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Statistical analyses of high-performance liquid chromatography (HPLC)-based chemical fingerprints of Bufo bufo gargarizans toad skin extracts were integrated with antitumor evaluations as a means to more effectively identify bioactive constituents. Specifically, chemical fingerprints of Bufo bufo gargarizans skin extract samples obtained from ten different regions in China were generated by HPLC, and then subjected to similarity analysis and principal component analysis (PCA). Concurrently, a MCF-7 breast cancer cell model was adopted to obtain the antitumor activity of different toad skin extracts. The chromatographic fingerprint-bioactivity relationships of the Bufo bufo gargarizans extracts were then evaluated by Pearson correlation analysis. The results demonstrated that all skin extract samples inhibited the proliferation of MCF-7 cells to some extent, and that there was a correlation between the chemical fingerprints and the anti-proliferative activity. Bufotalin, bufalin, and cinobufagin, three known components of Bufo bufo gargarizans that were identified in the chemical fingerprints, were highlighted as constituents responsible for the observed bioactivity. The activity of cinobufagin was confirmed by cell viability and colony formation assays using MCF-7 cells. The approach documented here provides a more effective means to associate chemical information on medicinal extracts with the principle components responsible for their bioactivity, and to therefore expedite drug discovery.

Chromatographic Fingerprint-Bioactivity Relationships of the Antitumor Properties of Bufo bufo gargarizans Skin Extracts