After launching the data analysis software, go to the my work column and click on import mass links data to open a new window. Click on the plus sign and select all five raw files from the positive mode acquired by high resolution mass spectrum metric analysis. Enter a group name and click on create unify sample set to start data import.
Double click on the analysis method file in the positive mode. Click on processing, followed by the target by mass button. Enter 5PPM for target match tolerance and fragment match tolerance.
Click on home, and then adducts. Select the rows of protylated and acetylated. Click on the right arrow-shaped button, and then save.
In the my work column, click on analysis. Choose the imported data and method, and then click on next. Enter the file name, select the folder for storing the analysis data, and click finish.
Then, click on the process button. Copy the result table into a blank form for analysis. Next, select a compound in the result table.
The corresponding secondary mass spectrum will appear in the lower right corner. Click on the table-shaped button, and select the high energy fragments option to display the table of secondary fragment ions. Select a secondary fragment ion in the table.
Move the cursor over the ion peak corresponding to the secondary mass spectrum, and the corresponding structural fracture will be displayed. Manually draw the fragmentation pattern in a drawing software. The chemical composition identification of BFH revealed 168 compounds based on the compound results of six extractants.
To understand the cleavage pathway in UPLC-TOF-MS for acids, 3-O-trans-coumaroylquinic acid was demonstrated. Cleavage via hydrolysis reaction led to the formation of intermediates. For esters, trans-O-glucosyl-methyl-trans-cinnamate showed conversion into intermediates via carbon-carbon cleavage.
For glycosides cintrucin C transformed into an intermediate via carbon-carbon bond cleavage in the alphalytic chain, or via the breakage of the carbon oxygen carbon bond on the benzene ring. Gene ontology enrichment analysis showed the probable gene functions of the identified compounds. Also, the KEGG pathway annotation analysis showed the pathways for differentially expressed genes.
