Abstract
To meet the need for the determination of the low activities of cytochrome P450-catalyzed 7-ethoxycoumarin deethylation (ECDE) in most extra-hepatic tissues and cultured cells, a reliable, sensitive, and accurate assay, which uses a modified column-switching high-performance liquid chromatographic system and is essentially free of sample pretreatment, has been developed. With this system, the recovery of the 7-hydroxycoumarin product added to the biological tissue-containing incubation medium was 87%. With ECDE activity as low as 1 pmol/mg of protein/min, replicate measurements with variation coefficients lower than 5% were easily obtainable. ECDE activities in several extrahepatic tissues and cultured cells were determined with the technique to illustrate its usefulness.
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