#METABOLOMICS WORKBENCH fangyuan_zhou_20200910_172015 DATATRACK_ID:2164 STUDY_ID:ST001511 ANALYSIS_ID:AN002504
VERSION                          	1
CREATED_ON                       	07-31-2023
#PROJECT
PR:PROJECT_TITLE                 	Delia antiqua bacteria-derived organic acids
PR:PROJECT_SUMMARY               	Metabolomic profiles of six bacteria and K. oxytoca were compared to identify
PR:PROJECT_SUMMARY               	candidate metabolites that may prevent B. bassiana infection of D. antiqua.
PR:INSTITUTE                     	Qilu University of Technology (Shandong Academy of Sciences)
PR:LAST_NAME                     	Zhou
PR:FIRST_NAME                    	Fangyuan
PR:ADDRESS                       	No. 28789 Jingshidong Road, Jinan, Shandong, 250103, China
PR:EMAIL                         	fangyuan_zhou@163.com
PR:PHONE                         	+8618511761347
PR:DOI                           	http://dx.doi.org/10.21228/M83T2V
#STUDY
ST:STUDY_TITLE                   	Symbiotic bacteria-derived organic acids
ST:STUDY_SUMMARY                 	Metabolomic profiles of six bacteria and K. oxytoca were compared to identify
ST:STUDY_SUMMARY                 	candidate metabolites that may prevent B. bassiana infection of D. antiqua.
ST:INSTITUTE                     	Qilu University of Technology (Shandong Academy of Sciences)
ST:LAST_NAME                     	Zhou
ST:FIRST_NAME                    	Fangyuan
ST:ADDRESS                       	No. 28789 Jingshidong Road, Jinan, Shandong, 250103, China
ST:EMAIL                         	fangyuan_zhou@163.com
ST:PHONE                         	+8618511761347
ST:SUBMIT_DATE                   	2020-09-10
#SUBJECT
SU:SUBJECT_TYPE                  	Bacteria
SU:SUBJECT_SPECIES               	Citrobacter freundii;Enterobacter ludwigii;Pseudomonas protegens;Serratia
SU:SUBJECT_SPECIES               	plymuthica;Sphingobacterium faecium;Stenotrophomonas maltophilia;Klebsiella
SU:SUBJECT_SPECIES               	oxytoca
SU:TAXONOMY_ID                   	546;299767;380021;82996;34087;40324;571
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	-	B505_1N	Sample Type:Citrobacter freundii	Sample_Data:RAW_FILE_NAME=B505_1N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B505_1P	Sample Type:Citrobacter freundii	Sample_Data:RAW_FILE_NAME=B505_1P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B505_2N	Sample Type:Citrobacter freundii	Sample_Data:RAW_FILE_NAME=B505_2N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B505_2P	Sample Type:Citrobacter freundii	Sample_Data:RAW_FILE_NAME=B505_2P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B505_3N	Sample Type:Citrobacter freundii	Sample_Data:RAW_FILE_NAME=B505_3N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B505_3P	Sample Type:Citrobacter freundii	Sample_Data:RAW_FILE_NAME=B505_3P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B505_4N	Sample Type:Citrobacter freundii	Sample_Data:RAW_FILE_NAME=B505_4N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B505_4P	Sample Type:Citrobacter freundii	Sample_Data:RAW_FILE_NAME=B505_4P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B505_5N	Sample Type:Citrobacter freundii	Sample_Data:RAW_FILE_NAME=B505_5N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B505_5P	Sample Type:Citrobacter freundii	Sample_Data:RAW_FILE_NAME=B505_5P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B505_6N	Sample Type:Citrobacter freundii	Sample_Data:RAW_FILE_NAME=B505_6N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B505_6P	Sample Type:Citrobacter freundii	Sample_Data:RAW_FILE_NAME=B505_6P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B424_1N	Sample Type:Enterobacter ludwigii	Sample_Data:RAW_FILE_NAME=B424_1N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B424_1P	Sample Type:Enterobacter ludwigii	Sample_Data:RAW_FILE_NAME=B424_1P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B424_2N	Sample Type:Enterobacter ludwigii	Sample_Data:RAW_FILE_NAME=B424_2N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B424_2P	Sample Type:Enterobacter ludwigii	Sample_Data:RAW_FILE_NAME=B424_2P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B424_3N	Sample Type:Enterobacter ludwigii	Sample_Data:RAW_FILE_NAME=B424_3N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B424_3P	Sample Type:Enterobacter ludwigii	