A new Monooxygenase Product from 7-Ethoxycoumarin and its Relation to the O-Dealkylation Reaction

Abstract
The widely used fluorometric microsomal monooxygenase test for 7-ethoxycoumarin O-dealkylation was reinvestigated with regard to other possible hydroxylation products. By HPLC(high-performance liquid chromatography)-analysis no .beta.-hydroxylation of the ethyl group and no 8-hydroxylation could be detected. Only a smalll percentage of 6-hydroxylation occurred, but as a new metaboite 7-ethoxy-3-hydroxycoumarin was found in quantities depending on the microsomal preparation used. The ratio of O-delakylation to 3-hydroxylation varied according to species, induction, buffer and pH, suggesting that different isozymes of cytochrome P450 were involved. The isozyme mainly responsible for 3-hydroxylation exhibited a great dependence on cytochrome b5 as the donor for the second electron. The fluorometric test does not include 3-hydroxylation due to the virtual absence of an emission spectrum above 450 nm.

This publication has 25 references indexed in Scilit: