RESUMO
A general global analysis of single-photon timing data is presented in which each fluorescence decay curve can be described by a different decay law. The model parameters can be held in common within one curve and/or between related curves. Any or all parameters can be kept fixed, or they may be variable to seek optimum values. This general analysis allows the determination of activation energies, frequency factors and quenching rate constants in one step. The construction of the global mapping table which relates parameters in one experiment to those in another is explained in detail. The use and performance of this general simultaneous analysis are examined using tryptophan fluorescence decays at pH 6.0 obtained at various emission wavelengths as a function of temperature and added solute quencher. The results show that tryptophan at pH 6.0 decays as a biexponential with decay times which are independent of the analysis wavelength. The decay component with the short lifetime has a deactivation rate constant of 1.4 x 10(9) s-1 independent of temperature. The decay component with the long lifetime has an activation energy of 28 kJ/mol and a frequency factor of 3 x 10(13) s-1; its temperature-independent decay rate constant equals 1 x 10(8) s-1. Recursion formulas for a computer program to estimate activation energies, frequency factors, and decay rate constants are provided.