Adriamycin and eight metabolites were separated and characterized with an ammonium formate/tetrahydrofuran solvent system on a muBondapak-Phenyl column. Danorubicin and seven daunorubicin metabolites were separated and characterized by the same method. Adriamycin and its major metabolite, adriamycinol, were extracted with 50-70% efficiency from human plasma with chloroform/2-propanol (1:1) and quantified against a daunorubicin internal standard. Urinary concentrations of parent drug, adriamycinol, and the 4-O-sulfate were determined as well. Daunorubicin and daunorubicinol were extracted from human plasma and quantified against an adriamycinol internal standard. The fluorescence detection system has a detection limit of 1.0 pmol, and levels as low as 25 mmol/ml are measurable in plasma.