In recent years, fluorescence spectrometry was widely used in quantitative determination of DNA. In this paper, a convenient synthesis of a new fluorescent 2-(2'- Chloro phenyl)- 5- (2'- hydroxyl phenyl)-1,3,4-oxadiazole (HOXD) was realized. Experimental data showed that fluorescence of HOXD could be quenched by DNA and the decreased fluorescence intensity of HOXD resulting from fluorescence quenching is proportional to DNA concentrations suggesting that HOXD could be used as a new fluorescent probe for quantitative determination of DNA. Optimal experimental conditions for DNA analysis were also studied in the paper.