[Comparison of chemical composition between fresh and processed Bufonis Venenum by UPLC-TQ-MS]

Area of Science:

• Pharmacology
• Analytical Chemistry
• Natural Products

Background:

• Toad venom, specifically Bufonis Venenum from Bufo species, contains bufadienolides with potential anti-cancer properties.
• Bufadienolides exist in both free (bufogenin) and conjugated (bufotoxin) forms, differing in structure and potentially activity.
• Understanding these structural differences is crucial for characterizing toad venom's therapeutic potential.

Purpose of the Study:

• To establish and validate an Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-TQ-MS) method.
• To identify and quantify major bufadienolides in Bufonis Venenum from B. bufo gargarizans.
• To differentiate between free-type (bufogenin) and conjugated-type bufadienolides and assess processing effects.

Main Methods:

• A UPLC-TQ-MS method was developed using a Waters ACQUITY UPLC BEH C column.
• Analysis was performed in positive ion mode with a mobile phase of aqueous formic acid and acetonitrile via gradient elution.
• Component identification was achieved through MS/MS analysis, comparing retention times and high-resolution mass data.

Main Results:

• The UPLC-TQ-MS method successfully identified 37 effective bufadienolide components in Bufonis Venenum.
• Twenty-six free-type bufadienolides (bufogenins) and 11 conjugated-type bufadienolides were identified.
• Significant compositional differences were observed between fresh and processed toad venom, with processed venom showing reduced conjugated-type content.

Conclusions:

• The established UPLC-TQ-MS method is effective for analyzing bufadienolides in toad venom.
• Processing toad venom drastically alters its chemical composition, particularly affecting the ratio of free to conjugated bufadienolides.
• Further research into these altered compositions may reveal new therapeutic applications or impact existing ones.