The ADx total serum tricyclic antidepressant (TCA) fluorescence polarization immunoassay (Abbott Diagnostics) for the semi-quantitation of imipramine or amitriptyline and their respective N-demethylated metabolites in cases of TCA overdose was evaluated. The assay is linear from 75-1000 ng/mL total TCA in serum, and flaggs as "HI" all results exceeding 300 ng/mL. The within and between run precision of the assay for patient serum containing imipramine or amitriptyline and their metabolites gave CV's of less than 5.5% and 8.9%, respectively. A good correlation between the results of patient serum containing imipramine and desipramine simultaneously analyzed by ADx and gas liquid chromatography (GC) was observed, r2 = 0.964, n = 32. Results of patient serum containing amitriptyline and nortriptyline or doxepin and desmethyldoxepin analyzed by ADx and GC or GC-mass spectrometry were not well correlated; r2 = 0.738, n = 44 and r2 = 0.695, n = 21, respectively. The assay consistently flagged as "HI" serum with total imipramine and desipramine concentrations above 300 ng/mL by GC, and serum with greater than 360 ng/mL of amitriptyline and nortriptyline by GC/MS. No significant cross-reactivity was observed for drugs other than the TCA.