Determination of clomipramine or imipramine and their mono-demethylated metabolites in human blood or plasma by high-performance liquid chromatography.
A high-performance liquid chromatographic method for the quantitative assay of clomipramine or imipramine and their mono-demethylated metabolites in human blood or plasma is described. After addition of the internal standards, imipramine for clomipramine, desipramine for desmethylclomipramine and conversely, the compounds are extracted from blood or plasma at pH 10 into heptane containing 1% isoamyl alcohol; they are then back-extracted into an acidic aqueous phase and re-extracted at a basic pH into heptane. After evaporation, the residue is dissolved in 300 microliters of mobile phase and 150 microliters is injected. Both drugs and their mono-demethylated metabolites are well separated from the blood or plasma components and the other metabolites on a silica gel column using ethanol--hexane-dichloromethane-diethylamine (30:62:8:5 . 10(-3)) as the mobile phase at a flow-rate of 1.5 ml/min. The limit of sensitivity is 5 ng/ml for clomipramine and imipramine and 10 ng/ml for the corresponding mono-demethylated metabolites. This method has been used to analyse plasma from subjects given therapeutic doses of clomipramine or imipramine.
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002997:Clomipramine / Q000097:blood* / Q000378:metabolism; D006801:Humans; D007099:Imipramine / Q000097:blood* / Q000378:metabolism
D002997:Clomipramine; D007099:Imipramine