NM:STANDARD_CONCENTRATION        	10mM pyrazine
NM:SPECTROMETER_FREQUENCY        	799.9
NM:NMR_PROBE                     	CP TXI CryoProbe
NM:NMR_SOLVENT                   	Deuterated chloroform
NM:NMR_TUBE_SIZE                 	1.5 mm O.D.
NM:SHIMMING_METHOD               	Topshim
NM:PULSE_SEQUENCE                	noesypr1d
NM:WATER_SUPPRESSION             	none
NM:PULSE_WIDTH                   	90-degree
NM:RECEIVER_GAIN                 	228
NM:OFFSET_FREQUENCY              	None
NM:CHEMICAL_SHIFT_REF_CPD        	CDCl3 at 7.26 ppm and pyrazine at 8.61 ppm
NM:TEMPERATURE                   	25 o C
NM:NUMBER_OF_SCANS               	128 scans
NM:DUMMY_SCANS                   	8
NM:ACQUISITION_TIME              	4 s
NM:RELAXATION_DELAY              	1 s
NM:SPECTRAL_WIDTH                	9523.8
NM:NUM_DATA_POINTS_ACQUIRED      	38094
NM:REAL_DATA_POINTS              	65536
NM:LINE_BROADENING               	0.22 Hz
NM:ZERO_FILLING                  	65,536 points
NM:APODIZATION                   	Exponential
NM:BASELINE_CORRECTION_METHOD    	Spline
NM:CHEMICAL_SHIFT_REF_STD        	7.26ppm for CDCl3
NM:BINNED_INCREMENT              	None
NM:BINNED_DATA_NORMALIZATION_METHOD	None
NM:BINNED_DATA_CHEMICAL_SHIFT_RANGE	None
NM:BINNED_DATA_EXCLUDED_RANGE    	None
#NMR_METABOLITE_DATA
NMR_METABOLITE_DATA:UNITS	A.U.
NMR_METABOLITE_DATA_START
Samples	Terence_MS18_Quad_Org	Terence_MS26_Quad_Org	Terence_MS27_Quad_Org	Terence_MS28_Quad_Org	Terence_MS29_Quad_Org	Terence_MS30_Quad_Org	Terence_MS36_Quad_Org	Terence_MS37_Quad_Org	Terence_MS39_Quad_Org	Terence_MS40_Quad_Org	Terence_MS6_Quad_Org	Terence_MS7_Quad_Org	Terence_MS8_Quad_Org	Terence_MS9_Quad_Org	Terence_MS10_Quad_Org	Terence_MS16_Quad_Org	Terence_MS17_Quad_Org	Terence_MS20_Quad_Org	Terence_MS14_Quad_Org	Terence_MS22_Quad_Org	Terence_MS23_Quad_Org	Terence_MS24_Quad_Org	Terence_MS25_Quad_Org	Terence_MS32_Quad_Org	Terence_MS33_Quad_Org	Terence_MS35_Quad_Org	Terence_MS1_Quad_Org	Terence_MS2_Quad_Org	Terence_MS3_Quad_Org	Terence_MS4_Quad_Org	Terence_MS5_Quad_Org	Terence_MS11_Quad_Org	Terence_MS12_Quad_Org	Terence_MS15_Quad_Org
Factors	Factor1***:CKD_AVF	Factor1***:CKD_AVF	Factor1***:CKD_AVF	Factor1***:CKD_AVF	Factor1***:CKD_AVF	Factor1***:CKD_AVF	Factor1***:CKD_AVF	Factor1***:CKD_AVF	Factor1***:CKD_AVF	Factor1***:CKD_AVF	Factor1***:CKD_SHAM	Factor1***:CKD_SHAM	Factor1***:CKD_SHAM	Factor1***:CKD_SHAM	Factor1***:CKD_SHAM	Factor1***:CKD_SHAM	Factor1***:CKD_SHAM	Factor1***:CKD_SHAM	Factor1***:CON_AVF	Factor1***:CON_AVF	Factor1***:CON_AVF	Factor1***:CON_AVF	Factor1***:CON_AVF	Factor1***:CON_AVF	Factor1***:CON_AVF	Factor1***:CON_AVF	Factor1***:CON_SHAM	Factor1***:CON_SHAM	Factor1***:CON_SHAM	Factor1***:CON_SHAM	Factor1***:CON_SHAM	Factor1***:CON_SHAM	Factor1***:CON_SHAM	Factor1***:CON_SHAM
