Determination of trimipramine and metabolites in plasma by liquid chromatography with electrochemical detection

A procedure for the determination of trimipramine, the demethyl, 2-hydroxy, and 2-hydroxy demethyl metabolites in plasma by liquid chromatography with electrochemical detection is described. A 1-mL plasma sample is made alkaline with a carbonate buffer (pH 9.8) and extracted with 20% ethyl acetate in n-heptane. After back-extraction into an acid phosphate buffer, an aliquot is injected onto a reverse-phase trimethylsilyl-packed column and eluted with a phosphate buffer-acetonitrile mobile phase (65:35) containing n-butylamine. The peaks were detected at +1.1 V versus the silver-silver chloride reference electrode. The method provides absolute recoveries of 60-91% and a day-to-day precision of less than 9% for all compounds. The minimum quantifiable level for all compounds was 3 ng/mL. Steady-state plasma concentration data for 29 depressed patients receiving either 75 mg or 150 mg/d is reported.