#METABOLOMICS WORKBENCH sysu604_20230916_011055 DATATRACK_ID:4322 STUDY_ID:ST002918 ANALYSIS_ID:AN004788 PROJECT_ID:PR001799
VERSION             	1
CREATED_ON             	September 27, 2023, 11:11 pm
#PROJECT
PR:PROJECT_TITLE                 	Metabolic Profiling in mouse Infected with Vibrio parahaemolyticus
PR:PROJECT_TYPE                  	MS quantitative analysis
PR:PROJECT_SUMMARY               	Multidrug-resistant bacteria present a major threat to public health. Therefore,
PR:PROJECT_SUMMARY               	new drugs or approaches are urgently needed to manage and mitigate this threat.
PR:PROJECT_SUMMARY               	Here, we screen the molecular candidates that allow the survival of mice upon
PR:PROJECT_SUMMARY               	multidrug-resistant Vibrio parahaemolyticus infection by integrated proteomic
PR:PROJECT_SUMMARY               	and metabolomics analysis, where L-Alanine metabolism and phagocytosis are
PR:PROJECT_SUMMARY               	highly correlated. The role of L-Alanine on boosting mouse survival is further
PR:PROJECT_SUMMARY               	confirmed with in vivo bacterial challenge studies on multidrug-resistant
PR:PROJECT_SUMMARY               	bacteria including V. parahaemolyticus, Escherichia coli, Pseudomonas
PR:PROJECT_SUMMARY               	aeruginosa, Klebsiella pneumoniae. Functional studies demonstrate that exogenous
PR:PROJECT_SUMMARY               	L-Alanine promotes phagocytosis to these different species of
PR:PROJECT_SUMMARY               	multidrug-resistant pathogens. The underlying mechanism involves two events that
PR:PROJECT_SUMMARY               	are L-Alanine-dependently increased TLR4 expression, and L-Alanine-enhanced TLR4
PR:PROJECT_SUMMARY               	signaling via increasing the biosynthesis and secretion of fatty acids such as
PR:PROJECT_SUMMARY               	palmitate. Palmitate enhances the binding of LPS to TLR4 and thereby promotes
PR:PROJECT_SUMMARY               	TLR4 dimmer formation and endocytosis for the subsequent activation of PI3K/Akt
PR:PROJECT_SUMMARY               	and NF-κB pathways and phagocytosis of bacteria. These results suggest that
PR:PROJECT_SUMMARY               	modulation of metabolic environment is a plausible approach for combating
PR:PROJECT_SUMMARY               	infection with multidrug-resistant bacteria.
PR:INSTITUTE                     	Sun Yat-sen University
PR:LAST_NAME                     	jiang
PR:FIRST_NAME                    	ming
PR:ADDRESS                       	No. 135, Xingang Xi Road, Guangzhou, 510275, P. R. China
PR:EMAIL                         	jiangm28@mail.sysu.edu.cn
PR:PHONE                         	13434283781
#STUDY
ST:STUDY_TITLE                   	Metabolic Profiling in mouse Infected with Vibrio parahaemolyticus
ST:STUDY_SUMMARY                 	mice were subject to intraperitoneal (i.p.) injection with the V.
ST:STUDY_SUMMARY                 	parahaemolyticus, and control mice were injected with saline. Data were
ST:STUDY_SUMMARY                 	collected and analyzed using unsupervised hierarchical clustering. In general,
ST:STUDY_SUMMARY                 	the abundance of metabolites increased more often than it decreased in animals
ST:STUDY_SUMMARY                 	with higher resistance to infection. Load weight analysis of biological pathways
ST:STUDY_SUMMARY                 	suggested that alanine, aspartate, glutamate metabolism could play roles in the
ST:STUDY_SUMMARY                 	capacity of mice to survive infection with V. parahaemolyticus.
ST:INSTITUTE                     	Sun Yat-sen University
ST:LAST_NAME                     	jiang
ST:FIRST_NAME                    	ming
ST:ADDRESS                       	No. 135, Xingang Xi Road, Guangzhou, 510275, P. R. China
ST:EMAIL                         	jiangm28@mail.sysu.edu.cn
ST:PHONE                         	13434283781
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
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           	-	20120522-zhxl-test-S1-1	Treatment:control	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-test-S1-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-test-S1-2	Treatment:control	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-test-S1-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-test-S2-1	Treatment:control	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-test-S2-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-test-S2-2	Treatment:control	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-test-S2-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-test-S3-1	Treatment:control	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-test-S3-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-test-S3-2	Treatment:control	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-test-S3-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A1-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A1-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A1-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A1-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A2-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A2-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A2-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A2-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A3-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A3-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A3-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A3-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A4-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A4-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A4-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A4-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A5-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A5-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A5-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A5-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A6-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A6-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A6-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A6-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A7-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A7-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A7-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A7-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A8-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A8-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A8-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A8-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A9-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A9-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A9-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A9-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A10-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A10-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A10-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A10-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B1-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B1-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B1-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B1-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B2-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B2-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B2-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B2-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B3-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B3-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B3-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B3-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B4-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B4-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B4-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B4-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B5-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B5-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B5-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B5-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B6-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B6-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B6-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B6-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B7-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B7-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B7-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B7-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B8-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B8-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B8-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B8-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B9-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B9-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B9-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B9-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B10-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B10-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B10-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B10-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C1-