#METABOLOMICS WORKBENCH cchlebek_20251027_100944 DATATRACK_ID:6600 STUDY_ID:ST004373 ANALYSIS_ID:AN007307 PROJECT_ID:PR002771
VERSION             	1
CREATED_ON             	November 20, 2025, 3:46 pm
#PROJECT
PR:PROJECT_TITLE                 	In C57BL/6J mice, weight loss in previously obese mice shifted the cortical bone
PR:PROJECT_TITLE                 	metabolome
PR:PROJECT_SUMMARY               	8-week-old male and female C57BL/6 J mice received 60 % kCal high-fat diet for
PR:PROJECT_SUMMARY               	12 weeks. From 20 to 30 weeks of age, mice either remained obese or lost weight
PR:PROJECT_SUMMARY               	through 30 % caloric restriction. Controls consumed 10 % kCal low-fat diet. Mice
PR:PROJECT_SUMMARY               	were euthanized at 30 weeks of age, and cortical bone was obtained for
PR:PROJECT_SUMMARY               	metabolomic analyses. Metabolites were extracted and underwent LC-MS analysis
PR:PROJECT_SUMMARY               	using Waters I-Class Ultra-High Performance Liquid Chromatography coupled to a
PR:PROJECT_SUMMARY               	Waters Synapt-XS Q-IMS-TOF in positive mode.
PR:INSTITUTE                     	Mainehealth
PR:LAST_NAME                     	Chlebek
PR:FIRST_NAME                    	Carolyn
PR:ADDRESS                       	81 Research Drive
PR:EMAIL                         	carolyn.chlebek@mainehealth.org
PR:PHONE                         	5085107184
#STUDY
ST:STUDY_TITLE                   	In C57BL/6J mice, weight loss in previously obese mice shifted the cortical bone
ST:STUDY_TITLE                   	metabolome
ST:STUDY_SUMMARY                 	Obesity is linked to increased fracture risk. Despite the negative effects of
ST:STUDY_SUMMARY                 	weight loss on the skeleton, patients with obesity are advised to lose weight
ST:STUDY_SUMMARY                 	via calorie restriction. Obesity and weight loss individually alter both
ST:STUDY_SUMMARY                 	whole-body and local metabolism. Little is known about changes to bone mass and
ST:STUDY_SUMMARY                 	metabolome following calorie restriction in obese preclinical models. We
ST:STUDY_SUMMARY                 	hypothesized that caloric restriction would worsen bone quality in obese mice by
ST:STUDY_SUMMARY                 	shifting the cortical bone metabolome. To induce obesity, 8-week-old male and
ST:STUDY_SUMMARY                 	female C57BL/6J mice received 60% kCal high-fat diet for 12 weeks. From 20 to 30
ST:STUDY_SUMMARY                 	weeks of age, mice either remained obese or lost weight through 30% caloric
ST:STUDY_SUMMARY                 	restriction. Controls consumed 10% kCal low-fat diet. Compared to obesity,
ST:STUDY_SUMMARY                 	calorie restriction elicited more bone loss in both cortical and trabecular
ST:STUDY_SUMMARY                 	compartments. Weight loss also reduced bone formation. Both obesity and
ST:STUDY_SUMMARY                 	subsequent calorie restriction altered the cortical bone metabolome in a
ST:STUDY_SUMMARY                 	sex-dependent manner. Metabolic pathways altered with diet generally mapped to
ST:STUDY_SUMMARY                 	amino acid or fatty acid metabolism. In males, weight loss was associated with a
ST:STUDY_SUMMARY                 	downregulation of pathways related to tryptophan, tyrosine, ubiquinone, and
ST:STUDY_SUMMARY                 	fatty acids. In females, calorie restriction downregulated taurine and
ST:STUDY_SUMMARY                 	hypotaurine metabolism but upregulated pyrimidine metabolism, nicotinate and
ST:STUDY_SUMMARY                 	nicotinamide metabolism, and pantothenate and CoA biosynthesis. Our findings
ST:STUDY_SUMMARY                 	highlight the negative effects of obesity and subsequent caloric restriction on
ST:STUDY_SUMMARY                 	the skeleton. Despite improvements in components of systemic metabolism, caloric
ST:STUDY_SUMMARY                 	restriction in obese preclinical models did not restore bone morphology or the
ST:STUDY_SUMMARY                 	cortical metabolome to control conditions.
