A method for the simultaneous analysis of cardiac steroids by high-pressure liquid chromatography has been developed. Reversed phase chromatography on a μ-Bondapak C18 column was particularly effective for the separation of sixteen bufadienolides and nine cardenolides. Determination of the principal bufogenins in the fresh venom of the Japanese toad was carried out by the internal standard method. The relationship between chemical structures and retention values is also discussed.