Sample_Data:RAW_FILE_NAME=B424_3P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B424_4N	Sample Type:Enterobacter ludwigii	Sample_Data:RAW_FILE_NAME=B424_4N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B424_4P	Sample Type:Enterobacter ludwigii	Sample_Data:RAW_FILE_NAME=B424_4P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B424_5N	Sample Type:Enterobacter ludwigii	Sample_Data:RAW_FILE_NAME=B424_5N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B424_5P	Sample Type:Enterobacter ludwigii	Sample_Data:RAW_FILE_NAME=B424_5P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B424_6N	Sample Type:Enterobacter ludwigii	Sample_Data:RAW_FILE_NAME=B424_6N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B424_6P	Sample Type:Enterobacter ludwigii	Sample_Data:RAW_FILE_NAME=B424_6P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B313_1N	Sample Type:Klebsiella oxytoca	Sample_Data:RAW_FILE_NAME=B313_1N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B313_1P	Sample Type:Klebsiella oxytoca	Sample_Data:RAW_FILE_NAME=B313_1P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B313_2N	Sample Type:Klebsiella oxytoca	Sample_Data:RAW_FILE_NAME=B313_2N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B313_2P	Sample Type:Klebsiella oxytoca	Sample_Data:RAW_FILE_NAME=B313_2P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B313_3N	Sample Type:Klebsiella oxytoca	Sample_Data:RAW_FILE_NAME=B313_3N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B313_3P	Sample Type:Klebsiella oxytoca	Sample_Data:RAW_FILE_NAME=B313_3P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B313_4N	Sample Type:Klebsiella oxytoca	Sample_Data:RAW_FILE_NAME=B313_4N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B313_4P	Sample Type:Klebsiella oxytoca	Sample_Data:RAW_FILE_NAME=B313_4P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B313_5N	Sample Type:Klebsiella oxytoca	Sample_Data:RAW_FILE_NAME=B313_5N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B313_5P	Sample Type:Klebsiella oxytoca	Sample_Data:RAW_FILE_NAME=B313_5P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B313_6N	Sample Type:Klebsiella oxytoca	Sample_Data:RAW_FILE_NAME=B313_6N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B313_6P	Sample Type:Klebsiella oxytoca	Sample_Data:RAW_FILE_NAME=B313_6P.wiff
SUBJECT_SAMPLE_FACTORS           	-	LB1N	Sample Type:LB medium	Sample_Data:RAW_FILE_NAME=LB1N.wiff
SUBJECT_SAMPLE_FACTORS           	-	LB1P	Sample Type:LB medium	Sample_Data:RAW_FILE_NAME=LB1P.wiff
SUBJECT_SAMPLE_FACTORS           	-	LB2N	Sample Type:LB medium	Sample_Data:RAW_FILE_NAME=LB2N.wiff
SUBJECT_SAMPLE_FACTORS           	-	LB2P	Sample Type:LB medium	Sample_Data:RAW_FILE_NAME=LB2P.wiff
SUBJECT_SAMPLE_FACTORS           	-	LB3N	Sample Type:LB medium	Sample_Data:RAW_FILE_NAME=LB3N.wiff
SUBJECT_SAMPLE_FACTORS           	-	LB3P	Sample Type:LB medium	Sample_Data:RAW_FILE_NAME=LB3P.wiff
SUBJECT_SAMPLE_FACTORS           	-	LB4N	Sample Type:LB medium	Sample_Data:RAW_FILE_NAME=LB4N.wiff
SUBJECT_SAMPLE_FACTORS           	-	LB4P	Sample Type:LB medium	Sample_Data:RAW_FILE_NAME=LB4P.wiff
SUBJECT_SAMPLE_FACTORS           	-	LB5N	Sample Type:LB medium	Sample_Data:RAW_FILE_NAME=LB5N.wiff
SUBJECT_SAMPLE_FACTORS           	-	LB5P	Sample Type:LB medium	Sample_Data:RAW_FILE_NAME=LB5P.wiff
SUBJECT_SAMPLE_FACTORS           	-	LB6N	Sample Type:LB medium	Sample_Data:RAW_FILE_NAME=LB6N.wiff
SUBJECT_SAMPLE_FACTORS           	-	LB6P	Sample Type:LB medium	Sample_Data:RAW_FILE_NAME=LB6P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B108_1N	Sample Type:Nseudomonas protegens	Sample_Data:RAW_FILE_NAME=B108_1N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B108_2N	Sample Type:Nseudomonas protegens	Sample_Data:RAW_FILE_NAME=B108_2N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B108_3N	Sample Type:Nseudomonas protegens	Sample_Data:RAW_FILE_NAME=B108_3N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B108_4N	Sample Type:Nseudomonas protegens	Sample_Data:RAW_FILE_NAME=B108_4N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B108_5N	Sample Type:Nseudomonas protegens	Sample_Data:RAW_FILE_NAME=B108_5N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B108_6N	Sample Type:Nseudomonas protegens	Sample_Data:RAW_FILE_NAME=B108_6N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B108_1P	Sample Type:Pseudomonas protegens	Sample_Data:RAW_FILE_NAME=B108_1P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B108_2P	Sample Type:Pseudomonas protegens	Sample_Data:RAW_FILE_NAME=B108_2P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B108_3P	Sample Type:Pseudomonas protegens	Sample_Data:RAW_FILE_NAME=B108_3P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B108_4P	Sample Type:Pseudomonas protegens	Sample_Data:RAW_FILE_NAME=B108_4P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B108_5P	Sample Type:Pseudomonas protegens	Sample_Data:RAW_FILE_NAME=B108_5P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B108_6P	Sample Type:Pseudomonas protegens	Sample_Data:RAW_FILE_NAME=B108_6P.