ST:INSTITUTE                     	Mainehealth
ST:LAST_NAME                     	Chlebek
ST:FIRST_NAME                    	Carolyn
ST:ADDRESS                       	81 Research Drive
ST:EMAIL                         	carolyn.chlebek@mainehealth.org
ST:PHONE                         	5085107184
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
SU:AGE_OR_AGE_RANGE              	30 wks
SU:GENDER                        	Male and female
SU:ANIMAL_LIGHT_CYCLE            	14 h light: 10 h dark
SU:SPECIES_GROUP                 	C57BL/6J
#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           	Blank 00	20240712_PB-Rosen_01_00_blank	Diet:NA - Blank | Sex:NA - Blank | Sample source:NA	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_00_blank.mzML
SUBJECT_SAMPLE_FACTORS           	Blank 01	20240712_PB-Rosen_01_01_blank	Diet:NA - Blank | Sex:NA - Blank | Sample source:NA	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_01_blank.mzML
SUBJECT_SAMPLE_FACTORS           	Blank 02	20240712_PB-Rosen_01_02_blank	Diet:NA - Blank | Sex:NA - Blank | Sample source:NA	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_02_blank.mzML
SUBJECT_SAMPLE_FACTORS           	Blank 03	20240712_PB-Rosen_01_03_blank	Diet:NA - Blank | Sex:NA - Blank | Sample source:NA	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_03_blank.mzML
SUBJECT_SAMPLE_FACTORS           	Blank 04	20240712_PB-Rosen_01_04_blank	Diet:NA - Blank | Sex:NA - Blank | Sample source:NA	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_04_blank.mzML
SUBJECT_SAMPLE_FACTORS           	Blank 05	20240712_PB-Rosen_01_05_blank	Diet:NA - Blank | Sex:NA - Blank | Sample source:NA	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_05_blank.mzML
SUBJECT_SAMPLE_FACTORS           	Blank 06	20240712_PB-Rosen_01_06_blank	Diet:NA - Blank | Sex:NA - Blank | Sample source:NA	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_06_blank.mzML
SUBJECT_SAMPLE_FACTORS           	Blank 07	20240712_PB-Rosen_01_07_blank	Diet:NA - Blank | Sex:NA - Blank | Sample source:NA	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_07_blank.mzML
SUBJECT_SAMPLE_FACTORS           	Neat 00	20240712_PB-Rosen_01_00_neat	Diet:NA - Neat | Sex:NA - Neat | Sample source:NA	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_00_neat.mzML
SUBJECT_SAMPLE_FACTORS           	Neat 01	20240712_PB-Rosen_01_01_neat	Diet:NA - Neat | Sex:NA - Neat | Sample source:NA	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_01_neat.mzML
SUBJECT_SAMPLE_FACTORS           	Neat 02	20240712_PB-Rosen_01_02_neat	Diet:NA - Neat | Sex:NA - Neat | Sample source:NA	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_02_neat.mzML
SUBJECT_SAMPLE_FACTORS           	Neat 03	20240712_PB-Rosen_01_03_neat	Diet:NA - Neat | Sex:NA - Neat | Sample source:NA	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_03_neat.mzML
SUBJECT_SAMPLE_FACTORS           	Neat 04	20240712_PB-Rosen_01_04_neat	Diet:NA - Neat | Sex:NA - Neat | Sample source:NA	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_04_neat.mzML
SUBJECT_SAMPLE_FACTORS           	Neat 05	20240712_PB-Rosen_01_05_neat	Diet:NA - Neat | Sex:NA - Neat | Sample source:NA	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_05_neat.mzML
SUBJECT_SAMPLE_FACTORS           	Neat 06	20240712_PB-Rosen_01_06_neat	Diet:NA - Neat | Sex:NA - Neat | Sample source:NA	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_06_neat.mzML
SUBJECT_SAMPLE_FACTORS           	Pool 1	20240712_PB-Rosen_01_P1	Diet:NA - Pooled samples | Sex:NA - Pooled samples | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_P1.mzML