C18-CH3 chol+V26C1C1:W25	70.655	27.752	27.048	44.38	47.43	28.337	23.29	72.172	32.084	66.42	25.232	25.967	25.427	25.043	24.077	25.408	24.809	18.766	28.188	29.088	42.952	45.541	46.372	30.963	77.07	71.476	23.017	21.865	24.685	24.524	23.878	24.87	17.8	26.693
CH3-protons	691.372	542.156	529.016	711.428	723.311	558.769	444.949	778.385	646.073	767.537	534.006	560.07	539.011	581.321	509.561	494.932	499.877	353.489	617.729	596.191	760.731	730.727	777.242	596.775	891.093	814.731	488.463	432.123	523.476	498.733	518.475	513.221	419.103	645.773
Unknown triplet	47.82	75.074	67.171	52.913	59.529	63.571	47.483	47.845	87.211	52.24	75.041	82.798	83.264	81.629	72.646	77.75	80.451	55.44	77.28	69.167	59.774	55.632	58.911	61.744	58.088	48.565	75.66	58.773	87.584	77.393	86.444	84.933	58.483	70.04
C19-CH3 Chol	78.603	31.821	30.808	49.717	51.737	32.289	26.034	80.11	37.522	73.017	29.471	30.227	29.436	29.603	27.176	29.267	28.894	22.459	32.59	32.717	47.49	49.111	50.294	35.813	84.989	80.154	26.171	24.64	28.081	27.101	27.76	28.292	19.524	32.397
CH2n chol	1564.606	1581.85	1515.987	1831.23	1885.353	1638.207	1288.852	1962.366	1906.215	1782.603	1563.808	1671.069	1611.632	1673.785	1596.773	1470.487	1504.582	997.995	1776.557	1723.287	1911.15	1869.333	2028.682	1641.192	2054.032	1795.856	1511.123	1285.858	1605.03	1532.11	1591.346	1570.263	1340.238	1745.25
CH2n aliphatic chains	1782.641	1543.131	1589.467	2017.276	2064.158	1550.354	1286.89	2044.771	1857.252	2101.739	1559.24	1576.19	1533.595	1730.72	1362.829	1373.817	1460.445	981.043	1811.215	1694.042	2288.391	2144.381	2243.717	1742.721	2477.924	2254.487	1310.006	1216.18	1511.576	1380.294	1464.627	1415.052	1153.951	1988.173
Cholesterol2	6.093	3.945	5.21	3.389	2.961	4.424	3.142	4.161	5.485	7.337	4.422	4.875	6.439	5.634	3.955	4.973	6.289	3.157	3.832	5.767	3.431	2.122	4.995	4.197	8.394	3.514	0.873	3.458	4.145	3.161	3.165	2.043	1.393	7.973
CH2-CH2-COO-beta	304.403	283.645	285.927	333.161	336.014	285.267	228.577	343.515	341.476	342.368	281.785	294.298	291.415	314.454	254.473	260.166	277.643	186.711	321.716	311.742	368.244	344.038	371.829	303.254	406.411	365.969	234.654	222.758	269.913	249.387	264.909	256.591	203.829	370.374
CH2-CH=CH-CH2-alpha	367.882	328.123	338.329	423.01	446.086	330.223	264.698	432.222	392.24	422.919	322.128	326.333	318.887	342.987	283.129	293.73	304.24	200.336	380.363	353.959	469.301	441.216	485.356	370.84	508.935	450.237	270.312	252.098	304.787	290.86	292.192	294.983	240.578	400.996
CH2COO-alpha	259.806	253.099	259.097	307.984	316.092	259.809	214.246	306.305	302.567	290.157	245.021	257.79	255.572	269.348	236.258	225.758	245.564	159.1	286.761	270.172	335.126	313.594	346.496	277.609	346.65	316.079	214.151	207.774	245.852	235.05	237.053	232.71	197.003	315.567
CH-CH2-CH=CH	260.089	421.392	404.766	409.118	396.197	439.408	302.859	305.085	515.694	317.249	426.946	474.88	474.251	457.874	464.247	433.241	445.757	288.517	469.834	447.438	425.059	374.912	466.109	411.871	376.158	250.229	426.963	353.089	483.579	463.532	470.212	462.938	365.807	372.556
