[Resolution of fluorescence spectra by degree of quenching accessibility]

The novel algorithm QUENCH is described which resolves fluorescence spectra onto two components differing in the accessibility (the Stern-Volmer constants) for small quenchers. In contrast to the known algorithms, the QUENCH first estimates Stern-Volmer constants using the total arrow of spectra measured at different quencher concentrations and, then, calculates contributions of each component into the emission intensities at every wavelength value. The component spectra, resolved with QUENCH from the tryptophan fluorescence of ribosomal S7 protein, were very similar to the log-normal components revealed using the SIMS program [16]. The QUENCH algorithm may be of use in the component analysis of fluorescence spectra of any system containing fluorophores with differing accessibilities for quenchers.