Targeted Submetabolome Profiling of Bufadienolides in Venenum Bufonis Using Scheduled Diagnostic Product Ion-Pair Filtering and Substructure Recognition-Based Strategy.
Xuelin Sun, Wei Ding, Zhanpeng Shang, Jiantao Qiu, Xueying Tan
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引用次数: 0
Abstract
Introduction: Venenum bufonis (VB), a traditional animal-derived medicine, exhibits significant antitumor, immunomodulatory, and analgesic activities that are primarily attributed to bufadienolides.
Objective: This study presented a novel targeted submetabolomics strategy for comprehensive profiling of bufadienolides in VB.
Material and methods: This targeted submetabolome mainly integrated scheduled diagnostic product ion (DPI) pair-based filtering and substructure recognition-based structural assembly based on the LC-MS platform. Key steps included 1) stepwise multiple ion monitoring (MIM) coupled with a dynamic exclusion function to enhance detection of minor peaks; 2) automated annotation of unconjugated bufadienolides via high-resolution extracted ion chromatograms (EICs) and a Microsoft Excel-based platform; 3) establishment of an extended tail database for conjugated bufadienolides (including dicarboxylic acid and dicarboxylic acid-amino acid conjugates) based on structural similarity and derivativity, enabling untargeted screening via validated or scheduled DPI pairs derived from MS/MS fragmentation patterns; and 4) potential conjugated bufadienolides were characterized via a substructure recognition-based strategy.
Results: The strategy identified 162 bufadienolides (39 primary, 39 secondary, and 84 tertiary) with a 96% annotation rate (155 characterized). Notably, 81 potential novel compounds were discovered, including 11 previously undescribed structural subclasses.
Conclusions: This targeted submetabolome strategy significantly expanded the diversity of bufadienolides in VB and also advanced the application of mass spectrometry in natural product discovery.
期刊介绍：
Phytochemical Analysis is devoted to the publication of original articles concerning the development, improvement, validation and/or extension of application of analytical methodology in the plant sciences. The spectrum of coverage is broad, encompassing methods and techniques relevant to the detection (including bio-screening), extraction, separation, purification, identification and quantification of compounds in plant biochemistry, plant cellular and molecular biology, plant biotechnology, the food sciences, agriculture and horticulture. The Journal publishes papers describing significant novelty in the analysis of whole plants (including algae), plant cells, tissues and organs, plant-derived extracts and plant products (including those which have been partially or completely refined for use in the food, agrochemical, pharmaceutical and related industries). All forms of physical, chemical, biochemical, spectroscopic, radiometric, electrometric, chromatographic, metabolomic and chemometric investigations of plant products (monomeric species as well as polymeric molecules such as nucleic acids, proteins, lipids and carbohydrates) are included within the remit of the Journal. Papers dealing with novel methods relating to areas such as data handling/ data mining in plant sciences will also be welcomed.
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