SUBJECT_SAMPLE_FACTORS           	Pool 2	20240712_PB-Rosen_01_P2	Diet:NA - Pooled samples | Sex:NA - Pooled samples | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_P2.mzML
SUBJECT_SAMPLE_FACTORS           	Pool 3	20240712_PB-Rosen_01_P3	Diet:NA - Pooled samples | Sex:NA - Pooled samples | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_P3.mzML
SUBJECT_SAMPLE_FACTORS           	Pool 4	20240712_PB-Rosen_01_P4	Diet:NA - Pooled samples | Sex:NA - Pooled samples | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_P4.mzML
SUBJECT_SAMPLE_FACTORS           	Pool 5	20240712_PB-Rosen_01_P5	Diet:NA - Pooled samples | Sex:NA - Pooled samples | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_P5.mzML
SUBJECT_SAMPLE_FACTORS           	Pool 6	20240712_PB-Rosen_01_P6	Diet:NA - Pooled samples | Sex:NA - Pooled samples | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_P6.mzML
SUBJECT_SAMPLE_FACTORS           	202	20240712_PB-Rosen_01_202	Diet:Ctrl | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_202.mzML
SUBJECT_SAMPLE_FACTORS           	207	20240712_PB-Rosen_01_207	Diet:Ctrl | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_207.mzML
SUBJECT_SAMPLE_FACTORS           	211	20240712_PB-Rosen_01_211	Diet:Ctrl | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_211.mzML
SUBJECT_SAMPLE_FACTORS           	213	20240712_PB-Rosen_01_213	Diet:Ctrl | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_213.mzML
SUBJECT_SAMPLE_FACTORS           	214	20240712_PB-Rosen_01_214	Diet:Ctrl | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_214.mzML
SUBJECT_SAMPLE_FACTORS           	215	20240712_PB-Rosen_01_215	Diet:Ctrl | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_215.mzML
SUBJECT_SAMPLE_FACTORS           	217	20240712_PB-Rosen_01_217	Diet:Ctrl | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_217.mzML
SUBJECT_SAMPLE_FACTORS           	218	20240712_PB-Rosen_01_218	Diet:Ctrl | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_218.mzML
SUBJECT_SAMPLE_FACTORS           	219	20240712_PB-Rosen_01_219	Diet:HFD-CR | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_219.mzML
SUBJECT_SAMPLE_FACTORS           	221	20240712_PB-Rosen_01_221	Diet:HFD | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_221.mzML
SUBJECT_SAMPLE_FACTORS           	223	20240712_PB-Rosen_01_223	Diet:HFD | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_223.mzML
SUBJECT_SAMPLE_FACTORS           	225	20240712_PB-Rosen_01_225	Diet:HFD-CR | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_225.mzML
SUBJECT_SAMPLE_FACTORS           	227	20240712_PB-Rosen_01_227	Diet:HFD | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_227.mzML
SUBJECT_SAMPLE_FACTORS           	228	20240712_PB-Rosen_01_228	Diet:HFD-CR | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_228.mzML
SUBJECT_SAMPLE_FACTORS           	230	20240712_PB-Rosen_01_230	Diet:HFD-CR | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_230.mzML
SUBJECT_SAMPLE_FACTORS           	232	20240712_PB-Rosen_01_232	Diet:HFD | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_232.mzML
SUBJECT_SAMPLE_FACTORS           	238	20240712_PB-Rosen_01_238	Diet:HFD-CR | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_238.mzML
SUBJECT_SAMPLE_FACTORS           	240	20240712_PB-Rosen_01_240	Diet:HFD-CR | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_240.mzML
SUBJECT_SAMPLE_FACTORS           	241	20240712_PB-Rosen_01_241	Diet:HFD-CR | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_241.mzML