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC1N	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC1N.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC1P	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC1P.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC2N	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC2N.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC2P	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC2P.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC3N	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC3N.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC3P	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC3P.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC4N	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC4N.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC4P	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC4P.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC5N	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC5N.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC5P	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC5P.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC6N	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC6N.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC6P	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC6P.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC7N	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC7N.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC7P	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC7P.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC8N	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC8N.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC8P	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC8P.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC9N	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC9N.wiff
SUBJECT_SAMPLE_FACTORS           	-	QC9P	Sample Type:Quality Control	Sample_Data:RAW_FILE_NAME=QC9P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B585_1N	Sample Type:Serratia plymuthica	Sample_Data:RAW_FILE_NAME=B585_1N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B585_1P	Sample Type:Serratia plymuthica	Sample_Data:RAW_FILE_NAME=B585_1P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B585_2N	Sample Type:Serratia plymuthica	Sample_Data:RAW_FILE_NAME=B585_2N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B585_2P	Sample Type:Serratia plymuthica	Sample_Data:RAW_FILE_NAME=B585_2P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B585_3N	Sample Type:Serratia plymuthica	Sample_Data:RAW_FILE_NAME=B585_3N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B585_3P	Sample Type:Serratia plymuthica	Sample_Data:RAW_FILE_NAME=B585_3P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B585_4N	Sample Type:Serratia plymuthica	Sample_Data:RAW_FILE_NAME=B585_4N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B585_4P	Sample Type:Serratia plymuthica	Sample_Data:RAW_FILE_NAME=B585_4P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B585_5N	Sample Type:Serratia plymuthica	Sample_Data:RAW_FILE_NAME=B585_5N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B585_5P	Sample Type:Serratia plymuthica	Sample_Data:RAW_FILE_NAME=B585_5P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B585_6N	Sample Type:Serratia plymuthica	Sample_Data:RAW_FILE_NAME=B585_6N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B585_6P	Sample Type:Serratia plymuthica	Sample_Data:RAW_FILE_NAME=B585_6P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B253_1N	Sample Type:Sphingobacterium faecium	Sample_Data:RAW_FILE_NAME=B253_1N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B253_1P	