Abstract: Ofloxacin was separated by capillary electrophoresis using β-cyclodextrin or its derivatives as chiral selectors. Not only the effect of the type and the concentration of cyclodextrins used on resolution, but also temperature and voltage, buffer of pH on separation was also studied. The quantitative of ability of CE linearity and precision was investigated, and the linearity coefficient was 0.998, and the RSD was no more than 4.3% respectively. The quantitation of Ofloxacin in Ofloxacin tablet and ophthalmic solution was tried and good results were obtained.