SUBJECT_SAMPLE_FACTORS           	242	20240712_PB-Rosen_01_242	Diet:HFD | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_242.mzML
SUBJECT_SAMPLE_FACTORS           	245	20240712_PB-Rosen_01_245	Diet:HFD | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_245.mzML
SUBJECT_SAMPLE_FACTORS           	247	20240712_PB-Rosen_01_247	Diet:HFD | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_247.mzML
SUBJECT_SAMPLE_FACTORS           	248	20240712_PB-Rosen_01_248	Diet:HFD-CR | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_248.mzML
SUBJECT_SAMPLE_FACTORS           	249	20240712_PB-Rosen_01_249	Diet:HFD-CR | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_249.mzML
SUBJECT_SAMPLE_FACTORS           	252	20240712_PB-Rosen_01_252	Diet:HFD | Sex:Male | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_252.mzML
SUBJECT_SAMPLE_FACTORS           	301	20240712_PB-Rosen_01_301	Diet:Ctrl | Sex:Female | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_301.mzML
SUBJECT_SAMPLE_FACTORS           	302	20240712_PB-Rosen_01_302	Diet:Ctrl | Sex:Female | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_302.mzML
SUBJECT_SAMPLE_FACTORS           	311	20240712_PB-Rosen_01_311	Diet:Ctrl | Sex:Female | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_311.mzML
SUBJECT_SAMPLE_FACTORS           	313	20240712_PB-Rosen_01_313	Diet:Ctrl | Sex:Female | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_313.mzML
SUBJECT_SAMPLE_FACTORS           	315	20240712_PB-Rosen_01_315	Diet:Ctrl | Sex:Female | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_315.mzML
SUBJECT_SAMPLE_FACTORS           	316	20240712_PB-Rosen_01_316	Diet:Ctrl | Sex:Female | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_316.mzML
SUBJECT_SAMPLE_FACTORS           	317	20240712_PB-Rosen_01_317	Diet:Ctrl | Sex:Female | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_317.mzML
SUBJECT_SAMPLE_FACTORS           	321	20240712_PB-Rosen_01_321	Diet:HFD | Sex:Female | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_321.mzML
SUBJECT_SAMPLE_FACTORS           	326	20240712_PB-Rosen_01_326	Diet:HFD-CR | Sex:Female | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_326.mzML
SUBJECT_SAMPLE_FACTORS           	332	20240712_PB-Rosen_01_332	Diet:HFD-CR | Sex:Female | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_332.mzML
SUBJECT_SAMPLE_FACTORS           	333	20240712_PB-Rosen_01_333	Diet:HFD | Sex:Female | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_333.mzML
SUBJECT_SAMPLE_FACTORS           	337	20240712_PB-Rosen_01_337	Diet:HFD | Sex:Female | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_337.mzML
SUBJECT_SAMPLE_FACTORS           	342	20240712_PB-Rosen_01_342	Diet:HFD-CR | Sex:Female | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_342.mzML
SUBJECT_SAMPLE_FACTORS           	344	20240712_PB-Rosen_01_344	Diet:HFD-CR | Sex:Female | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_344.mzML
SUBJECT_SAMPLE_FACTORS           	352	20240712_PB-Rosen_01_352	Diet:HFD | Sex:Female | Sample source:Cortical_Bone	RAW_FILE_NAME(Raw file name)=20240712_PB-Rosen_01_352.mzML
#COLLECTION
CO:COLLECTION_SUMMARY            	At euthanasia, femurs were cleaned of soft tissues and periosteum. Bone marrow
CO:COLLECTION_SUMMARY            	was removed via centrifugation from femurs in all animals. Trabecular bone was
CO:COLLECTION_SUMMARY            	removed from the femur and the cortical shell flash frozen in liquid nitrogen.