Sample Type:Sphingobacterium faecium	Sample_Data:RAW_FILE_NAME=B253_1P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B253_2N	Sample Type:Sphingobacterium faecium	Sample_Data:RAW_FILE_NAME=B253_2N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B253_2P	Sample Type:Sphingobacterium faecium	Sample_Data:RAW_FILE_NAME=B253_2P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B253_3N	Sample Type:Sphingobacterium faecium	Sample_Data:RAW_FILE_NAME=B253_3N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B253_3P	Sample Type:Sphingobacterium faecium	Sample_Data:RAW_FILE_NAME=B253_3P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B253_4N	Sample Type:Sphingobacterium faecium	Sample_Data:RAW_FILE_NAME=B253_4N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B253_4P	Sample Type:Sphingobacterium faecium	Sample_Data:RAW_FILE_NAME=B253_4P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B253_5N	Sample Type:Sphingobacterium faecium	Sample_Data:RAW_FILE_NAME=B253_5N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B253_5P	Sample Type:Sphingobacterium faecium	Sample_Data:RAW_FILE_NAME=B253_5P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B253_6N	Sample Type:Sphingobacterium faecium	Sample_Data:RAW_FILE_NAME=B253_6N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B253_6P	Sample Type:Sphingobacterium faecium	Sample_Data:RAW_FILE_NAME=B253_6P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B263_1N	Sample Type:Stenotrophomonas maltophilia	Sample_Data:RAW_FILE_NAME=B263_1N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B263_1P	Sample Type:Stenotrophomonas maltophilia	Sample_Data:RAW_FILE_NAME=B263_1P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B263_2N	Sample Type:Stenotrophomonas maltophilia	Sample_Data:RAW_FILE_NAME=B263_2N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B263_2P	Sample Type:Stenotrophomonas maltophilia	Sample_Data:RAW_FILE_NAME=B263_2P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B263_3N	Sample Type:Stenotrophomonas maltophilia	Sample_Data:RAW_FILE_NAME=B263_3N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B263_3P	Sample Type:Stenotrophomonas maltophilia	Sample_Data:RAW_FILE_NAME=B263_3P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B263_4N	Sample Type:Stenotrophomonas maltophilia	Sample_Data:RAW_FILE_NAME=B263_4N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B263_4P	Sample Type:Stenotrophomonas maltophilia	Sample_Data:RAW_FILE_NAME=B263_4P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B263_5N	Sample Type:Stenotrophomonas maltophilia	Sample_Data:RAW_FILE_NAME=B263_5N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B263_5P	Sample Type:Stenotrophomonas maltophilia	Sample_Data:RAW_FILE_NAME=B263_5P.wiff
SUBJECT_SAMPLE_FACTORS           	-	B263_6N	Sample Type:Stenotrophomonas maltophilia	Sample_Data:RAW_FILE_NAME=B263_6N.wiff
SUBJECT_SAMPLE_FACTORS           	-	B263_6P	Sample Type:Stenotrophomonas maltophilia	Sample_Data:RAW_FILE_NAME=B263_6P.wiff
#COLLECTION
CO:COLLECTION_SUMMARY            	Individual bacterial LB cultures (150 rpm/min at 28 °C for 72 h) for each
CO:COLLECTION_SUMMARY            	strain including K. oxytoca B313 were centrifuged at 3000 rpm/min for 5 min.
CO:SAMPLE_TYPE                   	Bacterial cells
#TREATMENT
TR:TREATMENT_SUMMARY             	20 μL of supernatant filtered with a 0.2 μm filter was mixed with 120 μL of
TR:TREATMENT_SUMMARY             	precooled 50% methanol (4 °C for 24 h), vortexed for 1 min, and incubated at
TR:TREATMENT_SUMMARY             	room temperature for 10 min.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	The mixture was then stored overnight at -20 °C and centrifuged at 4,000 ×g
SP:SAMPLEPREP_SUMMARY            	for 20 min, and the supernatants were transferred into a new glass vial for the
SP:SAMPLEPREP_SUMMARY            	LC-MS analysis.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	All samples from the nine groups were analyzed by a LC-MS system. First, all
CH:CHROMATOGRAPHY_SUMMARY        	chromatographic separations were performed using an ultra-performance liquid
CH:CHROMATOGRAPHY_SUMMARY        	chromatography (UPLC) system (SCIEX, UK) equipped with an ACQUITY UPLC BEH Amide
CH:CHROMATOGRAPHY_SUMMARY        	column (100 mm×2.1 mm, 1.7 µm, Waters, UK) for the reversed-phase separation.
