HPLC determination of doxorubicin, doxorubicinol and four aglycone metabolites in plasma of AIDS patients.

Abstract

A high-performance liquid chromatographic (HPLC) assay has been developed for the determination of the anticancer drug doxorubicin and the metabolites doxorubicinol, doxorubicinone, 7-deoxydoxorubicinone, doxorubicinolone and 7-deoxydoxorubicinolone in plasma of AIDS patients. Samples can be heated at 60 degrees C for 30 min to inactivate the human immunodeficiency virus. The sample pre-treatment involves a liquid-liquid extraction of the buffered plasma sample (pH 9) with a chloroform-1-propanol (4:1, v/v) mixture. The chromatographic analysis is performed on a Lichrosorb RP-8 (5 microns) column and by isocratic elution with a mobile phase of acetonitriletetrahydrofuran-phosphate buffer (pH 2.2) (800:5:200, w/w/w) with fluorescence detection (excitation wavelength: 460 nm; emission wavelength: 550 nm). The proposed method has been validated and, subsequently, implemented in a pharmacokinetic study of doxorubicin in AIDS patients with Kaposi's sarcoma who are treated with the combination regimen doxorubicin, vincristine and bleomycin.