CO:SAMPLE_TYPE                   	Bone
#TREATMENT
TR:TREATMENT_SUMMARY             	Male and female C57BL/6J mice aged 5 weeks (Jackson Laboratories, Bar Harbor,
TR:TREATMENT_SUMMARY             	ME) were acclimated for three weeks prior to the experiment. All mouse
TR:TREATMENT_SUMMARY             	procedures were performed under the approval of the Institutional Animal Care
TR:TREATMENT_SUMMARY             	and Use Committee (IACUC) at the MaineHealth Institute for Research. Throughout
TR:TREATMENT_SUMMARY             	the study, mice were housed in 14-hour light:10-hour dark cycles. At 8 weeks of
TR:TREATMENT_SUMMARY             	age, the High Fat Feeding Phase was initiated. During the High Fat Feeding
TR:TREATMENT_SUMMARY             	Phase, mice were housed in groups of two or three per cage. Cages were randomly
TR:TREATMENT_SUMMARY             	assigned to receive a 60% high fat diet, containing 60% of calories from fat or
TR:TREATMENT_SUMMARY             	a 10% kcal low fat diet, containing 10% of calories from fat (D12492i and
TR:TREATMENT_SUMMARY             	D12450Ji, respectively; Research Diets Inc, New Brunswick, NJ). High fat diet
TR:TREATMENT_SUMMARY             	was completely replaced every 2-3 days. Following obesity induction, obese mice
TR:TREATMENT_SUMMARY             	were randomly assigned to remain on the high fat diet (HFD) or were assigned to
TR:TREATMENT_SUMMARY             	the diet-induced weight loss (HFD-CR). At the conclusion of the High Fat Feeding
TR:TREATMENT_SUMMARY             	Phase, all mice were single housed. HFD animals continued to receive 60 % high
TR:TREATMENT_SUMMARY             	fat diet. HFD-CR mice received the low fat diet for 2 weeks (Stabilization
TR:TREATMENT_SUMMARY             	Phase) and were then transitioned to a 30 % calorie-restricted diet for 8 weeks
TR:TREATMENT_SUMMARY             	(Caloric Restriction Phase) (Fig. 1A). All mice were euthanized at the end
TR:TREATMENT_SUMMARY             	of the Caloric Restriction Phase, at 30 weeks of age.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Using a bead mill, bone was pulverized in a 70:30 methanol:acetone solution.
SP:SAMPLEPREP_SUMMARY            	Samples were centrifuged and supernatant containing metabolites was extracted.
SP:SAMPLEPREP_SUMMARY            	Supernatant was then dried in vacuum concentrator. Metabolites were then
SP:SAMPLEPREP_SUMMARY            	resuspended in 1:1 acetonitrile:water. All solvents used were high-performance
SP:SAMPLEPREP_SUMMARY            	liquid chromatography grade.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Waters I-Class Ultra-High Performance Liquid Chromatography
CH:COLUMN_NAME                   	MicroSolv Diamond Hydride (150 x 2.1mm, 2.2 um, 120A)
CH:SOLVENT_A                     	100% water; 0.1% formic acid
CH:SOLVENT_B                     	100% acetonitrile; 0.1% formic acid
CH:FLOW_GRADIENT                 	Linear: Time(min); Flow Rate; %A; %B. Initial; 0.4; 10; 90. 2; 0.4; 10; 90. 8;
CH:FLOW_GRADIENT                 	0.4; 50; 50. 9; 0.4; 50; 50. 9.1; 0.4; 10; 90. 15; 0.4; 10; 90.
CH:FLOW_RATE                     	0.4 mL/min
CH:COLUMN_TEMPERATURE            	50
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Waters Synapt-XS
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	Normalized in Progenesis
MS:MS_RESULTS_FILE               	ST004373_AN007307_Results.txt	UNITS:arbitrary units	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END