Analytical application of the co-fluorescence effect in detection of europium, terbium, samarium and dysprosium with time-resolved fluorimetry

Application of the co-fluorescence effect has been examined for the simultaneous detection of the lanthanide ions Eu(3+), Tb(3+), Sm(3+) and Dy(3+). In the presence of Y(3+) and 2,2'-bipyridine (BP), the fluorescence intensities of the pivaloyltrifluoroacetone chelates of these lanthanides were greatly enhanced by an inter-chelate energy transfer process. Under optimized conditions, the following detection limits were obtained; 0.019 pM for Eu(3+), 0.27 pM for Tb(3+), 3.8 pM for Sm(3+) and 20 pM for Dy(3+), when a sensitive time-resolved fluorometer was used for the measurement. A co-fluorescence based fluorescence enhancement solution was also tested in a dual-label time-resolved immunofluorometric assay of luteinizing and follicle stimulating hormones (LH and FSH) based on the use of Eu(3+) and Tb(3+) as the label ions. The present fluorometric detection system is particularly well suited for multilabel time-resolved fluorometric immunoassays utilizing two, three or four ion labels simultaneously.