This paper reports the assay of ofloxacin by fluorescence spectrophotometry. The fluorescence strength of ofloxacin is proportional to its concentration within the range from 0.500-25.0 ppm. The linear regression coefficient equals to 0.999 9, and the recovery raito is 99.0%. The results of determination for the samples of ofloxacin capsule from China and Japan are very close to that determined by the ultraviolet spectrophotometry, there isn't any obviously difference which was examined by Student's t test. This method is both simplicity and accuracy,also it needs only one kind of reagent. It's believable that this method would be suitable for the use in clinical medicine and pharmaceutical industry.