Resibufogenin can be eluted out with silica gel column chromatography, using a 5:1 ratio of cyclohexane to acetone for the solvent in the mobile phase.

[citation needed] In human adrenocortical cells, cinobufagin inhibits the secretion of aldosterone and cortisol.

Cinobufagin first binds to a Ca2+/K+ plasma membrane ATPase, subsequently inducing the phosphorylation of extracellular signal-regulated kinases (ERK).

It is thought to trigger increased synthesis of β-END and the up-regulation of the mu opioid receptor in mouse tumor tissue thereby leading to pain relief.

[5] C. elegans can catabolize cinobufagin into five distinct metabolites, each of which has been shown to have cytotoxic effects to HeLa cancer cells.