CH:CHROMATOGRAPHY_SUMMARY        	The column oven was maintained at 35 °C. The flow rate was 0.4 mL/min and the
CH:CHROMATOGRAPHY_SUMMARY        	mobile phase consisted of solvent A (25 mM ammonium acetate) and solvent B
CH:CHROMATOGRAPHY_SUMMARY        	(isopropanol: acetonitrile =9:1 + 0.1% formic acid). Gradient elution conditions
CH:CHROMATOGRAPHY_SUMMARY        	were set as follows: 0-0.5 min, 95% B; 0.5-9.5 min, 95% to 65% B; 9.5-10.5 min,
CH:CHROMATOGRAPHY_SUMMARY        	65%-40% B; 10.5-12 min, 40% B; 12-12.2 min, 40%-95% B; and 12.2-15 min, 95% B.
CH:CHROMATOGRAPHY_SUMMARY        	The injection volume for each sample was 4 µL.
CH:INSTRUMENT_NAME               	Unspecified
CH:COLUMN_NAME                   	Waters Acquity BEH Amide (150 x 2.1mm,1.7um)
CH:COLUMN_TEMPERATURE            	35
CH:FLOW_GRADIENT                 	0-0.5 min, 95% B; 0.5-9.5 min, 95% to 65% B; 9.5-10.5 min, 65%-40% B; 10.5-12
CH:FLOW_GRADIENT                 	min, 40% B; 12-12.2 min, 40%-95% B; and 12.2-15 min, 95% B.
CH:FLOW_RATE                     	0.4 mL/min
CH:SOLVENT_A                     	100% water; 25 mM ammonium acetate
CH:SOLVENT_B                     	90% isopropanol/10% acetonitrile; 0.1% formic acid
CH:CHROMATOGRAPHY_TYPE           	HILIC
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	ABI Sciex 5600+ TripleTOF
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	EI
MS:MS_COMMENTS                   	A high-resolution tandem mass spectrometer TripleTOF 5600 plus (SCIEX, UK) was
MS:MS_COMMENTS                   	used to detect metabolites eluted from the column in both positive and negative
MS:MS_COMMENTS                   	ion modes with the curtain gas set to 30 psi, ion source gas1 set to 60 psi, ion
MS:MS_COMMENTS                   	source gas2 set to 60 psi, and an interface heater temperature set to 650 °C.
MS:MS_COMMENTS                   	The ionspray floating voltage was set to 5000 V for positive ion mode and -4500
MS:MS_COMMENTS                   	V for negative ion mode. The mass spectrometry data were acquired in IDA mode,
MS:MS_COMMENTS                   	and the TOF mass range was from 60 to 1200 Da. Ion survey scans were completed
MS:MS_COMMENTS                   	in 150 ms, and 12 product ions were collected if they exceeded a threshold of
MS:MS_COMMENTS                   	100 counts per second (counts/s) and with a 1+ charge-state. The total cycle
MS:MS_COMMENTS                   	time was set to 0.56 s. Four bins were summed during each scan at a pulser
MS:MS_COMMENTS                   	frequency of 11 kHz through the monitoring of the 40 GHz multichannel TDC
MS:MS_COMMENTS                   	detector with four-anode/channel detection. Dynamic exclusion was set for 4 s.
MS:MS_COMMENTS                   	During mass spectrometry data acquisition, the mass accuracy was calibrated
MS:MS_COMMENTS                   	every 10 samples. Furthermore, to maintain the stability of the LC-MS system
MS:MS_COMMENTS                   	during the whole analysis, all samples from the nine groups were analyzed in a
MS:MS_COMMENTS                   	random sequence, and a QC sample was analyzed every 10 samples.
MS:ION_MODE                      	POSITIVE
MS:MS_RESULTS_FILE               	ST001511_AN002504_Results.txt	UNITS:peak area	Has m/z:Yes	Has RT:Yes	RT units:Seconds
#END