#METABOLOMICS WORKBENCH fangyuan_zhou_20200910_172015 DATATRACK_ID:2164 STUDY_ID:ST001511 ANALYSIS_ID:AN002505 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 NEGATIVE MS:MS_RESULTS_FILE ST001511_AN002505_Results.txt UNITS:peak area Has m/z:Yes Has RT:Yes RT units:Seconds #END