Luminescence conditional stability constants

The control of pH in a solution containing a luminescent analyte is also of great importance for sensitive and reliable analysis. For aromatic compounds with acidic or basic substituents, excitation and emission wavelengths of the ionized and free forms are likely to differ. In the case of fluorescence from metal chelates, the pH must be controlled to ensure that the conditional stability constant for the complex is optimal for the particular analytical situation. 

Under stoichiometric conditions, fluorexon and its derivatives form 1 1 complexes with Ln ions. However when the ratio Ln Fx is increased, complexes with other stoichiometries are observed, the exact nature of which has not been determined. On the other hand, luminescence data of solutions with a ratio Yb Fx < 1 clearly indicate the presence of only one luminescent species, the 1 1 complex. Monoexponential luminescence decays are observed corresponding to a lifetime of 1.9 ps, whereas multi-exponential decays are measured when the Yb Fx ratio is increased. Further proof of the existence of 1 1 complexes has been brought by mass spectrometry. Competitive titration with edta has been followed by monitoring the Yb luminescence, since the edta complex is non-luminescent, contrary to the chelate formed with Fx. After addition of 5 equivalents of edta to a solution of [Yb(fx)] in Tris-HCl buffer, the Yb luminescence intensity decreases to 12% of its initial value. The thermodynamic stability of the fluorexon chelate is, therefore, comparable to [Yb(edta)] . In addition, the luminescence decay after addition of edta aliquots is relatively slow, the estimated rate constant being 7.1 x 10" s indicating a reasonably high kinetic stability of the fluorexon chelate. 

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