ABSTRACT
The measurement of intracellular concentrations of the anti-cancer drug doxorubicin was performed by the application of a simple cell extraction technique combined with a rapid high-performance liquid chromatographic separation. Quantitation was done by fluorescence detection. The extraction procedure was non-degradative and the mean recovery of drug was 95%. A high drug extraction efficiency was confirmed with radiolabeled [3H] doxorubicin. The method is applicable to normal and neoplastic tissue.
Subject(s)
Daunorubicin/analysis , Doxorubicin/analysis , Cell Line , Chromatography, High Pressure Liquid/methods , Daunorubicin/pharmacology , Doxorubicin/pharmacology , Humans , Leukemia/blood , Lymphocytes/analysis , Monocytes/analysisSubject(s)
Benzopyrenes/metabolism , Kidney/metabolism , Lung/metabolism , Microsomes, Liver/metabolism , Animals , Cricetinae , Humans , In Vitro Techniques , Kinetics , Male , Mesocricetus , Methylcholanthrene/pharmacology , Microsomes/metabolism , Organ Specificity , Phenobarbital/pharmacology , RatsABSTRACT
Optical spectral repetitive scan analysis during the oxidative metabolism of benzo(a)pyrene by liver microsomal suspensions reveals the time-dependent formation of an intermediate(s) of which the visible spectra resemble those of several benzo(a)pyrene phenols. Liver microsomes from 3-methylcholanthrene-treated rats showed a greater rate of formation of the phenols than did microsomes from control animals; the rate of formation catalyzed by liver microsomes from phenobarbital-pretreated rats was intermediate. When 3-hydroxybenzo(a)pyrene was used as a standard for comparison of activity, the rates of formation of phenols were compared when measured by fluorometric, spectrophotometric, or high-pressure liquid chromatographic analytical techniques. An epoxide hydrase inhibitor, 1,1,1-trichloropropene-2,3-oxide, enhanced phenol formation regardless of the source of liver microsomes, and 7,8-benzoflavone inhibited control and 3-methylcholanthrene-induced microsomal metabolism of benzo(a)pyrene, 7,8-Benzoflavone did not effect benzo(a)pyrene metabolism by liver microsomes from phenobarbital-pretreated rats. The effect of inhibitors on the spectrophotometric assay correlates well with the results obtained from benzo(a)pyrene metabolite analysis using high-pressure liquid chromatography.