Monomerizing effect of caffeine, o-phenanthroline, and tannin on cationic dyes: a model system to analyze spectral characteristics of the intercalative binding to nucleic acids.

If used as co-solutes in concentrated solutions of cationic planar dyes, caffeine, o-phenanthroline, and tannin induce striking hyperchromic and bathochromic shifts in their absorption spectra. Likewise, the fluorescence of acridine orange at high concentration greatly increases in the presence of caffeine, the emission peak appearing at a shorter wave-length. These spectral changes, which are similar to those produced by organic solvents, detergents, and alpha-cyclodextrin, reflect the disaggregating (monomerizing) capacity of the co-solutes on stacked chromophores. After washing with saturated solutions of caffeine or o-phenanthroline, the chromatin fluorescence by intercalating fluorochromes is reduced or abolished, which suggests competition effects for intercalative binding modes. These results support the use of caffeine, o-phenanthroline, and tannin in spectroscopic and histochemical studies of dye-stuff interactions with DNA and chromatin.