OBJECTIVE ： To determine the concentrations of adriamycin in rabbit plasma and lymph nodes. METHOD ： After addition of the internal standard (IS； daunorubicin), a single-step extraction was performed with chloroform-methyl alcohol (4:1). The organic layer was evaporated and the remains was redissolved with 100 methyl alcohol. The drug and IS were chromatographed on C18 column with a mobile phase consisting, of methyl alcohol ：acetonitrile ：0.01mol/L ammonium dihydric phosphate ：acetic acid(50:22:28: 0.5) at a flow rate of 1.2 ml/min and detected by fluorescence detector at excitation and emission wavelengths of 479 and 587 nm,(respectively. RESULTS ： A better linearity was obtained from 10 to 1 000 ng of adriamycin per ml for plasma and 0.05 to 5.0 μg/g for lymph nodes with a good correlation coefficient(r = 0. 999 9,n = 6). The intra and interday coefficients of variation were always<6.15%. The mean recovery of adriamycin from plasma and lymph nodes was 100. 11 % and 100.5% ,respectively. The improved method was very sensitive; the minimum detectable concentration was 2 ng/ml for plasma and 0. 025 μg/g for lymph nodes, respectively. CONCLUSION ： This method is suitable for pharmacokinetics studies in rabbits.