Simultaneous determination of clomipramine and its N-desmethyl metabolite in human whole blood by capillary gas chromatography with mass-selective detection

A method for the simultaneous determination of clomipramine and its N-desmethyl metabolite at concentrations down to ca. 2 nmol/l in human whole blood is described. After addition of a known amount of deuterium-labelled internal standards, compounds are extracted into n-heptane-isoamyl alcohol (99:1, v/v) at basic pH, back-extracted into an acidic aqueous solution and re-extracted at basic pH into n-heptane. N-Desmethylclomipramine and the internal standard are derivatized with pentafluoropropionic anhydride. The compounds are determined by capillary gas chromatography with mass-selective detection. The technique was applied to determine the human blood concentrations of clomipramine and its N-desmethyl metabolite after oral administration of Anafranil; mean blood concentrations are reported.