N+CH33	290.602	377.11	376.386	395.683	431.415	384.813	371.402	346.969	426	327.774	367.71	389.653	387.283	379.97	379.428	351.756	367.06	254.952	396.836	383.829	418.687	377.84	479.084	368.557	366.897	342.778	339.941	345.516	372.931	383.694	367.85	358.979	340.011	375.678
Cholesterol3	176.249	23.677	28.865	30.184	29.496	26.062	18.015	69.495	33.374	71.925	36	33.377	35.951	38.699	23.965	33.39	71.33	73.49	35.08	27.213	32.069	34.518	28.236	178.727	72.578	72.398	26.208	24.145	28.47	23.046	32.443	28.568	15.108	139.923
3CH2 glycerophospholipids	159.703	390.705	299.169	521.087	323.473	354.212	423.084	256.175	737.816	272.034	730.579	426.256	525.06	459.44	328.038	525.033	230.624	142.813	481.928	401.955	461.532	488.404	300.495	199.278	324.327	141.221	463.648	299.565	431.398	376.185	434.754	472.286	283.451	174.167
CH2_TG1	80.742	191.836	265.475	163.196	367.929	105.731	91.465	108.482	174.529	115.361	203.523	253.803	192.363	169.573	195.274	206.53	103.18	64.007	197.763	389.637	242.58	158.593	508.118	95.809	135.575	99.394	256.763	95.593	147.68	145.921	135.275	155.663	153.085	109.781
CH2_TG2	103.596	142.689	172.35	138.291	175.207	116.452	107.064	129.631	151.589	131.148	151.458	172.66	153.245	151.889	143.835	147.78	118.207	77.828	161.58	184.118	169.01	142.677	246.452	113.234	147.385	125.369	158.058	105.854	132.362	128.93	127.403	130.778	119.292	138.053
1CH phospholipids+TG	61.344	94.668	111.991	97.022	111.716	85.102	68.66	75.602	103.694	81.995	98.745	110.027	100.203	96.253	92.999	95.831	81.336	52.531	105.559	112.585	111.284	93.4	139.386	81.768	88.542	55.38	100.056	69.76	93.547	88.351	88.787	87.458	74.907	70.232
CH phospholipid	58.588	77.897	87.112	88.629	93.522	82.132	67.261	74.864	92.572	72.712	78.673	88.096	86.723	86.733	82.438	77.335	80.645	50.472	89.631	83.075	94.214	85.861	104.952	83.439	83.628	54.324	74.39	68.32	82.922	85.615	74.479	76.459	68.638	71.64
CH TG	6.267	3.157	5.119	3.606	2.619	2.396	1.739	5.628	5.543	7.087	6.41	6.308	5.901	8.837	3.889	4.81	4.634	2.422	7.198	3.871	5.737	5.556	2.483	4.941	9.876	17.41	4.094	2.661	4.18	4.358	3.868	3.197	2.122	13.844
CH=CH	493.792	662.154	644.024	696.462	708.134	663.782	491.166	575.366	787.899	599.451	664.536	719.98	711.923	714.538	672.533	653.552	662.933	436.193	744.946	693.399	740.627	677.652	797.293	666.744	698.554	541.781	643	534.038	717.566	681.181	686.89	687.097	547.395	626.471
NMR_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name
C18-CH3 chol+V26C1C1:W25
CH3-protons
Unknown triplet
C19-CH3 Chol
CH2n chol
CH2n aliphatic chains
Cholesterol2
CH2-CH2-COO-beta
CH2-CH=CH-CH2-alpha
CH2COO-alpha
CH-CH2-CH=CH
N+CH33
Cholesterol3
3CH2 glycerophospholipids
CH2_TG1
CH2_TG2
1CH phospholipids+TG
CH phospholipid
CH TG
CH=CH
METABOLITES_END
#END