Comparative Analysis of the Bufonis Venenum by Using TLC, HPLC, and LC-MS for Different Extraction Methods.

Area of Science:

• Pharmacology
• Natural Product Chemistry
• Traditional Oriental Medicine

Background:

• Toad venom (Chan-Su) is a traditional Oriental medicine used for pain, heart conditions, and cancer.
• It is sourced from Bufo species and widely used in Asian countries.
• Key chemical constituents include serotonin and bufodienolides.

Purpose of the Study:

• To quantify main chemical constituents in commercial toad venom.
• To compare the efficacy of different extraction methods on constituent concentrations.
• To understand how solvent choice affects the enrichment of active compounds.

Main Methods:

• Extraction using water, ethanol (EtOH), methanol (MeOH), or ethyl acetate (EtOAc).
• Fractionation via precipitation or reflux.
• Analysis using thin layer chromatography (TLC), high-performance liquid chromatography (HTLC), and liquid chromatography-mass spectrometry (LC-MS).

Main Results:

• Serotonin was highest in water extracts (75.7 mg/g); bufodienolides were low (3.8 mg/g).
• Methanol or ethanol extracts showed 5-26 times higher bufodienolide concentrations with minimal serotonin.
• Ethanol extracts yielded the highest total bufodienolides; EtOAc precipitation yielded the lowest.

Conclusions:

• Toad venom contains serotonin and bufodienolides.
• Solvent selection is crucial for enriching specific active components.
• Optimizing extraction can enhance toad venom's therapeutic potential for various conditions.