DICARBOXYLATE AND TETRACARBOXYLATE DERIVATIVES OF PYRIDINES, BIPYRIDINES AND TERPYRIDINES AS LUMINOGENIC REAGENTS FOR TIME-RESOLVED FLUOROMETRIC-DETERMINATION OF TERBIUM AND DYSPROSIUM

A group of compounds composed of pyridine, 2,2'-bipyridine and 2,2':6' ,2''-terpyridine as the energy absorbing, triplet sensitizing moieties and two or four carboxylic acids as the chelating groups were synthes ized and their ability to enhance terbium ion luminescence was elucida ted. The dicarboxylic acid derivatives form highly luminescent Tb3+ ch elates with luminescence quantum yields approaching 50%. Additional gr oups, such as methoxy groups or additional pyridine rings in the aroma tic structure can be utilized to shift the excitation maxima towards l onger wavelengths in order to produce more practicable excitation for practical use. The ligands giving the highest Tb3+ emissions were furt her evaluated in a two-step fluorescence enhancement system constructe d for specific binding assays based on the simultaneous use of three o r four ions as the labels (Eu3+, Tb3+, Sm3+ and Dy3+). In the two-step procedure, the first step comprises dissociative fluorescence enhance ment based on an acidic solution of an aromatic beta-diketone used as the luminogenic ligand, whereafter Eu3+ and Sm3+ are quantitated. In t he second step, the lanthanide ions form new chelates with-the added l igands. These ligands are able to transfer the excitation energy they have absorbed with high efficiency to the chelated ions and produce hi gh ion luminescence for Tb3+ and Dy3+.