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High pressure liquid chromatography procedures are reported for the determination of amitriptyline and its metabolite, nortriptyline, and for imipramine and its metabolite, desipramine. Separations and quantitations were carried out at ambient temperature using a silica column with a variable wavelength UV detector. The procedures have been used successfully on clinical samples (serum) and on forensic samples (hemolyzed blood). If need be, analysis can be performed on 0.25 mL serum. Several potential interferences, related compounds and drugs, have been evaluated.