The interactions of fish sperm deoxyribonucleic acid (DNA) with anthraquinones, such as chrysophanol, physcion and 1,8-dihydroxy anthraquinone, were investigated by using ethidium bromide (EB) as fluorescence probe. The binding constants of anthraquinones and DNA were obtained by the fluorescence quenching technique. Further, the binding mechanisms on the reaction of the three anthraquinones with DNA and effect of ionic strength on the fluorescence property of the system have also been investigated. The results of the assay indicate that the binding modes of chrysophanol, physcion and 1,8-dihydroxy anthraquinone with DNA were evaluated to be groove binding. And the binding constants of chrysophanol, physcion and 1,8-dihydroxy anthraquinone with DNA-EB complex were 1.64x10(4), 3.04x10(4) and 2.88x10(5) l mol(-1), respectively.