Abbott Laboratories Corporation

Daryl E Hartter

Maciej B Adamczyk

Jeffrey R Fishpaugh

Donald D Johnson

Robert E Hruska

Charles A Harrington

Immunoassay methods and reagents for the specific quantification of amitriptyline or nortriptyline in a test sample are disclosed employing antibodies prepared with amitriptyline or nortriptyline derivatives of the Formula III: ##STR1## wherein for amitriptyline, R is CH.sub.3, and for nortriptyline, R is H. The present invention also describes the synthesis of unique fluorescein tracers of the structure of Formula IV and Formula V: ##STR2## wherein for a specific amitriptyline immunoassay, W.sub.1 is a heteroatom linked to the aromatic ring at the 2 or 3 position, and for a specific nortriptyline immunoassay, W.sub.2 is two heteroatoms linked together and attached to the aromatic ring at the 2 or 3 position, and wherein Q is a detectable moiety, preferably fluorescein or a fluorescein derivative.

Reagents and methods for the quantification of amitriptyline or

Method and reagents for determining a compound of interest present in a test sample also containing one or more interfering compounds having substantially similar chemical structures, and otherwise analytically indistinguishable from each other, employing a pretreatment reagent capable of selectively modifying the chemical structure of one of the compounds without significantly modifying or altering the chemical structure of the other one of the compounds. The selective modification results in the modified compound having a chemical structure which is substantially dissimilar to the chemical structure of the other one of said compounds wherein the compounds are substantially distinguishable from each other to permit the analytical determination of one or the other of such compounds by immunoassay, high pressure liquid chromatography, and thin layer chromatography techniques, especially for the fluorescent polarization immunoassay determination of tricyclic antidepressant drugs in the presence of phenothiazines.

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Mundelein, IL, United States of America

Daryl E Hartter