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C1-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C1-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C1-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C2-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C2-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C2-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C2-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C3-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C3-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C3-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C3-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C4-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C4-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C4-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C4-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C5-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C5-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C5-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C5-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C6-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C6-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C6-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C6-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C7-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C7-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C7-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C7-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C8-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C8-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C8-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C8-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C9-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C9-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C9-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C9-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C10-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C10-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C10-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C10-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A1-L-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A1-L-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A1-L-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A1-L-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A2-L-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A2-L-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A2-L-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A2-L-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A3-L-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A3-L-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A3-L-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A3-L-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A4-L-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A4-L-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A4-L-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A4-L-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A5-L-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A5-L-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A5-L-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A5-L-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A6-L-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A6-L-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-A6-L-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-A6-L-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B1-L-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B1-L-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B1-L-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B1-L-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B2-L-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B2-L-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B2-L-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B2-L-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B3-L-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B3-L-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-B3-L-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-B3-L-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C1-L-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C1-L-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C1-L-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C1-L-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C2-L-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C2-L-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C2-L-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C2-L-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C3-L-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C3-L-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C3-L-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C3-L-2.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C4-L-1	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C4-L-1.raw
SUBJECT_SAMPLE_FACTORS           	-	20120522-zhxl-C4-L-2	Treatment:infection	Genotype=Wild-type; RAW_FILE_NAME=20120522-zhxl-C4-L-2.raw
#COLLECTION
CO:COLLECTION_SUMMARY            	Samples were counted, washed with cold PBS and then flash-frozen in liquid N2
CO:SAMPLE_TYPE                   	Spleen
#TREATMENT
TR:TREATMENT_SUMMARY             	Spleen tissues were weighed and homogenized with the first solvent (the mixture
TR:TREATMENT_SUMMARY             	of chloroform, methanol and water (1:2:1, v/v/v)) for 30 s at 4 0C and then
TR:TREATMENT_SUMMARY             	centrifuged at 12,000 rpm for 10 min at 4 0C. The supernatant was collected and
TR:TREATMENT_SUMMARY             	deposit was re-homogenized with the second solvent (methanol alone) before a
TR:TREATMENT_SUMMARY             	second centrifugation. The two supernatants were mixed, and aliquot of sample
TR:TREATMENT_SUMMARY             	was transferred to a GC sampling vial containing 5 μL 0.1 mg/mL ribitol (Sigma)
TR:TREATMENT_SUMMARY             	as an analytical internal standard and then dried in a vacuum centrifuge
TR:TREATMENT_SUMMARY             	concentrator before the subsequent derivatization. Two technical replicates were
TR:TREATMENT_SUMMARY             	prepared for each sample.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Spleen tissues were weighed and homogenized with the first solvent (the mixture
SP:SAMPLEPREP_SUMMARY            	of chloroform, methanol and water (1:2:1, v/v/v)) for 30 s at 4 0C and then
SP:SAMPLEPREP_SUMMARY            	centrifuged at 12,000 rpm for 10 min at 4 0C. The supernatant was collected and
SP:SAMPLEPREP_SUMMARY            	deposit was re-homogenized with the second solvent (methanol alone) before a
SP:SAMPLEPREP_SUMMARY            	second centrifugation. The two supernatants were mixed, and aliquot of sample
SP:SAMPLEPREP_SUMMARY            	was transferred to a GC sampling vial containing 5 μL 0.1 mg/mL ribitol (Sigma)
SP:SAMPLEPREP_SUMMARY            	as an analytical internal standard and then dried in a vacuum centrifuge
SP:SAMPLEPREP_SUMMARY            	concentrator before the subsequent derivatization. Two technical replicates were
SP:SAMPLEPREP_SUMMARY            	prepared for each sample.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Low pH polar (LC/MS Pos early)
CH:CHROMATOGRAPHY_TYPE           	GC
CH:INSTRUMENT_NAME               	Thermo Scientific Trace GC Ultra with DSQ II GC/MS
CH:COLUMN_NAME                   	Agilent DB5-MS (30m x 0.25mm, 0.25um)
CH:SOLVENT_A                     	none
CH:SOLVENT_B                     	none
CH:FLOW_GRADIENT                 	none
CH:FLOW_RATE                     	1.0 mL/min
CH:COLUMN_TEMPERATURE            	270 °C
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Scientific Trace GC Ultra with DSQ II GC/MS
MS:INSTRUMENT_TYPE               	Triple quadrupole
MS:MS_TYPE                       	EI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	samples was derivatized and then used to firstly protect carbonyl moieties
MS:MS_COMMENTS                   	through methoximation, through a 90 min 37 ℃ reaction with 40 μL of 20 mg/mL
MS:MS_COMMENTS                   	methoxyamine hydrochloride (Sigma-Aldrich) in pyridine, followed by
MS:MS_COMMENTS                   	derivatization of acidic protons through a 30 min 37 0C reaction with the
MS:MS_COMMENTS                   	addition of 80 μL N-methyl-N-trimethylsilyltrifluoroace-tamide (MSTFA,
MS:MS_COMMENTS                   	Sigma-Aldrich). The derivatized sample of 1 μL was injected into a 30m × 250
MS:MS_COMMENTS                   	μm i.d. × 0.25 μm DBS-MS column using splitless injection and analysis was
MS:MS_COMMENTS                   	carried out by Trace DSQ II (Thermo Scientific). The initial temperature of the
MS:MS_COMMENTS                   	GC oven was held at 85 0C for 5 min followed by an increase to 330 0C at a rate
MS:MS_COMMENTS                   	of 15 0C min-1 then held for 5 min. Helium was used as carrier gas and flow was
MS:MS_COMMENTS                   	kept constant at 1 mL min-1. The MS was operated in a range of 50-600 m/z.
MS:MS_RESULTS_FILE               	ST002918_AN004788_Results.txt	UNITS:Peak area 	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END