Re-evaluation of the steady-state self-quenching constant of quinine bisulphate from fluorescence measurements in transmission geometry

Abstract

In the present work we show that a recent methodology developed by us to acquire emission spectra and fluorescence quantum yields of highly absorbing samples in transmission configuration, constitutes a very simple and robust alternative to determine self-quenching constants, KSQ. We measured the absorption and the steady-state emission spectra of quinine bisulphate, QBS, solutions ranging between 1.5 × 10−5 and 1.5 × 10−1 M. From these data, we calculated the expected emission spectra, affected by re-absorption, for all QBS concentrations. For higher concentrations, the re-absorption in the excitation/detection direction reaches values up to 6% of the total emitted intensity. The KSQ of the dye was re-evaluated from the concentration dependence of the quotients between the calculated and the experimental integrated emission spectra. The obtained value, KSQ = 18.4 ± 0.1 M−1, shows no significant differences with those obtained from steady-state and average lifetimes by other authors, pointing out the diffusional nature of the self-quenching phenomenon. The present work helps clarify some ambiguous aspects concerning the photophysics of QBS, stressing that re-absorption phenomena must be considered in QBS concentrated solutions for accuracy measurements.